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interacts_with DB01238?

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Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . Novel phenolic antioxidants as multifunctional inhibitors of inducible P19320 expression for use in atherosclerosis . A series of novel phenolic compounds has been discovered as potent inhibitors of P01375 -inducible expression of vascular cell adhesion molecule-1 ( P19320 ) with concurrent antioxidant and lipid-modulating properties . Optimization of these multifunctional agents led to the identification of 3a ( DB05399 ) as a clinical candidate with demonstrated efficacies in animal models of atherosclerosis and hyperlipidemia . Functional autoradiography of neuropeptide Y Q03519 and P49146 subtypes in rat brain using agonist stimulated [35S]GTPgammaS binding . Neuropeptide Y , one of the most abundant brain peptides , has been found to modulate several important biological functions via a family of G-protein coupled receptors . To investigate the localization of functional P01303 receptor subtypes in the rat brain , we performed agonist-induced [35S]GTPgammaS autoradiography . The Q03519 /Y4/Y5 agonist DB00149 (31) , Pro(34)- P01303 increased [35S]GTPgammaS binding in several brain areas with a regional distribution consistent with that produced when labeling adjacent sections with [ 125I ] - DB00149 (31) , Pro(34)- P10082 . The Q03519 selective antagonist BIBP3226 antagonized the DB00149 (31) , Pro(34)- P01303 stimulated increase in [35S]GTPgammaS binding in all areas examined . The P28062 agonist P06681 - P01303 stimulated [35S]GTPgamma binding in numerous brain areas with a regional distribution similar to the binding observed with [ 125I ] - DB05004 . No increase in [35S]GTPgammaS binding above basal was observed in any brain area evaluated using Y4 and Y5 selective agonists . This study demonstrates abundant Q03519 and P49146 activation in the rat brain , while evidence for functional Y4 and Y5 receptors was not observed . Structural basis for P01133 receptor inhibition by the therapeutic antibody DB05774 . Therapeutic anticancer strategies that target and inactivate the epidermal growth factor receptor ( P00533 ) are under intense study in the clinic . Here we describe the mechanism of P00533 inhibition by an antibody drug DB05774 . DB05774 is a fully human antibody that has similar antitumor potency as the chimeric cetuximab/Erbitux and might represent a safer therapeutic alternative . We report the X-ray crystal structure of the Fab fragment of DB05774 ( Fab11F8 ) in complex with the entire extracellular region and with isolated domain III of P00533 . We compare this to our previous study of the cetuximab/ P00533 interaction . Fab11F8 interacts with a remarkably similar epitope , but through a completely different set of interactions . Both the similarities and differences in binding of these two antibodies have important implications for the development of inhibitors that could exploit this same mechanism of P00533 inhibition . 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 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 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 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 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 -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 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 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 receptors located on neurons and endothelial cells . Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . Comparative molecular profiling of the PPARα/γ activator aleglitazar : Q07869 selectivity , activity and interaction with cofactors . Peroxisome proliferator-activated receptors ( PPARs ) are a family of nuclear hormone receptors that control the expression of genes involved in a variety of physiologic processes , through heterodimerization with retinoid X receptor and complex formation with various cofactors . Drugs or treatment regimens that combine the beneficial effects of PPARα and γ agonism present an attractive therapeutic strategy to reduce cardiovascular risk factors . DB08915 is a dual PPARα/γ agonist currently in phase III clinical development for the treatment of patients with type 2 diabetes mellitus who recently experienced an acute coronary event . The potency and efficacy of aleglitazar was evaluated in a head-to-head comparison with other PPARα , γ and δ ligands . A comprehensive , 12-concentration dose-response analysis using a cell-based assay showed aleglitazar to be highly potent , with EC(50) values of 5 nM and 9 nM for PPARα and PPARγ , respectively . Cofactor recruitment profiles confirmed that aleglitazar is a potent and balanced activator of PPARα and γ . The efficacy and potency of aleglitazar are discussed in relation to other dual PPARα/γ agonists , in context with the published X-ray crystal structures of both PPARα and γ . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Inhibition of tumor cell growth , invasion , and metastasis by EXEL-2880 ( DB05030 , GSK1363089 ) , a novel inhibitor of P14210 and P15692 receptor tyrosine kinases . The DB00134 receptor tyrosine kinase and its ligand , hepatocyte growth factor ( P14210 ) , are overexpressed and/or activated in a wide variety of human malignancies . Vascular endothelial growth factor ( P15692 ) receptors are expressed on the surface of vascular endothelial cells and cooperate with DB00134 to induce tumor invasion and vascularization . EXEL-2880 ( DB05030 , GSK1363089 ) is a small-molecule kinase inhibitor that targets members of the P14210 and P15692 receptor tyrosine kinase families , with additional inhibitory activity toward P10721 , Flt-3 , platelet-derived growth factor receptor beta , and Tie-2 . Binding of EXEL-2880 to DB00134 and P15692 receptor 2 ( P35968 ) is characterized by a very slow off-rate , consistent with X-ray crystallographic data showing that the inhibitor is deeply bound in the DB00134 kinase active site cleft . EXEL-2880 inhibits cellular P14210 -induced DB00134 phosphorylation and P15692 -induced extracellular signal-regulated kinase phosphorylation and prevents both P14210 -induced responses of tumor cells and P14210 / P15692 -induced responses of endothelial cells . In addition , EXEL-2880 prevents anchorage-independent proliferation of tumor cells under both normoxic and hypoxic conditions . In vivo , these effects produce significant dose-dependent inhibition of tumor burden in an experimental model of lung metastasis . Collectively , these data indicate that EXEL-2880 may prevent tumor growth through a direct effect on tumor cell proliferation and by inhibition of invasion and angiogenesis mediated by P14210 and P15692 receptors . Differential regulation of the serotonin 1 A transcriptional modulators five prime repressor element under dual repression-1 and nuclear-deformed epidermal autoregulatory factor by chronic stress . Chronic stress is known to affect brain areas involved in learning and emotional responses . These changes , thought to be related to the development of cognitive deficits are evident in major depressive disorder and other stress-related pathophysiologies . The serotonin-related transcription factors ( Q6P1N0 / Q6P1N0 ; five prime repressor element under dual repression/coiled-coil P06681 domain 1a , and O75398 /Deaf-1 ; nuclear-deformed epidermal autoregulatory factor ) are two important regulators of the P08908 receptor . Using Western blotting and quantitative real-time polymerase chain reaction ( qPCR ) we examined the expression of mRNA and proteins for Q6P1N0 , O75398 , and the P08908 receptor in the prefrontal cortex ( P27918 ) of male rats exposed to chronic restraint stress ( CRS ; 6 h/day for 21 days ) . After 21 days of CRS , significant reductions in both Q6P1N0 mRNA and protein were observed in the P27918 ( 36.8 % and 32 % , respectively ; P < 0.001 ) , while the levels of both O75398 protein and mRNA did not change significantly . Consistent with reduced Q6P1N0 protein , P08908 receptor mRNA levels were equally upregulated in the P27918 , while protein levels actually declined , suggesting post-transcriptional receptor downregulation . The data suggest that CRS produces distinct alterations in the serotonin system specifically altering Q6P1N0 and the P08908 receptor in the P27918 of the male rat while having no effect on O75398 . These results point to the importance of understanding the mechanism for the differential regulation of Q6P1N0 and O75398 in the P27918 as a basis for understanding the related effects of chronic stress on the serotonin system ( serotonin-related transcription factors ) and stress-related disorders like depression . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Complement deposition and microglial activation in the outer retina in light-induced retinopathy : inhibition by a P08908 agonist . PURPOSE : Increasing evidence supports a role for complement in the pathogenesis of age-related macular degeneration ( AMD ) . This study evaluated retinal microglia , T-lymphocytes , and complement deposition in a light-induced retinopathy model . The effect of a serotonin ( 5-hydroxytryptamine , 5-HT(1A) ) agonist on these processes was investigated . METHODS : Rats were dark adapted for 24 hours before a 6-hour blue light exposure . Some animals were predosed subcutaneously with AL-8309A . Retinas were evaluated at different times after light exposure . Paraffin sections were stained with antibody for a microglial marker ( Iba1 ) , a T-lymphocyte marker ( CD3 ) , and complement components C1q , P01024 , factor B , factor H , and membrane attack complex ( MAC ) . RESULTS : Light exposure resulted in substantial photoreceptor and Q96AT9 loss . Robust microglia activation and migration to the outer retina occurred rapidly . Substantial T-lymphocyte recruitment did not occur . Complement alternative pathway was strongly activated , resulting in the deposition of P01024 , factor B , factor H , and MAC in the area of photic lesions . Dosing with AL-8309A prevented retinal lesions and decreased microglia activation/recruitment and complement deposition in the outer retina . CONCLUSIONS : In blue light exposed retinas , microglia were activated and migrated toward the outer retina , whereas a T-lymphocyte response was minimal . The innate immune system was markedly activated , with substantial complement deposition in the outer retina after light exposure . This complement deposition was prevented by AL-8309A . This model may be useful in the evaluation of complement inhibitors and other neuroprotectants intended for ocular use . AL-8309 is under evaluation in the clinic and may be useful in the treatment of AMD . Examination of ceramic restorative material interfacial debonding using acoustic emission and optical coherence tomography . OBJECTIVE : CAD/ P62158 ceramic restorative material is routinely bonded to tooth substrates using adhesive cement . This study investigates micro-crack growth and damage in the ceramic/dentin adhesive interface under fatigue shear testing monitored using the acoustic emission ( AE ) technique with optical coherence tomography ( O75051 ) . METHODS : Ceramic/dentin adhesive samples were prepared to measure the shear bond strength ( SBS ) under static load . Fatigue shear testing was performed using a modified ISO14801 method . Loads in the fatigue tests were applied at 80 % , 70 % , and 60 % of the SBS to monitor interface debonding . The AE technique was used to detect micro-crack signals in static and fatigue shear bond tests . RESULTS : The results showed that the average SBS value in the static tests was 10.61±2.23MPa ( mean±standard deviation ) . The average number of fatigue cycles in which ceramic/dentin interface damage was detected in 80 % , 70 % and 60 % of the SBS were 152 , 1962 and 9646 , respectively . The acoustic behavior varied according to the applied load level . Events were emitted during 60 % and 70 % fatigue tests . A good correlation was observed between crack location in O75051 images and the number of AE signal hits . SIGNIFICANCE : The AE technique and O75051 images employed in this study could potentially be used as a pre-clinical assessment tool to determine the integrity of cemented load bearing restored ceramic material . Sustainable cyclic load stresses in ceramic/dentin-bonded specimens were substantially lower than the measured SBS . Predicted S-N curve showed that the maximum endured load was 4.18MPa passing 10(6) fatigue cyclic . LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Dose selection of siltuximab for multicentric Castleman 's disease . PURPOSE : DB09036 is a monoclonal antibody that binds to interleukin ( IL ) -6 with high affinity and specificity ; P02741 ( CRP ) is an acute-phase protein induced by P05231 . CRP suppression is an indirect measurement of P05231 activity . Here , modeling and simulation of the pharmacokinetic ( PK ) /pharmacodynamic ( PD ) relationship between siltuximab and CRP were used to support dose selection for multicentric Castleman 's disease ( CD ) . METHODS : PK/PD modeling was applied to explore the relationship between siltuximab PK and CRP suppression following intravenous siltuximab infusion in 47 patients with B cell non-Hodgkin 's lymphoma ( n = 17 ) , multiple myeloma ( n = 13 ) , or CD ( n = 17 ) . DB09036 was administered as 2.8 , 5.5 , or 11 mg/kg q2wks , 11 mg/kg q3wks , or 5.5 mg/kg weekly . Simulations of studied or hypothetical siltuximab dosage regimens ( 15 mg/kg q4wks ) were also performed to evaluate maintenance of CRP suppression below the cutoff value of 1 mg/L . RESULTS : A two-compartment PK model and an inhibitory indirect response PD model adequately described the serum siltuximab and CRP concentration-time profiles simultaneously . PD parameter estimates were physiologically plausible . For all disease types , simulations showed that 11 mg/kg q3wks or 15 mg/kg q4wks would reduce serum CRP to below 1 mg/L after the second dose and throughout the treatment period . CONCLUSIONS : PK/PD modeling was used to select doses for further development of siltuximab in multicentric CD . The dosing recommendation was also supported by the observed efficacy dose-response relationship . CRP suppression in the subsequent randomized multicentric CD study was in agreement with the modeling predictions . ABC-cassette transporter 1 ( O95477 ) expression in epithelial cells in Chlamydia pneumoniae infection . DB00171 -binding cassette transporter A1 ( O95477 ) mediates reverse cholesterol transport and innate immunity response in different cell types . We have investigated the regulation of O95477 expression in response to intracellular Chlamydia pneumoniae infection in A549 epithelial lung carcinoma cells. C. pneumoniae infection decreased O95477 expression in A549 cells , and the activity of the O95477 promoter was decreased . The decreased promoter activity was dependent on its E-box and GnT-box elements of the promoter . Chlamydial growth was decreased in O95477 -silenced epithelial lung carcinoma cells . These data indicate an important role for O95477 in intracellular bacterial infection . P62158 interacts with DB00171 binding cassette transporter A1 to protect from calpain-mediated degradation and upregulates high-density lipoprotein generation . OBJECTIVE : To investigate the interaction of DB00171 -binding cassette transporter A1 ( O95477 ) with calmodulin in relation to its calpain-mediated degradation because many calpain substrates bind calmodulin to regulate cellular functions . METHODS AND RESULTS : The activity of O95477 is regulated through proteolysis by calpain . An immunoprecipitation and glutathione S-transferase pull-down assay revealed that O95477 directly binds calmodulin in a Ca(2+)-dependent manner . The cytoplasmic loop of O95477 contains a typical calmodulin binding sequence of 1-5-8-14 motifs ( 1245 to 1257 amino acids ) . The peptide of this region showed binding to calmodulin , and deletion of the 1-5-8-14 motif abolished this interaction . This motif is located near the O95477 Pro- DB00142 - DB00133 - DB00156 sequence , and the presence of calmodulin/Ca(2+) protected the peptides from proteolysis by calpain . The knockdown of calmodulin by a specific small and interfering RNA increased the degradation of O95477 and decreased O95477 protein and apolipoprotein A-I-mediated lipid release . Surprisingly , calmodulin inhibitor W7 increased calmodulin binding to O95477 and protected it from calpain-mediated degradation , consistent with our previous finding that this compound increased apolipoprotein A-I-mediated cell cholesterol release . CONCLUSIONS : P62158 directly binds and stabilizes O95477 in the presence of Ca(2+) and increases the generation of high-density lipoprotein . Structure and function of eritadenine and its 3-deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d- DB03769 ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 ) and has hypocholesterolemic activity . We have hypothesized that 3-deaza-DEA ( P01024 -DEA ) and its analogues retain high level of P23526 inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 -DEA analogues would have much higher hypocholesterolemic activity . P01024 -DEA , and its methyl ester ( P01024 -OMeDEA ) and its methyl amido ( P01024 -NMeDEA ) were synthesized to examine their P23526 inhibitory and hypocholesterolemic activities . A crystal structure of P23526 containing P01024 -DEA was determined and confirmed that DEA and P01024 -DEA bound to the same site of P23526 with the same binding mode . The P23526 inhibitory activities of P01024 -DEA ( K(I)=1.5 microM ) and P01024 -OMeDEA ( K(I)=1.5 microM ) are significantly lower than that of DEA ( K(I)=30 nM ) , while rats fed by P01024 -DEA and P01024 -OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 -NMeDEA lost most of the P23526 inhibitory activity ( K(I)=30 microM ) and dietary P01024 -NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 -DEA analogues are neither substrates nor inhibitors of adenosine deaminase .

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interacts_with DB00333?

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P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . DB09559 , a fully human IgG1 mAb directed against the P00533 for the potential treatment of cancer . DB09559 ( DB05774 ) , under development by ImClone Systems in collaboration with Bristol-Myers Squibb , is a fully human IgG1 mAb targeting the epidermal growth factor receptor ( P00533 ) , for the potential intravenous treatment of cancer , in particular NSCLC . In vitro studies demonstrate that necitumumab inhibits downstream targets in the P00533 pathway ( eg , MAPK ) , which are important for cellular proliferation , differentiation , invasion and metastasis . Furthermore , because necitumumab is an IgG1 construct , it has the potential to induce antibody-dependent cell-mediated cytotoxicity against tumor cells . Preclinical studies indicated that the antitumor activity of necitumumab is either comparable with or superior to that of ImClone 's chimeric anti- P00533 mAb cetuximab . In a phase I clinical trial in patients with advanced solid malignancies , necitumumab displayed nonlinear pharmacokinetic behavior . The toxicity profile of necitumumab is acceptable , with skin toxicity being the most frequently reported adverse event in the phase I and II clinical trials conducted to date . Preliminary data from a phase II clinical trial of necitumumab in combination with chemotherapy for the first-line treatment of advanced colon cancer are promising . Success in the ongoing phase III clinical trials in patients with advanced NSCLC would lead to necitumumab becoming a valuable addition to future therapeutic strategies in oncology . P14416 signaling modulates mutant ataxin-1 S776 phosphorylation and aggregation . Spinocerebellar ataxia 1 ( P54253 ) is a dominantly inherited neurodegenerative disease associated with progressive ataxia resulting from the loss of cerebellar Purkinje cells ( PCs ) and neurons in the brainstem . In PCs of P54253 transgenic mice , the disease causing ataxin-1 protein mediates the formation of P04271 containing cytoplasmic vacuoles and further self-aggregates to form intranuclear inclusions . The exact function of the ataxin-1 protein is not fully understood . However , the aggregation and neurotoxicity of the mutant ataxin-1 protein is dependent on the phosphorylation at serine 776 ( S776 ) . Although protein kinase A ( PKA ) has been implicated as the S776 kinase , the mechanism of PKA/ataxin-1 regulation in P54253 is still not clear . We propose that a dopamine D(2) receptor ( D2R ) / P04271 pathway may be involved in modulating PKA activity in PCs . Using a D2R/ P04271 P29320 stable cell line transiently transfected with GFP-ataxin-1[82Q] , we demonstrate that stimulation of the D2R/ P04271 pathway caused a reduction in mutant ataxin-1 S776 phosphorylation and ataxin-1 aggregation . Activation of PKA by forskolin resulted in an enhanced S776 phosphorylation and increased ataxin-1 nuclear aggregation , which was suppressed by treatment with D2R agonist bromocriptine and PKA inhibitor H89 . Furthermore , treating P54253 transgenic PC slice cultures with forskolin induced neurodegenerative morphological abnormalities in PC dendrites consistent with those observed in vivo . Taken together our data support a mechanism where PKA dependent mutant ataxin-1 phosphorylation and aggregation can be regulated by D2R/ P04271 signaling . Clinical and pathogenic aspects of candidate genes for lithium prophylactic efficacy . A number of candidate genes for lithium prophylactic efficacy have been proposed , some of them being also associated with a predisposition to bipolar illness . The aim of the present study was to investigate a possible association between polymorphisms of 14 common genes with the quality of prophylactic lithium response in patients with bipolar mood disorder , in relation to the putative role of these genes in the pathogenesis of this disorder . Some association with lithium prophylactic efficacy was found for the polymorphisms of P31645 , P21728 , P21964 , P23560 and P06241 genes , but not for 5HT2A , 5HT2C , P14416 , P35462 , P21917 , GSK-3 , Q16620 , Q13224 and P14780 . Possible aspects of these genes with regard to the mechanism of lithium activity and pathogenesis of bipolar mood disorder are discussed . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . P35372 and P00533 contribute to skin pigmentation differences between Indigenous Americans and Europeans . Contemporary variation in skin pigmentation is the result of hundreds of thousands years of human evolution in new and changing environments . Previous studies have identified several genes involved in skin pigmentation differences among African , Asian , and European populations . However , none have examined skin pigmentation variation among Indigenous American populations , creating a critical gap in our understanding of skin pigmentation variation . This study investigates signatures of selection at 76 pigmentation candidate genes that may contribute to skin pigmentation differences between Indigenous Americans and Europeans . Analysis was performed on two samples of Indigenous Americans genotyped on genome-wide SNP arrays . Using four tests for natural selection -- locus-specific branch length ( LSBL ) , ratio of heterozygosities ( lnRH ) , Tajima 's D difference , and extended haplotype homozygosity ( EHH ) -- we identified 14 selection-nominated candidate genes ( SNCGs ) . SNPs in each of the SNCGs were tested for association with skin pigmentation in 515 admixed Indigenous American and European individuals from regions of the Americas with high ground-level ultraviolet radiation . In addition to Q71RS6 and Q9UMX9 , genes previously associated with European/non-European differences in skin pigmentation , P35372 and P00533 were associated with variation in skin pigmentation in New World populations for the first time . 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 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D 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 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Lectin-like oxidized P01130 -1 ( P78380 ) acts as a receptor for remnant-like lipoprotein particles ( RLPs ) and mediates RLP-induced migration of vascular smooth muscle cells . OBJECTIVE : Remnant-like lipoprotein particles ( RLPs ) have been implicated in atherogenesis especially by diabetic dyslipidemia ; however , their receptor(s) and effects on vascular smooth muscle cells ( VSMCs ) remain unclear . In this study , we examined if lectin-like oxidized P01130 -1 ( P78380 ) acts as a receptor for RLPs and its biological effects in VSMCs . METHODS AND RESULTS : RLPs were isolated from human plasma by immunoaffinity gel containing anti-apolipoprotein A-I and anti-apolipoprotein B-100 monoclonal antibodies . DiI-labeled RLPs were taken up by CHO- P04264 cells stably expressing P78380 but not by wild-type CHO- P04264 cells . RLPs induced P78380 expression and cell migration in bovine VSMCs ( BVSMCs ) , which were significantly suppressed by transfection with P78380 specific siRNAs . Inhibitors of metalloproteinases , epidermal growth factor ( P01133 ) receptor tyrosine kinase , heparin-binding P01133 -like growth factor ( HB- P01133 ) , p38 mitogen-activated protein kinase ( p38 MAPK ) , MAPK kinase ( Q02750 ) and phosphoinositide 3-kinase ( PI3K ) significantly blocked RLP-induced P78380 expression and cell migration of BVSMCs . CONCLUSIONS : The present study provides direct evidence that P78380 is a novel receptor for RLPs in VSMCs . P78380 -mediated uptake of RLPs may thus play important roles in atherogenesis by inducing P78380 expression and VSMC migration especially in the settings of postprandial hyperlipidemia , diabetes and metabolic syndrome . Q08462 selectively couples to E prostanoid type 2 receptors , whereas adenylyl cyclase 3 is not receptor-regulated in airway smooth muscle . Adenylyl cyclases ( ACs ) are important regulators of airway smooth muscle function , because β-adrenergic receptor ( βAR ) agonists stimulate AC activity and DB02527 production . We have previously shown in a number of cell types that AC6 selectively couples to βAR and these proteins are coexpressed in lipid rafts . We overexpressed AC2 , O60266 , and AC6 in mouse bronchial smooth muscle cells ( mBSMCs ) and human embryonic kidney ( P29320 ) -293 cells by using recombinant adenoviruses and assessed their localization and regulation by various G protein-coupled receptors ( GPCRs ) . O60266 and AC6 were expressed primarily in caveolin-rich fractions , whereas AC2 expression was excluded from these domains . AC6 expression enhanced DB02527 production in response to isoproterenol but did not increase responses to butaprost , reflecting the colocalization of AC6 with β(2)AR but not E prostanoid type 2 receptor ( EP(2)R ) in lipid raft fractions . AC2 expression enhanced butaprost-stimulated DB02527 production but had no effect on the β(2)AR-mediated response . O60266 did not couple to any GPCR tested . DB02587 -induced arborization of mBSMCs was assessed as a functional readout of DB02527 signaling . Arborization was enhanced by overexpression of AC6 and O60266 , but AC2 had no effect . GPCR-stimulated arborization mirrored the selective coupling observed for DB02527 production . With the addition of the phosphodiesterase 4 ( DB05876 ) inhibitor rolipram AC2 accelerated forskolin-stimulated arborization . Thus , AC2 selectively couples to EP(2)R , but signals from this complex are limited by DB05876 activity . O60266 does not seem to couple to GPCR in either mBSMCs or P29320 -293 cells , so it probably exists in a distinct signaling domain in these cells . DB05387 ( AEterna ) . AEterna is developing DB05387 , an angiogenesis inhibitor derived from the ultrafiltration of liquid shark cartilage , with matrix metalloprotease ( MMP ) -2 , P14780 and vascular endothelial growth factor inhibitory properties , for the potential treatment of non-small-cell lung cancer . Cytokinins control endocycle onset by promoting the expression of an P25054 /C activator in Arabidopsis roots . Plant roots respond to various internal and external signals and adjust themselves to changes of environmental conditions . In the root meristem , stem cells produce daughter cells that continue to divide several times . When these latter cells reach the transition zone , they stop dividing and enter the endocycle , a modified cell cycle in which DNA replication is repeated without mitosis or cytokinesis . The resultant DNA polyploidization , named endoreduplication , is usually associated with an increase of nuclear and cell volume and with cell differentiation . At the transition zone , cytokinin signaling activates two transcription factors , type-B ARABIDOPSIS RESPONSE REGULATOR 1 ( P49407 ) and ARR12 , and induces SHY2/IAA3 , a member of the Aux/IAA family of auxin signaling repressors . This inhibits auxin signaling and reduces the expression of auxin efflux carriers , resulting in cell division arrest . Such counteracting actions of two hormones are assumed to determine meristem size . However , it remains unknown whether cytokinins additionally control meristem size through an auxin-independent pathway . Here we show that , in Arabidopsis , the cytokinin-activated P32121 directly upregulates the expression of CCS52A1 , which encodes an activator of an E3 ubiquitin ligase , anaphase-promoting complex/cyclosome ( P25054 /C ) , thereby promoting the onset of the endocycle and restricting meristem size . Our genetic data revealed that CCS52A1 function is independent of SHY2-mediated control of auxin signaling , indicating that downregulation of auxin signaling and P25054 /C-mediated degradation of cell-cycle regulators cooperatively promote endocycle onset , and thus fine tune root growth . Arundic acid ( DB05343 ) ameliorates delayed ischemic brain damage by preventing astrocytic overproduction of P04271 . After focal cerebral ischemia , the infarct volume increases rapidly within acute infarct expansion ( initial 12 to 24 h ) and continues slowly during delayed infarct expansion ( 25 to 168 h ) . While acute infarct expansion represents progressive necrosis within the ischemic core , delayed infarct expansion starts as disseminated apoptotic cell death in a narrow rim surrounding the infarct border , which gradually coalesces to form a larger infarct . Discovery of a distinct correlation between reactive astrogliosis along the infarct border and delayed infarct expansion in the rodent ischemia model led us to investigate the possible causal relationship between the two events . Specifically , the calcium binding protein P04271 exerts detrimental effects on cell survival through activation of various intracellular signaling pathways , resulting in altered protein expression . Arundic acid [ ( R ) -(-)-2-propyloctanoic acid , DB05343 ] is a novel agent that inhibits P04271 synthesis in cultured astrocytes . In the rodent ischemia model , this agent was shown to inhibit both the astrocytic overexpression of P04271 and the subsequent activation of signaling pathways in the peri-infarct area . Concurrently , delayed infarct expansion was prevented , and neurologic deficits were promptly ameliorated . The results of subsequent studies suggest that the efficacy of arundic acid is mediated by restoring the activity of astroglial glutamate transporters via enhanced genetic expression . Eosinophil cysteinyl leukotriene synthesis mediated by exogenous secreted phospholipase A2 group X . Secreted phospholipase A(2) group X ( sPLA(2)-X ) has recently been identified in the airways of patients with asthma and may participate in cysteinyl leukotriene ( CysLT ; C(4) , D(4) , and E(4) ) synthesis . We examined CysLT synthesis and arachidonic acid ( AA ) and lysophospholipid release by eosinophils mediated by recombinant human sPLA(2)-X . We found that recombinant sPLA(2)-X caused marked AA release and a rapid onset of CysLT synthesis in human eosinophils that was blocked by a selective sPLA(2)-X inhibitor . Exogenous sPLA(2)-X released lysophospholipid species that arise from phospholipids enriched in AA in eosinophils , including phosphatidylcholine , phosphatidylinositol , and phosphatidylethanolamine as well as plasmenyl phosphatidylcholine and phosphatidylethanolamine . CysLT synthesis mediated by sPLA(2)-X but not AA release could be suppressed by inhibition of cPLA(2)α . Exogenous sPLA(2)-X initiated DB00133 (505) phosphorylation of cPLA(2)α , an intracellular Ca(2+) flux , and translocation of cPLA(2)α and P09917 in eosinophils . Synthesis of CysLTs in response to sPLA(2)-X or lysophosphatidylcholine was inhibited by p38 or JNK inhibitors but not by a Q02750 /2 inhibitor . A further increase in CysLT synthesis was induced by the addition of sPLA(2)-X to eosinophils under conditions of N-formyl-methionyl-leucyl-phenylalanine-mediated cPLA(2)α activation . These results indicate that sPLA(2)-X participates in AA and lysophospholipid release , resulting in CysLT synthesis in eosinophils through a mechanism involving p38 and JNK MAPK , cPLA(2)α , and P09917 activation and resulting in the amplification of CysLT synthesis during cPLA(2)α activation . Transactivation of eosinophils by sPLA(2)-X may be an important mechanism leading to CysLT formation in the airways of patients with asthma . Association study of 45 candidate genes in nicotine dependence in Han Chinese . Numerous genetic linkages , association studies have been performed in different ethnic groups and revealed many susceptibility loci and genes for nicotine dependence . However , limited similar researches were performed in Han Chinese . This study was designed to investigate the association of candidate genes with nicotine dependence in Han Chinese . We genotyped 384 SNPs within 45 candidate genes with nicotine dependence in a Han Chinese population consisting 223 high nicotine dependent subjects and 257 low nicotine dependent subjects by employing GoldenGate genotyping assay ( Illumina ) . Following association analysis was performed using PLINK software . Individual SNP-based association analysis revealed that nine SNPs located in P35462 ( rs2630351 ) , P21918 ( rs1967550 ) , Q9Y6R4 ( rs2314378 ) , DDC ( rs11575461 ) , Q05901 ( rs4954 ) , O75899 ( rs2779562 ) , P14416 ( rs11214613 and rs6589377 ) and P43681 ( rs2236196 ) were significantly associated with FTND after correction for multiple testing with the p values from 2.59×10(-7) to 9.99×10(-5) . Haplotype-based association analysis revealed haplotype G-A-A formed by rs2630351 , rs167771 and rs324032 and haplotype G-G-G-A formed by rs3773678 , rs2630349 , rs2630351 and rs167771 in P35462 ; haplotype of G-A formed by rs2779562 and rs2808566 in O75899 and haplotype of T-T-A-G-A formed by rs6832644 , rs4057797 , rs9764 , rs4552421 and rs10033119 in P25929 are associated with FTND ( p=3.61×10(-7)-8.78×10(-6) ) . Our results provided confirmation of the previous findings that P14416 , P35462 , DDC , Q05901 , O75899 and P43681 are associated with nicotine dependence . Furthermore , we for the first time report a significant association between nicotine dependence and P21918 , Q9Y6R4 and P25929 . These findings need independent replication in the future studies . DB04725 ( Merckle ) . EuroAlliance ( a consortium of Alfa Wassermann SpA , Lacer SA and Merckle GmbH ) is developing licofelone , a dual cyclooxygenase and P09917 inhibitor for the potential treatment of inflammatory disorders including osteoarthritis . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . DB04958 , a P20273 -targeting recombinant humanized antibody with a different mode of action from rituximab . DB04958 is a humanized anti- P20273 monoclonal antibody currently in clinical trials for treatment of non-Hodgkin lymphoma ( Q9NZ71 ) and certain autoimmune diseases . Here we report the results of investigations of epratuzumab 's mode of action in comparison to and in combination with the anti- P11836 mAb , rituximab . In vitro cell growth inhibition , induction of apoptosis , and the ability of the mAbs to mediate complement-dependent cytotoxicity ( CDC ) and antibody-dependent cellular cytotoxicity ( ADCC ) were evaluated . We also investigated the potential activity of epratuzumab in the regulation of B-cell antigen receptor ( P11274 ) activation . DB04958 and rituximab displayed very distinct modes of action ; epratuzumab acts as an immunomodulatory agent , while rituximab is an acutely cytotoxic therapeutic antibody . DB04958 has distinct effects on cell growth from rituximab . For example , rituximab+anti-human IgG Fcgamma yielded marked inhibition of proliferation in human Q9NZ71 cell lines , while epratuzumab had little or no effect in this assay . However , when cells were immobilized and stimulated with anti-IgM , epratuzumab , but not rituximab , caused a significant antiproliferative effect . Unlike rituximab , no CDC could be detected , and ADCC was modest but significant with epratuzumab . Importantly , combining rituximab and epratuzumab did not decrease rituximab 's ability to induce apoptosis , CDC , and ADCC . In fact , the combination is more effective than rituximab alone in inhibiting proliferation of Daudi Burkitt lymphoma cells in the presence of second antibody , and at least equally effective to rituximab in the absence of crosslinking . These observations suggest that it may be possible to enhance clinical efficacy by combination therapy comprised of anti- P11836 and anti- P20273 mAbs . DB00055 suppresses tissue factor expression on U937 cells in the endothelial protein C receptor-dependent manner . The new functional role of activated protein C ( P25054 ) in the regulation of tissue factor ( TF ) expression was investigated using the cultured human monoblastic leukemia U937 cell line . A flow cytofluorometric analysis demonstrated that treatment with P25054 resulted in time- and dose-dependent decrease in TF expression in unstimulated and phorbol ester-stimulated cells . The effect was antagonized by the monoclonal antibody ( mAb ) to endothelial protein C/ Q9UNN8 ( Q9UNN8 ) , 252 , which strongly inhibited the interaction between P25054 and Q9UNN8 . In contrast , mAbs 49 and 379 , which bind to Q9UNN8 without blocking P25054 binding , had no or only a modest effect . It is concluded that culturing U937 cells in the presence of P25054 caused down-regulation of TF expression through the Q9UNN8 -dependent mechanism , independent of whether induction was triggered by phorbol ester . Growth modulation of low density lipoprotein receptor sterol sensitivity in cultured human fibroblasts . Sterols regulate low density lipoprotein ( LDL ) receptor and 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase gene expressions by end product repression . Studies on cultured cells have shown that growing cells have a higher LDL uptake than quiescent cells and that incubation of cells with growth factors or mitogenic compounds leads to sterol-resistant upregulation of P01130 gene expression . The recent finding that elevated P01130 activity in acute myelogenous leukemia cells was characterized by a decreased sensitivity to downregulation by sterols raises the possibility that the mechanism behind this is related to the cellular growth rate . By using cultured human fibroblasts as a model system we therefore studied whether growth modulation of sterol sensitivity takes place in normal actively growing cells . Judging from the ability of sterols ( 25-hydroxycholesterol + cholesterol ) to inhibit 125I-LDL degradation , we found that the sensitivity to sterols varied markedly between cells of different densities . The lowest sensitivity to sterols and highest 125I-LDL degradation rate were found in subconfluent cells , whereas sparse and confluent cells were the most sensitive ones . In contrast to the P01130 , P04035 sterol sensitivity did not appear to be growth regulated . We conclude that growth-dependent modulation of sterol sensitivity and P01130 activity takes place in normal human fibroblasts . Modulation of sterol sensitivity may be an important mechanism to ensure an adequate cholesterol supply in growing cells .

MH_train_1202

MH_train_1202

MH_train_1202

interacts_with DB00783?

multiple_choice

Activation of intestinal peroxisome proliferator-activated receptor-α increases high-density lipoprotein production . AIMS : Peroxisome proliferator-activated receptor ( Q07869 ) -α is a transcription factor controlling lipid metabolism in liver , heart , muscle , and macrophages . Peroxisome proliferator-activated receptor-α activation increases plasma HDL cholesterol and exerts hypotriglyceridaemic actions via the liver . However , the intestine expresses Q07869 -α , produces HDL and chylomicrons , and is exposed to diet-derived Q07869 -α ligands . Therefore , we examined the effects of Q07869 -α activation on intestinal lipid and lipoprotein metabolism . METHODS AND RESULTS : The impact of Q07869 -α activation was evaluated in term of HDL-related gene expression in mice , ex vivo in human jejunal biopsies and in Caco-2/TC7 cells . Apolipoprotein-AI/HDL secretion , cholesterol esterification , and trafficking were also studied in vitro . In parallel to improving plasma lipid profiles and increasing liver and intestinal expression of fatty acid oxidation genes , treatment with the dual Q07869 -α/δ ligand DB05187 resulted in a more pronounced increase in plasma HDL compared with fenofibrate in mice . DB05187 , but not fenofibrate , increased the expression of HDL production genes such as apolipoprotein-AI and DB00171 -binding cassette A1 transporter in murine intestines . A similar increase was observed upon Q07869 -α activation of human biopsies and Caco-2/TC7 cells . Additionally , HDL secretion by Caco-2/TC7 cells increased . Moreover , Q07869 -α activation decreased the cholesterol esterification capacity of Caco-2/TC7 cells , modified cholesterol trafficking , and reduced apolipoprotein-B secretion . CONCLUSION : Peroxisome proliferator-activated receptor-α activation reduces cholesterol esterification , suppresses chylomicron , and increases HDL secretion by enterocytes . These results identify the intestine as a target organ of Q07869 -α ligands with entero-hepatic tropism to reduce atherogenic dyslipidaemia . P25021 mediated relaxation of buffalo ( Bubalus bubalus ) ureter . On the buffalo ureter , histamine did not elicit any direct effect . However , it caused concentration-dependent relaxation of the tissues precontracted by carbachol , phenylephrine , norepinephrine , KCI or BaCl2 and also inhibited the contractile effect of carbachol . DB08805 selectively antagonised the relaxation and inhibition of contractile response but mepyramine did not show this effect . Isoprenaline , dobutamine , salbutamol , verapamil and papaverine neither produced any direct effect nor relaxed the carbachol-contracted tissues ; norepinephrine and epinephrine had contractile effects . Hence , the histamine-induced relaxation was mediated through the activation of H2 receptors and not through adrenergic mechanisms or blockade of Ca(2+)-channels or inhibition of cyclic nucleotide phosphodiesterase . Discovery of ( 2E ) -3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide ( DB05223 ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 enzyme and COLO 205 cellular IC(50) ) , liver microsomal stability ( t(1/2) ) , cytochrome P450 inhibitory ( 3A4 IC(50) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT-116 , PC-3 , A2780 , MV4-11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials . P08246 inhibitors as treatment for P48444 . Chronic obstructive pulmonary disease , characterised by a slowly progressive , irreversible airways limitation , is a major worldwide cause of chronic morbidity and mortality . The imbalance between human neutrophil elastase and endogenous antiproteases may cause excess human neutrophil elastase in pulmonary tissues , which may be considered a major pathogenic factor in chronic obstructive pulmonary disease . Great effort has been devoted to finding a method to restore the balance , resulting in the discovery of potent two-typed small-molecular-weight human neutrophil elastase inhibitors . In the application of chronic obstructive pulmonary disease therapy , the human neutrophil elastase inhibitors mainly focused upon include ONO-5046 , MR-889 , L-694,458 , CE-1037 , GW-311616 and TEI-8362 as the acyl-enzyme inhibitors ; and DB03925 , AE-3763 , FK-706 , ICI-200,880 , ZD-0892 and ZD-8321 as the transition-state inhibitors . In this review , various problems that remain to be solved in the clinical use of human neutrophil elastase inhibitors are discussed . iSubgraph : integrative genomics for subgroup discovery in hepatocellular carcinoma using graph mining and mixture models . The high tumor heterogeneity makes it very challenging to identify key tumorigenic pathways as therapeutic targets . The integration of multiple omics data is a promising approach to identify driving regulatory networks in patient subgroups . Here , we propose a novel conceptual framework to discover patterns of miRNA-gene networks , observed frequently up- or down-regulated in a group of patients and to use such networks for patient stratification in hepatocellular carcinoma ( HCC ) . We developed an integrative subgraph mining approach , called iSubgraph , and identified altered regulatory networks frequently observed in HCC patients . The miRNA and gene expression profiles were jointly analyzed in a graph structure . We defined a method to transform microarray data into graph representation that encodes miRNA and gene expression levels and the interactions between them as well . The iSubgraph algorithm was capable to detect cooperative regulation of miRNAs and genes even if it occurred only in some patients . Next , the miRNA-mRNA modules were used in an unsupervised class prediction model to discover HCC subgroups via patient clustering by mixture models . The robustness analysis of the mixture model showed that the class predictions are highly stable . Moreover , the Kaplan-Meier survival analysis revealed that the HCC subgroups identified by the algorithm have different survival characteristics . The pathway analyses of the miRNA-mRNA co-modules identified by the algorithm demonstrate key roles of Myc , Q01094 , let-7 , P01137 , P01375 and P00533 in HCC subgroups . Thus , our method can integrate various omics data derived from different platforms and with different dynamic scales to better define molecular tumor subtypes . iSubgraph is available as MATLAB code at http://www.cs.umd.edu/~ozdemir/isubgraph/ . Functional promoter SNPs in cell cycle checkpoint genes . A substantial number of genes mutated in human cancers encode components of the cell cycle processes . As the P55008 /S transition in the cell cycle is a finely regulated biological process , we hypothesized that sequence variations in the promoter region of the related genes might indeed lead to abnormal expression , thus predisposing the individuals carrying these genetic variants to cancer . In this report , we screened the promoter regions of 16 cell cycle checkpoint genes for DNA variants and assessed the functional impact of these promoter region single nucleotide polymorphisms ( pSNPs ) by combining in silico analysis and in vitro functional assays . We identified 127 pSNPs including 90 with predicted impact on putative binding sites of known transcription factors . Eleven pSNPs were selected for electrophoresis mobility shift assays because of their association with predicted gains of binding sites , and nine pSNPs showed differential allelic shifts in at least one cell line tested . Following the subcloning of the promoter regions into a gene reporter system , we found that at least four promoter haplotypes associated with P24385 , Q01094 , Q13547 and P06400 significantly influenced transcriptional activity in an allele-specific manner . Although the biological significance of these observations still remains to be demonstrated , the expected variability of expression levels in key cell cycle components might influence individual 's risk of cancer . [ A molecular study of methylmalonic aciduria : structure-function correlations ] . Cobalamin ( DB00115 ) non-responsive methylmalonic acidemia is caused by mutations in the P22033 locus on chromosome 6p21 encoding the enzyme methylmalonyl DB01992 mutase ( EC 5.4.99.2 ) . This disorder has been extensively studied by biochemical , somatic cell genetic and molecular techniques . Mutations have been identified which cause classic mut(o) phenotypes in which there is no detectable enzymatic activity , as well as mut- phenotypes in which there is residual cobalamin-dependent activity . Mutations which exhibit interallelic complementation have been identified within both of these groups . These mutations illustrate the position , structure , and function of critical domains within this cobalamin binding enzyme and provide new insights into the biochemical and clinical consequences of enzyme deficiency . The homology of the cobalamin binding region has allowed mutations of the mutase to be mapped onto the x-ray structure of methionine synthase ( EC 2.1.1.13 ) . The role of P38936 Waf1/Cip1 in large airway epithelium in smokers with and without P48444 . Airway epithelium alterations , including squamous cell metaplasia , characterize smokers with and without chronic obstructive pulmonary disease ( P48444 ) . The P38936 regulates cell apoptosis and differentiation and its role in P48444 is largely unknown . Molecules regulating apoptosis ( cytoplasmic P38936 , caspase-3 ) , cell cycle ( nuclear P38936 ) , proliferation ( Ki67/ P12004 ) , and metaplasia ( survivin ) in central airways from smokers ( S ) , smokers- P48444 ( s- P48444 ) and non-smokers ( Controls ) were studied . The role of cigarette smoke extracts ( CSE ) in P38936 , survivin , apoptosis ( caspase-3 and annexin-V binding ) and proliferation was assessed in a bronchial epithelial cell line ( 16HBE ) . Immunohistochemistry , image analysis in surgical samples and flow-cytometry and carboxyfluorescein succinimidyl ester proliferative assay in 16HBE with/without CSE were applied . Cytoplasmic and nuclear P38936 , survivin , and Ki67 expression significantly increased in large airway epithelium in S and in s- P48444 in comparison to Controls . P42574 was similar in all the studied groups . P38936 correlated with epithelial metaplasia , P12004 , and Ki67 expression . CSE increased cytoplasmic P38936 and survivin expression but not apoptosis and inhibited the cell proliferation in 16HBE . In large airway epithelium of smokers with and without P48444 , the cytoplasmic P38936 inhibits cell apoptosis , promotes cell proliferation and correlates with squamous cell metaplasia thus representing a potential pre-oncogenic hallmark . Estrogen upregulates endothelial nitric oxide synthase gene expression in fetal pulmonary artery endothelium . NO , produced by endothelial NO synthase ( P29474 ) , is a key mediator of pulmonary vasodilation during cardiopulmonary transition at birth . The capacity for NO production is maximal at term because pulmonary P29474 expression increases during late gestation . Since fetal estrogen levels rise markedly during late gestation and there is indirect evidence that the hormone enhances nonpulmonary NO production in adults , estrogen may upregulate P29474 in fetal pulmonary artery endothelium . Therefore , we studied the direct effects of estrogen on P29474 expression in ovine fetal pulmonary artery endothelial cells ( PAECs ) . DB00783 caused a 2.5-fold increase in NOS enzymatic activity in PAEC lysates . This effect was evident after 48 hours , and it occurred in response to physiological concentrations of the hormone ( 10(-10) to 10(-6) mol/L ) . The increase in NOS activity was related to an upregulation in P29474 protein expression , and P29474 mRNA abundance was also enhanced . P03372 antagonism with DB00947 completely inhibited estrogen-mediated P29474 upregulation , indicating that estrogen receptor activation is necessary for this response . In addition , immunocytochemistry revealed that fetal PAECs express estrogen receptor protein . Furthermore , transient transfection assays with a specific estrogen-responsive reporter system have demonstrated that the endothelial estrogen receptor is capable of estrogen-induced transcriptional transactivation . Thus , estrogen upregulates P29474 gene expression in fetal PAECs through the activation of PAEC estrogen receptors . This mechanism may be responsible for pulmonary P29474 upregulation during late gestation , thereby optimizing the capacity for NO-mediated pulmonary vasodilation at birth . Oncogenic activity of P33993 transforming cluster . The miniature chromosome maintenance ( P22033 ) complex is a group of proteins that are essential for DNA replication licensing and control of cell cycle progression from P55008 to S phase . Recent studies suggest that P33993 is overexpressed and amplified in a variety of human malignancies . P33993 genome sequence contains a cluster of miRNA that has been shown to downregulate expression of several tumor suppressors including P38936 , Q01094 , O43521 and pTEN . The oncogenic potential of P33993 and its embedded miRNA has been demonstrated vigorously in in vitro experiments and in animal models , and they appear to cooperate in initiation of cancer . P33993 protein also serves as a critical target for oncogenic signaling pathways such as androgen receptor signaling , or tumor suppressor pathways such as integrin α7 or retinoblastoma signaling . This review analyzes the transforming activity and signaling of P33993 , oncogenic function of miRNA cluster that is embedded in the P33993 genome , and the potential of gene therapy that targets P33993 . The interplay between NF-kappaB and Q01094 coordinately regulates inflammation and metabolism in human cardiac cells . DB11194 dehydrogenase kinase 4 ( Q16654 ) inhibition by nuclear factor-κB ( NF-κB ) is related to a shift towards increased glycolysis during cardiac pathological processes such as cardiac hypertrophy and heart failure . The transcription factors estrogen-related receptor-α ( ERRα ) and peroxisome proliferator-activated receptor ( Q07869 ) regulate Q16654 expression through the potent transcriptional coactivator PPARγ coactivator-1α ( P20142 -1α ) . NF-κB activation in AC16 cardiac cells inhibit ERRα and PPARβ/δ transcriptional activity , resulting in reduced P20142 -1α and Q16654 expression , and an enhanced glucose oxidation rate . However , addition of the NF-κB inhibitor parthenolide to these cells prevents the downregulation of Q16654 expression but not ERRα and PPARβ/δ DNA binding activity , thus suggesting that additional transcription factors are regulating Q16654 . Interestingly , a recent study has demonstrated that the transcription factor Q01094 , which is crucial for cell cycle control , may regulate Q16654 expression . Given that NF-κB may antagonize the transcriptional activity of Q01094 in cardiac myocytes , we sought to study whether inflammatory processes driven by NF-κB can downregulate Q16654 expression in human cardiac AC16 cells through Q01094 inhibition . Protein coimmunoprecipitation indicated that Q16654 downregulation entailed enhanced physical interaction between the p65 subunit of NF-κB and Q01094 . Chromatin immunoprecipitation analyses demonstrated that p65 translocation into the nucleus prevented the recruitment of Q01094 to the Q16654 promoter and its subsequent Q01094 -dependent gene transcription . Interestingly , the NF-κB inhibitor parthenolide prevented the inhibition of Q01094 , while Q01094 overexpression reduced interleukin expression in stimulated cardiac cells . Based on these findings , we propose that NF-κB acts as a molecular switch that regulates Q01094 -dependent Q16654 gene transcription . Differential gene expression underlying the functional distinctions of primary human P28906 + hematopoietic stem and progenitor cells from peripheral blood and bone marrow . The restorative capacity of human P28906 (+) hematopoietic cells is clinically used in the autologous and allogeneic transplant setting to support cytotoxic therapy . We examined gene expression patterns of highly enriched bone marrow P28906 (+) ( BM- P28906 (+) ) or G- P04141 -mobilized peripheral blood P28906 (+) ( PB- P28906 (+) ) cells by cDNA array technology , quantitative real-time RT-PCR , and flow cytometry , to identify molecular causes underlying the functional differences between circulating and sedentary hematopoietic stem and progenitor cells . The greater cell cycle and DNA synthesis activity of BM- P28906 (+) compared to PB- P28906 (+) cells was reflected by the 2- to 5-fold higher expression of 9 genes involved in cell cycle , 11 genes regulating DNA synthesis , and the cell cycle-initiating transcription factor Q01094 . The 2- to 3-fold greater expression of 5 pro-apoptotic genes in PB- P28906 (+) cells indicated a higher apoptotic activity , which could functionally be corroborated by apoptosis assays . P25116 ( PAR1 ) , known to play a role in trafficking of malignant cells , was 3.6-fold higher expressed in circulating P28906 (+) cells than in BM- P28906 (+) cells . Guidance via thrombin receptor might molecularly mediate stem cell migration . In summary , our study provides gene expression profiles of primary human P28906 (+) hematopoietic cells of blood and marrow . Our data molecularly confirm and explain the finding that P28906 (+) cells residing in the bone marrow are cycling more rapidly , whereas circulating P28906 (+) cells consist of a higher number of quiescent stem and progenitor cells . Moreover , our data give novel molecular insights into stem cell migration and differentiation . DB11320 inhibits cell proliferation and modulates the expression of Bcl-2 family proteins via the H2 receptor in human pancreatic cancer cells . In this study , the mechanisms involved in the inhibitory effect of histamine ( HA ) on PANC-1 cell proliferation were investigated . The action of HA on cell growth was evaluated by determining the cell doubling time from experimental growth curves and analysing the cell cycle using a flow cytometer . The expression of proteins related to cell death and proliferation ( P12004 , p53 , c-Fos and Bcl-2 family proteins ) was studied using Western blot , immunocytochemistry and flow cytometric analysis . The results indicated that HA produced an accumulation of PANC-1 cells in GO/Gl-phase and increased the doubling time via H2HA ( P25021 ) stimulation . Expression of p53 , c-Fos and Bcl-2 were not modulated by HA . However , HA decreased P12004 and Bax expression , while it increased the Bcl-x level . In summary , the antiproliferative effect exerted by HA was associated with a G0/ P55008 -phase arrest and a modulation of the Bcl-2 family proteins . DB08810 protects against ethanol-induced gastric mucosal injury in rats : role of 5-hydroxytryptamine , prostaglandins and sulfhydryl compounds . This study was designed to determine the gastroprotective properties of cinitapride ( CNT ) , a novel prokinetic benzamide derivative agonist of Q13639 and 5-HT1 receptors and 5-HT2 antagonist , on mucosal injury produced by 50 % ( v/v ) ethanol . Results were compared with those for 5-hydroxytryptamine ( 5-HT : 10 mg kg-1 ) . The possible involvements of gastric mucus secretion , endogenous prostaglandins ( PGs ) and sulfhydryl compounds ( SH ) in the protection mediated by CNT were also examined . Intraperitoneal administration of CNT ( 0.50 and 1 mg kg-1 ) , 30 min before ethanol , significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus . CNT ( 1 mg kg-1 ) also produced a significant increase in gastric mucosal levels of DB00917 , but did not induce any significant changes in SH values . On the contrary , pretreatment with 5-HT worsened ethanol-induced erosions , however , did not affect gastric mucus secretion , glycoprotein content or DB00917 levels , although the non-protein SH fraction was significantly decreased . The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism . We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved . Inhibition of the T790M gatekeeper mutant of the epidermal growth factor receptor by EXEL-7647 . PURPOSE : Agents inhibiting the epidermal growth factor receptor ( P00533 ) have shown clinical benefit in a subset of non-small cell lung cancer patients expressing amplified or mutationally activated P00533 . However , responsive patients can relapse as a result of selection for P00533 gene mutations that confer resistance to DB00171 competitive P00533 inhibitors , such as erlotinib and gefitinib . We describe here the activity of EXEL-7647 ( DB05007 ) , a novel spectrum-selective kinase inhibitor with potent activity against the P01133 and vascular endothelial growth factor receptor tyrosine kinase families , against both wild-type ( WT ) and mutant P00533 in vitro and in vivo . EXPERIMENTAL DESIGN : The activity of P00533 inhibitors against WT and mutant EGFRs and their effect on downstream signal transduction was examined in cellular assays and in vivo using A431 and MDA-MB-231 ( WT P00533 ) and H1975 ( L858R and T790M mutant P00533 ) xenograft tumors . RESULTS : EXEL-7647 shows potent and long-lived inhibition of the WT P00533 in vivo . In addition , EXEL-7647 inhibits cellular proliferation and P00533 pathway activation in the erlotinib-resistant H1975 cell line that harbors a double mutation ( L858R and T790M ) in the P00533 gene . In vivo efficacy studies show that EXEL-7647 substantially inhibited the growth of H1975 xenograft tumors and reduced both tumor P00533 signaling and tumor vessel density . Additionally , EXEL-7647 , in contrast to erlotinib , substantially inhibited the growth and vascularization of MDA-MB-231 xenografts , a model which is more reliant on signaling through vascular endothelial growth factor receptors . CONCLUSIONS : These studies provide a preclinical basis for clinical trials of DB05007 in solid tumors and in patients bearing tumors that are resistant to existing P00533 -targeted therapies . DB00114 values in cerebrospinal fluid : reference values and diagnosis of Q9NVS9 deficiency in paediatric patients . Our aim was to establish reference values for cerebrospinal fluid ( P04141 ) pyridoxal 5'-phosphate ( PLP ) in a paediatric population for the diagnosis of pyridox(am)ine 5'-phosphate oxidase ( Q9NVS9 ) deficiency . For reference values , P04141 samples from 113 paediatric controls ( age range : 1 day-18 years ) from Barcelona and London were analysed . Cerebrospinal fluid PLP and biogenic amine concentrations were analysed by HPLC with fluorescence and electrochemical detection . DB00114 concentrations in 4 patients with Q9NVS9 deficiency were determined . A negative correlation between P04141 PLP values and age of controls was observed in both populations ( r=-0.503 ; p < 0.0001 and r=-0.542 ; p=0.002 ) . Reference values were stratified into 4 ( Barcelona ) and 3 age groups ( London ) . For the newborn period , P04141 PLP reference intervals were 32-78 and 44-89 nmol/L for the Barcelona and London centers , respectively ) . No correlation was observed in the different age groups between PLP values and biogenic amines metabolites . PLP values in neonates with Q9NVS9 deficiency were clearly decreased ( PLP=3.6 , 12.0 , 14.0 and 18.0 nmol/L ) compared with our reference ranges . In conclusion , reference values for P04141 PLP should be stratified according to age . No association was observed between PLP values and biogenic amines metabolites . In our 4 cases with Q9NVS9 deficiency , P04141 PLP values were clearly below the reference values . 5-hydroxytryptamine and its receptors in systemic vascular walls . 5-hydroxytryptamine ( 5-HT ) in the bloodstream is largely contained in platelets and circulates throughout the entire vascular system . 5-HT released from activated platelets dramatically changes the function of vascular smooth muscle cells ( VSMCs ) and endothelial cells ( ECs ) . In VSMCs , 5-HT induces proliferation and migration via 5- Q13049 receptors . These effects are further enhanced by vasoactive substances such as thromboxane A2 and angiotensin II . 5- Q13049 receptor activation in VSMCs also causes both enhancement of prostaglandin I2 production by inducing cyclooxygenase-2 and reduction of nitric oxide ( NO ) by suppressing inducible NO synthase . Evidence showing that 5-HT in ECs plays a principal role in angiogenesis now exists . Stimulation of 5-HT1 and/or 5-HT2 receptors has been implicated in the angiogenic effect of 5-HT . The extracellular signal-regulated kinase and endothelial NO synthase ( P29474 ) activation-dependent pathways are involved in the mechanisms . Moreover , Q13639 receptors in ECs have been shown to also regulate angiogenesis . Recent reports show sarpogrelate , a selective antagonist of the 5- Q13049 receptor , indirectly enhances the function of P28222 receptors in ECs via inhibition of 5- Q13049 receptors in VSMCs or platelets . This indirect action of P28222 receptors in ECs may increase NO production derived from P29474 and a vasodilator response . Furthermore , sarpogrelate and other 5- Q13049 receptor antagonists have been shown to reduce the constitutive activity of 5- Q13049 receptors . It is believed that increasing evidence on the role of 5-HT receptors will contribute to the expansion of the clinical application of existing therapeutic drugs such as sarpogrelate , and to the development of new 5-HT receptor-related drugs for treating cardiovascular diseases . Engineering the response to vascular injury : divergent effects of deregulated Q01094 expression on vascular smooth muscle cells and endothelial cells result in endothelial recovery and inhibition of neointimal growth . P01375 -alpha ( P01375 ) is expressed locally in the vessel wall after angioplasty and induces growth arrest and apoptosis in endothelial cells ( ECs ) , thereby delaying reendothelialization . Prior studies have shown that direct antagonism of P01375 , using a systemically administered soluble receptor , can enhance endothelial recovery and reduce neointimal thickening . These studies have also shown that downregulation of the transcription factor Q01094 was a key mechanism of P01375 's effect on ECs . We now show that Ad- Q01094 overexpression at sites of balloon injury accelerates functional endothelial recovery , consistent with the prior in vitro findings . Moreover these studies also reveal divergent effects of P01375 and overexpression of Q01094 on ECs versus VSMCs . P01375 exposure of VSMCs had no affect on proliferation or apoptosis , in contrast to the effect seen in ECs . In Ad- Q01094 -transduced VSMCs , however , P01375 -induced marked apoptosis in contrast to the survival effect seen in ECs . Finally , these studies suggest that differential activation of NF-kappaB may play a key role in mediating these opposing effects . Nuclear translocation and transcriptional activity of NF-kappaB was markedly attenuated in Ad- Q01094 -transduced VSMCs , whereas it remained active in similarly treated ECs when the cells were exposed to P01375 . These studies reveal that overexpression of Ad- Q01094 primes VSMCs to P01375 -induced apoptosis . Furthermore , Q01094 potentiates VSMC death by blocking antiapoptotic signaling pathway through inhibition of NF-kappaB activation . The divergent responses of VSMCs and ECs to Q01094 overexpression provide unique therapeutic possibilities : simultaneously targeting the cell cycle of two different cell types , within same tissue microenvironment resulting in opposite and biologically complimentary effects . [ Therapeutic use of anti- P01375 agents in spondyloarthropathies ] . CLASSICAL DATA : Spondyloarthropathies regroup several rheumatological entities ( ankylosing spondylitis , reactive arthritis , psoriatic rheumatism , entero-colopathic disease rheumatism , undifferentiated spondyloarthropathies ) with validated diagnosis criteria . Drug therapy is based upon NSAIDs ( non-steroidal antiinflammatories ) . Refractory forms may lead to severe functional impairment , raising the need of more effective treatments . IN FAVOUR OF ANTI- P01375 -ALPHA AGENTS : Several arguments ( P01375 serum levels , elevated levels of mRNA , P01375 messengers , in sacro iliac biopsies , efficacy of anti P01375 agents in Crohn 's disease ) justify the use of anti- P01375 agents in the treatment of spondyloarthropathies . Two biologic agents have been assessed in these circumstances : a monoclonal antibody ( DB00065 ) and a soluble form of the P01375 receptor ( DB00005 ) . EFFICACY AND SAFETY : Results of open and controlled studies , although on small series , demonstrated the significant efficacy of anti- P01375 agents on the various clinical , biological , functional and quality of life parameters , and confirmed by imaging ( Q9BWK5 ) . Tolerance is fair , but two cases of diffuse tuberculosis have been reported with DB00065 . THERAPEUTIC PROGRESS : Even if additional studies are required to answer some questions ( long term efficacy and safety , treatment modalities ) , anti P01375 agents appear as a therapeutic progress in refractory spondyloarthropathies , for which few validated options have existed up till now . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis .

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interacts_with DB00951?

multiple_choice

[ Toxicogenetics of antiretroviral treatment ( II ) : neurotoxicity , hepatotoxicity , lactic acidosis , kidney damage , and other adverse effects of antiretroviral drugs ] . Several pharmacogenetics studies have analyzed the influence of specific genetic polymorphisms on the toxicity of antiretroviral treatment . The present review describes some of the adverse effects of antiretroviral drugs in which a genetic predisposition may be involved : efavirenz-induced neurological toxicity , generally associated with the 516G > T polymorphism of liver enzyme cytochrome P450 2B6 ( P20813 ) ; hypersensitivity reactions to nevirapine , associated with specific alleles of major histocompatibility complex , mainly the HLA- Q8IUH3 *0101 allele , which , in combination with a high P01730 lymphocyte count , has been associated with systemic reactions and hepatitis in Caucasians , and the HLA-Cw8 allele , which is associated with hypersensitivity reactions in persons from the Italian island of Sardinia and from Japan ; nevirapine-induced hepatotoxicity associated with the C > T polymorphism in position 3435T of the P08183 ( MDR-1 ) gene codifying for glycoprotein P ( lower risk ) ; hyperbilirubinemia in patients exposed to atazanavir or indinavir carrying the P22309 *28 polymorphism ; peripheral neuropathy with nucleoside analogues associated with haplogroup T of the mitochondrial genome ( higher risk ) and with the Q30201 C282Y genotype of the hemochromatosis gene ( lower risk ) ; the mutation in codon 964 ( R964C ) of the P54098 gene that codifies the mitochondrial polymerase DNA gamma described in a Thai patient with lactic acidosis ; the Q92887 gene haplotypes associated with tenofovir-induced proximal tubulopathy , and the risk of pancreatitis in persons with mutations in the P13569 and SPINK-1 genes . p38 Q96HU1 kinase regulates stem cell apoptosis in human hematopoietic failure . Myelodysplastic syndromes ( P43034 ) are clonal stem cell disorders that lead to ineffective hematopoiesis and are common causes of low blood counts in the elderly . The exact molecular mechanisms regulating increased stem apoptosis in these disorders are not well defined . p38 MAPK activation is important in regulating the growth inhibitory signals of P01375 , TGF-beta and Interferons on human hematopoiesis . Our findings show that p38 MAPK is overactivated in myelodysplasia bone marrows and regulates hematopoietic stem cell apoptosis . Inhibition of p38 MAPK by genetic or pharmacologic means decreases apoptosis and stimulates in vitro hematopoiesis from primary P43034 hematopoietic progenitors . These studies point to the potential efficacy of selective p38alpha inhibitor , DB05412 , in human bone marrow failure . Immunomodulatory influence of bone marrow-derived DB05914 on neuroinflammation in astrocyte cultures . The therapeutic benefits associated with DB05914 ( MSCs ) largely result from their immunomodulatory and neurotrophic properties . In this study , we evaluated the effects of MSCs on astrocyte cultures exposed to lipopolysaccharide . In response to this inflammatory trigger , astrocytes showed an increased expression of pro-inflammatory genes ( IL-1β , TNFα , P05231 ) , which was attenuated by pre-exposure to O60682 conditioned medium . Furthermore , mediators released by MSCs increased cell proliferation and altered the regulation of intermediate filaments ( P14136 , vimentin ) , pro-inflammatory enzymes ( P35228 , P35354 ) and receptors ( O00206 , P08571 , Q14832 , P41594 ) . These data demonstrate that MSCs influence diverse cell types participating in the response to neuroinflammation . Renal changes induced by a cyclooxygenase-2 inhibitor during normal and low sodium intake . P35354 ( P35354 ) has been identified in renal tissues under normal conditions , with its expression enhanced during sodium restriction . To evaluate the role of P35354 -derived metabolites in the regulation of renal function , we infused a selective inhibitor ( nimesulide ) in anesthetized dogs with normal or low sodium intake . The renal effects elicited by nimesulide and a non-isozyme-specific inhibitor ( meclofenamate ) were compared during normal sodium intake . In ex vivo assays , meclofenamate , but not nimesulide , prevented the platelet aggregation elicited by arachidonic acid . During normal sodium intake , nimesulide infusion ( n=6 ) had no effects on arterial pressure or renal hemodynamics but did reduce urinary sodium excretion , urine flow rate , and fractional lithium excretion . In contrast , nimesulide administration increased arterial pressure and decreased renal blood flow , urine flow rate , and fractional lithium excretion during low sodium intake ( n=6 ) . P35354 inhibition reduced urinary prostaglandin E(2) excretion in both groups but did not modify plasma renin activity in dogs with low ( 8.1+/-1.1 ng angiotensin I. mL(-1). h(-1) ) or normal ( 1.8+/-0.4 ng angiotensin I. mL(-1). h(-1) ) sodium intake . DB00939 infusion in dogs with normal sodium intake ( n=8 ) induced a greater renal hemodynamic effect than nimesulide infusion . These results suggest that P35354 -derived metabolites ( 1 ) are involved in the regulation of sodium excretion in dogs with normal sodium intake , ( 2 ) play an important role in the regulation of renal hemodynamic and excretory function in dogs with low sodium intake , and ( 3 ) are not involved in the maintenance of the high renin levels during a long-term decrease in sodium intake . Genetic polymorphisms , the metabolism of estrogens and breast cancer : a review . Breast cancer is the most common female cancer and the second cause of cancer death in women . Despite recent breakthroughs , much of the etiology of this disease is unknown and the most important risk factor , i.e. , exposure to endogenous and exogenous estrogen throughout life can not explain the heterogeneity of prognosis nor clinical features of patients . Recently , many gene polymorphisms in the metabolism of breast cancer have been described as possible neoplasm etiologic factors . This review is an attempt to summarize the current knowledge about these polymorphisms and to determine new target genes for diagnosis and treatment of the disease . Polymorphisms in the genes P05093 , P11511 , P04798 , P05177 , Q16678 , P22309 , P50225 , 17-hydroxysteroid-dehydrogenase , P21964 , Q86UG4 , P03372 , and Q92731 are described . Pharmacology of recombinant low-voltage activated calcium channels . Several types of voltage- or ligand-activated calcium channels contribute to the excitability of neuronal cells . Low-voltage-activated ( LVA ) , T-type calcium channels are characterised by relatively negative threshold of activation and therefore they can generate low-threshold spikes , which are essential for burst firing . At least three different proteins form T-type calcium current in neurons : Ca(v)3.1 , Ca(v)3.2 and Q9P0X4 . Expression of these proteins in various brain regions is complementary . Individual channel types could be distinguished by different sensitivity towards inorganic cations . This inhibition can contribute to the toxicity of some heavy metals . Selective inhibition of T-type calcium channels by organic blockers may have clinical importance in some forms of epilepsy . DB01388 inhibits the expressed Ca(v2)3.1 , Ca(v)3.2 and Q9P0X4 channels in nanomolar concentrations with Q9P0X4 channel having lowest affinity . The sensitivity of the expressed Ca(v)3.1 channel to the antiepileptic drugs , valproate and ethosuximide , is low . Ca(v)3.1 channel is moderately sensitive to phenytoin . The Ca(v)3.2 channel is sensitive to ethosuximide , amlodipine and amiloride . All three LVA calcium channels are moderately sensitive to active metabolites of methosuximide , i.e. alpha-methyl-alpha-phenylsuccinimide . Several neuroleptics inhibit all three LVA channels in clinically relevant concentrations . All three channels are also inhibited by the endogenous cannabinoid anandamide . A high affinity peptide blocker for these Ca channels is the scorpion toxin kurtoxin which inhibits the Ca(v)3.1 and Ca(v)3.2 , but not the Q9P0X4 channel in nanomolar concentrations . DB06690 selectively inhibits the Ca(v)3.2 , but not the Ca(v)3.1 channel . The Ca(v)3.2 , but not the Ca(v)3.1 channel is potentiated by stimulation of Ca(2+)/ P62158 -dependent protein kinase . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) 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 ) -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 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 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 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- 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 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . Battle of the kinases : integration of adrenal responses to DB02527 , DG and Ca2+ at the level of steroidogenic cytochromes P450 and 3betaHSD expression in H295R cells . While DB01285 receptors ( activating the protein kinase A pathway ) are expressed throughout the human/bovine/ovine zona glomerulosa ( zg ) and zona fasciculata ( zf ) , there are clear zonal differences in AII Type-1 receptor levels ( activating protein kinase C/Ca2+ ) , as well as resting membrane potential . Thus zg is most responsive to AII and K+ ( Ca2+ signalling ) , while zf is less responsive to AII with no K+ response . Zonal function in turn requires differential expression of P05093 /3betaHSD and P19099 / P15538 . We have used the H295R cell to study how differential activation of kinase A , kinase C and Ca2+/calmodulin ( P62158 ) kinases may alter the relative expression of the steroidogenic P450s and 3betaHSDII . While P05108 , P05093 , 3betaHSDII , P08686 , and P15538 are all induced by increases in DB02527 , studies with TPA alone or in combination with forskolin reveal subsets of steroidogenic enzymes regulated either positively ( P08686 , 3betaHSDII ) or negatively ( P05093 , P05108 ) by protein kinase C . Thus adrenal 3betaHSDII and P08686 expression is high in zg and zf , but P05093 is not expressed in the zg where AII activation of kinase C is highest . In turn both K+ and AII-induced elevation of Ca2+ strongly induces P19099 but not P15538 , consistent with preferential expression of P19099 in the zg . We conclude that differential signaling through kinase C and P62158 kinases in addition to kinase A underlies zonal differences in both the early and late pathways involved in steroid hormone production within the adrenocortical zones . Randomised clinical trial : effects of monotherapy with DB05070 , a P41594 inhibitor , on symptoms and reflux events in patients with gastro-oesophageal reflux disease . BACKGROUND : DB05070 , a metabotropic glutamate receptor 5 ( P41594 ) negative allosteric modulator , has been shown to reduce gastro-oesophageal reflux events and oesophageal acid exposure in patients with gastro-oesophageal reflux disease ( GERD ) and healthy subjects . AIM : To evaluate the effects of DB05070 monotherapy for 2 weeks on symptom control in patients with GERD . METHODS : This was a double-blind , placebo-controlled , multi-centre trial in GERD patients who were responders to proton pump inhibitors ( PPIs ) . Following PPIs withdrawal , a 2-week baseline washout period was followed by 2-week treatment with either DB05070 120 mg or placebo b.d . The primary clinical efficacy endpoint was the number of GERD symptom-free days in treatment week 2 compared with the last 7 days of baseline . The effect on reflux events using 24-h impedance-pH monitoring was also determined in a subset of 24 patients . RESULTS : The full analysis set comprised 103 patients DB05070 ( N= 50 ) , Placebo ( N=53 ) . In treatment week 2 , DB05070 significantly increased GERD symptom-free days ( P=0.045 ) and heartburn-free days ( P=0.037 ) , reduced antacid use ( P=0.017 ) , improved total symptom score ( P=0.048 ) including subscale heartburn/regurgitation ( P=0.007 ) and sleep disturbance because of GERD ( P= 0.022 ) . DB05070 significantly reduced total ( P=0.034 ) and acidic reflux events ( P=0.003 ) . DB05070 was well tolerated . Most common adverse events for DB05070 were mild to moderate dizziness 16 % and vertigo 12 % ( placebo 4 % and 2 % ) . CONCLUSIONS : Inhibition of P41594 with DB05070 monotherapy reduces reflux events and improves symptoms in GERD patients . This mechanism has promise for the management of GERD . Effect of resveratrol in experimental osteoarthritis in rabbits . OBJECTIVE : DB02709 ( DB02709 ) is a phytoalexin found in high concentration in the skins of grapes and red wines which has been shown to have antiinflammatory , anticancerogen and antioxidant properties . DB02709 is a potent and specific inhibitor of nuclear factor kappa B ( NF-kappaB ) . DB02709 also inhibits P35354 gene expression and enzyme activity . We aimed to determine the in vivo effects of intra-articular injections of resveratrol on cartilage and synovium in an experimental osteoarthritis ( OA ) model in rabbits . METHODS : As OA model , rabbits underwent unilateral anterior cruciate ligament transection ( ACLT ) . Five weeks after test group was injected with 10 microMol/kg resveratrol in dimethylsulphoxide ( DB01093 ) in the knees once daily for two weeks and as the control group at the same time DB01093 was injected into the knees . All rabbits were killed one week after the last injection . Cartilage tissue and synovium were evaluated with a histological scoring system . RESULTS : Histological evaluation of cartilage tissue by H & E staining revealed a significantly reduced average cartilage tissue destruction score of 1.7 in the resveratrol group versus 2.8 in the control group ( p = 0.016 ) . Loss of matrix proteoglycan content in cartilage was also much lower , as determined by safranin O staining . Scores of synovial inflammation did n't show difference between groups ( 1.3 vs 2.2 ; p = 0.057 ) . CONCLUSION : A characteristic parameter in arthritis is the progressive loss of articular cartilage . This study suggests that intraarticular injections of resveratrol starting at the onset of disease may protect cartilage against the development of experimentally induced OA . Decreasing Poly(ADP- DB01936 ) Polymerase Activity Restores ΔF508 P13569 Trafficking . Most cystic fibrosis is caused by mutations in P13569 that prevent its trafficking from the ER to the plasma membrane and is associated with exaggerated inflammation , altered metabolism , and diminished responses to oxidative stress . P09874 is activated by oxidative stress and causes energy depletion and cell dysfunction . Inhibition of this enzyme protects against excessive inflammation and recent studies have also implicated it in intracellular protein trafficking . We hypothesized that P09874 activity is altered in CF and affects trafficking and function of the most common CF mutant ΔF508 P13569 . Indeed , P09874 activity was 2.9-fold higher in CF ( ΔF508/ΔF508 ) human bronchial epithelial primary cells than in non-CF cells , and similar results were obtained by comparing CF vs. non-CF bronchial epithelial cell lines ( 2.5-fold higher in CFBE41o(-) vs. 16HBE14o(-) , P < 0.002 ) . A P09874 inhibitor ( ABT-888 , DB07232 ) partially restored P13569 channel activity in CFBE41o(-) cells overexpressing ΔF508 P13569 . Similarly , reducing P09874 activity by 85 % in ileum from transgenic CF mice ( Cftr(tm1)Eur ) partially rescued ΔF508 P13569 activity to 7 % of wild type mouse levels , and similar correction ( 7.8 % ) was observed in vivo by measuring salivary secretion . Inhibiting P09874 with ABT-888 or siRNA partially restored ΔF508 P13569 trafficking in cell lines , and most ΔF508 P13569 was complex glycosylated when heterologously expressed in P09874 (-/-) mouse embryonic fibroblasts . Finally , levels of the mature glycoform of P13569 were reduced by peroxynitrite , a strong activator of P09874 . These results demonstrate that P09874 activity is increased in CF , and identify a novel pathway that could be targeted by proteostatic correctors of P13569 trafficking . Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 , CYP2B1/2 , P05181 , CYP3A , catechol-O-methyltransferase ( P21964 ) and microsomal glutathione transferase 1 ( P10620 ) . All but actin and soluble P21964 were positively detected at ≥1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 . The binding correlated well with inhibition of CYP activities , except for P05181 whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 antibodies showed a significant reduction in binding at ≥1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors . Transcriptional modulation of monoaminergic neurotransmission genes by the histone deacetylase inhibitor trichostatin A in neuroblastoma cells . Histone deacetylase inhibitors are promising anti-tumor agents partly due to their ability to disrupt the hypoxic signaling pathway in human malignancies . However , little is known about any effects of these drugs on the central nervous system . The aim of the present study was to analyze the effects of trichostatin A ( P32119 ) -- a broad-spectrum histone deacetylase inhibitor -- on the transcriptional regulation of several genes involved in dopamine- and serotonergic neurotransmission . To this end , short-term parallel cultures of SK-NF-I neuroblastoma cells were treated with P32119 either alone or in combination with hypoxia , and mRNA levels of dopamine receptor D3 ( P35462 ) and D4 ( P21917 ) , dopamine transporter ( Q01959 ) , dopamine hydroxylase ( P09172 ) , dopamine receptor regulating factor ( DRRF ) , catechol-O-methyltransferase ( P21964 ) , serotonin receptor 1A ( P08908 ) , monoamino oxidase A ( P21397 ) , serotonin transporter ( P31645 ) and tryptophan hydroxylase 2 ( Q8IWU9 ) were determined by quantitative PCR . We found that P32119 did not antagonize the hypoxia-induced activation of D3 and D4 dopamine receptor genes , implying that induction of these genes is not mediated directly by hypoxia inducible factor-1alpha . On the other hand , P32119 dramatically upregulated the expression of Q01959 and P31645 ( 45-fold and 15-fold , respectively ) , while transcript levels of P21397 and P21964 were significantly reduced ( by 70 % and by more than 90 % , respectively ) . Induction of Q01959 protein expression was detected by western blotting . These results suggest that inhibition of histone deacetylases might help restore presynaptic monoamine pools via suppression of catecholamine breakdown and facilitation of monoamine reuptake in neurons . The uremic toxin 3-indoxyl sulfate is a potent endogenous agonist for the human aryl hydrocarbon receptor . The aryl hydrocarbon receptor ( P35869 ) is a ligand-activated transcription factor involved in the regulation of multiple cellular pathways , such as xenobiotic metabolism and Th17 cell differentiation . Identification of key physiologically relevant ligands that regulate P35869 function remains to be accomplished . Screening of indole metabolites has identified indoxyl 3-sulfate ( I3S ) as a potent endogenous ligand that selectively activates the human P35869 at nanomolar concentrations in primary human hepatocytes , regulating transcription of multiple genes , including P04798 , P05177 , Q16678 , P22309 , P19224 , P05231 , and P0DJI8 . Furthermore , I3S exhibits an approximately 500-fold greater potency in terms of transcriptional activation of the human P35869 relative to the mouse P35869 in cell lines . Structure-function studies reveal that the sulfate group is an important determinant for efficient P35869 activation . This is the first phase II enzymatic product identified that can significantly activate the P35869 , and ligand competition binding assays indicate that I3S is a direct P35869 ligand . I3S failed to activate either CAR or O75469 . The physiological importance of I3S lies in the fact that it is a key uremic toxin that accumulates to high micromolar concentrations in kidney dialysis patients , but its mechanism of action is unknown . I3S represents the first identified relatively high potency endogenous P35869 ligand that plays a key role in human disease progression . These studies provide evidence that the production of I3S can lead to P35869 activation and altered drug metabolism . Our results also suggest that prolonged activation of the P35869 by I3S may contribute to toxicity observed in kidney dialysis patients and thus represent a possible therapeutic target . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . Monoclonal antibodies targeting P01584 reduce biomarkers of atherosclerosis in vitro and inhibit atherosclerotic plaque formation in P02649 -deficient mice . OBJECTIVE : Atherosclerosis is a condition that is increasingly contributing to worldwide mortality through complications such as stroke and myocardial infarction . IL-1β plays multiple direct , local roles in the formation and stability of the atheroma by eliciting the production of additional cytokines and proteolytic enzymes from macrophages , endothelial cells ( EC ) and smooth muscle cells ( SMC ) . We therefore tested whether an anti-IL-1β antibody , DB06062 , might inhibit the secretion of pro-atherogenic cytokines from macrophages in vitro and affect a positive outcome in the P02649 -deficient mouse ( ApoE(-/-) ) model of atherosclerosis in vivo . METHODS AND RESULTS : In an in vitro co-culture model , DB06062 inhibited macrophage-induced secretion of key atherogenic cytokines from EC and SMC , including P05231 , P10145 , P13500 and TNFα . The release of degradative enzymes , such as the matrix metalloproteinases P08254 and P14780 , was also decreased by DB06062 . In addition , DB06062 inhibited the secretion of P13232 from EC and P05112 from SMC , cytokines not previously reported to be driven by IL-1β in this context . In vivo , XMA052 MG1K , a chimeric murine version of DB06062 , inhibited the formation of atherosclerotic lesions in the ApoE(-/-) model at all three doses tested . This effect was comparable to that reported for complete genetic ablation of IL-1β or IL-1R1 on an ApoE(-/-) background and was associated with decreases in plasma non-HDL/HDL cholesterol ratio and plaque lipid content and macrophage infiltration . CONCLUSIONS : These results demonstrate for the first time that an antibody targeting IL-1β can inhibit the progression of atherosclerosis in vivo , highlighting the importance of this key cytokine in cardiovascular disease . Clozapine treatment causes oxidation of proteins involved in energy metabolism in lymphoblastoid cells : a possible mechanism for antipsychotic-induced metabolic alterations . There is increasing concern about the serious metabolic side effects and neurotoxicity caused by atypical ( second-generation ) antipsychotics . In a previous study by our group ( Walss-Bass et al. Int J Neuropsychopharmacol 2008;11:1097-104 ) , using a novel proteomic approach , we showed that clozapine treatment in SKNSH cells induces oxidation of proteins involved in energy metabolism , leading us to hypothesize that protein oxidation could be a mechanism by which atypical antipsychotics increase the risk for metabolic alterations . In this study , the same proteomic approach was used to identify specific proteins oxidized after clozapine treatment in lymphoblastoid cell lines from patients with schizophrenia and normal controls . Cells were treated with 0 and 20 μM clozapine for 24 hours and protein extracts were labeled with 6-iodoacetamide fluorescein ( 6-IAF ) . The lack of incorporation of 6-IAF into the thiol group of cysteine residues is an indicator of protein oxidation . Labeled proteins were exposed to two dimensional electrophoresis , and differential protein labeling was assessed . Increased oxidation after clozapine treatment was observed in 9 protein spots ( P < 0.05 ) . The following 7 proteins were identified by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry ( HPLC- P19957 -MS/MS ) in those 9 spots : enolase , triosephosphate isomerase ( P60174 ) , glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , Rho GDP dissociation inhibitor ( GDI ) , cofilin , uridine monophosphate/ cytidine monophosphate ( UMP- P21941 ) kinase , and translation elongation factor . Several of these proteins play important roles in energy metabolism and mitochondrial function . These results further support the hypothesis that oxidative stress may be a mechanism by which antipsychotics increase the risk of metabolic syndrome and diabetes . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . P19957 kinase/AKT pathway as a therapeutic target in multiple myeloma . The development of novel therapies for multiple myeloma depends on a comprehensive understanding of the events leading to cellular proliferation and survival . Controlling pathways that regulate growth signals is an emerging and complementary approach to myeloma treatment . The PI3K/Akt pathway is a central gatekeeper for crucial cellular functions including adhesion , angiogenesis , migration and development of drug resistance . Established proteins and genes such as P42345 , p53 , NF-kappaB and Q92934 are all regulated through PI3K and Akt activation , making them attractive targets for broad downstream effects . Direct PI3K inhibition has demonstrated impressive tumor inhibition and regression in cell-line and animal models , and multiple agents including DB05210 are currently in clinical trials . Drugs such as perifosine that are specific for Akt are also in development . Combinations of these agents with existing therapies are rational approaches on the path to improving myeloma treatment .

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The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Comparison of effects of P11473 versus O75469 , Q96RI1 and GR ligands on the regulation of CYP3A isozymes in rat and human intestine and liver . In this study , we compared the regulation of CYP3A isozymes by the vitamin D receptor ( P11473 ) ligand 1 alpha,25-dihydroxyvitamin D(3) ( 1,25(OH)(2)D(3) ) against ligands of the pregnane X receptor ( O75469 ) , the glucocorticoid receptor ( GR ) and the farnesoid X receptor ( Q96RI1 ) in precision-cut tissue slices of the rat jejunum , ileum , colon and liver , and human ileum and liver . In the rat , 1,25(OH)(2)D(3) strongly induced CYP3A1 mRNA , quantified by qRT-PCR , along the entire length of the intestine , induced CYP3A2 only in ileum but had no effect on CYP3A9 . In contrast , the O75469 /GR ligand , dexamethasone ( DEX ) , the O75469 ligand , pregnenolone-16 alpha carbonitrile ( Q15149 ) , and the Q96RI1 ligand , chenodeoxycholic acid ( DB06777 ) , but not the GR ligand , budesonide ( BUD ) , induced CYP3A1 only in the ileum , none of them influenced CYP3A2 expression , and Q15149 , DEX and BUD but not DB06777 induced CYP3A9 in jejunum , ileum and colon . In rat liver , CYP3A1 , CYP3A2 and CYP3A9 mRNA expression was unaffected by 1,25(OH)(2)D(3) , whereas DB06777 decreased the mRNA of all CYP3A isozymes ; Q15149 induced CYP3A1 and CYP3A9 , BUD induced CYP3A9 , and DEX induced all three CYP3A isozymes . In human ileum and liver , 1,25(OH)(2)D(3) and DEX induced P08684 expression , whereas DB06777 induced P08684 expression in liver only . In conclusion , the regulation of rat CYP3A isozymes by P11473 , O75469 , Q96RI1 and GR ligands differed for different segments of the rat and human intestine and liver , and the changes did not parallel expression levels of the nuclear receptors . Effect of lipopolysaccharide on the xenobiotic-induced expression and activity of hepatic cytochrome P450 in mice . Infection-associated inflammation can alter the expression levels and functions of cytochrome P450s ( CYPs ) . Cyp gene expression is regulated by the activation of several nuclear receptors , including pregnane X receptor ( O75469 ) , constitutive androstane receptor ( CAR ) , and aryl hydrocarbon receptor ( P35869 ) . These receptors can be activated by xenobiotics , including medicines . Here , to study the xenobiotic-induced fluctuations in CYP during inflammation , we examined the effect of lipopolysaccharide ( LPS ) treatment on the level of mRNAs encoding hepatic CYPs induced by xenobiotic-activated nuclear receptors , in mice . Both the mRNA induction of Cyp genes and the metabolic activities of CYP proteins were examined . LPS treatment caused a significant decrease in the induced expression of the mRNAs for Cyp3a11 , 2c29 , 2c55 , and 1a2 , but not for Cyp2b10 . To assess the CYP enzymatic activities , CYP3A-mediated testosterone 6β-hydroxylation and the intrinsic clearance ( CL(int) ) of nifedipine in liver microsomes were measured in mice treated with the xenobiotic pregnenolone-16alpha-carbonitrile ( Q15149 ) with or without LPS administration . Both assays revealed that the CYP3A activity , which was induced by Q15149 , declined significantly after LPS treatment , and this decline correlated with the Cyp3a11 mRNA level . In addition , we found that the mRNAs for interleukin ( IL ) -1β and tumor necrosis factor ( P01375 ) α were increased after treatment with LPS plus xenobiotics . Our findings demonstrated that LPS treatment reduces the O75469 - and P35869 -mediated , and possibly CAR-mediated Cyp gene expression and further suggest that these decreases are dependent on inflammatory cytokines in the liver . Q9BYW2 -α and survivin involved in the anti-apoptotic effect of DB02342 after global ischemia in rats . Survivin is an anti-apoptotic gene that decreases the apoptosis by depressing the expression of caspase-3 . Hypoxia-inducible factor-1-alpha ( Q16665 ) is a transcription factor specifically activated by hypoxia . 2-methoxyestradiol ( DB02342 ) is an estradiol derivative and a known Q16665 inhibitor . DB02342 decreased apoptosis by inhibiting Q16665 . The aim of the present study was to investigate if survivin is involved in the anti-apoptotic effect of DB02342 . Male adult rats were used to make the global ischemia ( GI ) model . Ten minutes after GI , DB02342 was injected intraperitoneally ( 16 mg/kg weight ) . Rats were killed at 6 hours , 12 hours , 24 hours , 48 hours , 96 hours , and 7 days . GI produced a marked increase in Q16665 expressions in the hippocampus at 6 hours and peaked at 48-96 hours . The expressions of survivin and caspase-3 were increased lightly in a similar time course . These molecular changes were accompanied by massive cell loss and apoptosis in the hippocampal regions . DB02342 treatment reduced the expression of Q16665 , increased survivin expression , and decreased the expression of caspase-3 . These results indicate that survivin and Q16665 were involved in the anti-apoptotic effect of DB02342 treated following GI . DB02342 may decrease the Q16665 expression , up-regulate the survivin expression , inhibit the expression of caspase-3 , and finally reduce apoptosis after GI . Prevention of hemorrhagic shock-induced lung injury by heme arginate treatment in rats . Hemorrhagic shock followed by resuscitation ( HSR ) induces oxidative stress , which leads to acute lung injury . Heme oxygenase ( HO ) -1 ( EC 1.14.99.3 ) , the rate-limiting enzyme in heme catabolism , is inducible by oxidative stress and is thought to play an important role in the protection from oxidative tissue injuries . In this study , we examined expression of P09601 as well as tissue injuries in the lung , liver , and kidney after HSR in rats . We also pretreated animals with heme arginate ( HA ) , a strong inducer of P09601 , and examined its effect on the HSR-induced lung injury . P09601 expression significantly increased in the liver and kidney following HSR , while its expression in the lung was very low and unchanged after HSR . In contrast to P09601 expression , tissue injury and tumor necrosis factor-alpha ( P01375 ) gene expression was more prominent in the lung compared with those in the liver and kidney . HA pretreatment markedly induced P09601 in pulmonary epithelial cells , and ameliorated the lung injury induced by HSR as judged by the improvement of histological changes , while it decreased P01375 and inducible nitric oxide synthase gene expression , lung wet weight to dry weight ratio , and myeloperoxidase activity . In contrast , inhibition of P09601 by DB04912 administration abolished the beneficial effect of HA pretreatment . These findings suggest that tissues with higher P09601 may be better protected than those with lower P09601 from oxidative tissue injury induced by HSR . Our findings also indicate that HA pretreatment can significantly suppress the HSR-induced lung injury by virtue of its ability to induce P09601 . A possible mechanism of the nucleus accumbens and ventral pallidum P28222 receptors underlying the antidepressant action of ketamine : a PET study with macaques . DB01221 is a unique anesthetic reagent known to produce various psychotic symptoms . DB01221 has recently been reported to elicit a long-lasting antidepressant effect in patients with major depression . Although recent studies provide insight into the molecular mechanisms of the effects of ketamine , the antidepressant mechanism has not been fully elucidated . To understand the involvement of the brain serotonergic system in the actions of ketamine , we performed a positron emission tomography ( PET ) study on non-human primates . Four rhesus monkeys underwent PET studies with two serotonin ( 5-HT ) -related PET radioligands , [(11)C]AZ10419369 and [(11)C]DASB , which are highly selective for the P28222 receptor and serotonin transporter ( P31645 ) , respectively . Voxel-based analysis using standardized brain images revealed that ketamine administration significantly increased P28222 receptor binding in the nucleus accumbens and ventral pallidum , whereas it significantly reduced P31645 binding in these brain regions . DB00574 , a 5-HT releaser , significantly decreased P28222 receptor binding , but no additional effect was observed when it was administered with ketamine . Furthermore , pretreatment with 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)quinoxaline ( NBQX ) , a potent antagonist of the glutamate α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ( AMPA ) receptor , blocked the action of ketamine on the P28222 receptor but not P31645 binding . This indicates the involvement of AMPA receptor activation in ketamine-induced alterations of P28222 receptor binding . Because NBQX is known to block the antidepressant effect of ketamine in rodents , alterations in the serotonergic neurotransmission , particularly upregulation of postsynaptic P28222 receptors in the nucleus accumbens and ventral pallidum may be critically involved in the antidepressant action of ketamine . DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . Immunocytochemical localization of Q9UBS5 receptor subunits in the basolateral amygdala . Gamma-aminobutyric acid B ( GABAB ) receptors ( GBRs ) are G-protein-coupled receptors that mediate a slow , prolonged form of inhibition in the basolateral amygdala ( P00519 ) and other brain areas . Recent studies indicate that this receptor is a heterodimer consisting of Q9UBS5 ( GBR1 ) and O75899 subunits . In the present investigation , antibodies to the Q9UBS5 subunit were used to study the neuronal localization of GBRs in the rat P00519 . GBR immunoreactivity was mainly found in spine-sparse interneurons and astrocytes at the light microscopic level . Very few pyramidal neurons exhibited perikaryal staining . Dual-labeling immunofluorescence analysis indicated that each of the four main subpopulations of interneurons exhibited GBR immunoreactivity . Virtually 100 % of large CCK+ neurons in the basolateral and lateral nuclei were GBR+ . In the basolateral nucleus 72 % of somatostatin ( Q8TE85 ) , 73 % of parvalbumin ( PV ) and 25 % of P01282 positive interneurons were GBR+ . In the lateral nucleus 50 % of somatostatin , 30 % of parvalbumin and 27 % of P01282 positive interneurons were GBR+ . Electron microscopic ( EM ) analysis revealed that most of the light neuropil staining seen at the light microscopic level was due to the staining of dendritic shafts and spines , most of which probably belonged to spiny pyramidal cells . Very few axon terminals ( Ats ) were GBR+ . In summary , this investigation demonstrates that the distal dendrites of pyramidal cells , and varying percentages of each of the four main subpopulations of interneurons in the P00519 , express GBRs . Because previous studies suggest that GBR-mediated inhibition modulates DB01221 -dependent EPSPs in the P00519 , these receptors may play an important role in neuronal plasticity related to emotional learning . Systemic safety of anti- P15692 drugs : a commentary . INTRODUCTION : P15692 is a mediator of angiogenesis . Thus , concerns have been expressed following the use of P15692 inhibitors for the treatment of neovascular age-related macular degeneration ( nAMD ) . DB01270 , and more recently aflibercept , are P15692 inhibitors licensed for the treatment of nAMD . DB00112 is also used but unlicensed for this application . AREAS COVERED : A non-systematic review of nAMD trials was undertaken to investigate four outcomes : all-cause mortality , all systemic serious adverse events ( SSAEs ) , arteriothrombotic events ( ATEs ) and gastrointestinal ( GI ) complications . Differences in event rates with injections of ranibizumab compared to bevacizumab , aflibercept , photodynamic therapy ( PDT ) and sham were explored and quantified using fixed-effect meta-analyses . EXPERT OPINION : Anti- P15692 agents can influence vascular health ; however , the data suggest no difference in the risk of an ATE or death between anti- P15692 agents . Clinical trials are limited in their size and eligibility criteria and databases of patients treated in routine practice should also be scrutinized . P35858 : cytokine profile in cerebrospinal fluid T-cell clones . T -cells are present in the spinal cord from patients with amyotrophic lateral sclerosis ( P35858 ) , and could attack neurons or activate microglia through secretion of cytokines . We report that interferon ( IFN ) -gamma , tumour necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -2 , P05112 , P05113 and P22301 could not be detected in cerebrospinal fluid ( P04141 ) samples from 15 P35858 patients and 23 out of 25 controls with a multiplexed cytometric bead assay . In vivo activated T-cell clones were established from P04141 ( n = 26 ) and blood ( n = 21 ) of one P35858 patient . The proliferative capacity of P04141 T-cell clones was lower than that of T-cell clones from blood ( p = 0.0007 ) . All P01730 + P04141 T-cell clones produced P01579 , compatible with a predominant T helper ( h ) 1 phenotype , but several T-cell clones also produced Th2 cytokines . These data suggest that in vivo activated intrathecal T-cells can be induced to secrete cytokines which may play a role in P35858 . Epidermal growth factor as a local mediator of the neurotrophic action of vitamin B(12) ( cobalamin ) in the rat central nervous system . We have recently demonstrated that the myelinolytic lesions in the spinal cord ( SC ) of rats made deficient in vitamin B(12) ( cobalamin ) ( Cbl ) through total gastrectomy ( TG ) are tumor necrosis factor-alpha ( P01375 ) -mediated . We investigate whether or not permanent Cbl deficiency , induced in the rat either through TG or by chronic feeding of a Cbl-deficient diet , might modify the levels of three physiological neurotrophic factors-epidermal growth factor ( P01133 ) , vasoactive intestinal peptide ( P01282 ) , and somatostatin ( SS ) -in the cerebrospinal fluid ( P04141 ) of these rats . We also investigated the ability of the central nervous system ( CNS ) in these Cbl-deficient rats to synthesize P01133 mRNA and of the SC to take up labeled Cbl in vivo . Cbl-deficient rats , however the vitamin deficiency is induced , show a selective decrease in P01133 P04141 levels and an absence of P01133 mRNA in neurons and glia in various CNS areas . In contrast , radiolabeled Cbl is almost exclusively taken up by the SC white matter , but to a much higher degree in totally gastrectomized ( O43548 ) rats . Chronic administration of Cbl to O43548 rats restores to normal both the P01133 P04141 level and P01133 mRNA expression in the various CNS areas examined . This in vivo study presents the first evidence that the neurotrophic action of Cbl in the CNS of O43548 rats is mediated by stimulation of the P01133 synthesis in the CNS itself . It thus appears that Cbl inversely regulates the expression of P01133 and P01375 genes in the CNS of O43548 rats . Isoflurane attenuates LPS-induced acute lung injury by targeting miR-155- Q16665 . Isoflurane alleviates the inflammatory response in endotoxin-induced acute lung injury ( ALI ) . In this study , we investigated the protective mechanism of isoflurane postconditioning in lipopolysaccharide (LPS)induced ALI . Exposure to isoflurane decreased miR-155 and upregulated Q9BYW2 alpha and P09601 mRNA and protein . The effects of isoflurane on Q9BYW2 alpha mRNA and protein could be inhibited by overexpression of miR-155 . Furthermore , mice overexpressing miR-155 had higher levels of P01375 and P01584 in BALF when exposed to isoflurane after LPS challenge.Conversely , downregulation of miR-155 promoted isoflurane effects on Q9BYW2 alpha expression . These results suggest that isoflurane posttreatment hr alleviates LPS-induced ALI and cell injury by triggering miR-155- Q9BYW2 alpha pathway , leading to upregulation of P09601 . MnSOD drives neuroendocrine differentiation , androgen independence , and cell survival in prostate cancer cells . An increase in neuroendocrine ( NE ) cell number has been associated with progression of prostate tumor , one of the most frequent cancers among Western males . We previously reported that mitochondrial manganese superoxide dismutase ( MnSOD ) increases during the NE differentiation process . The goal of this study was to find whether MnSOD up-regulation is enough to induce NE differentiation . Several human prostate cancer LNCaP cell clones stably overexpressing MnSOD were characterized and two were selected ( MnSOD-S4 and MnSOD- P28222 ) . MnSOD overexpression induces NE morphological features as well as coexpression of the NE marker synaptophysin . Both MnSOD clones exhibit lower superoxide levels and higher H(2)O(2) levels . MnSOD-overexpressing cells show higher proliferation rates in complete medium , but in steroid-free medium MnSOD- P28222 cells are still capable of proliferation . MnSOD up-regulation decreases androgen receptor and prevents its nuclear translocation . MnSOD also induces up-regulation of Bcl-2 and prevents docetaxel- , etoposide- , or P01375 -induced cell death . Finally , MnSOD-overexpressing cells enhance growth of androgen-independent PC-3 cells but reduce growth of androgen-dependent cells . These results indicate that redox modulation caused by MnSOD overexpression explains most NE-like features , including morphological changes , NE marker expression , androgen independence , inhibition of apoptosis , and enhancement of cell growth . Many of these events can be associated with the androgen dependent-independent transition during prostate cancer progression . Antiinflammatory effect of lactic acid bacteria : inhibition of cyclooxygenase-2 by suppressing nuclear factor-kappaB in Raw264.7 macrophage cells . Lactobacillus casei 3260 ( L. casei 3260 ) was evaluated in relation to the inflammatory response mediated by lipopolysaccharide ( LPS ) -induced nuclear factor-kappaB ( NF-kappaB ) and cyclooxygenase-2 ( P35354 ) expression in Raw264.7 macrophage cells . The treatment of Raw264.7 cells with L. casei 3260 significantly inhibited the secretion of tumor necrosis factor-alpha ( P01375 ) and prostaglandins E2 ( DB00917 ) , followed by suppression of P35354 . To clarify the molecular mechanism , the inhibitory effect of L. casei 3260 on the NF-kappaB signaling pathway was examined based on the luciferase reporter activity . Although the treatment of Raw264.7 cells with L. casei 3260 did not affect the transcriptional activity of NF-kappaB , it did inhibit NF-kappaB activation , as determined by the cytosolic p65 release and degradation of I-kappaBalpha . Therefore , these findings suggest that the suppression of P35354 through inhibiting the NF-kappaB activation by LPS may be associated with the antiinflammatory effects of L. casei 3260 on Raw264.7 cells . The glutamate/neutral amino acid transporter family Q99705 : molecular , physiological and pharmacological aspects . The solute carrier family 1 ( Q99705 ) includes five high-affinity glutamate transporters , P43005 , GLT-1 , P43003 , P48664 and O00341 ( P43005 , P43004 , P43003 , P48664 , and O00341 , respectively ) as well as the two neutral amino acid transporters , P43007 and Q15758 ( P43007 and ALC1A5 , respectively ) . Although each of these transporters have similar predicted structures , they exhibit distinct functional properties which are variations of a common transport mechanism . The high-affinity glutamate transporters mediate transport of l- DB00142 , l- DB00128 and d- DB00128 , accompanied by the cotransport of 3 Na(+) and 1 H(+) , and the countertransport of 1 K(+) , whereas ASC transporters mediate Na(+)-dependent exchange of small neutral amino acids such as Ala , DB00133 , DB00151 and DB00156 . The unique coupling of the glutamate transporters allows uphill transport of glutamate into cells against a concentration gradient . This feature plays a crucial role in protecting neurons against glutamate excitotoxicity in the central nervous system . During pathological conditions , such as brain ischemia ( e.g. after a stroke ) , however , glutamate exit can occur due to " reversed glutamate transport " , which is caused by a reversal of the electrochemical gradients of the coupling ions . Selective inhibition of the neuronal glutamate transporter P43005 ( P43005 ) may be of therapeutic interest to block glutamate release from neurons during ischemia . On the other hand , upregulation of the glial glutamate transporter P43004 ( P43004 ) may help protect motor neurons in patients with amyotrophic lateral sclerosis ( P35858 ) , since loss of function of P43004 has been associated with the pathogenesis of certain forms of P35858 . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients .

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interacts_with DB00472?

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DB09372 mitigates the DMBA/croton oil-induced skin cancer progression in mice . Skin cancer is the most common malignancy in the world and also one of the major causes of death worldwide . The toxic environmental pollutant 7,12-dimethylbenz[a]anthracene ( DMBA ) is a skin-specific carcinogen . DB09372 ( TA ) is reported to be effective against various types of chemical-induced toxicities and carcinogenesis as well . In the present study , we have evaluated the therapeutic potential of tannic acid in DMBA + croton oil-induced skin cancer in Swiss albino mice . Protective effect of TA against skin cancer was evaluated in terms of antioxidant enzymes activities , lipid peroxidation , histopathological changes and expression of inflammation and early tumour markers . DMBA + croton oil causes depletion of antioxidant enzymes ( p < 0.001 ) and elevation of early inflammatory and tumour promotional events . TA prevents the DMBA + croton oil-induced toxicity through a protective mechanism that involves the reduction of oxidative stress as well as P35354 , i-NOS , P12004 protein expression and level of proinflammatory cytokine such as P05231 release at a very significant level ( p < 0.001 ) . It could be concluded from our results that TA attenuates DMBA + croton oil-induced tumour promotional potential possibly by inhibiting oxidative and inflammatory responses and acts as antioxidant , anti-inflammatory and antiproliferative agent . [ Effect of NF-kappaB on inhibition of non-small cell lung cancer cell cyclooxygenase-2 by brucine ] . OBJECTIVE : To study the molecular mechanism of cyclooxygenase-2 ( P35354 ) , one of effective ingredient of brucine , in inducing non-small cell lung cancer cell apoptosis . METHOD : P35354 promoter , transcription factor deletion mutants and P35354 mRNA 3'-UTR-containing report plasmids were transfected with Renillia to non-small cell lung cancer A549 cell , in order to detect the activity of report gene luciferase and minimum cis-acting element of P35354 promoter inhibited by brucine . The influence of brucine on IkappaB phosphorylation and the nuclear translocation of p65 were detected by immunoblotting assay . RESULT : Brucine significantly suppressed LPS-induced P35354 promoter activation , but revealed minor impact on P35354 mRNA stability . NF-kappaB in the vicinity of P35354 promoter-262 was an important cis-acting element of brucine for inhibiting the activity of P35354 promoter . Brucine was found to inhibit the phosphorylation of P25963 as well as the nuclear translocation of p65 . CONCLUSION : Brucine can improve A549 cells apoptosis by inhibiting the activity of NF-kappaB and the subsequent P35354 gene expression . Solubilized active pituitary and ovarian gonadotropin-releasing hormone receptors retain binding properties for adenosine 3',5'-cyclic monophosphate derivatives . The non-denaturing zwitterionic detergent , ( 3 ( 3- cholamidopropyl ) -dimethyl- ammonio ) -1-propane sulfonate ( CHAPS ) , has been used to solubilize membrane gonadotropin-releasing hormone ( DB00644 ) receptors from rat ovaries . The solubilized receptors retain a high affinity ( Ka = 1.85 +/- 0.3 nM-1 ) , comparable to the affinity measured in membrane particles ( Ka = 3.25 +/- 0.7 nM-1 ) , and a preserved specificity for several analogs and fragments of DB00644 . At millimolar concentrations , cyclic AMP derivatives inhibit [ 125I ] - DB00644 analog binding to both membrane particles and soluble receptors from pituitary and ovary . These results support the hypothesis that cyclic AMP may play the role of an extracellular messenger by interacting with the P30968 itself . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . Vitamin D increases plasma renin activity independently of plasma Ca2+ via hypovolemia and β-adrenergic activity . 1 , 25-Dihydroxycholechalciferol ( calcitriol ) and 19-nor-1 , 25-dihydroxyvitamin D2 ( paricalcitol ) are vitamin D receptor ( P11473 ) agonists . Previous data suggest P11473 agonists may actually increase renin-angiotensin activity , and this has always been assumed to be mediated by hypercalcemia . We hypothesized that calcitriol and paricalcitol would increase plasma renin activity ( P06703 ) independently of plasma Ca(2+) via hypercalciuria-mediated polyuria , hypovolemia , and subsequent increased β-adrenergic sympathetic activity . We found that both calcitriol and paricalcitol increased P06703 threefold ( P < 0.01 ) . Calcitriol caused hypercalcemia , but paricalcitol did not . Both calcitriol and paricalcitol caused hypercalciuria ( 9- and 7-fold vs. control , P < 0.01 ) and polyuria ( increasing 2.6- and 2.2-fold vs. control , P < 0.01 ) . Paricalcitol increased renal calcium-sensing receptor ( P41180 ) expression , suggesting a potential cause of paricalcitol-mediated hypercalciuria and polyuria . Volume replacement completely normalized calcitriol-stimulated P06703 and lowered plasma epinephrine by 43 % ( P < 0.05 ) . β-Adrenergic blockade also normalized calcitriol-stimulated P06703 . P35354 inhibition had no effect on calcitriol-stimulated P06703 . Our data demonstrate that vitamin D increases P06703 independently of plasma Ca(2+) via hypercalciuria , polyuria , hypovolemia , and increased β-adrenergic activity . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV-1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE3-tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e.g. , all-trans-retinoic acid ( RA ) , 13-cis-retinoic acid ( 13- DB00982 ) , arotinoid acid ( DB02877 ) and m-carboxy-arotinoid acid ( m-carboxy- DB02877 , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 ( 24 nM ) greater than P10826 ( 4.0 nM ) greater than P13631 ( 1.3 nM ) , while the ED50 values for DB02877 and 13- DB00982 decreased in the order of P10276 ( 6.5 nM , 190 nM ) greater than P13631 ( 2.3 nM , 140 nM ) greater than P10826 ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy- DB02877 , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 greater than P10826 greater than P13631 . P08183 , Q9UNQ0 , and P60484 determine the response of glioblastoma to temozolomide and ABT-888 therapy . PURPOSE : Little is known about the optimal clinical use of ABT-888 ( veliparib ) for treatment of glioblastoma . ABT-888 is a PARP inhibitor undergoing extensive clinical evaluation in glioblastoma , because it may synergize with the standard-of-care temozolomide ( DB00853 ) . We have elucidated important factors controlling ABT-888 efficacy in glioblastoma . EXPERIMENTAL DESIGN : We used genetically engineered spontaneous glioblastoma mouse models and allograft models that were orthotopically transplanted into wild-type ( WT ) and Abcb1/Abcg2-deficient ( KO ) recipients . RESULTS : ABT-888/ DB00853 is not efficacious against p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR allografts in wild-type recipients , indicating inherent resistance . Abcb1/Abcg2 mediated efflux of ABT-888 at the blood-brain barrier ( BBB ) causes a 5-fold reduction of ABT-888 brain penetration ( P < 0.0001 ) that was fully reversible by elacridar . Efficacy studies in WT and KO recipients and/or concomitant elacridar demonstrate that Abcb1/Abcg2 at the BBB and in tumor cells impair DB00853 /ABT-888 combination treatment efficacy . DB04881 also markedly improved DB00853 /ABT-888 combination treatment in the spontaneous p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR glioblastoma model . Importantly , ABT-888 does enhance DB00853 efficacy in Pten deficient glioblastoma allografts and spontaneous tumors , even in Abcb1/Abcg2 proficient wild-type mice . Loss of P60484 occurs frequently in glioblastoma ( 36 % ) and in silico analysis on patient with glioblastoma samples revealed that it is associated with a worse overall survival ( 310 days vs. 620 days , n = 117 ) . CONCLUSIONS : The potential of ABT-888 in glioblastoma can best be demonstrated in patients with P60484 null tumors . Therefore , clinical trials with ABT-888 should evaluate these patients as a separate group . Importantly , inhibition of P08183 and Q9UNQ0 ( by elacridar ) may improve the efficacy of DB00853 /ABT-888 therapy in all glioblastoma patients . Tat-3L4F does not significantly affect spatial learning and memory . We recently found that the interfering peptide Tat-3L4F is able not only to disrupt the protein-protein interaction of P60484 ( phosphatase and tensin homologue deleted on chromosome 10 ) with the serotonin P28335 receptor in the rat ventral tegmental area ( VTA ) but also to suppress the conditioned place preference induced by cannabinoid and nicotine without significant effects on anxiety , feeding behavior and motor activity . It is unknown , however , whether Tat-3L4F affects learning and memory . Using a Morris water maze test , we show here that while the cannabinoid HU210 significantly inhibited the performance of spatial learning without significant effects on long-term memory , Tat-3L4F did not induce significant effects on the acquisition and retrieval of spatial memory . These results indicate that Tat-3L4F can suppress the rewarding effects of abused drugs without significant effects on learning and memory . Expression of type I P01148 receptor and in vivo and in vitro P01148 -I effects in corpora lutea of pseudopregnant rabbits . The expression of type I P01148 receptor ( P30968 -I ) and the direct role of P01148 -I on corpora lutea ( CL ) function were studied in the pseudopregnant rabbit model . Immunohistochemistry evidenced P30968 -I and P01148 -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 -I mRNA and protein at the three luteal stages . DB06719 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 (2α) ) secretion and cyclo-oxygenase-2 ( P35354 ) and nitric oxide synthase ( NOS ) activities at days 9 and 13 , and decreased PGE₂ at day 13 . Co-incubation with antagonists for P01148 -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 , compound 48/80 ) , and protein kinase C ( PKC , staurosporine ) counteracted the buserelin effects . DB06719 co-incubated with P36551 inhibitor ( acetylsalicylic acid ) increased progesterone and decreased P49763 (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 -I is constitutively expressed in rabbit CL independently of luteal stage , whereas P01148 -I down-regulates directly CL progesterone production via P49763 (2α) at mid- and late-luteal stages of pseudopregnancy , utilizing its cognate type I receptor with a post-receptorial mechanism that involves P98160 , IP₃ , DAG , PKC , P35354 , and NOS . Novel pyrrolyllactone and pyrrolyllactam indolinones as potent cyclin-dependent kinase 2 inhibitors . P12004 -dependent kinases ( CDKs ) are essential in the control of cell cycle progression . Inhibition of CDKs represents a new approach for pharmacological intervention in the treatment of a variety of proliferative diseases , especially cancer . Based on the crystal structure of P24941 in complex with an imidazole indolinone compound 1 ( DB03428 ) , lead optimization through modeling , synthesis , and SAR studies has led to the discovery of a novel series of pyrrolyllactone and pyrrolyllactam indolinones as potent P24941 inhibitors . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . The effects of a cyclooxygenase-2 ( P35354 ) expression and inhibition on human uveal melanoma cell proliferation and macrophage nitric oxide production . BACKGROUND : P35354 ( P35354 ) expression has previously been identified in uveal melanoma although the biological role of P35354 in this intraocular malignancy has not been elucidated . This study aimed to investigate the effect of a P35354 inhibitor on the proliferation rate of human uveal melanoma cells , as well as its effect on the cytotoxic response of macrophages . METHODS : Human uveal melanoma cell lines were transfected to constitutively express P35354 and the proliferative rate of these cells using two different methods , with and without the addition of Amfenac , was measured . DB00435 production by macrophages was measured after exposure to melanoma-conditioned medium from both groups of cells as well as with and without Amfenac , the active metabolite of DB06802 . RESULTS : Cells transfected to express P35354 had a higher proliferation rate than those that did not . The addition of Amfenac significantly decreased the proliferation rate of all cell lines . DB00435 production by macrophages was inhibited by the addition of melanoma conditioned medium , the addition of Amfenac partially overcame this inhibition . CONCLUSION : Amfenac affected both P35354 transfected and non-transfected uveal melanoma cells in terms of their proliferation rates as well as their suppressive effects on macrophage cytotoxic activity . Induction of retinoic acid receptor-beta suppresses cyclooxygenase-2 expression in esophageal cancer cells . Since retinoic acid receptor ( RAR ) -beta mRNA is frequently lost during esophageal carcinogenesis and esophageal cancer cells that do not express P10826 are resistant to retinoic acid ( RA ) , we stably transfected P10826 expression vector into an esophageal cancer cell line TE-8 and an antisense P10826 into TE-3 cells . Transfection of P10826 decreased cell growth and colony formation and induced apoptosis in TE-8 cells . Antisense P10826 -transfected TE-3 cells had a shorter doubling time and became resistant to RA . Induction of P10826 decreased P35354 expression in P10826 transfected TE-8 cells , whereas antisense P10826 transfected TE-3 cells increased P35354 expression . The inhibitory effect of P10826 on P35354 expression was further enhanced in the presence of RA , which was blocked by an RAR antagonist . The synthetic retinoid N-(4-hydroxyphenyl)retinamide , which does not bind effectively to P10826 , had no effect on P35354 suppression . Furthermore , RA blocked bile acid-induced P35354 expression and prostaglandin E(2) production only in the P10826 positive cells . Our data demonstrated that anticancer effect of P10826 may be related to its ability to suppress P35354 expression and support that the loss of P10826 expression may contribute to esophageal carcinogenesis . DB04998 inhibits activation of nuclear factor-kappaB ( NF-kappaB ) by forming a complex with NF-kappaB essential modulator ( Q9Y6K9 ) and nucleolin . DB04998 , also known as DB04998 , is an experimental anticancer drug that recently entered human clinical trials . It is a member of a novel class of antiproliferative agents known as G-rich oligonucleotides ( P09341 ) , which are non-antisense , guanosine-rich phosphodiester oligodeoxynucleotides that form stable G-quadruplex structures . The biological activity of GROs results from their binding to specific cellular proteins as aptamers . One important target protein of GROs has been previously identified as nucleolin , a multifunctional protein expressed at high levels by cancer cells . Here , we report that DB04998 also associates with nuclear factor-kappaB ( NF-kappaB ) essential modulator ( Q9Y6K9 ) , which is a regulatory subunit of the inhibitor of kappaB ( IkappaB ) kinase ( IKK ) complex , and also called IKKgamma . In the classic NF-kappaB pathway , the IKK complex is required for phosphorylation of P25963 and subsequent activation of the transcription factor NF-kappaB . We found that treatment of cancer cells with DB04998 inhibits IKK activity and reduces phosphorylation of P25963 in response to tumor necrosis factor-alpha stimulation . Using a reporter gene assay , we showed that DB04998 blocks both tumor necrosis factor-alpha-induced and constitutive NF-kappaB activity in human cancer cell lines derived from cervical , prostate , breast , and lung carcinomas . In addition , we showed that , in DB04998 -treated cancer cells , Q9Y6K9 is coprecipitated by nucleolin , indicating that both proteins are present in the same complex . Our studies suggest that abrogation of NF-kappaB activity may contribute to the anticancer effects of DB04998 and that nucleolin may play a previously unknown role in regulating the NF-kappaB pathway . DB00163 , P09917 and oxidative stress in haemodialysis patients : facts , not fancies . Clinical utility of ramucirumab in advanced gastric cancer . Gastric cancer is currently the third most common cause of cancer deaths worldwide . Prognosis remains poor with most patients presenting with advanced or metastatic disease . A better understanding of angiogenesis has led to the investigation of drugs that inhibit the vascular endothelial growth factor ( P15692 ) pathway including anti- P15692 antibody therapy ( eg , bevacizumab ) , inhibitors of angiogenic receptor tyrosine kinases ( eg , sunitinib , sorafenib , apatinib , regorafenib ) , and inhibitors of vascular endothelial growth factor receptors ( VEGFRs ) ( eg , ramucirumab ) . DB05578 , a P35968 inhibitor , is the first anti-angiogenic agent approved by the US Food and Drug Administration for use in the treatment of advanced gastric cancers . This review will focus on the clinical utility and potential use of ramucirumab in advanced gastric cancer .

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[ Circumventing multidrug resistance in human cancer by anti-ribozyme ] . The demonstration tha RNA can be cleavaged by cis-ribozyme ( catalytic RNAs , RNA enzyme ) has potentially important therapeutic implications . Ribozymes are effective for modulation of gene expression because of their simple structure , site-specific cleavage activity and catalytic potential . The targets of ribozyme-mediated gene modulation have ranged from cancer cells to foreign genes that cause infectious diseases . Additional target sites for ribozymes are in initial phases of development and design . Ribozymes have been targeted against myriad genes , including oncogenes ( ras , P11274 - P00519 ) and drug resistance genes ( MDR-1 , c-fos , P00374 ) . These ribozymes have cleaved the target RNAs in culture system ( in vitro ) and developed in vivo system . We reported that anti-fos ribozyme has altered the expression of c-fos and DNA repair genes in cisplatin-resistance cancer cells , and reversed the sensitivity to ciaplatin . Furthermore , we have developed high efficiency by the transfer system using an electroporation in vivo . DB00619 inhibition effect on KITAsn822Lys-mediated signal transduction cascade . OBJECTIVE : Alterations in growth factor signaling pathways may be a frequent collaborating event in Q01196 - Q06455 -mediated leukemogenesis . Gain-of-function P10721 receptor mutations have been reported in adult AML patients , especially those with core binding factor leukemia ( CBFL ) . We have previously reported a new gain-of-function P10721 (Asn822Lys) mutation that is constitutively expressed in the Kasumi-1 CBFL cell line , and has recently been described in two childhood AML patients . To explore the molecular basis of the effects of this mutation in the appropriate context of hemopoietic dysregulation , we investigated P10721 downstream signaling in the Kasumi-1 cell line by means of DB00619 ( Imatinib , Gleevec ) pharmacological inhibition . MATERIALS AND METHODS : We investigated P10721 (Asn822Lys) mutant-initiated signaling in Kasumi-1 cell line , and characterized the inhibitory effect of the DB00619 protein tyrosine kinase inhibitor on downstream signaling . RESULTS : The use of DB00619 -mediated inhibition impaired the tyrosine phosphorylation of P10721 (Asn822Lys) and its association with the p85 subunit of phosphatidylinositol 3'-kinase ( p85PI3K ) . The downstream constitutive phosphorylation of P45983 /2 and P40763 was also significantly inhibited , but DB00619 had no effect on the constitutive activation of Akt , thus suggesting that it is due to other signaling in Kasumi-1 cells . DB00619 inhibited the P10721 -mediated proliferation of Kasumi-1 cells in a dose-dependent manner . CONCLUSIONS : These findings show the role of PI3K in P10721 (Asn822Lys)-mediated constitutive activation through the Akt-independent downstream signaling pathway of JNK , and also demonstrate the mutant 's susceptibility to DB00619 , which may therefore have therapeutic potential in CBFL patients with susceptible P10721 mutations . Sequential activation of JAKs , STATs and xanthine dehydrogenase/oxidase by hypoxia in lung microvascular endothelial cells . P47989 ( P47989 /XO ) is associated with various pathological conditions related to the endothelial injury . However , the molecular mechanism underlying the activation of P47989 /XO by hypoxia remains largely unknown . In this report , we determined whether the Janus kinases ( JAKs ) and signal transducers and activators of transcription ( STATs ) signaling pathway is involved in hypoxia-induced activation of P47989 /XO in primary cultures of lung microvascular endothelial cells ( LMVEC ) . We found that hypoxia significantly increased interleukin 6 ( P05231 ) production in a time-dependent manner in LMVEC . Hypoxia also markedly augmented phosphorylation/activation of JAKs ( P23458 , O60674 and P52333 ) and the JAK downstream effectors STATs ( P40763 and P42229 ) . Hypoxia-induced activation of P40763 was blocked by P05231 antibodies , the JAK inhibitor AG490 and the suppressor of cytokine signaling 3 ( O14543 ) , implying that hypoxia-promoted P05231 secretion activates the JAK/ P35610 pathway in LMVEC . Phosphorylation and DNA-binding activity of P40763 were also inhibited by the p38 MAPK inhibitor SB203580 and the phosphatidylinositol 3-kinase inhibitor LY294002 , suggesting that multiple signaling pathways involved in P35610 activation by hypoxia . Importantly , hypoxia promoted P47989 /XO activation in LMVEC , which was markedly reversed by inhibiting the JAK- P35610 pathway using P05231 antibodies , AG490 and O14543 . These data demonstrated that JAKs , STATs and P47989 /XO were sequentially activated by hypoxia . These data provide the first evidence indicating that the JAK- P35610 pathway is involved in hypoxia-mediated P47989 /XO activation in LMVEC . Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . Mobilization of Ph chromosome-negative peripheral blood stem cells in chronic myeloid leukaemia patients with imatinib mesylate-induced complete cytogenetic remission . Imatinib mesylate ( IM , DB00619 , Glivec ) can induce a high rate of complete cytogenetic response ( CCR ) in chronic myeloid leukaemia ( CML ) patients , although to date the majority of patients continue to have detectable disease by sensitive reverse transcription polymerase chain reaction ( RT-PCR ) . It is therefore possible that these patients may ultimately relapse and require treatment such as autologous peripheral blood stem cell transplant ( APBSCT ) . We attempted mobilization of haemopoietic progenitor cells from 58 patients in CCR using recombinant human granulocyte colony-stimulating factor [ rHu- DB00099 ; 10 micro g/kg/d subcutaneously ( s.c. ) for at least 4 d ] alone , while continuing IM treatment . The median d 5 ( peak ) P28906 + count was 11.5/ microl ( range 0-108/ microl ) , and 43/58 ( 74 % ) patients underwent a median of two ( range 1-3 ) apheresis procedures . A median dose of 2.1 x 10(6)/kg P28906 + cells ( range 0.1-6.5 x 10(6)/kg ) was collected . Some 84 % of 31 collections analysed were negative for the Philadelphia ( Ph ) chromosome or breakpoint cluster region and Abelson murine leukaemia viral oncogene homologue ( P11274 - P00519 ) translocation by cytogenetics or fluorescent in situ hybridization respectively . No toxicity was reported with the regimen . Overall , the target P28906 + dose ( 2 x 10(6)/kg P28906 + ) was attained in 23/58 ( 40 % ) patients who entered the study . In summary , we have demonstrated that successful mobilization of Ph- P28906 + cells from IM-treated patients in CCR is possible using rHu-G- P04141 alone . ICE/ P29466 inhibitors as novel anti-inflammatory drugs . In recent years , several strategies that selectively inhibit pro-inflammatory cytokines , have yielded effective protein-based therapies for inflammatory disorders , validating the therapeutic hypothesis that intervention in cytokine signalling can provide clinical benefit . However , these protein-based products must be administered by injection , a constraint associated with inconvenience , adverse effects and expense for patients , caregivers and insurers . Besides interfering with the effects of cytokines such as P01375 or IL-1beta that have already been produced , inhibition of pro-inflammatory cytokine production or signalling with low-molecular weight orally-active drugs would combine the convenience of conventional pharmaceuticals with the focused efficacy of the protein therapies . Reducing IL-1beta and Q14116 production by inhibition of IL-1beta converting enzyme ( ICE , caspase-1 ) is one promising strategy because of the key roles of these cytokines in many inflammatory diseases . DB04875 , the first orally available , potent and selective ICE inhibitor to enter clinical trials , is currently under investigation in rheumatoid arthritis . Effect of the hemoregulatory peptide (pEEDCK)2 (pyroGlu- DB00142 - DB00128 - DB00151 -Lys)2 and MIP-1alpha is reduced in bone marrow cultures from patients with chronic myeloid leukemia ( CML ) . The granulocyte-derived hemoregulatory peptide pyroGlu- DB00142 - DB00128 - DB00151 -Lys = pEEDCK is known to keep hematopoietic cells quiescent . When oxidized to its dimeric form (pEEDCK)2 , it activates growth of hematopoietic progenitors in association with stroma-derived cytokines . (pEEDCK)2 has a DB00151 - DB00151 motif which is also a typical feature of the macrophage inflammatory protein ( MIP-1alpha ) . The present study was designed to analyze differences between the response of normal and leukemic progenitor cells to (pEEDCK)2 or MIP-1alpha . When long-term bone marrow cultures ( LTBMCs ) were incubated with (pEEDCK)2 or MIP-1alpha and/or cytokines , the stimulatory effect on colony-forming units-granulocyte/erythroid/macrophage/megakaryocyte of LTBMC from chronic myeloid leukemia ( CML ) patients was less than 50 % compared to LTBMC from healthy humans . No difference in oncogene expression could be observed in LTBMC from CML patients regarding reduction of Philadelphia chromosome-associated transcription of the P11274 - P00519 gene . With respect to the expression of growth and differentiation-associated genes ( Galpha16 , P09917 , phospholipaseA2 , c-kit , and P28906 ) , which were analyzed from LTBMC by semiquantitative reverse transcriptase-polymerase chain reaction , the same transcription rate was observed in CML patients and healthy donors . However , two isoforms of a key enzyme of oxidative metabolism , carnitine palmitoyltransferase ( P50416 and Q92523 ) , showed 50-fold higher expression rates in LTBMC cells of healthy donors compared to CML patients . It is known that a decrease in oxidative metabolism is associated with an increase in redox equivalents in malignancy . This might result in a reduction of disulphide bonds in (pEEDCK)2 or MIP-1alpha , thus inducing a downregulation of these factors in bone marrow from CML patients . Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) -positive leukemia cell line with common-B cell phenotype , designated TMD5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD5 cells expressed 190 kDa P11274 / P00519 chimeric protein and 145 kDa P00519 protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM- P04141 , P08700 , P05231 , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia . The P11274 / P00519 -inhibitors imatinib , nilotinib and dasatinib differentially affect NK cell reactivity . In chronic myeloid leukemia ( CML ) , P11274 / P00519 -mediated oncogenic signaling can be targeted with the P11274 / P00519 -inhibitors Imatinib , DB04868 and Dasatinib . However , these agents may also affect anti-tumor immunity . Here , we analyzed the effects of the 3 P11274 / P00519 -inhibitors on natural killer ( NK ) cell reactivity . Exposure of CML cells ( K562 , Meg-01 ) to pharmacological concentrations of Imatinib , DB04868 and Dasatinib diminished expression of ligands for the activating immunoreceptor P26718 to a similar extent . This resulted in comparably reduced NK cell cytotoxicity and P01579 production . When direct effects on NK cell responses to K562 and primary CML cells as well as activating cytokines were studied , Dasatinib was found to abrogate NK cytotoxicity and cytokine production . DB04868 did not alter cytotoxicity but , at high levels , impaired NK cytokine production , while Imatinib had no direct influence on NK cell reactivity . Of note , DB04868 , but not the other P11274 / P00519 -inhibitors increased cell death within the preferentially cytokine-secreting CD56(bright)CD16(-) NK cell subset , which may , at least in part , serve to explain the effect of DB04868 on NK cytokine production . Analysis of NK cell signaling revealed that Dasatinib inhibited proximal signaling events leading to decreased phosphorylation of PI3K and P29323 that are crucial for NK cell reactivity . Imatinib and DB04868 , in contrast , showed no relevant effect on NK cell PI3K or P29323 activity . In light of the potential role of NK cells in the immunesurveillance of residual leukemia and for future combinatory immunotherapeutic approaches , our data indicate that choice and dosing of the most suitable P11274 / P00519 -inhibitor for a given patient require careful consideration . DB03886 -dependent hyperphenylalaninemia due to deficiency of 6-pyruvoyl tetrahydropterin synthase . We describe the clinical , neurologic , and biochemical findings in 10 patients with 6-pyruvoyl tetrahydropterin synthase ( 6- Q03393 ) deficiency from seven families , all of whom originate from one large tribe in Saudi Arabia . This deficiency presents with severe , early onset of failure to thrive , neurologic deterioration , and morbidity and mortality secondary to repeated episodes of bronchopneumonia or cardiorespiratory abnormalities . The urinary pterin excretion pattern indicates deficient activity of 6- Q03393 , which has been confirmed by direct enzyme assay in red blood cells of three patients . We treated our patients with combined use of tetrahydrobiopterin 20 mg/kg/d , L-dihydroxyphenylalanine 15 mg/kg/d , carbidopa 3.75 mg/kg/d , and L-5-hydroxytryptophan 5 mg/kg/d . Neurologic findings improved significantly in all after 5 to 24 months . Although head circumference and weight returned to the lower limit of normal in four , height normalized only in one of seven patients . Despite an unrestricted diet during combined therapy , blood phenylalanine and urinary excretion of neopterin and biopterin returned to normal . DB01892 is a dual inhibitor of cyclooxygenase-1 and P09917 . The acylphloroglucinol derivative hyperforin is the major lipophilic constituent in the herb Hypericum perforatum ( St. John 's wort ) . The aim of the present study was to investigate if hyperforin as well as extracts of H. perforatum can suppresses the activities of P09917 ( P09917 ) and cyclooxygenases ( P36551 ) , key enzymes in the formation of proinflammatory eicosanoids from arachidonic acid ( AA ) . In freshly isolated human polymorphonuclear leukocytes stimulated with Ca(2+) ionophore A23187 , hyperforin inhibited P09917 product formation with IC(50) values of about 1-2 microM , in the absence or presence of exogenous AA ( 20 microM ) , respectively , being almost equipotent to the well-documented P09917 inhibitor zileuton ( IC(50) = 0.5-1 microM ) . Experiments with purified human P09917 demonstrate that hyperforin is a direct P09917 inhibitor ( IC(50) approximately 90 nM ) , acting in an uncompetitive fashion . In thrombin- or ionophore-stimulated human platelets , hyperforin suppressed P23219 product ( 12(S)-hydroxyheptadecatrienoic acid ) formation with an IC(50) of 0.3 and 3 microM , respectively , being about 3- to 18-fold more potent than aspirin . At similar concentrations , hyperforin suppressed P23219 activity in platelets in presence of exogenous AA ( 20 microM ) as well as in cell-free systems . DB01892 could not interfere with P35354 product formation and did not significantly inhibit 12- or 15-LO in platelets or leukocytes , respectively . We conclude that hyperforin acts as a dual inhibitor of P09917 and P23219 in intact cells as well as on the catalytic activity of the crude enzymes , suggesting therapeutic potential in inflammatory and allergic diseases connected to eicosanoids . Antifolate resistance due to new and known Plasmodium falciparum dihydrofolate reductase mutations expressed in yeast . Two new dihydrofolate reductase ( P00374 ) mutations were recently discovered in Plasmodium falciparum samples from an area of Bolivia with high rates of in vivo resistance to pyrimethamine-sulfadoxine : a DB00151 --> DB00125 point mutation in codon 50 and a five amino acid insertion after codon 30 , termed the Bolivia repeat . We used a yeast expression system to screen these new P00374 mutants , as well as all of the other known P00374 mutant genotypes , against four antifolates : pyrimethamine , cycloguanil , chlorcycloguanil , and WR99210 . The prodrug proguanil was also evaluated . The primary 108- DB00174 mutation , the known secondary mutations 51- DB00167 , 59- DB00125 and 164- DB00149 , as well as the 50- DB00125 mutation , all progressively enhanced pyrimethamine resistance in naturally observed combinations with one another , with the presence of 164- DB00149 most significantly increasing resistance . Cycloguanil and chlorcycloguanil resistance were most impacted by 164- DB00149 and the paired 16- DB00161 /108- DB00156 . DB01131 had no effect on malaria P00374 . All DHFRs analyzed were sensitive to WR99210 . The Bolivia repeat did not markedly affect drug sensitivity . We conclude that malaria P00374 can be reliably , rapidly and inexpensively analyzed in yeast for activity against a broad spectrum of antifolates . This system may be useful for initially characterizing newly discovered genotypes before proceeding to P. falciparum transfection ; for large-scale geographic surveys of drug resistance ; and for screening new antifolates or new antifolate combinations for their effectiveness against a large panel of P00374 mutants . P04141 -1 ( P09603 ) delivers a proatherogenic signal to human macrophages . P09603 / P09603 supports the proliferation and differentiation of monocytes and macrophages . In mice , P09603 also promotes proinflammatory responses in vivo by regulating mature macrophage functions , but little is known about the acute effects of this growth factor on mature human macrophages . Here , we show that in contrast to its effects on mouse bone marrow-derived macrophages , P09603 did not induce expression of urokinase plasminogen activator mRNA , repress expression of apolipoprotein E mRNA , or prime LPS-induced P01375 and P05231 secretion in human monocyte-derived macrophages ( HMDM ) from several independent donors . Instead , we show by expression profiling that P09603 modulates the HMDM transcriptome to favor a proatherogenic environment . P09603 induced expression of the proatherogenic chemokines P02778 /IFN-inducible protein 10 , P13500 , and P80098 but repressed expression of the antiatherogenic chemokine receptor P61073 . P09603 also up-regulated genes encoding enzymes of the cholesterol biosynthetic pathway ( P04035 , P53602 , Q13907 , P14324 , Q14534 , Q16850 , EBP , Q15738 , Q9UBM7 , and Q15392 ) , and expression of P45844 , encoding a cholesterol efflux transporter , was repressed . Consistent with these effects , P09603 increased levels of free cholesterol in HMDM , and the selective P07333 kinase inhibitor GW2580 ablated this response . These data demonstrate that P09603 represents a further link between inflammation and cardiovascular disease and suggest two distinct mechanisms by which P09603 , which is known to be present in atherosclerotic lesions , may contribute to plaque progression . DB00399 -induced IPP/ApppI production in vivo . Bisphosphonates are currently the most important class of anti-resorptive drugs used for the treatment of diseases involving excess bone resorption . Recently we discovered a new mechanism of action for bisphosphonates . Previously it has been shown that nitrogen-containing bisphosphonates ( N-BPs ) are not metabolized . However , our studies revealed that N-BPs induce formation of a novel pro-apoptotic DB00171 analog ( ApppI ) , as a consequence of the inhibition of P14324 in the mevalonate pathway , and the subsequent accumulation of isopentenyl pyrophosphate ( IPP ) in vitro . The primary aim of the current study was to determine whether zoledronic acid ( a N-BP ) induces IPP/ApppI formation in vivo . Mass spectrometry was used to identify whether in vivo administration of zoledronic acid-induced IPP/ApppI production by mouse peritoneal macrophages or bone marrow cells . IPP/ApppI could be detected in extracts from peritoneal macrophages isolated from zoledronic acid-treated animals . Increasing IPP/ApppI accumulation was determined up to 7 days after drug injection , indicating prolonged P14324 inhibition by zoledronic acid . Importantly , this is the first report of in vivo production of ApppI , supporting the biological significance of this molecule . Atomic-absorption determination of beryllium in geological materials by use of electrothermal atomization . A method is described for the atomic-absorption determination of beryllium in geological materials , that utilizes electrothermal atomization after a separation by solvent extraction . Samples are decomposed with hydrofluoric acid and nitric acid in Teflon-lined pressure decomposition vessels . Beryllium is isolated by its extraction as beryllium acetylacetonate at pH 8 into xylene and back-extraction in 3M hydrochloric acid . The method has been successfully applied to the determination of beryllium in 14 U.S . Geological Survey standard rocks . Four subsamples from four bottles of each standard sample were analysed in random order . The mean beryllium contents ( ppm ) are : AGV-1 , 1.98 ; DB09058 -1 , 0.024 ; Q53EU6 , 2.84 ; BHVO-1 , 0.90 ; Q99075 -1 , 0.026 ; SCo-1 , 1.74 ; P18827 -1 , 2.52 ; P11274 -1 , 1.44 ; P63092 -1 , 1.22 ; SGR-1 , 0.86 ; QLO-1 , 1.83 ; Q96B86 -1 , 2.21 ; P0DMM9 -1 , 8.75 ; G-2 , 2.29 . An analysis of variance shows that all the samples may be considered homogeneous at F(0.95) except AGV-1 and Q99075 -1 which may be considered homogeneous at F(0.99) . OVAREX MAb- DB04964 : P01579 could improve the ovarian tumor cell sensitivity to Q8WXI7 -specific allogenic cytotoxic T cells . Various immunological parameters were studied in 100 ovarian cancer patients injected with the OVAREX therapeutic vaccine ( the functional component of which is anti- Q8WXI7 MAb- DB04964 ) to explain the serendipitous observation of prolonged survival after such treatment . In addition to Q8WXI7 -specific humoral and cellular responses , interferon-gamma ( P01579 ) was also found to be induced in those patients receiving the vaccine . In vitro studies indicated that the expression of MHC I , MHC II , and ICAM I in ovarian tumor cells were upregulated in response to P01579 . Such tumor cells were also found to be more sensitive to Q8WXI7 -specific cytotoxic T cells compared to cells that were not incubated with P01579 . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Painless burn injury caused by post-traumatic syringomyelia . BACKGROUND : Syringomyelia , which is generally related to congenital malformations and tumors , may lead to paresthesia and dysfunctions in thermo-algesic perception . Post-traumatic syringomyelia ( Q03393 ) is a rare type of this disease characterised by the development of a cystic formation containing cerebrospinal fluid ( P04141 ) , which develops inside the spinal cord after spinal trauma . AIM : The description of a case diagnosed as painless burn injury caused by Q03393 . METHOD : One case report . CONCLUSION : Although there are a number of reports regarding the formation of neuropathic ulcers related to syringomyelia , painless burn injury descriptions are very rare . Despite available limited data related to this subject , it is important to warn patients about traumas-especially burns-after a diagnosis of Q03393 . The role of the [ 2Fe-2s ] cluster centers in xanthine oxidoreductase . DB02134 oxidoreductases ( XOR ) , xanthine dehydrogenase ( P47989 , EC1.1.1.204 ) and xanthine oxidase ( XO , EC1.2.3.2 ) , are the best-studied molybdenum-containing iron-sulfur flavoproteins . The mammalian enzymes exist originally as the dehydrogenase form ( P47989 ) but can be converted to the oxidase form ( XO ) either reversibly by oxidation of sulfhydryl residues of the protein molecule or irreversibly by proteolysis . The active form of the enzyme is a homodimer of molecular mass 290 kDa . Each subunit contains one molybdopterin group , two non-identical [ 2Fe-2S ] centers , and one flavin adenine dinucleotide ( DB03147 ) cofactor . This review focuses mainly on the role of the two iron-sulfur centers in catalysis , as recently elucidated by means of X-ray crystal structure and site-directed mutagenesis studies . The arrangements of cofactors indicate that the two iron-sulfur centers provide an electron transfer pathway from molybdenum to DB03147 . However , kinetic and thermodynamic studies suggest that these two iron-sulfur centers have roles not only in the pathway of electron flow , but also as an electron sink to provide electrons to the DB03147 center so that the reactivity of DB03147 with the electron acceptor substrate might be thermodynamically controlled by way of one-electron-reduced or fully reduced state . Distribution of polyamines and their biosynthetic enzymes in intestinal adaptation . P11926 ( ODC ) and the polyamines have been shown to be important for growth processes in the intestinal mucosa . The highest activity of ODC is found in the differentiated , nonproliferating villus-tip cells rather than in the rapidly proliferating undifferentiated crypt cells . During poststarvation refeeding and lactation , we now show that increases in ODC activity paralleled the time course of mucosal hyperplasia and thymidine incorporation . Increases in ODC ( threefold ) were similar in villus and crypt cells , and the villus-crypt gradient of decreasing ODC activity ( 40:1 ) was maintained . The activity of the other polyamine biosynthetic enzyme , S-adenosylmethionine decarboxylase ( P18827 ) , was highest in the crypt cells in the basal state and increased throughout the entire villus-crypt axis during refeeding and lactation , preserving a villus-crypt gradient opposite to that of ODC . During hyperplasia , all three polyamines increased . DB01917 was highest in the villus-tip cells , paralleling ODC activity , whereas spermidine and spermine were highest in the crypt cells and paralleled the distribution of P18827 activity . Thus P18827 activity and spermidine and spermine content may play a more important role than ODC and putrescine in regulation of intestinal mucosal proliferation . It is also possible that the threefold increases in the low levels of ODC in the crypt cells are adequate to trigger cell proliferation , whereas the higher ODC levels in villus cells may represent an association with the differentiation of the enterocytes . The anti-interleukin-6 antibody siltuximab down-regulates genes implicated in tumorigenesis in prostate cancer patients from a phase I study . BACKGROUND : P05231 ( P05231 ) is associated with prostate cancer morbidity . In several experimental models , P05231 has been reported to have anti-apoptotic and pro-angiogenic effects . DB09036 ( CNTO 328 ) is a monoclonal anti- P05231 antibody which has been successfully applied in several models representing prostate cancer . This study was designed to assess preliminary safety of siltuximab in patients with early prostate cancer . PATIENTS AND METHODS : Twenty patients scheduled to undergo radical prostatectomy received either no drug or siltuximab ( 6 mg/kg , five patients per group with administration once , two times , and three times prior to surgery ) . Blood samples were collected for pharmacokinetic and pharmacodynamic analyses . Expression of elements of P05231 signaling pathways was analyzed in tumor tissue by immunohistochemistry . Gene analysis in tumor specimens was performed with the DASL array . RESULTS : No adverse events related to siltuximab were observed . Patients treated with siltuximab presented with higher levels of proliferation and apoptosis markers . Following a single dose , serum concentrations of siltuximab declined in a biexponential manner . This study revealed a decrease in phosphorylation of Stat3 and Q8TCB0 / Q8NFH3 mitogen-activated protein kinases . In addition , gene expression analyses indicate down-regulation of genes immediately downstream of the P05231 signaling pathway and key enzymes of the androgen signaling pathway . CONCLUSIONS : Preliminary safety of siltuximab is favorable . Future studies in which siltuximab could be combined with androgen-deprivation therapy and experimental therapies in advanced prostate cancer are justified .

MH_train_1207

MH_train_1207

MH_train_1207

interacts_with DB01050?

multiple_choice

P49190 -mediated inhibitory effect of parathyroid hormone and Q96A98 on cell proliferation . The parathyroid hormone ( PTH ) has dual mitogenic and inhibitory effects on cell proliferation , depending on the cell type and experimental conditions . PTH can signal via two different receptors , both positively coupled to the adenylyl cyclase/cyclic AMP pathway which can mimic some of the proliferative effects of PTH . We evaluated the role of the type-2 PTH ( PTH2 ) receptor on cell proliferation in clonal human embryonic kidney HEK293 cells stably expressing the human P49190 . Using a cyclic AMP-responsive gene-reporter , we confirmed that the tuberoinfundibular peptide ( Q96A98 ) and various human ( h ) PTH fragments including DB05829 -(1-34) were potent agonists ( EC(50) in the range of 0.01-0.3 nM ) whereas the bovine ( b ) PTH peptides b( DB00135 (34))PTH-(7-34) and its tryptophan derivative b[D- DB00150 (12), DB00135 (34)]PTH-(7-34) behaved as antagonists ( IC(50)=117 and 249 nM , respectively ) . DB05829 -(1-34) produced a dose-dependent inhibition of cell proliferation ( EC(50)=8.5+/-0.4 nM ) after 3 days and this effect was fully reversed by the tryptophan derivative antagonist . The same effect was observed with Q96A98 which caused a 30 % maximal inhibition . These findings reveal that P49190 activation can inhibit cell proliferation and might explain the dual functionality of PTH on cell proliferation . Diverse effects of inhibition of 3-hydroxy-3-methylglutaryl- DB01992 reductase on the expression of P19320 and P16581 in endothelial cells . The expression of monocyte adhesion molecules , such as P19320 ( vascular cell adhesion molecule-1 ) and P16581 , on the surface of the endothelium is an important step in the initiation and progression of atherosclerotic lesions . We hypothesized that the inhibition of 3-hydroxy-3-methylglutaryl- DB01992 ( HMG- DB01992 ) reductase in endothelial cells could influence the expression of P19320 and P16581 . Using cultured human umbilical vein endothelial cells , we found that mevastatin ( 0.1-1 microM ) significantly reduced the expression of P19320 protein in cells activated by tumour necrosis factor-alpha ( P01375 ) for 7 h . In contrast , P01375 -induced P16581 protein expression was augmented after mevastatin treatment . DB06693 inhibited the mRNA expression of both P19320 and P16581 in P01375 -stimulated endothelial cells . The activity of the transcription factor nuclear factor-kappa B , which is known to regulate the transcription of P19320 and P16581 , was significantly reduced after incubation with mevastatin . Analysis of the time-dependent variation in the P01375 -induced expression of P16581 , and estimation of the rate of surface disappearance of P16581 together with measurement of the amounts of P16581 molecules secreted , indicated that mevastatin inhibited the surface removal of P16581 . This is compatible with the observed increase in P16581 expression after statin treatment . All observed effects of mevastatin were reversed by mevalonate , the product of the P04035 reaction . In conclusion , inhibition of P04035 in endothelial cells attenuates P19320 expression , but increases P16581 expression , after cytokine induction . These diverse effects are associated with changes in the transcriptional regulation of the two adhesion molecule genes and modulation of the surface removal of P16581 . Inhibition of human steroid 5beta-reductase ( P51857 ) by finasteride and structure of the enzyme-inhibitor complex . The Delta(4)-3-ketosteroid functionality is present in nearly all steroid hormones apart from estrogens . The first step in functionalization of the A-ring is mediated in humans by steroid 5alpha- or 5beta-reductase . DB01216 is a mechanism-based inactivator of 5alpha-reductase type 2 with subnanomolar affinity and is widely used as a therapeutic for the treatment of benign prostatic hyperplasia . It is also used for androgen deprivation in hormone-dependent prostate carcinoma , and it has been examined as a chemopreventive agent in prostate cancer . The effect of finasteride on steroid 5beta-reductase ( P51857 ) has not been previously reported . We show that finasteride competitively inhibits P51857 with low micromolar affinity but does not act as a mechanism-based inactivator . The structure of the P51857 .NADP(+)*finasteride complex determined at 1.7 A resolution shows that it is not possible for NADPH to reduce the Delta(1-2)-ene of finasteride because the cofactor and steroid are not proximal to each other . The P01024 -ketone of finasteride accepts hydrogen bonds from the catalytic residues DB00135 -58 and DB00142 -120 in the active site of P51857 , providing an explanation for the competitive inhibition observed . This is the first reported structure of finasteride bound to an enzyme involved in steroid hormone metabolism . Defective postnatal endochondral bone development by chondrocyte-specific targeted expression of parathyroid hormone type 2 receptor . The human parathyroid hormone type 2 receptor ( P49190 ) is activated by PTH and by tuberoinfundibular peptide of 39 residues ( Q96A98 ) , the latter likely acting as its natural ligand . Although the receptor is expressed at highest levels in the nervous system , we have observed that both P49190 and Q96A98 are expressed in the newborn mouse growth plate , with the receptor localizing in the resting zone and the ligand Q96A98 localizing exclusively in prehypertrophic and hypertrophic chondrocytes . To address the role of P49190 in postnatal skeletal growth and development , Col2a1-hPTH2R ( P49190 -Tg ) transgenic mice were generated . The mice were viable and of nearly normal size at birth . Expression of the transgene in the growth plate was limited to chondrocytes . We found that chondrocyte proliferation was decreased , as determined by in vivo BrdU labeling of proliferating chondrocytes and P11802 and P38936 expression in the growth plate of Col2a1-hPTH2R transgenic mice . Similarly , the differentiation and maturation of chondrocytes was delayed , as characterized by decreased Sox9 expression and weaker immunostaining for the chondrocyte differentiation markers collagen type II and type X and proteoglycans . As well , there was altered expression of Gdf5 , Wdr5 , and β-catenin , factors implicated in chondrocyte maturation , proliferation , and differentiation.These effects impacted on the process of endochondral ossification , resulting in delayed formation of the secondary ossification center , and diminished trabecular bone volume . The findings substantiate a role for P49190 signaling in postnatal growth plate development and subsequent bone mass acquisition . Concanavalin-A triggers inflammatory response through JAK/ P40763 signalling and modulates P50281 regulation of P35354 in mesenchymal stromal cells . Pharmacological targeting of inflammation through P40763 and NF-κB signaling pathways is , among other inflammatory biomarkers , associated with cyclooxygenase ( P36551 ) -2 inhibition and is believed to play a crucial role in prevention and therapy of cancer . Recently , inflammatory factors were found to impact on mesenchymal stromal cells ( O60682 ) contribution to tumor angiogenesis . Given O60682 chemotaxis and cell survival are regulated , in part , by the membrane type-1 matrix metalloproteinase ( P50281 ) , an MMP also involved in transducing NF-κB intracellular signaling pathways , we tested whether P40763 regulation by P50281 may also contribute to the expression balance of P35354 in O60682 . We demonstrate that P40763 phosphorylation was triggered in O60682 treated with the P50281 inducer lectin Concanavalin-A ( ConA ) , and that this phosphorylation was abrogated by the O60674 inhibitor AG490 . P50281 gene silencing significantly inhibited ConA-induced P40763 phosphorylation and this was correlated with reduced proMMP-2 activation and P35354 expression . On the other hand , P40763 gene silencing potentiated ConA-induced P35354 expression , providing evidence for a new P50281 /JAK/ P40763 signaling axis that may , in part , explain how P50281 contributes to proinflammatory intracellular signaling . Given that O60682 are avidly recruited within inflammatory microenvironments and within experimental vascularizing tumors , these mechanistic observations support a possible dual control of cell adaptation to inflammation by P50281 and that may enable O60682 to be active participants within inflamed tissues . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Comparison of expression profiles induced by dust mite in airway epithelia reveals a common pathway . BACKGROUND : Airway epithelial cells have shown to be active participants in the defense against pathogens by producing signaling and other regulatory molecules in response to the encounter . METHODS : In previous manuscripts , we have studied the effect of house dust mite ( HDM ) extract on both an epithelial cell-line ( H292 ) and primary nasal epithelial cell . When we compare these responses we conclude that the H292 cells more closely resemble nasal epithelium of healthy controls ( share 107 probe-sets ) than of allergic individuals ( share 17 probe-sets ) . RESULTS : Interestingly , probably because of an absent intraindividual variation between samples , more probe-sets ( 8280 ) change expression significantly in H292 than in either healthy ( 555 ) or allergic ( 401 ) epithelium . CONCLUSIONS : A direct comparison of all the responses in these epithelial cells reveals a core-response to HDM of just 29 genes . These genes ( P78556 , P10145 , P19875 , P09341 , IL-1B , P15514 , P21580 , Q99075 , P35354 , P12643 , P01130 , Q03405 , P00749 , Q00653 , P19838 , P05412 , P18847 , P18146 , O15118 , Q8IUC6 , P29317 , P29279 , P28562 , O43609 , TLR-3 , complement factor P01024 , Q9Y6Y0 , SerpinB3 , and Q9Y617 ) have described links with allergy or inflammation and may even describe the well-established relationship between viral infections and allergic exacerbations or allergy development . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Identification and quantification of concentration-dependent biomarkers in MCF-7/BOS cells exposed to 17β-estradiol by 2-D DIGE and label-free proteomics . This paper reports the identification of biomarkers resulting from the exposure of MCF-7/BOS cells to 17β-estradiol ( E(2) ) . The biomarkers were identified using 2 independent and complementary techniques , 2-D DIGE/MALDI-TOF peptide mass fingerprint , and 2-D UPLC- P19957 MS/MS . They were identified from the cytosolic fractions of cells treated for 24h with mitogenic concentrations of 1 , 30 and 500 pM of 17β-estradiol . Five biomarkers were up-regulated proteins , namely HSP 74 , P13639 , Q02790 , EF1 and GDIB and one was a down-regulated protein , namely K2C8 . Three of these proteins , P13639 , Q02790 and K2C8 are implicated in a network centered on the estrogen receptors P03372 and Q92731 as well as on P31749 . After the discovery phase , three biomarkers were selected to test the presence of estrogens using selected reaction monitoring ( P19623 ) . They were monitored using P19623 after incubation of MCF-7/BOS in the presence of E(2) for confirmation or selected xenoestrogens . Daidzein , coumestrol and enterolactone induced an up-regulation of P13639 and FKPB4 proteins , while tamoxifen and resveratrol induced a down-regulation . The exposure of all phytoestrogens induced the down-regulation of K2C8 . These markers form a preliminary molecular signature that can be used when testing the estrogenic activity of xenobiotics , either pure or in mixtures . Estrogen receptors and function in the male reproductive system . A substantial advance in our understanding on the estrogen signaling occurred in the last decade . DB00286 interact with two receptors , P03372 and Q92731 , also known as ERalpha and ERbeta , respectively . P03372 and Q92731 belong to the nuclear receptor family of transcription factors . In addition to the well established transcriptional effects , estrogens can mediate rapid signaling , triggered within seconds or minutes . These rapid effects can be mediated by ESRs or the G protein-coupled estrogen receptor Q99527 , also known as Q99527 . The effects of estrogen on cell proliferation , differentiation and apoptosis are often mediated by growth factors . The understanding of the cross-talk between androgen , estrogen and growth factors signaling pathways is therefore essential to understand the physiopathological mechanisms of estrogen action . In this review we focused on recent discoveries about the nature of the estrogen receptors , and on the signaling and function of estrogen in the male reproductive system . DB04956 . Tumour necrosis factor-alpha ( P01375 ) is well established as a key mediator in the inflammatory response seen in various disease processes including sepsis . P01375 is involved in virtually all features of septic shock and multiple organ failure . Anti- P01375 strategies are thus appealing and have been effective at reducing inflammation and morbidity in certain conditions including rheumatoid arthritis and Crohn 's disease . DB04956 is the F(ab')2 fragment of a murine anti- P01375 antibody , and has been evaluated in clinical trials in septic patients . The results suggest that the drug is well tolerated , and may be of benefit in certain groups of patients with sepsis . A large , randomised , clinical trial of afelimomab in patients with severe sepsis has recently been completed and the results are eagerly awaited . More work is necessary to identify a means of selecting which patients are most likely to benefit from this type of therapy in sepsis . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . Opposite effects of tamoxifen on metabolic syndrome-induced bladder and prostate alterations : a role for Q99527 / Q99527 ? BACKGROUND : BPH and LUTS have been associated to obesity , hypogonadism , and metabolic syndrome ( MetS ) . MetS-induced prostate and bladder alterations , including inflammation and tissue remodeling , have been related to a low-testosterone and high-estrogen milieu . In addition to ERs , Q99527 / Q99527 is able to mediate several estrogenic non-genomic actions . METHODS : Supplementing a subgroup of MetS rabbits with tamoxifen , we analyzed the in vivo effects on MetS-induced prostate and bladder alterations . The effects of selective ER/ Q99527 ligands and Q99527 silencing on prostate inflammation were also studied in vitro using hBPH cells . RESULTS : ERα , ERβ , and PR expression was upregulated in MetS bladder , where tamoxifen decreased ERα and PR expression , further stimulating ERβ . In addition , tamoxifen-dosing decreased MetS-induced overexpression of inflammatory and tissue remodeling genes . In prostate , sex steroid receptors , pro-inflammatory and pro-fibrotic genes were upregulated in MetS . However , tamoxifen did not affect them and even increased P35354 . In hBPH cells , 17β-estradiol increased P10145 secretion , an effect blunted by co-treatment with Q99527 antagonist Q99943 but not by ER antagonist DB00947 , which further increased it . Q99527 agonist P55008 dose-dependently ( IC50 = 1.6 nM ) induced P10145 secretion . In vitro analysis demonstrated that Q99527 silencing reverted these stimulatory effects . CONCLUSIONS : Q99527 can be considered the main mediator of estrogen action in prostate , whereas in bladder the mechanism appears to rely on ERα , as indicated by in vivo experiments with tamoxifen dosing . Limiting the effects of the MetS-induced estrogen action via Q99527 could offer new perspectives in the management of BPH/LUTS , whereas tamoxifen dosing showed potential benefits in bladder . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . Activation of cellular invasion by trefoil peptides and src is mediated by cyclooxygenase- and thromboxane A2 receptor-dependent signaling pathways . We have investigated the possible functional relationships between cellular invasion pathways induced by trefoil factors ( TFFs ) , src , and the cyclooxygenases P23219 and P35354 . Pharmacological inhibitors of the Rho small GTPase ( P01024 exoenzyme ) , phospholipase C ( U-73122 ) , cyclooxygenases ( SC-560 , NS-398 ) , and the thromboxane A2 receptor ( P21731 ) antagonist SQ-295 completely abolished invasion induced by intestinal trefoil factor , pS2 , and src in kidney and colonic epithelial cells MDCKts.src and PCmsrc . In contrast , invasion was induced by the P21731 mimetic U-46619 , constitutively activated forms of the heterotrimeric G-proteins Galphaq ( AGalphaq ) , Galpha12 , Galpha13 ( AGalpha12/13 ) , which are signaling elements downstream of P21731 . Ectopic overexpression of pS2 cDNA and protein in MDCKts.src-pS2 cells and human colorectal cancer cells HCT8/S11-pS2 initiate distinct invasion signals that are Rho independent and P36551 and P21731 dependent . We detected a marked induction of P35354 protein and accumulation of the stable PGH2/TXA2 metabolite TXB2 in the conditioned medium from cells transformed by src . This led to activation of the P21731 -dependent invasion pathway , which is monitored via a Rho- and Galpha12/Galpha13-independent mechanism using the Galphaq/PKC signaling cascade . These findings identify a new intracrine/paracrine loop that can be monitored by TFFs and src in inflammatory diseases and progression of colorectal cancers . Inhibition of the invasion capacity of carcinoma cells by DB05476 , a novel synthetic inhibitor of the urokinase-type plasminogen activator system . The overall survival rate of patients suffering from carcinomas has remained poor and nearly unchanged over the last decades . This is mainly due to the so-called minimal residual disease , i.e. , remaining tumor cells that overcome surgery and/or radiotherapy and are the cause of locoregional and distant metastases . To metastasize , tumor cells take advantage of proteases to invade and remodel surrounding tissues . Here , we analyzed the efficiency of DB05476 , a novel 3-amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 expression was characterized in different carcinoma cell lines , including SCCHN , breast and cervical carcinoma . Thereafter , the invasive potential of these cell lines was determined using Matrigel invasion chambers and a spheroid cocultivation model with human fibroblasts . Q03405 expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 . A decrease of tumor cell invasion by up to 50 % was achieved in both models with the SCCHN line FaDu and the cervical carcinoma line HeLa after treatment with DB05476 . Thus , our results demonstrate the potential of DB05476 in vitro as a promising adjuvant antimetastatic therapy of carcinomas . Prevention and treatment of pancreatic cancer by curcumin in combination with omega-3 fatty acids . Pancreatic cancer BxPC-3 cells were exposed to curcumin , docosahexaenoic acid ( DB01708 ) , or combinations of both and analyzed for proliferation and apoptosis . Pancreatic tumor xenografts were established by injecting BxPC-3 cells into each flank of nude mice . After the tumors reached a size of approximately 190-200 mm(3) , animals were fed diets with or without 2,000 ppm curcumin in 18 % corn oil or 15 % fish oil + 3 % corn oil for 6 more wk before assessing the tumor volume and expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygeanse-2 ( P35354 ) , 5-lipoxinase ( 5- P28300 ) , and P38936 . A synergistic effect was observed on induction of apoptosis ( approximately sixfold ) and inhibition of cell proliferation ( approximately 70 % ) when cells were treated with curcumin ( 5 microM ) together with the DB01708 ( 25 microM ) . Mice fed fish oil and curcumin showed a significantly reduced tumor volume , 25 % ( P < 0.04 ) and 43 % ( P < 0.005 ) , respectively , and importantly , a combination of curcumin and fish oil diet showed > 72 % ( P < 0.0001 ) tumor volume reduction . Expression and activity of P35228 , P35354 , and 5- P28300 are downregulated , and P38936 is upregulated in tumor xenograft fed curcumin combined with fish oil diet when compared to individual diets . The preceding results evidence for the first time that curcumin combined with omega-3 fatty acids provide synergistic pancreatic tumor inhibitory properties . Biosynthetic labeling of diphthamide in Saccharomyces cerevisiae . DB03223 , the post-translational amino acid derivative in the diphtheria toxin-modification site of protein synthesis elongation factor 2 ( P13639 ) , has the proposed structure 2-[3-carboxyamido-3-(trimethylammonio)propyl]histidine ( Van Ness , B. G. , Howard , J. B. , and Bodley , J. W. ( 1980 ) J. Biol. Chem. 255 , 10710-10716 ) . The identification of the biosynthetic precursors of diphthamide would provide a means of evaluating its proposed structure and determining if the amino acid occurs in proteins other than P13639 . Toward this end , yeast were grown on potential radiolabeled precursors and the resulting radiolabeled protein was hydrolyzed in acid . The acid hydrolysates were subjected to amino acid analysis with a program optimized to resolve diphthine , the acid hydrolysis product of diphthamide , from the common amino acids . Radiolabel from [beta-3H]histidine , [alpha-3H]methionine , and [ Me-3H ] methionine was found to be incorporated into diphthine in a molar ratio of 1:1:3 while that of [35S]methionine was not incorporated . These results are in accord with the proposed structure of diphthamide and suggest that in its biosynthesis the backbone and 3 methyl groups of methionine are added to a histidine residue in the peptide chain of P13639 . These labeling experiments show that diphthine ( diphthamide ) constitutes approximately 6 X 10(-6) mol fraction of the total amino acids in yeast protein hydrolysates . Estimates of the amount of diphthamide present in the diphtheria toxin-modification site of P13639 indicate that it constitutes from 4.5 to 9 X 10(-6) mol fraction of the total amino acids in yeast protein . The present evidence suggests that diphthamide occurs only in P13639 . SAR and in vivo evaluation of 4-aryl-2-aminoalkylpyrimidines as potent and selective O60674 ( O60674 ) inhibitors . We report the discovery of a series of 4-aryl-2-aminoalkylpyrimidine derivatives as potent and selective O60674 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 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 ) it was advanced into clinical trials .

MH_train_1208

MH_train_1208

MH_train_1208

interacts_with DB00035?

multiple_choice

DB09210 defines a new binding site for allosteric modulators of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid ( AMPA ) receptors . Glutamate receptors are the most prevalent excitatory neurotransmitter receptors in the vertebrate central nervous system and are important potential drug targets for cognitive enhancement and the treatment of schizophrenia . Allosteric modulators of AMPA receptors promote dimerization by binding to a dimer interface and reducing desensitization and deactivation . The pyrrolidine allosteric modulators , piracetam and aniracetam , were among the first of this class of drugs to be discovered . We have determined the structure of the ligand binding domain of the AMPA receptor subtypes P42262 and P42263 with piracetam and a corresponding structure of P42263 with aniracetam . Both drugs bind to P42262 and P42263 in a very similar manner , suggesting little subunit specificity . However , the binding sites for piracetam and aniracetam differ considerably . DB04599 binds to a symmetrical site at the center of the dimer interface . DB09210 binds to multiple sites along the dimer interface with low occupation , one of which is a unique binding site for potential allosteric modulators . This new site may be of importance in the design of new allosteric regulators . Role of chronic inhibition of dopamine-metabolizing enzymes in the regulation of renal sodium and phosphate excretion in the rat remnant kidney . BACKGROUND/AIMS : The present study examined the effects of chronic selective or combined inhibition of type A monoamine oxidase ( MAO ) and catechol-O-methyltransferase ( P21964 ) on daily urinary excretion of dopamine and metabolites and on natriuresis and phosphaturia in 3/4 nephrectomized ( 3/4nx ) and Sham rats . METHODS : The 3/4nx and Sham rats were placed in metabolic cages and received the P21397 -selective inhibitor Ro-411049 ( 7.5 mg x kg(-1) bid ) and/or the P21964 -selective inhibitor DB03336 3-202 ( 30 mg x kg(-1) bid ) orally for 3 days during high sodium diet . RESULTS : Selective P21964 inhibition increased the urinary excretion of the deaminated metabolite ( 3,4-dihydroxyphenylacetic acid , DOPAC ) and decreased the urinary excretion of the methylated ( 3-methoxytyramine , 3-MT ) and deaminated plus methylated metabolite ( homovanillic acid , HVA ) in both groups . Selective P21397 inhibition increased the urinary excretion of 3-MT and reduced the urinary excretion of both DOPAC and HVA in either 3/4nx or Sham rats . Combined inhibition of P21397 and P21964 did not significantly change the urinary excretion of DOPAC and markedly decreased the urinary excretion of 3-MT and HVA in both groups . Selective or combined inhibition of P21397 and P21964 did not alter the daily urinary excretion of dopamine , sodium or phosphate in either 3/4nx or Sham rats . CONCLUSIONS : Chronic selective or combined inhibition of P21397 and P21964 is not of major importance in regulating the dopamine-dependent natriuresis and phosphaturia in either 3/4nx or Sham rats . Ectopic DB01285 syndrome caused by bronchial carcinoid tumor indistinguishable from Cushing 's disease . A 75-year-old woman was admitted to our hospital because of a poor glycemic control . She was found to have Cushingoid feature and dynamic endocrine tests showed elevated plasma DB01285 and cortisol levels , lack of their circadian rhythm , non-suppressibility to high-dose dexamethasone , responsiveness to P06850 , but not to DB00035 , and suppression to octreotide . Pituitary Q9BWK5 showed an equivocal small lesion . CT scan of the chest showed two nodular lesions in the right lung ( S5 , S7 ) , while a mild uptake was noted only in S5 lesion by DB09150 -PET , but positive uptake was only in S7 lesion by somatostatin receptor scintigraphy ( SRS ) . Inferior petrosal sinus sampling revealed a gradient of plasma DB01285 after P06850 stimulation , consistent with the diagnosis of Cushing ' s disease . She underwent middle and inferior lobectomy of the right lung . The resected tumor in S7 was consistent with the diagnosis of a bronchial carcinoid tumor with positive DB01285 immunoreactivity , while that of S5 was cryptococcal granuloma . RT-PCR revealed abundant expressions of P01189 and SSTR ( -1 , -2 , -5 ) , but not of P34998 and P47901 . Postoperatively , abnormal endocrine data were normalized along with improvement of hypertension and diabetes . This was a diagnostic challenging case with ectopic DB01285 syndrome indistinguishable from Cushing ' s disease by various endocrine and imaging tests , among which SRS successfully localized the tumor responsible for ectopic DB01285 secretion . A randomized , placebo-controlled study of the effects of the p38 MAPK inhibitor SB- DB05250 on blood biomarkers of inflammation in P48444 patients . The p38 mitogen-activated protein kinase ( MAPK ) signaling upregulates inflammation and is known to be increased in chronic obstructive pulmonary disease ( P48444 ) . The authors assessed the pharmacology of the novel p38 MAPK inhibitor SB- DB05250 using blood biomarkers in P48444 . Seventeen P48444 patients ( forced expiratory volume in 1 second 50 % -80 % predicted ) using short-acting bronchodilators participated in a double-blind , double-dummy , randomized , crossover study . Patients received single oral doses of SB- DB05250 7.5 mg and 25 mg , prednisolone 10 mg and 30 mg , and placebo . Blood was obtained predose and at 1 , 2 , 6 , and 24 hours postdose . Whole-blood sorbitol-induced phosphorylated ( p ) heat shock protein ( HSP ) 27 levels as a marker of p38 pathway activation and lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -alpha production were assessed . Both doses of SB- DB05250 , but not prednisolone , significantly ( P < .0001 ) reduced weighted mean ( WM ) pHSP27 ( 0-6 hours ) by 58 % compared with placebo . WM P01375 production ( 0-24 hours ) was significantly reduced compared with placebo by SB- DB05250 25 mg ( 40 % , P = .005 ) and 7.5 mg ( 33.4 % , P = .02 ) , while prednisolone 30 mg and 10 mg caused 81.5 % and 58.2 % suppression , respectively ( both P < .0001 ) . SB- DB05250 inhibited the p38 MAPK pathway to a greater degree than prednisolone did . SB- DB05250 inhibited P01375 production . SB- DB05250 is a potent p38 MAPK inhibitor that potentially suppresses inflammation in P48444 . cDNA cloning and characterization of a novel calmodulin-like protein from pearl oyster Pinctada fucata . DB01373 metabolism in oysters is a very complicated and highly controlled physiological and biochemical process . However , the regulation of calcium metabolism in oyster is poorly understood . Our previous study showed that calmodulin ( P62158 ) seemed to play a regulatory role in the process of oyster calcium metabolism . In this study , a full-length cDNA encoding a novel calmodulin-like protein ( CaLP ) with a long C-terminal sequence was identified from pearl oyster Pinctada fucata , expressed in Escherichia coli and characterized in vitro . The oyster CaLP mRNA was expressed in all tissues tested , with the highest levels in the mantle that is a key organ involved in calcium secretion . In situ hybridization analysis reveals that CaLP mRNA is expressed strongly in the outer and inner epithelial cells of the inner fold , the outer epithelial cells of the middle fold , and the dorsal region of the mantle . The oyster CaLP protein , with four putative Ca(2+)-binding domains , is highly heat-stable and has a potentially high affinity for calcium . CaLP also displays typical Ca(2+)-dependent electrophoretic shift , Ca(2+)-binding activity and significant Ca(2+)-induced conformational changes . Ca(2+)-dependent affinity chromatography analysis demonstrated that oyster CaLP was able to interact with some different target proteins from those of oyster P62158 in the mantle and the gill . In summary , our results have demonstrated that the oyster CaLP is a novel member of the P62158 superfamily , and suggest that the oyster CaLP protein might play a different role from P62158 in the regulation of oyster calcium metabolism . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Role of monoamine oxidases in the exaggerated 5-hydroxytryptamine-induced tension development of human isolated preeclamptic umbilical artery . We investigated the role(s) of monoamine oxidases ( MAOs ) on the altered 5-hydroxytryptamine ( 5-HT , serotonin ) -induced tension development of the isolated umbilical artery of preeclamptic pregnancy of Chinese women . An enhanced 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy was observed when compared with that of normal pregnancy . The enhanced component of 5-HT-induced tension development was eradicated by clorgyline ( a P21397 inhibitor ) . Blockade of P29474 ( endothelial isoform nitric oxide synthase ) ( N(omega)-nitro-L-arginine methyl ester ) , 5-HT transporter ( citalopram ) , 5-HT receptor subtypes ( 5HT2B , SB 204741 ; P28335 , RS 102221 ; P34969 , SB 269970 ) , and endothelium denudation of the umbilical artery of normal pregnancy mimicked the enhanced 5-HT-induced tension development as observed in the preeclamptic tissues . In contrast , no apparent changes in 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy were observed with the same pharmacological manipulations . A decreased protein expression levels of P21397 and P29474 ( no P35228 and P27338 expression was detected ) and no change in caveolin-1 and 5-HT transporter expression were demonstrated in the umbilical artery ( endothelium intact ) lysate of preeclamptic pregnancy , compared to that of the umbilical artery of normal pregnancy . Thus , in the umbilical artery of preeclamptic pregnancy , a decrease of P21397 and P29474 protein expression levels are probably associated with , or responsible for , the exaggerated 5-HT-induced tension development . Bacterial translocation in cirrhotic rats stimulates P29474 -derived NO production and impairs mesenteric vascular contractility . DB00435 ( NO ) has been implicated in the arterial vasodilation and associated vascular hyporesponsiveness to vasoconstrictors observed in liver cirrhosis . Bacteria , potent activators of NO and P01375 synthesis , are found in the mesenteric lymph nodes ( MLNs ) of ascitic cirrhotic rats . Here , we investigated the impact of bacterial translocation ( BT ) to MLNs on P01375 production , vascular NO release , and contractility in the mesenteric vasculature of ascitic cirrhotic rats . Vascular response to the alpha-adrenoagonist methoxamine , which is diminished in the superior mesenteric arterial beds of cirrhotic rats , is further blunted in the presence of BT . BT promoted vascular NO release in cirrhotic rats , an effect that depended on pressure-induced shear stress and was blocked by the NO inhibitor N(omega)-nitro-L-arginine . Removing the endothelium had the same effect . Endothelial NO synthase ( P29474 ) , but not the inducible isoform ( P35228 ) , was present in mesenteric vasculature of cirrhotic rats with and without BT , and its expression was enhanced compared with controls . P01375 was induced in MLNs by BT and accumulated in parallel in the serum . This P01375 production was associated with elevated levels of tetrahydrobiopterin ( BH(4) ) , a P01375 -stimulated cofactor and enhancer of P29474 -derived NO biosynthesis and NOS activity in mesenteric vasculature . These findings establish a link between BT to MLNs and increased P01375 production and elevated BH(4) levels enhancing P29474 -derived NO overproduction , further impairing contractility in the cirrhotic mesenteric vasculature . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . Neuromedin U-induced anorexigenic action is mediated by the corticotropin-releasing hormone receptor-signaling pathway in goldfish . Our recent research has indicated that neuromedin U ( P48645 ) orthologs exist in goldfish , and that P48645 consisting of 21 amino acid residues ( P48645 -21 ) can potently inhibit food intake in goldfish , as is the case in rodents . However , the anorexigenic pathway of P48645 -21 has not yet been clarified in this species . P06850 ( P06850 ) , P06850 -related peptides and alpha-melanocyte-stimulating hormone ( alpha-MSH ) , which exert potent anorexigenic effects , are important mediators involved in feeding regulation in fish . We examined whether P06850 or alpha-MSH mediates P48645 -21-induced anorexigenic action in goldfish . We first investigated the effect of intracerebroventricular ( ICV ) administration of P48645 -21 at 100 pmol/g body weight ( BW ) , which is enough to suppress food intake , on expression levels of mRNA for P06850 and proopiomelanocortin ( P01189 ) in the hypothalamus . ICV-injected P48645 -21 induced a significant increase in the expression level of P06850 mRNA , but not that of P01189 mRNA . We also examined the effects of ICV administration of the P06850 1/2 receptor antagonist , alpha-helical P06850 ( ( 9-41 ) ) , and the melanocortin 4 receptor antagonist , HS024 , on the anorexigenic action of ICV-injected P48645 -21 . The anorexigenic effect of P48645 -21 was blocked by treatment with alpha-helical P06850 ( ( 9-41 ) ) at 400 pmol/g BW , but not HS024 at 200 pmol/g BW . These results suggest that the anorexigenic action of P48645 -21 is mediated by the P06850 1 or 2 receptor-signaling pathway in goldfish . Cellular mechanisms of the hemostatic effects of desmopressin ( DB00035 ) . The synthetic analog of vasopressin desmopressin ( DB00035 ) is widely used for the treatment of patients with von Willebrand disease ( VWD ) , hemophilia A , several platelet disorders , and uremic bleeding . DB00035 induces an increase in plasma levels of P04275 ( P04275 ) , coagulation factor VIII ( FVIII ) , and tissue plasminogen activator ( t-PA ) . It also has a vasodilatory action . In spite of its extensive clinical use , its cellular mechanism of action remains incompletely understood . Its effect on P04275 and t-PA as well as its vasodilatory effect are likely explained by a direct action on the endothelium , via activation of endothelial vasopressin P30518 receptor and DB02527 -mediated signaling . This leads to exocytosis from Weibel Palade bodies where both P04275 and t-PA are stored , as well as to nitric oxide ( NO ) production via activation of endothelial NO synthase . The mechanism of action of DB00035 on FVIII plasma levels remains to be elucidated . The hemostatic effect of DB00035 likely involves additional cellular effects that remain to be discovered . PP2Cdelta ( Ppm1d , O15297 ) , an endogenous inhibitor of p38 MAPK , is regulated along with Trp53 and Cdkn2a following p38 MAPK inhibition during mouse preimplantation development . Preimplantation embryos utilize mitogen-activated protein kinase signaling ( MAPK ) pathways to relay signals from the external environment to prepare appropriate responses and adaptations to a changing milieu . It is therefore important to investigate how MAPK pathways are regulated during preimplantation development . This study was conducted to investigate whether PP2Cdelta ( Ppm1d , O15297 ) is expressed during mouse preimplantation development and to determine the influences of p38 MAPK inhibition on expression of Trp53 ( p53 ) , Ppm1d , ( O15297 ) , and Cdkn2a ( p16 ) during mouse preimplantation development . Our results indicate that Trp53 , Ppm1d , and Cdkn2a mRNAs and TRP53 and PP2Cdelta proteins are expressed throughout mouse preimplantation development . Treatment of 2-cell embryos with DB04338 ( potent inhibitor of p38 MAPK alpha/beta/ Q16539 /11 ) significantly increased Trp53 , Ppm1d and Cdkn2a and Mapk14 mRNA levels at 12 and 24 hr . Treatment of 8-cell embryos with DB04338 for 12 hr increased Trp53 , Ppm1d , and Cdkn2a mRNA levels , but not Mapk14 mRNA levels . Treatment of 8-cell embryos for 24 hr increased Trp53 , and Ppm1d mRNA levels , but decreased Cdkn2a and Mapk14 mRNA levels . Therefore , blockade of p38 MAPK activity is associated with embryo stage specific influences on Trp53 , Ppm1d , Cdkn2a , and Mapk14 expression during mouse preimplantation development . These results define downstream targets of p38 MAPK during preimplantation development and indicate that the p38 MAPK pathway regulates Trp53 , Ppm1d , and Cdkn2a expression . This study increases our understanding of the mechanisms controlling preimplantation development and of the interactions between preimplantation embryos and their culture environments . Antihistamine effects on prefrontal cortex activity during working memory process in preschool children : a near-infrared spectroscopy ( NIRS ) study . P35367 antagonists ( antihistamines ) are widely used for the treatment of allergic disorders in young children . This study examined the effects of antihistamine on prefrontal cortex activity in preschool children using near-infrared spectroscopy ( NIRS ) , an emerging brain-imaging method suitable for psychological experiments , especially in young children . We examined the changes of oxygenated hemoglobin concentration in the prefrontal cortex while children performed a spatial working memory task , 3h after taking a first-generation antihistamine ( ketotifen ) , second-generation antihistamine ( epinastine ) , or placebo . Fifteen healthy preschool children ( mean age , 5.5 years ) participated . DB00920 significantly impaired behavioral performance and cortical activation at the lateral prefrontal cortex in the working memory task , compared with epinastine and placebo . There were no sedative effects on neural response or behavioral performance after epinastine administration . This paper demonstrates for the first time differential sedation effects of first- and second-generation antihistamines on brain hemodynamic response in young children . Also discussed is the utility of the NIRS technique in neuropsychopharmacological studies of children . P35354 in newborn hyperoxic lung injury . Supraphysiological O2 concentrations , mechanical ventilation , and inflammation significantly contribute to the development of bronchopulmonary dysplasia (BPD).Exposure of newborn mice to hyperoxia causes inflammation and impaired alveolarization similar to that seen in infants with BPD.Previously , we demonstrated that pulmonary cyclooxygenase-2 ( P35354 ) protein expression is increased in hyperoxia-exposed newborn mice.The present studies were designed to define the role of P35354 in newborn hyperoxic lung injury.We tested the hypothesis that attenuation of P35354 activity would reduce hyperoxia-induced inflammation and improve alveolarization.Newborn C3H/HeN micewere injected daily with vehicle , aspirin ( nonselective P35354 inhibitor ) , or celecoxib ( selective P35354 inhibitor ) for the first 7 days of life.Additional studies utilized wild-type ( C57Bl/6 , P35354 (+/+) ) , heterozygous ( P35354 (+/-) ) , and homozygous ( P35354 (-/-) ) transgenic mice.Micewere exposed to room air ( 21 % O2 ) or hyperoxia ( 85 % O2 ) for 14 days. DB00945 -injected and P35354 (-/-) pups had reduced levels of monocyte chemoattractant protein ( P13500 ) in bronchoalveolar lavage fluid (BAL).Both aspirin and celecoxib treatment reduced macrophage numbers in the alveolar walls and air spaces. DB00945 and celecoxib treatment attenuated hyperoxia-induced P36551 activity , including altered levels of prostaglandin (PG)D2 metabolites.Decreased P36551 activity , however , did not prevent hyperoxia-induced lung developmental deficits.Our data suggest thatincreased P35354 activity may contribute to proinflammatory responses , including macrophage chemotaxis , during exposure to hyperoxia.Modulation of P35354 activity may be a useful therapeutic target to limit hyperoxia-induced inflammation in preterm infants at risk of developing BPD . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . A population-based association study of candidate genes for depression and sleep disturbance . The clinical manifestation of depression comprises a variety of symptoms , including early morning awakenings and fatigue , features also indicating disturbed sleep . The presence or absence of these symptoms may reflect differences in neurobiological processes leading to prolonged depression . Several neurobiological mechanisms have been indicated in the induction of depression , including disturbances in serotonergic and glutamatergic neurotransmission and in the action of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . The same transmitters have also been linked to sleep regulation . We hypothesized that depression without simultaneous symptoms of disturbed sleep would partly have a different genetic background than depression with symptoms of disturbed sleep . We tested this hypothesis using a systematic population-based association study of 14 candidate genes related to depression and disturbed sleep . Association of genetic variants with either depression alone , depression with early morning awakenings , or depression with fatigue was investigated using permutation-based allelic association analysis of a sample of 1,654 adults recruited from Finland 's population-based program . The major findings were associations of Q8IWU9 ( rs12229394 ) with depression accompanied by fatigue in women and P16220 ( rs11904814 ) with depression alone in men . We also found suggestive associations in women for Q99259 , P42263 , and P23560 with depression accompanied by fatigue , and for P34998 with depression accompanied by early morning awakenings . The results indicate sex-dependent and symptom-specific differences in the genetic background of depression . These differences may partially explain the broad spectrum of depressive symptoms , and their systematic monitoring could potentially be used for diagnostic purposes . [ O6-benzylguanine stimulates regulatory functions of the Ada protein in Escherichia coli ] . In vitro experiments showed that O6-benzylguanine ( O6-benzG , 0.2 microM ) fully inhibited the repair activity of human O6-alkylguanine-DNA alkyltransferase ( P16455 ) due to the formation of S-benzylcytosine in the protein acceptor site . O6-benzG at concentrations increased many times ( up to 800 microM ) failed to inhibit the repair activity of the Escherichia coli Ada protein , the structural and functional analog of P16455 . It has been shown for the first time that O6-benzG stimulates the regulatory activity of the Ada protein . In experiments with N-nitroso-N-methylurea ( P48645 ) , the pretreatment of Escherichia coli cells with O6-benzG at a sublethal concentration of 10 microM led to a twofold enhancement of transcription at the Ada-dependent promoter of the alkA gene in control cells and ensured transcription enhanced 1.6-1.7 times at alkA and alkB promoters in cells with the induced " classical " Ada response . Apparently , an increase in the regulatory activity of the Ada protein was associated with the formation of the stable protein molecule having the strong affinity for alkA and alkB promoters after transfer of the benzyl group from O6-benzG to the acceptor site DB00151 -69 in the N-terminal domain of Ada protein . O6-benzG did not affect the regulative activity of Ada in alternative quasi-adaptive responses to P48645 . Expression of metabotropic glutamate receptors in rat meningeal and brain microvasculature and choroid plexus . This study investigated the distribution of metabotropic glutamate receptors ( mGluRs ) in meningeal and parenchymal microvasculature and in choroid plexus by means of Western blot analysis and immunohistochemistry . Western blot analysis demonstrated mGluR expression in both rat and human leptomeningeal tissues . In the rat , mGluR expression was developmentally regulated , with only Q14416 /3 showing expression at the embryonic day 19 developmental stage . In contrast , Q13255 alpha , Q14416 /3 , mGluR4a , and Q14831 were expressed in leptomeninges from adult rats . Immunohistochemical analyses showed intense Q13255 alpha immunoreactivity in the pia mater and blood vessels in the subarachnoid space and in the arachnoid layer of the meninges . Q14416 /3 , mGluR4a , P41594 , and Q14831 were also expressed in meningeal microvasculature . In addition , the parenchymal microvasculature and choroid plexus were strongly immunoreactive for Q13255 alpha , Q14416 /3 , mGluR4a , P41594 , and Q14831 . We used antibodies specific for phenotypic markers of microvascular and glial cells to characterize the cell type(s) immunopositive for mGluRs . Comparison of staining with anti- P04275 antibody and anti-mGluR antibodies revealed that mGluR immunoreactivity was present in cells that surrounded the luminal surface labeled by the endothelial cell marker . In these cells , smooth muscle actin and mGluR immunoreactivity overlapped , suggesting that , in addition to endothelial cells , pericytes within the microvasculature also express mGluRs . Furthermore , expression of Q13255 alpha was also observed in pure pericyte cultures isolated from bovine retina . These data suggest that glutamate by means of activation of mGluRs may have a broad sphere of physiological influence in the brain which in addition to modulating synaptic transmission may also have a role in determining microvascular function and dysfunction . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . DB06698 ameliorates olanzapine-induced weight gain through modulation of histaminergic , P01303 and AMPK pathways . Olanzapine is widely used to treat schizophrenia and other disorders , but causes adverse obesity and other metabolic side-effects . Both animal and clinical studies have shown that co-treatment with betahistine ( a histaminergic H1 receptor agonist and H3 receptor antagonist ) is effective for ameliorating olanzapine-induced weight gain/obesity . To reveal the mechanisms underlying these effects , this study investigated the effects of co-treatment of olanzapine and betahistine ( O+B ) on expressions of histaminergic H1 receptor ( P35367 ) , AMP-activated protein kinase ( AMPK ) , neuropeptide Y ( P01303 ) , and proopiomelanocortin ( P01189 ) in the hypothalamus associated with reducing olanzapine-induced weight gain . Olanzapine significantly upregulated the mRNA and protein expressions of P35367 , while O+B co-treatment significantly downregulated the P35367 levels , compared to the olanzapine-only treatment group . The P01303 mRNA expression was significantly enhanced by olanzapine , but it was significantly reversed by O+B co-treatment . The hypothalamic P35367 expression was positively correlated with total food intake , and P01303 expression . Olanzapine also increased AMPKα activation measured by the AMPKα phosphorylation ( pAMPKα ) /AMPKα ratio compared with controls , whereas O+B co-treatment decreased the pAMPKα/AMPKα ratio , compared with olanzapine only treatment . The pAMPKα/AMPKα ratio was positively correlated with total food intake and P35367 expression . Although olanzapine administration decreased the P01189 mRNA level , this level was not affected by O+B co-treatment . Therefore , these results suggested that co-treatment with betahistine may reverse olanzapine-induced body weight gain via the P35367 - P01303 and P35367 -pAMPKα pathways . P62158 -sensitive adenylyl cyclases mediate AVP-dependent DB02527 production and Cl- secretion by human autosomal dominant polycystic kidney cells . In autosomal dominant polycystic kidney disease ( ADPKD ) , binding of AVP to the V2 receptor ( P30518 ) increases DB02527 and accelerates cyst growth by stimulating cell proliferation and Cl(-)-dependent fluid secretion . Basal DB02527 is elevated in human ADPKD cells compared with normal human kidney ( NHK ) cells . P30518 mRNA levels are elevated in ADPKD cells ; however , AVP caused a greater increase in global DB02527 in NHK cells , suggesting an intrinsic difference in DB02527 regulation . Expression , regulatory properties , and receptor coupling of specific adenylyl cyclases ( ACs ) provide temporal and spatial regulation of the DB02527 signal . ADPKD and NHK cells express mRNAs for all nine ACs . Ca(2+)-inhibited ACs 5 and 6 are increased in ADPKD cells , while Ca(2+)/ P62158 -stimulated ACs 1 and 3 are downregulated . ACs 1 , 3 , 5 , and 6 were detected in cyst cells in situ , and codistribution with aquaporin-2 suggests that these cysts were derived from collecting ducts . To determine the contribution of P62158 -sensitive ACs to AVP signaling , cells were treated with W-7 , a P62158 inhibitor . W-7 decreased AVP-induced DB02527 production and Cl(-) secretion by ADPKD cells . CaMKII inhibition increased AVP-induced DB02527 , suggesting that DB02527 synthesis is mediated by O60266 . In contrast , P62158 and CaMKII inhibition in NHK cells did not affect AVP-induced DB02527 production . Restriction of intracellular Ca(2+) switched the response in NHK cells , such that P62158 inhibition decreased AVP-induced DB02527 production . We suggest that a compensatory response to decreased Ca(2+) in ADPKD cells switches P30518 coupling from Ca(2+)-inhibited ACs 5/6 to Ca(2+)/ P62158 -stimulated O60266 , to mitigate high DB02527 levels in response to continuous AVP stimulation . O00206 and Q99836 -dependent signaling mechanisms of the innate immune system are essential for the response to lipopolysaccharide by epithelial and stromal cells of the bovine endometrium . Infection of the bovine endometrium with Gram-negative bacteria commonly causes uterine disease . O00206 ( O00206 ) on cells of the immune system bind Gram-negative bacterial lipopolysaccharide ( LPS ) , stimulating the secretion of the proinflammatory cytokines interleukin 1B ( P01584 ) and P05231 , and the chemokine P10145 . Because the endometrium is the first barrier to infection of the uterus , the signaling cascade triggered by LPS and the subsequent expression of inflammatory mediators were investigated in endometrial epithelial and stromal cells , and the key pathways identified using short interfering RNA ( siRNA ) and biochemical inhibitors . Treatment of endometrial cells with ultrapure LPS stimulated an inflammatory response characterized by increased P01584 , P05231 , and P10145 mRNA expression , and P05231 protein accumulation in epithelial cells , and by increased P01584 and P10145 mRNA expression , and P05231 and P10145 protein accumulation in stromal cells . Treatment of endometrial cells with LPS also induced the degradation of IKB and the nuclear translocation of NFKB , as well as rapid phosphorylation of mitogen-activated protein kinase 3/1 ( P27361 /1 ) and Q16539 . Knockdown of O00206 or its signaling adaptor molecule , myeloid differentiation factor 88 ( Q99836 ) , using siRNA reduced the inflammatory response to LPS in epithelial and stromal cells . Biochemical inhibition of P27361 /1 , but not JNK or Q16539 , reduced LPS-induced P01584 , P05231 , and P10145 expression in endometrial cells . In conclusion , epithelial and stromal cells have an intrinsic role in innate immune surveillance in the endometrium , and in the case of LPS this recognition occurs via O00206 - and Q99836 -dependent cell signaling pathways . DB00659 : recent findings and future research directions . This article explores the mechanisms of action and the potential responder profile of acamprosate , a compound efficacious in relapse prevention of alcoholism . New evidence at the molecular and cellular level suggests that acamprosate attenuates hyper-glutamatergic states that occur during early abstinence and involves iono ( DB01221 ) - and metabotrotropic ( P41594 ) glutamate receptors along with augmented intracellular calcium release and electrophysiological changes . Thus mutant mice with enhanced glutamate levels exhibit higher alcohol consumption than wild type mice and respond better to acamprosate , demonstrating that acamprosate acts mainly on a hyper-glutamatergic system . This mode of action further suggests that acamprosate exhibits neuroprotective properties . In rats , cue-induced reinstatement behavior is significantly reduced by acamprosate treatment whereas cue-induced craving responses in alcohol-dependent patients seem not to be affected by this treatment . An ongoing study ( " Project Predict " ) defines specific responder profiles for an individualized use of acamprosate and naltrexone . Neurophysiological as well as psychometric data are used to define 2 groups of patients : " reward cravers " and " relief cravers " . While naltrexone should work better in the first group , acamprosate is hypothesized to be efficacious in the latter where withdrawal associated and/or cue induced hyper-glutamatergic states are thought to trigger relapse . Further research should target the definition of subgroups applying endophenotypic approaches , e.g. by detecting a hyperglutamatergic syndrome using MR spectroscopy .

MH_train_1209

MH_train_1209

MH_train_1209

interacts_with DB01197?

multiple_choice

DB01197 inhibits neutrophil synthesis of leukotriene B4 in vitro and in vivo . The aim of this investigation was to determine the effects of metalloproteinase inhibitors on leukotriene ( LT ) A4 hydrolase in human neutrophil cytosol and to examine the effects of captopril on intact neutrophils in vitro and in vivo . Cytosolic fractions were assayed for P09960 hydrolase and P09917 activity in the presence or absence of inhibitors . Only bestatin , 1,10-O-phenanthroline , captopril and fosinoprilat demonstrated significant effects . The IC50 of captopril and fosinoprilat for P09960 hydrolase activity were 500 microM and 1 mM , respectively . No inhibition of P09917 activity in cytosolic fractions was detected . The effect of captopril was only minimally reversed by ZnSO4 . The IC50 of captopril for inhibition of LTB4 synthesis in intact neutrophils was 63 microM . Furthermore , 5-HETE production in intact cells was diminished 25.3 +/- 8.5 % in the presence of 1 mM captopril . Oral captopril inhibited stimulated LTB4 release by subsequently isolated neutrophils by 48.1 +/- 5.6 % and 5-HETE release by 43.2 +/- 5.5 % . Thus , captopril is an inhibitor of LTB4 synthesis in neutrophils in vitro and in vivo . However , there are differences between the potency of this drug as assessed in cytosol and intact cell studies . This study significantly extends previous reports in that it demonstrates that captopril is a more potent inhibitor of LTB4 synthesis in intact neutrophils than in cytosol and in that it demonstrates an inhibitory effect of captopril on synthesis of LTB4 by neutrophils exposed to captopril in vivo . P49238 receptor is up-regulated in monocytes of coronary artery diseased patients : impact of pre-inflammatory stimuli and renin-angiotensin system modulators . P78423 / P49238 pathway is considered a major modulator of atherosclerosis . In the present study , expression of P49238 on PBMCs/monocytes of healthy individuals and coronary artery diseased patients was initially assessed by flow cytometry . Effects of pre-inflammatory cytokines interferon ( P27352 ) -gamma and tumor necrosis factor ( P01375 ) -alpha on expression of P49238 and a single representative of each major chemokine family ( P51681 and P61073 ) were further assessed in three cell models : THP-1 monocytes , Jurkat T lymphocytes and primary monocytes isolated from healthy donors . Finally , effects of angiotensin-converting enzyme ( P12821 ) inhibitors captopril , lisinopril and angiotensin receptor blocker ( ARB ) losartan on chemokine receptor expression were evaluated in the same cell models either in a naive or stimulated state . P27352 -gamma significantly affected the chemokine receptor phenotype of THP-1 cells by increasing the rate of P49238 -positive cells . Pre-treatment with the P12821 inhibitors , captopril and lisinopril , and the ARB , losartan , did not influence these effects . DB01197 and lisinopril similarly had no effect on either stimulated or naive primary monocytes . Yet , a small but repeatable increase in P49238 expression after treatment with losartan was noted . Nevertheless , the latter observation did not retain statistical significance after applying the Bonferroni correction . In conclusion , our data did not indicate any significant effect of the P12821 inhibitors on the chemokine receptor phenotype of monocytes . DB02010 -induced growth inhibition of glioma cells is accompanied by altered expression of cyclins , CDKs and CDK inhibitors . DB02010 was found to bring about complete growth inhibition of human glioma cell lines . U87 MG cells were arrested in S phase while U373 MG cells in G2/M phase on staurosporine treatment . Consistent with this observation , no change in P55008 phase regulators viz. , P12004 D1 , D3 and P11802 was seen on staurosporine treatment . The levels of P24941 , P06493 , P12004 A and P12004 B proteins decreased , while the levels of CDK inhibitors viz. , P38936 and p27 were found to increase on staurosporine treatment . The mRNA levels of P24941 and P06493 genes were also found to decrease on staurosporine treatment . Thus apart from staurosporine 's known direct inhibitory effect on P24941 and P06493 activities , staurosporine was found to down-regulate activities of these two kinases by modulating the expression of the kinases themselves as well that of their activating partners ( Cyclins ) and their inhibitors . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . Human parainfluenza virus-associated respiratory tract infection among children and genetic analysis of HPIV-3 strains in Beijing , China . The relevance of human parainfluenza viruses ( HPIVs ) to the epidemiology of acute respiratory infections ( Q9Y4X5 ) in China is unclear . From May 2008 to September 2010 , 443 nasopharyngeal aspirates ( NPAs ) from hospitalized pediatric patients ( age from 1 to 93 months ) in Beijing were collected and screened for HPIVs and other common respiratory viruses by real-time RT-PCR . Sixty-two of 443 samples were positive for HPIVs with 4 positive for HPIV-2 and 58 positive for HPIV-3 , indicating that HPIV-3 was the predominant virus present during the study period . A phylogenetic tree based on all the available HN ( hemagglutinin-neuraminidase ) sequences of HPIV-3 indicated that three distinct clusters ( A,B , and C ) were circulating with some temporal and regional clustering . Cluster C was further divided into sub-clusters , C1 , P06681 , P01024 and C4 . HPIV-3 from Beijing isolates belonged to sub-cluster P01024 , and were grouped with the isolates from two Provinces of China and the neighboring country of Japan . Genetic analysis based on entire HN gene revealed that the HPIV-3 isolates from Beijing were highly similar with 97.2 % -100 % identity at the nucleotide level and these could be divided into two closely related lineages , C3a and C3b . These findings suggested that there was co-circulation of multiple lineages of HPIV-3 in the Beijing region during the study period . This is the first study to describe the epidemiology and molecular characterization of HPIVs in China . Comparative expression analysis of the renin-angiotensin system components between white and brown perivascular adipose tissue . Recent studies have demonstrated that the rat adipose tissue expresses some of the components necessary for the production of angiotensin II ( Ang II ) and the receptors mediating its actions . The aim of this work is to characterize the expression of the renin-angiotensin system ( DB01367 ) components in perivascular adipose tissue and to assess differences in the expression pattern depending on the vascular bed and type of adipose tissue . We analyzed Ang I and Ang II levels as well as mRNA levels of DB01367 components by a quantitative RT-PCR method in periaortic ( PAT ) and mesenteric adipose tissue ( P24752 ) of 3-month-old male Wistar-Kyoto rats . PAT was identified as brown adipose tissue expressing uncoupling protein-1 ( P25874 -1 ) . It had smaller adipocytes than those from P24752 , which was identified as white adipose tissue . All DB01367 components , except renin , were detected in both PAT and P24752 . Levels of expression of angiotensinogen , Ang-converting enzyme ( P12821 ) , and Q9BYF1 were similar between PAT and P24752 . O75787 expression was five times higher , whereas expression of chymase , AT(1a) , and AT(2) receptors were significantly lower in PAT compared with P24752 respectively . In addition , three isoforms of the AT(1a) receptor were found in perivascular adipose tissue . The AT(1b) receptor was found at very a low expression level . Ang II levels were higher in P24752 with no differences between tissues in Ang I . The results show that the DB01367 is differentially expressed in white and brown perivascular adipose tissues implicating a different role for the system depending on the vascular bed and the type of adipose tissue . Selective inhibition of the tumor marker O60218 by antiinflammatory N-phenylanthranilic acids and glycyrrhetic acid . A human aldose reductase-like protein , O60218 in the aldo-keto reductase ( AKR ) superfamily , was recently identified as a tumor marker of several types of cancer . DB02383 , an aldose reductase inhibitor ( Q9Y4X5 ) , is known to be the most potent inhibitor of the enzyme . In this study , we compared the inhibitory effects of other ARIs including flavonoids on O60218 and aldose reductase to evaluate their specificity . However , ARIs showed lower inhibitory potency for O60218 than for aldose reductase . In the search for potent and selective inhibitors of O60218 from other drugs used clinically , we found that non-steroidal antiinflammatory N-phenylanthranilic acids , diclofenac and glycyrrhetic acid competitively inhibited O60218 , showing K(i) values of 0.35-2.9 microM and high selectivity to this enzyme ( 43-57 fold versus aldose reductase ) . Molecular docking studies of mefenamic acid and glycyrrhetic acid in the O60218 -nicotinamide adenine dinucleotide phosphate ( NADP(+) ) complex and site-directed mutagenesis of the putative binding residues suggest that the side chain of Val301 and a hydrogen-bonding network among residues Val301 , Gln114 and Ser304 are important for determining the inhibitory potency and selectivity of the non-steroidal antiinflammatory drugs . Thus , the potent and selective inhibition may be related to the cancer chemopreventive roles of the drugs , and their structural features may facilitate the design of new anti-cancer agents targeting O60218 . Enhanced therapeutic effects of doxorubicin and paclitaxel in combination with liposome-entrapped ends-modified raf antisense oligonucleotide against human prostate , lung and breast tumor models . P04049 protein kinase plays an important role in cell growth , proliferation and cell survival . We have previously described the use of liposome-entrapped antisense raf oligonucleotide ( DB04973 ) to inhibit P04049 expression resulting in tumor growth inhibition and radiosensitization . The present study was undertaken to evaluate the chemosensitization effects of DB04973 in combination with doxorubicin or paclitaxel on a panel of human tumor xenografts . DB04973 ( 25.0 mg/kg i.v. x 10 ) displayed significant antitumor activity ( P < 0.05 ) when administered as a single agent in prostate ( PC-3 ) , lung ( A549 ) and breast ( MDA-MB 231 ) carcinoma models . Doxorubicin ( 1.0-4.0 mg/kg i.v. per week x 3 ) and paclitaxel ( 1.0-4.0 mg/kg i.v. on alternate days x 3 ) were administered as single agents at non-toxic doses that led to only minimal to moderate antitumor activity . However , a combination of DB04973 with doxorubicin or paclitaxel led to significantly enhanced antitumor activity in all the tumor types tested ( PC-3 , P < 0.03 ; A549 , P < 0.035 ; MDA-MB 231 , P < 0.045 ) as compared with DB04973 alone or chemotherapeutic agents alone treated groups . This effect of chemosensitization appeared to be sequence-specific because a mismatch control oligonucleotide continued to show significant tumor growth . Additionally , no inhibition in P04049 expression in MDA-MB 231 tumor tissue was observed with mismatch oligonucleotide treatment . On the other hand , DB04973 treatment led to > 75 % inhibition of P04049 expression in tumor tissue . These preclinical observations support the use of DB04973 in combination with chemotherapeutic agents to improve the treatment of human cancers . P09917 -activating protein inhibitors : development of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) . The potent and selective P09917 -activating protein leukotriene synthesis inhibitor 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( 11j ) is described . Lead optimization was designed to afford compounds with superior in vitro and in vivo inhibition of leukotriene synthesis in addition to having excellent pharmacokinetics and safety in rats and dogs . The key structural features of these new compounds are incorporation of heterocycles on the indole N-benzyl substituent and replacement of the quinoline group resulting in compounds with excellent in vitro and in vivo activities , superior pharmacokinetics , and improved physical properties . The methoxypyridine derivative 11j has an IC(50) of 4.2 nM in a P09917 -activating protein ( P20292 ) binding assay , an IC(50) of 349 nM in the human blood Q06643 (4) inhibition assay , and is efficacious in a murine ovalbumin model of allergen-induced asthma . Compound 11j was selected for clinical development and has successfully completed phase 1 trials in healthy volunteers . Suppression of P05121 expression through inhibition of the P00533 -mediated signaling cascade in rat kidney fibroblast by ascofuranone . Fibrosis in glomerulosclerosis causes progressive loss of renal function . Transforming growth factor ( TGF ) -beta , one of the major profibrotic cytokines , induces the synthesis of plasminogen activator inhibitor ( P05121 ) -1 , a factor that plays a crucial role in the development of fibrosis . Here , we found that an isoprenoid antibiotic , ascofuranone , suppresses expression of profibrotic factors including matrix proteins and P05121 induced by TGF-beta in renal fibroblasts . Ascofuranone selectively inhibits phosphorylation of epidermal growth factor receptor ( P00533 ) , and downstream kinases such as P04049 , MEK-1/2 , and P27361 /2 . P05121 transcription also is suppressed by treatment with kinase inhibitors for MEK-1/2 or P00533 , and with small interfering RNA for P00533 . Ascofuranone inhibits cellular metalloproteinase activity , and an inhibitor of metalloproteinases suppresses P00533 phosphorylation and P05121 transcription . These results suggest that ascofuranone suppresses expression of profibrotic factors through the inhibition of an P00533 -dependent signal transduction pathway activated by metalloproteinases . P12821 inhibition down-regulates the pro-atherogenic chemokine receptor 9 ( P51686 ) -chemokine ligand 25 ( O15444 ) axis . Many experimental and clinical studies suggest a relationship between enhanced angiotensin II release by the angiotensin-converting enzyme ( P12821 ) and the pathophysiology of atherosclerosis . The atherosclerosis-enhancing effects of angiotensin II are complex and incompletely understood . To identify anti-atherogenic target genes , we performed microarray gene expression profiling of the aorta during atherosclerosis prevention with the P12821 inhibitor , captopril . Atherosclerosis-prone apolipoprotein E ( apoE ) -deficient mice were used as a model to decipher susceptible genes regulated during atherosclerosis prevention with captopril . Microarray gene expression profiling and immunohistology revealed that captopril treatment for 7 months strongly decreased the recruitment of pro-atherogenic immune cells into the aorta . DB01197 -mediated inhibition of plaque-infiltrating immune cells involved down-regulation of the C-C chemokine receptor 9 ( P51686 ) . Reduced cell migration correlated with decreased numbers of aorta-resident cells expressing the P51686 -specific chemoattractant factor , chemokine ligand 25 ( O15444 ) . The O15444 - P51686 axis was pro-atherogenic , because inhibition of P51686 by RNA interference in hematopoietic progenitors of apoE-deficient mice significantly retarded the development of atherosclerosis . Analysis of coronary artery biopsy specimens of patients with coronary artery atherosclerosis undergoing bypass surgery also showed strong infiltrates of P51686 -positive cells in atherosclerotic lesions . Thus , the C-C chemokine receptor , P51686 , exerts a significant role in atherosclerosis . Design and synthesis of 3,5-disubstituted-1,2,4-oxadiazoles as potent inhibitors of phosphodiesterase4b2 . A series of 3,5-disubstituted-1,2,4-oxadiazoles has been prepared and evaluated for phosphodiesterase inhibition ( PDE4B2 ) . Among the prepared 3,5-disubstituted-1,2,4-oxadiazoles , compound 9a is the most potent inhibitor ( PDE4B2 IC(50) = 5.28 μm ) . Structure-activity relationship studies of 3,5-disubstituted-1,2,4-oxadiazoles revealed that substituents 3-cyclopentyloxy-4-methoxyphenyl group at 3-position and cyclic ring bearing heteroatoms at 5-position are important for activity . Molecular modeling study of the 3,5-disubstituted-1,2,4-oxadiazoles with Q07343 has shown similar interactions of 3-cyclopentyloxy-4-methoxyphenyl group ; however , heteroatom ring is slightly deviating when compared to DB01791 . 3-(3-Cyclopentyloxy-4-methoxyphenyl)-5-(piperidin-4-yl)-1,2,4-oxadiazole ( 9a ) exhibited good analgesic and antiinflammatory activities in formalin-induced pain in mice and carrageenan-induced paw edema model in rat . P00797 -angiotensin-aldosterone system in brain infarction and vascular death . The renin-angiotensin-aldosterone system has functions that may contribute to brain infarction ( BI ) . In 459 matched pairs of white patients and control subjects , we measured plasma angiotensin-converting enzyme ( P12821 ) levels , seven polymorphisms ( angiotensinogen T174M and M235T , P12821 I/D and 4656 2/3CT repeat [ rpt ] , angiotensin II type 1 receptor A1166C and A153G , and aldosterone synthase P19099 ) , and evaluated 5-year poststroke mortality . Mean plasma P12821 levels ( +/-standard error ) were significantly greater in patients than control subjects ( 37.5 +/- 0.9 vs 33.9 +/- 0.9 ) , in patients with lacunar stroke , and in patients with no previous vascular ( cerebrovascular or cardiovascular ) history . The risk for BI increased with tertiles of plasma P12821 , without an interaction with hypertension . After adjustments , the association disappeared except among patients with cardioembolic BI and those without previous vascular events . Among the polymorphisms , there was a weak association of BI with angiotensin II type 1 receptor 1166C , a weak protective effect with angiotensinogen 174M , and a strong association of angiotensinogen 235T with 5-year vascular mortality . These results suggest that renin-angiotensin-aldosterone system activity and genes contribute to cerebrovascular disease and poststroke vascular death in white patients . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . Synthesis , biological evaluation , and molecular modeling studies of methylene imidazole substituted biaryls as inhibitors of human 17alpha-hydroxylase-17,20-lyase ( P05093 ) -- part II : Core rigidification and influence of substituents at the methylene bridge . Thirty-five novel substituted imidazolyl methylene biphenyls have been synthesized as P05093 inhibitors for the potential treatment of prostate cancer . Their activities have been tested with recombinant human P05093 expressed in Escherichia coli . Promising compounds were tested for selectivity against P15538 , P19099 , and hepatic CYP enzymes 3A4 , 1A2 , 2B6 and 2D6 . The core rigidified compounds ( 30-35 ) were the most active ones , being much more potent than Ketoconazole and reaching the activity of DB05812 . However , they were not very selective . Another rather potent and more selective inhibitor ( compound 23 , IC(50)=345 nM ) was further examined in rats regarding plasma testosterone levels and pharmacokinetic properties . Compared to the reference DB05812 , 23 was more active in vivo , showed a longer plasma half-life ( 10h ) and a higher bioavailability . Using our P05093 homology protein model , docking studies with selected compounds were performed to study possible interactions between inhibitors and amino acid residues of the active site . Modulation of cytokine production and enhancement of cell viability by Q9NYK1 and Q9NR96 ligands during anthrax infection of macrophages . Inhalation of Bacillus anthracis , a bioterrorism agent , results in a high mortality rate despite appropriate antibiotic therapy . Macrophages appear to be a key factor in B. anthracis pathogenesis . The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax . However , preactivation of Toll-like receptors ( TLRs ) could modify the host response . TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens . We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli . We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines P05231 , P01375 , P22301 , and IL-12 p40 . Our studies showed that pretreatment of mouse macrophages with the Q9NR96 ligand DB05463 , or the Q9NYK1 ligands R-848 and IT-37 , results in a substantial decrease in the subsequent secretion of P05231 and P01375 in response to B. anthracis infection of macrophages . Furthermore , the Q9NYK1 and Q9NR96 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages . These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability . [ The effect of blood pressure-reducing therapy with captopril on tubular marker excretion in type-1 diabetics with nephropathy ] . A prospective open clinical trial was carried out with 23 hypertensive type I diabetics ( 13 men , ten women , mean age 49 +/- 9.1 years , duration of diabetes 18 +/- 9.1 years ) with early nephropathy . Glomerular and tubular renal function and metabolic parameters were monitored during 8 months ' treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , captopril , in addition to previous antihypertensive treatment with one or more drugs . Blood pressure control tended to improve on captopril ( systolic pressures 152 +/- 13 vs 140 +/- 13 mm Hg , P < 0.05 ; diastolic pressures 89 +/- 10 vs 87 +/- 10 mm Hg , not significant ) . Proteinuria ( > 0.5 g/24 hours ) fell into the microalbuminuria range ( albumin excretion 2-20 mg/mmol creatinine ) in four out of 13 patients , and microalbuminuria disappeared in four out of ten patients . Urinary levels of the brush border enzyme O60502 ( NAG ) , a marker of tubular dysfunction , were initially raised and fell significantly after 8 months ' treatment with captopril ( 20.3 +/- 14.4 vs 8.8 +/- 8.1 U/g creatinine ; P < 0.01 ) . DB01197 did not affect metabolic control ( HbA1 , total , HDL and LDL cholesterol , triglycerides , apolipoproteins A1 and B ) or the insulin dosage . These results show that long-term treatment with captopril may favourably influence both albumin excretion and NAG activity , a marker of tubular dysfunction , in type I diabetics with nephropathy . DB02426 effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 and P05141 may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 -/- mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 -independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 -/- brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 -independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 -/- mice . However , in liver , only Ant2 mRNA was found , whereas in brown adipose tissue , Ant1 and Ant2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 isoform mediates fatty-acid-induced uncoupling , whereas the P12235 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria . Migration potential and gene expression profile of human DB05914 induced by O15444 . Recruitment of DB05914 ( O60682 ) to tissue damages is a promising approach for in situ tissue regeneration . The physiological mechanisms and regulatory processes of O60682 trafficking to injured tissue remain poorly understood . However , the pivotal role of chemokines in O60682 recruitment has already been shown . The aim of this study was to determine the migratory potential and the gene expression profile of O60682 stimulated with the CC chemokine O15444 ( O15444 ) . Bone marrow derived human O60682 were exposed to different doses of O15444 in a standardized chemotaxis assay . Microarray gene expression profiling and pathway analysis were performed for O15444 stimulated O60682 . Maximum migration of O60682 towards O15444 was observed at 10(3) nM . Microarray analysis revealed an induction of molecules directly involved in chemotaxis and homing of bone marrow cells ( P09341 -3 , P10145 , Q07343 ) , cytoskeletal and membrane reorganisation ( P10145 , Q13393 , P08833 ) , cellular polarity ( Q13393 ) , and cell movement ( P09341 -3 , P80162 , P10145 , P35354 , Q07343 , P21980 ) . Respective chemokine secretion was confirmed by protein membrane-array analysis . The activation of P25025 ligands ( P09341 -3 , P42830 -6 , P10145 ) and a P15018 -receptor/ P40189 ligand ( P15018 ) indicated an involvement of the respective signaling pathways during initiation of chemotaxis and migration . These results suggest O15444 as a new potential candidate for further in situ regeneration approaches . Rapid mobilization of functional donor hematopoietic cells without G- P04141 using DB06809 , an antagonist of the P61073 / P48061 interaction . Allografts from HLA-matched sibling donors were mobilized and collected without granulocyte colony-stimulating factor ( DB00099 ) using DB06809 , a direct antagonist of P61073 /stromal-derived factor 1 ( P48061 / P48061 ) . Donors ( N = 25 ) were treated with DB06809 at a dose of 240 mug/kg by subcutaneous injection , and leukapheresis was then initiated just 4 hours later . Two-thirds of the donors collected an allograft with a P28906 (+) cell dose sufficient for transplantation after just one dose of DB06809 . No donor experienced more than grade 1 toxicity . After a myeloablative regimen , 20 patients with hematologic malignancies received allografts collected after DB06809 alone . All patients engrafted neutrophils ( median day 10 ) and platelets ( median day 12 ) promptly . Acute graft-versus-host disease ( GVHD ) grades 2 through 4 occurred in 35 % of patients . One patient died due to complications related to acute GVHD . No unexpected adverse events were observed in any of the recipients . All 14 patients surviving in remission have robust trilineage hematopoiesis and are transfusion-free with a median follow-up of 277 days ( range , 139-964 days ) . Direct antagonism of P61073 by DB06809 may provide a more rapid and possibly less toxic and cumbersome alternative to traditional G- P04141 -based mobilization in normal donors . This trial was registered as no . NCT00241358 at www.ClinicalTrials.gov . Inhibitors of angiotensin-converting enzyme modulate mitosis and gene expression in pancreatic cancer cells . The angiotensin-converting enzyme ( P12821 ) inhibitor captopril inhibits mitosis in several cell types that contain P12821 and renin activity . In the present study , we evaluated the effect of the P12821 inhibitors captopril and CGS 13945 ( 10(-8) to 10(-2) M ) on proliferation and gene expression in hamster pancreatic duct carcinoma cells in culture . These cells lack renin and P12821 activity . Both P12821 inhibitors produced a dose-dependent reduction in tumor cell proliferation within 24 hr . DB01197 at a concentration of 0.36 mM and CGS 13945 at 150 microM decreased cellular growth rate to approximately half that of the control . Neither drug influenced the viability or the cell cycle distribution of the tumor cells . Slot blot analysis of mRNA for four genes , proliferation associated cell nuclear antigen ( P12004 ) , K-ras , protein kinase C-beta ( P05771 ) and carbonic anhydrase II ( CA II ) was performed . Both P12821 inhibitors increased K-ras expression by a factor of 2 , and had no effect on CA II mRNA levels . DB01197 also lowered P12004 by 40 % and CGS 13945 lowered P05771 gene expression to 30 % of the control level . The data demonstrate that P12821 inhibitors exhibit antimitotic activity and differential gene modulation in hamster pancreatic duct carcinoma cells . The absence of renin and P12821 activity in these cells suggests that the antimitotic action of captopril and CGS 13945 is independent of renin-angiotensin regulation . The growth inhibition may occur through downregulation of growth-related gene expression .

MH_train_1210

MH_train_1210

MH_train_1210

interacts_with DB08815?

multiple_choice

Melatonin receptor potentiation of cyclic AMP and the cystic fibrosis transmembrane conductance regulator ion channel . We have used the cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channel as a model system to study the DB02527 signal transduction pathways coupled to the Xenopus melatonin receptor . During forskolin ( Fsk ) stimulation , melatonin reduced the amplitude of the P13569 currents in oocytes injected with in vitro transcribed cRNAs for the Xenopus melatonin receptor and P13569 . Pertussis toxin ( Ptx ) treatment eliminated melatonin inhibition of Fsk stimulated P13569 currents . In oocytes injected with cRNA for melatonin receptors , serotonin receptors ( P34969 ) , and P13569 Cl- channels , application of melatonin together with serotonin ( 5-HT ) activated an additional inward current showing potentiation of adenylyl cyclases by melatonin receptors . Subthreshold activation of P34969 receptors was sufficient and necessary to permit activation of P13569 channels by melatonin . Preexposure to melatonin desensitized the melatonin receptor mediated response . Therefore , based on this model system , the effects of melatonin in vivo could be either positive or negative modulation of other neuronal inputs , depending on the mode of adenylyl cyclase stimulation . Effects of lurasidone on executive function in common marmosets . Cognitive impairment is one of the major symptoms of schizophrenia , and is considered largely due to dysfunctions in the prefrontal cortex ( P27918 ) . DB08815 , a novel atypical antipsychotic agent with high binding affinity for dopamine D2 , serotonin P34969 , 5- Q13049 and P08908 receptors has been reported to have superior efficacy in rodents ' models of cognitive impairment . However , the beneficial effect of lurasidone on cognitive impairment has not been evaluated in non-human primates . In this study , we investigated the effect of lurasidone on executive function , which is one of the cognitive domains , in common marmosets and compared the results to those of other antipsychotics . The effects of lurasidone , haloperidol , olanzapine , risperidone , quetiapine and clozapine on executive function were evaluated in naïve marmosets using the object retrieval with detours ( ORD ) task . Before drug treatment , marmosets ' success rates in the easy trial of the test were almost 90 % . However , maximum success in the difficult trial of the task reached only 50 % after 8 days of training . DB00502 , olanzapine and risperidone decreased correct performance even in the easy trial of the task . All drugs , except lurasidone , impaired success rate in the difficult trial . On the other hand , lurasidone dose-dependently increased marmosets ' success rates in the difficult trial with significant effect at 10mg/kg . In conclusion , we have shown in this study that lurasidone , unlike conventional antipsychotics , improves cognition associated with executive function in common marmosets . These findings suggest that lurasidone would be more useful for treatment of schizophrenia cognitive impairment than other antipsychotics . Chemical ablation of androgen receptor in prostate cancer cells by the histone deacetylase inhibitor LAQ824 . P10275 plays a critical role in the development of primary as well as advanced hormone-refractory prostate cancer . Therefore , ablation of androgen receptor from prostate cancer cells is an interesting concept for developing a new therapy not only for androgen-dependent prostate cancer but also for metastatic hormone-refractory prostate cancer , for which there is no effective treatment available . We report here that LAQ824 , a cinnamyl hydroxamatic acid histone deacetylase inhibitor currently in human clinical trials , effectively depleted androgen receptor in prostate cancer cells at nanomolar concentrations . LAQ824 seemed capable of depleting both the mutant and wild-type androgen receptors in either androgen-dependent and androgen-independent prostate cancer cells . Although LAQ824 may exert its effect through multiple mechanisms , several lines of evidence suggest that inactivation of the heat shock protein-90 ( Hsp90 ) molecular chaperone is involved in LAQ824-induced androgen receptor depletion . Besides androgen receptor , LAQ824 reduced the level of Hsp90 client proteins HER-2 ( ErbB2 ) , Akt/ P31749 , and P04049 in LNCaP cells . Another Hsp90 inhibitor , 17-allyamino-17-demethoxygeldanamycin ( 17- P29372 ) , also induced androgen receptor diminution . LAQ824 induced Hsp90 acetylation in LNCaP cells , which resulted in inhibition of its DB00171 -binding activity , dissociation of Hsp90-androgen receptor complex , and proteasome-mediated degradation of androgen receptor . Consequently , LAQ824 blocked androgen-induced prostate-specific antigen production in LNCaP cells . LAQ824 effectively inhibited cell proliferation and induced apoptosis of these prostate cancer cells . These results reveal that LAQ824 is a potent agent for depletion of androgen receptor and a potential new drug for prostate cancer . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Signal regulatory protein alpha ( SIRPalpha ) cells in the adaptive response to ESAT-6/ P27918 -10 protein of tuberculous mycobacteria . BACKGROUND : Early secretory antigenic target-6 ( ESAT-6 ) and culture filtrate protein-10 ( P27918 -10 ) are co-secreted proteins of Mycobacterium tuberculosis complex mycobacteria ( includes M. bovis , the zoonotic agent of bovine tuberculosis ) involved in phagolysosome escape of the bacillus and , potentially , in the efficient induction of granulomas . Upon tuberculosis infection , multi-nucleate giant cells are elicited , likely as a response aimed at containing mycobacteria . In tissue culture models , signal regulatory protein ( P78324 )alpha ( also referred to as macrophage fusion receptor or CD172a ) is essential for multi-nucleate giant cell formation . METHODOLOGY/PRINCIPAL FINDINGS : In the present study , ESAT-6/ P27918 -10 complex and SIRPalpha interactions were evaluated with samples obtained from calves experimentally infected with M. bovis . Peripheral blood CD172a(+) ( SIRPalpha-expressing ) cells from M. bovis-infected calves proliferated upon in vitro stimulation with ESAT-6/ P27918 -10 ( either as a fusion protein or a peptide cocktail ) , but not with cells from animals receiving M. bovis strains lacking ESAT-6/ P27918 -10 ( i.e , M. bovis BCG or M. bovis DeltaRD1 ) . Sorted CD172a(+) cells from these cultures had a dendritic cell/macrophage morphology , bound fluorescently-tagged rESAT-6: P27918 -10 , bound and phagocytosed live M. bovis BCG , and co-expressed CD11c , O60449 , P16070 , MHC II , P33681 /86 ( a subset also co-expressed CD11b or CD8alpha ) . Intradermal administration of rESAT-6: P27918 -10 into tuberculous calves elicited a delayed type hypersensitive response consisting of CD11c(+) , CD172a(+) , and CD3(+) cells , including CD172a-expressing multi-nucleated giant cells . CONCLUSIONS/SIGNIFICANCE : These findings demonstrate the ability of ESAT-6/ P27918 -10 to specifically expand CD172a(+) cells , bind to CD172a(+) cells , and induce multi-nucleated giant cells expressing CD172a . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . Ca2+ response of rat mesangial cells to DB00171 analogues . The aim of this investigation was to characterise the effects of DB00171 analogues and UTP on the single cell intracellular Ca2+ concentration ( [Ca2+]i ) in cultured rat mesangial cells . Typically , there were two phases in the Ca2+ response to the agonists , an initial fast transient peak and a subsequent slower decline , or plateau , phase . For the peak amplitude in [Ca2+]i the agonists had about equal effect . But when taking in consideration the percentage of responding cells and the integrated Ca2+ response over 1 min , the order of efficacy of nucleotide agonists ( 100 microM ) was UTP = DB00171 > ATPgammaS > ADP = 2MeS- DB00171 ( 2-methylthio- DB00171 ) . DB00640 , AMP and beta,gamma-Me- DB00171 ( 100 microM ) had no effect . DB04786 ( 100 microM ) and reactive blue ( 50 microM ) decreased the number of responding cells . Removing Ca2+ from the bath diminished neither the peak in [Ca2+]i nor the percentage of responding cells , but the average [Ca2+]i increase in 1 min was significantly reduced . The results indicate that P41231 receptors are present in rat mesangial cells but it can not be excluded that there are receptors distinct from P41231 which also mediate a rise in [Ca2+]i . DB08890 -a novel secretagogue in the treatment of irritable bowel syndrome with constipation and chronic idiopathic constipation . Irritable bowel syndrome with constipation ( IBS-C ) and chronic idiopathic constipation ( Q96RK0 ) are highly prevalent gastrointestinal disorders . Traditional symptoms based therapies had somewhat limited success and efficacy in addressing the disorders . Recently , linaclotide emerged as novel peptide capable of improving abdominal symptoms in patients suffering from IBS-C and Q96RK0 . Guanylate cyclase C ( P25092 ) receptor a multi domain protein , found to be molecular target for linaclotide which acts by activating P25092 receptor on the apical surface of intestinal epithelial cells . Binding of linaclotide to P25092 receptor triggers the elevation of second messenger cGMP that elicits fluid secretion into intestinal cells which play a critical role in maintaining homeostasis through cystic fibrosis transmembrane conductance regulator ( P13569 ) . Data from Phase II and III clinical trials demonstrated that linaclotide seems to produce a statistically significant increase in stool frequency , improved straining , decreased abdominal pain and discomfort . Constitutive interaction of the P41231 receptor with the hematopoietic cell-specific G protein G(alpha16) and evidence for receptor oligomers . Hematopoietic cells uniquely express G(alpha16) , a G protein alpha-subunit of the G(q)-type . G(alpha16) is obligatory for P41231 receptor-dependent Ca2+-mobilization in human erythroleukemia cells and induces hematopoietic cell differentiation . We tested whether P41231 receptors physically interact with G(alpha16) . Receptor and G protein were fused to cyan ( P27918 ) and yellow ( YFP ) variants of the green fluorescent protein ( GFP ) , respectively . When expressed in K562 leukemia cells , the fusion proteins were capable of triggering a Ca2+-signal upon receptor stimulation , demonstrating their functional integrity . In fluorescence resonance energy transfer ( FRET ) measurements using confocal microscopy , a strong FRET signal from the plasma membrane region of fixed , resting cells was detected when the receptor was co-expressed with the G protein as the FRET acceptor , as well as when the P27918 -tagged receptor was co-expressed with receptor fused to YFP . We conclude that , under resting conditions , G(alpha16) and P41231 receptors form constitutive complexes , and that the P41231 receptor is present as an oligomer . [ Is DB08901 ( DB08901 ) the next treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia ? ] . Distinct clinicopathologic acute lymphoblastic leukemia ( ALL ) entities have been identified , resulting in the adoption of risk-oriented treatment approaches . In Philadelphia chromosome-positive ( Ph(+) ) ALL , the optimal treatment requires the addition of P11274 - P00519 tyrosine kinase inhibitors , as imatinib . However , the outcome remains poor in absence of allogeneic stem cell transplantation , and novel agents are desperately required . Resistance attributable to kinase domain mutations can lead to relapse despite the development of second-generation compounds , including dasatinib and nilotinib . Despite these therapeutic options , the cross-resistant P11274 - P00519 ( T315I ) mutation remains a major clinical challenge . The first evaluations of DB08901 present this drug as a potent multi-targeted kinase inhibitor active against T315I and all other P11274 - P00519 mutants . DB08901 could be the next treatment of choice in hematological malignancies with Philadelphia-positive chromosome , particularly Ph(+) ALL known for its frequent occurrence of T315I mutation . DB00435 modulates the frog heart ventricle morphodynamics . The aim of this work was to investigate in the avascular heart of the frog Rana esculenta the influence of nitric oxide ( NO ) on ventricular systolic and diastolic functions by using a novel image analysis technique . The external volume variations of the whole ventricle were monitored during the heart cycle by video acquisition ( visible light ) and analysed by an appropriately developed software with a specific formula for irregular convex solids . The system , which measures the rate of volume changes and the ejection fraction , directly determined the volumetric behaviour of the working frog heart after stimulation or inhibition of NOS-NOcGMP pathway . End-diastolic volume ( EDVext ) , end-systolic volume ( ESVext ) , contraction and relaxation velocities ( dV/dtsys and dV/dtdia , respectively ) , stroke volume ( SV ) and ejection fraction ( EF ) , were measured before and after perfusion with NOS substrate ( L-arginine ) , NO donor ( SIN-1 ) , cGMP analogue (8-Br-cGMP),NOS inhibitors ( NG-monomethyl-L-arginine , L-NMMA ; L-N(5)-(1-iminoethyl)-ornithine , L-NIO ; DB02207 ,7-NI ) and guanylyl cyclase inhibitor ( ODQ ) . The results showed that NO reduces ventricular systolicfunction improving diastolic filling , while NOS inhibition increases contractility impairing ventricular filling capacity . The presence of activated P29474 ( p- P29474 ) was morphologically documented , further supporting that the mechanical activity of the ventricular pump in frog is influenced by a tonic release of NOS-generated NO . Effects of antihistamines on the function of human α7-nicotinic acetylcholine receptors . Effects of the histamine H₁ receptor ( P35367 ) antagonists ( antihistamines ) , promethazine ( PMZ ) , orphenadrine ( ORP ) , chlorpheniramine ( CLP ) , DB06691 ( PYR ) , diphenhydramine ( DPH ) , citerizine ( CTZ ) , and triprolidine ( TRP ) on the functional properties of the cloned α7 subunit of the human nicotinic acetylcholine receptor expressed in Xenopus oocytes were investigated . Antihistamines inhibited the α7-nicotinic acetylcholine receptor in the order PYR > CLP > TRP > PMZ > ORP≥DPH≥CTZ . Among the antihistamines , PYR showed the highest reversible inhibition of acetylcholine ( 100 µM ) -induced responses with IC₅₀ of 6.2 µM . PYR-induced inhibition was independent of the membrane potential and could not be reversed by increasing the concentration of acetylcholine . Specific binding of [ ¹²⁵I ] α-bungarotoxin , a selective antagonist for α7-nicotinic acetylcholine receptor , was not changed in the presence of PYR suggesting a non-competitive inhibition of nicotinic receptors . In line with functional experiments , docking studies indicated that PYR can potentially bind allosterically with the α7 transmembrane domain . Our results indicate that the H₂-H₄ receptor antagonists tested in this study ( 10 µM ) showed negligible inhibition of α7-nicotinic acetylcholine receptors . On the other hand , H₁ receptor antagonists inhibited the function of human α7-nicotinic acetylcholine receptor , with varying potencies . These results emphasize the importance of α7-nicotinic acetylcholine receptor for future pharmacological/toxicological profiling . Irritable bowel syndrome neuropharmacology . A review of approved and investigational compounds . Anticholinergics and prokinetics are mainstays of therapy for Irritable Bowel Syndrome ( IBS ) patients despite their limited efficacy and troublesome side-effect profile . The clinical limitations of these drugs are a result of their relative broad and nonspecific pharmacologic interaction with various receptors . Recent advances in gut physiology have led to the identification of various receptor targets that may play a pivotal role in the pathogenesis of IBS . Medicinal chemists searching for safe and effective IBS therapies are now developing compounds targeting many of these specific receptors . The latest generation of anticholinergics , such as zamifenacin , darifenacin , and YM-905 , provide selective antagonism of the muscarinic type-3 receptor . DB01079 , a selective Q13639 partial agonist , tested in multiple clinical trials , is effective in reducing the symptoms of abdominal pain , bloating , and constipation . Ezlopitant and nepadudant , selective antagonists for neurokinin receptors type 1 and type 2 , respectively , show promise in reducing gut motility and pain . DB00836 , a mu ( mu ) opioid receptor agonist , is safe and effective for IBS patients with diarrhea ( IBS-D ) as the predominant bowel syndrome . Fedotozine , a kappa ( kappa ) opioid receptor agonist , has been tried as a visccral analgesic in various clinical trials with conflicting results . DB00969 , a 5- Q9H205 receptor antagonist , has demonstrated efficacy in IBS-D patients but incidents of ischemic colitis seen in post-marketing follow-up resulted its removal from the market . Compounds that target cholecystokinin . A , N-methyl-D-aspartate , alpha 2-adrenergic , and DB05394 receptors are also examined in this review . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . Presynaptic serotonergic inhibition of GABAergic synaptic transmission in mechanically dissociated rat basolateral amygdala neurons . 1. The basolateral amygdala ( P00519 ) nuclei contribute to the process of anxiety . GABAergic transmission is critical in these nuclei and serotonergic inputs from dorsal raphe nuclei also significantly regulate GABA release . In mechanically dissociated rat P00519 neurons , spontaneous miniature inhibitory postsynaptic currents ( mIPSCs ) arising from attached GABAergic presynaptic nerve terminals were recorded with the nystatin-perforated patch method and pharmacological isolation . 2 . 5-HT reversibly reduced the GABAergic mIPSC frequency without affecting the mean amplitude . The serotonergic effect was mimicked by the P08908 specific agonist 8-OH DPAT ( 8-hydroxy-2-(di-n-propylamino)tetralin ) and blocked by the P08908 antagonist spiperone . 3 . The GTP-binding protein inhibitor N-ethylmaleimide removed the serotonergic inhibition of mIPSC frequency . In either K+-free or Ca2+-free external solution , 5-HT could inhibit mIPSC frequency . 4 . High K+ stimulation increased mIPSC frequency and 8-OH DPAT inhibited this increase even in the presence of Cd2+ . 5 . DB02587 , an activator of adenylyl cyclase ( AC ) , significantly increased synaptic GABA release frequency . Pretreatment with forskolin prevented the serotonergic inhibition of mIPSC frequency in both the standard and high K+ external solution . 6 . Ruthenium Red ( RR ) , an agent facilitating the secretory process in a Ca2+-independent manner , increased synaptic GABA release . 5-HT also suppressed RR-facilitated mIPSC frequency . 7 . We conclude that 5-HT inhibits GABAergic mIPSCs by inactivating the AC- DB02527 signal transduction pathway via a G-protein-coupled P08908 receptor and this intracellular pathway directly acts on the GABA-releasing process independent of K+ and Ca2+ channels in the presynaptic nerve terminals . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . P10275 repression of gonadotropin-releasing hormone gene transcription via enhancer 1 . DB00644 ( DB00644 ) plays a major role in the hypothalamic-pituitary-gonadal ( HPG ) axis , and synthesis and secretion of DB00644 are regulated by gonadal steroid hormones . Disruptions in androgen levels are involved in a number of reproductive defects , including hypogonadotropic hypogonadism and polycystic ovarian syndrome . Androgens down-regulate DB00644 mRNA synthesis in vivo and in vitro via an androgen receptor ( AR ) -dependent mechanism . DB02998 ( R1881 ) , a synthetic AR agonist , represses DB00644 expression through multiple sites in the proximal promoter . In this study , we show AR also represses DB00644 transcription via the major enhancer ( DB00644 -E1 ) . A multimer of the -1800/-1766 region was repressed by R1881 treatment . Mutation of two bases , -1792 and -1791 , resulted in decreased basal activity and a loss of AR-mediated repression . AR bound to the -1796/-1791 sequence in electrophoretic mobility shift assays , indicating a direct interaction with DNA or other transcription factors in this region . We conclude that AR repression of DB00644 -E1 acts via multiple AR-responsive regions , including the site at -1792/-1791 . DB08818 - P16070 interaction stimulates Rac1 signaling and Q16513 kinase activation leading to cytoskeleton function and cell migration in astrocytes . Both hyaluronan [ HA , the major glycosaminoglycans in the extracellular matrix ( Q13201 ) ] and P16070 ( a primary HA receptor ) are associated with astrocyte activation and tissue repair following central nervous system ( CNS ) injury . In this study we investigated the question of whether HA- P16070 interaction influences astrocyte signaling and migration . Our data indicated that HA binding to the cultured astrocytes stimulated Rac1 signaling and cytoskeleton-mediated migration . To determine the cellular and molecular basis of these events , we focused on Q16513 , a Rac1-activated serine/threonine kinase in astrocytes . We determined that HA binding to astrocytes stimulated Rac1-dependent Q16513 kinase activity which , in turn , up-regulated the phosphorylation of the cytoskeletal protein , cortactin , and attenuated the ability of cortactin to cross-link F-actin . Further analyses indicated that the N-terminal antiparallel coiled-coil ( ACC ) domains of Q16513 interacted with Rac1 , and transfection of astrocytes with Q16513 -ACCcDNA inhibited Q16513 activity . Over-expression of the Q16513 -ACC domain also functions as a dominant-negative mutant to block HA/ P16070 -mediated Q16513 activation of cortactin and astrocyte migration . Taken together , these findings strongly suggest that hyaluronan/ P16070 interaction with Rac1- Q16513 plays a pivotal role in cytoskeleton activation and astrocyte migration . These newly discovered HA/ P16070 -induced astrocyte function may provide important insight into novel therapeutic treatments for tissue repair following CNS injury . Pulmonary arterial dysfunction in insulin resistant obese Zucker rats . BACKGROUND : P01308 resistance and obesity are strongly associated with systemic cardiovascular diseases . Recent reports have also suggested a link between insulin resistance with pulmonary arterial hypertension . The aim of this study was to analyze pulmonary vascular function in the insulin resistant obese Zucker rat . METHODS : Large and small pulmonary arteries from obese Zucker rat and their lean counterparts were mounted for isometric tension recording . mRNA and protein expression was measured by RT-PCR or Western blot , respectively . KV currents were recorded in isolated pulmonary artery smooth muscle cells using the patch clamp technique . RESULTS : Right ventricular wall thickness was similar in obese and lean Zucker rats . Lung Q13873 , KV1.5 and 5- Q13049 receptor mRNA and protein expression and KV current density were also similar in the two rat strains . In conductance and resistance pulmonary arteries , the similar relaxant responses to acetylcholine and nitroprusside and unchanged lung P29474 expression revealed a preserved endothelial function . However , in resistance ( but not in conductance ) pulmonary arteries from obese rats a reduced response to several vasoconstrictor agents ( hypoxia , phenylephrine and 5-HT ) was observed . The hyporesponsiveness to vasoconstrictors was reversed by L-NAME and prevented by the P35228 inhibitor 1400W . CONCLUSIONS : In contrast to rat models of type 1 diabetes or other mice models of insulin resistance , the obese Zucker rats did not show any of the characteristic features of pulmonary hypertension but rather a reduced vasoconstrictor response which could be prevented by inhibition of P35228 . Further characterization of a somatic cell hybrid panel : ten new assignments to the bovine genome . Thirty-six partially characterized hamster-bovine hybrid cell lines were used for the determination of synteny groups . Sixteen additional reference loci , selected for their coverage of the bovine genome , were analysed on these hybrid cells . This increases to 25 the number of synteny groups detected . This panel was then used to make synteny assignments for 10 additional loci , eight by Southern blotting ( P02452 , P08123 , FAS , P07858 , P07711 , P07510 , P07686 and P08908 ) and two by polymerase chain reaction ( PCR ) amplification ( P35367 and ETH1112 ) . These loci were assigned to international synteny groups U12 ( P35367 ) , U13 ( P08123 ) , U17 ( P07510 ) , U21 ( P02452 , FAS ) , U29 ( ETH1112 ) , to chromosome 20 ( U14 or U25 ) for P07686 and P08908 , and to the same local synteny group ( A ) , which is probably U18 , for P07858 and P07711 . For three loci already mapped in humans ( P02452 , P08123 and P07510 ) , the present results are in accordance with the predictions based on comparative mapping between the human and bovine species .

MH_train_1211

MH_train_1211

MH_train_1211

interacts_with DB09053?

multiple_choice

P10275 inducing bladder cancer progression by promoting an epithelial-mesenchymal transition . The study investigated the role of androgen receptor ( AR ) as a potential target for the treatment of bladder cancer in regulating epithelial-mesenchymal transition or transformation ( EMT ) . Cell proliferation , and migration capacity were determined in bladder cancer T24 cells treated with small interfering RNA directed against AR , and expression levels of P12830 , β-catenin and N- cadherin were assessed using quantitative reverse transcription PCR ( qRT-PCR ) . Tumour cell growth was evaluated in vivo in T24 tumour-bearing nude mice receiving electroporation-assisted administration of anti-AR small interfering RNA . It was found that low AR expression decreased proliferation and migration of bladder cancer cells . In vivo experiments showed that silencing AR expression significantly suppressed AR-positive bladder tumour growth with decreased cell proliferation . Low AR level of T24 bladder cancer cells treated with DB01541 ( DB02901 ) decreased expression of P12830 , β-catenin and P19022 expression , indicating a strong sensitivity to the EMT and In cells with low AR content , TGF-β induced down-regulation of P12830 and β-catenin . It is concluded that suppression of AR expression decreased the production of TGF-β , inhibiting EMT and bladder cancer cell growth in vitro and in vivo , implying that its use might be a potential therapeutic target for the treatment of bladder cancer . Inhibition of PI3K/ P42345 pathways in glioblastoma and implications for combination therapy with temozolomide . Due to its molecular heterogeneity and infiltrative nature , glioblastoma multiforme ( GBM ) is notoriously resistant to traditional and experimental therapeutics . To overcome these hurdles , targeted agents have been combined with conventional therapy . We evaluated the preclinical potential of a novel , orally bioavailable PI3K/ P42345 dual inhibitor ( DB05241 ) in in vitro and in vivo studies . In vivo serially passaged human GBM xenografts that are more genetically stable than GBM cell lines in culture were used for all experiments . Biochemical downstream changes were evaluated by immunoblot and cytotoxicity by colorimetric DB00171 -based assay . For in vivo experiments , human xenograft GBM 39 grown intracranially in nude mice was altered to express luciferase to monitor tumor burden by optical imaging . DB05241 resulted in concentration-dependent decreases in cell viability in vitro . Cytotoxic doses resulted in specific inhibition of PI3K signaling . Combining DB05241 with temozolomide ( DB00853 ) resulted in additive toxicity in 4 of 5 xenografts . In vivo , DB05241 administered by oral gavage resulted in greater than 12-fold reduction in median tumor bioluminescence compared with control ( Mann-Whitney test p = 0.001 ) and improvement in median survival ( logrank p = 0.05 ) . DB00853 alone showed a 30-fold decrease in median bioluminescence , but the combination DB05241 + DB00853 yielded a 140-fold reduction in median bioluminescence ( Mann-Whitney test p = 0.05 ) with a trend toward improvement in median survival ( logrank p = 0.09 ) compared with DB00853 alone . DB05241 shows activity as monotherapy and in combination with conventional therapeutics in a range of genetically diverse GBM xenografts . Inhibitors of P11274 signalling interrupt the survival signal mediated by the micro-environment in mantle cell lymphoma . Several studies provide evidences for mantle cell lymphoma ( Q8WXI8 ) cell survival relying on B-cell receptor ( P11274 ) -mediated signalling pathways , whereas the nature of this activation is unknown . Significant progress in Q8WXI8 treatment is achieved through therapies targeting P11274 -associated kinases , i.e. , DB09053 and Fostamatinib , inhibitors of Q06187 and P43405 , respectively . Our study addresses survival signals emanating from the P11274 or the tumour environment and how inhibiting P11274 signalling effectors might impact these survival signals . We found that Q06187 was constitutively activated and that P43405 phosphorylation was highly increased and sustained upon P11274 activation of primary Q8WXI8 cells . Moreover , Q8WXI8 cells from leukaemic patients secreted high amount of IL-1β , P05231 , P10145 and P13501 . Activation of the P11274 induced ( i ) cell survival , ( ii ) P40763 activation and ( iii ) increased autocrine secretion of IL-1β , P05231 , P10145 , P13501 , P22301 , TNFα and P15692 . Specific inhibition of Q06187 by DB09053 or P43405 by Fostamatinib ( R406 ) reversed these protective effects and decreased both basal and P11274 -induced autocrine cytokine secretions associated with P40763 phosphorylation . Interestingly , targeting Q06187 and P43405 prevented and inhibited P11274 -induced Q8WXI8 cell adhesion to human bone marrow stromal cells ( HMSCs ) in short- and long-term co-culture . We demonstrated that P11274 -induced survival relies on autocrine secretion of IL-1β , TNFα and P13501 that might facilitate adhesion of Q8WXI8 cells to HMSC . Treatment with DB09053 or Fostamatinib blocked the chemotactic signal thus increasing apoptosis . Characterization of ibrutinib-sensitive and -resistant mantle lymphoma cells . DB09053 inhibits Q06187 ( Q06187 ) , a key component of early B-cell receptor ( P11274 ) signalling pathways . A multicentre phase 2 trial of ibrutinib in patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) demonstrated a remarkable response rate . However , approximately one-third of patients have primary resistance to the drug while other patients appear to lose response and develop secondary resistance . Understanding the molecular mechanisms underlying ibrutinib sensitivity is of paramount importance . In this study , we investigated cell lines and primary Q8WXI8 cells that display differential sensitivity to ibrutinib . We found that the primary cells display a higher Q06187 activity than normal B cells and Q8WXI8 cells show differential sensitivity to Q06187 inhibition . Genetic knockdown of Q06187 inhibits the growth , survival and proliferation of ibrutinib-sensitive but not resistant Q8WXI8 cell lines , suggesting that ibrutinib acts through Q06187 to produce its anti-tumour activities . Interestingly , inhibition of P27361 /2 and AKT , but not Q06187 phosphorylation per se , correlates well with cellular response to Q06187 inhibition in cell lines as well as in primary tumours . Our study suggests that , to prevent primary resistance or to overcome secondary resistance to Q06187 inhibition , a combinatory strategy that targets multiple components or multiple pathways may represent the most effective approach . The centrality of P08397 expression levels on DB00855 efficacy . Successful 5-aminolevulinic acid-based photodynamic therapy ( DB00855 ) is dependent on efficient porphyrin synthesis in the inflicted cancer tissue , which is regulated by several enzymes . Irradiation of the tumor excites the light-sensitive porphyrins and results in ROS production and cell death . In this study we investigated the effect of the expression levels of two main enzymes in heme biosynthesis , ALA dehydratase ( P13716 ) and porphobilinogen deaminase ( P08397 ) , on the capacity of K562 cells to undergo cell death following DB00855 . We manipulated P08397 and P13716 expression levels by shRNAs and P08397 overexpressing plasmid . P08397 down-regulation induced an elevation in P13716 activity , while overexpression of P08397 reduced P13716 activity , indicating a novel regulation feedback of P08397 on P13716 activity . This feedback mechanism enabled partial PpIX synthesis under P08397 silencing , whereas P13716 silencing reduced PpIX production to a minimum . DB00855 efficacy was directly correlated to PpIX levels . Thus , only P13716 -silenced cells were not affected by ALA+ irradiation , while following P08397 silencing , the accumulated PpIX , though decreased , was sufficient for successful DB00855 . The alterations in P13716 activity level initiated by changes in P08397 expression indicates P08397 's central role in heme synthesis . This enables efficient DB00855 , even when P08397 is not fully active . Conversely , P13716 loss resulted in reduced PpIX synthesis and consequently failure in DB00855 , due to the absence of compensation mechanism for P13716 . Effects of novel safrole oxide derivatives , 1-propyl-3-(3,4-methylenedioxyphenyl)-2-propanol and 1-isopropoxy-3-(3,4-methylenedioxyphenyl)-2-propanol , on apoptosis induced by deprivation of survival factors in vascular endothelial cells . Two safrole oxide derivatives , 1-propoxy-3-(3,4-methylenedioxyphenyl)-2-propanol ( FOD ) and 1-isopropoxy-3-(3,4-methylenedioxyphenyl)-2-propanol ( GOD ) , were newly synthesized as promoters of apoptosis in vascular endothelial cells . The purpose of this study was to investigate the effects of these two safrole oxide derivatives on cell growth and apoptosis induced by deprivation of survival factors ( serum and fibroblast growth factors , P05230 and P09038 ) in vascular endothelial cells ( VECs ) . MTT ( 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium ) method , agarose gel electrophoresis , laser scanning confocal microscopy , flow cytometry ( DB00828 ) , and immunofluorescence assay were used . The cells deprived of FGF and serum were exposed to FOD or GOD 30 to 90 mg x L(-1) for 24 h , cell growth was suppressed ( p < .05 ) , whereas detachment and DNA fragmentation of these cells were promoted ( p < .01 ) . When the cells were treated with FOD 90 mg x L(-) for 24 h , apoptosis rate was 14.99 % ( p < .01 ) . There were more cells in G2-M phase and less cells in S phase . At 90 mg x L(-1) concentration , GOD blocked 77.03 % of the cells at G0- P55008 phase. , P04637 level in VEC exposed to FOD or GOD was increased ( p < .01 ) . The data suggested that FOD and GOD might promote apoptosis of VEC by affect the cell cycle distribution , whereas P04637 was involved in this pathway . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . DB08870 followed by allogeneic transplantation as salvage regimen in patients with relapsed and/or refractory Hodgkin 's lymphoma . Patients with relapsed or refractory Hodgkin lymphoma ( RR-HL ) have poor outcomes . DB08870 ( BV ) , an antibody-drug conjugate comprising an anti- P28908 antibody conjugated to the potent anti-microtubule agent , monomethyl auristatin E , induces high tumour responses with moderate adverse effects . In a retrospective study , we describe objective response rates and subsequent allogeneic stem cell transplantation ( allo- P09683 ) in patients with RR-HL treated by BV in a named patient program in two French institutions . Twenty-four adult patients with histologically proven P28908 (+) RR-HL treated with BV were included from July 2009 to November 2012 . Response to BV treatment was evaluated after four cycles . Eleven patients were in complete response ( 45.8 % ) , while five patients were in partial response ( 20.8 % ) , with an overall response rate of 66.6 % . Eight patients failed to respond to BV ( 33.3 % ) . All of the responding patients could receive consolidation treatment after BV : three patients underwent autologous stem cell transplantation ( auto- P09683 ) , three patients received a tandem auto- P09683 /allo- P09683 , nine patients received allo- P09683 and one patient was treated with donor lymphocyte infusion . We found no treatment-related mortality at day 100 among the 12 patients who underwent BV following by allogeneic transplantation . With a median follow-up of 20 months ( range 10.5-43.2 ) , none of them relapsed or died . BV followed by allo- P09683 represents an effective salvage regimen in patients with RR-HL . 1p21 deletions are strongly associated with 1q21 gains and are an independent adverse prognostic factor for the outcome of high-dose chemotherapy in patients with multiple myeloma . Deletions involving chromosome 1p are frequent events in multiple myeloma ( MM ) . As karyotyping and single nucleotide polymorphism-based mapping analysis identify a minimal common deletion region involving the 1p21 locus , we investigated the prevalence and prognostic significance of del(1p21) in 203 MM patients undergoing high-dose therapy and autologous P09683 . 1p21 status was also evaluated in 16 patients with monoclonal gammopathy of undetermined significance ( MGUS ) and 41 patients with plasma cell leukemia ( PCL ) . Q5TCZ1 combined with cytoplasmic light chain detection ( cIg- Q5TCZ1 ) detected hemizygous 1p21 deletions in 18 % of the MM , 34 % of PCL but none of the MGUS cases . The presence of 1p21 deletions was correlated with 1q21( P61024 ) amplification ( P=0.01 ) , and del17p( P04637 ) ( P=0.05 ) but not with del(13q) , t(11;14) or t(4;14) . Patients with 1p21 deletions had significantly shorter progression-free survival ( PFS ; median 14.2 vs 25.4 months , P < 0.001 ) and overall survival ( OS ; median 39.4 vs 82.3 months , P=0.001 ) than those without such deletions . In multivariate analysis , del(1p21) was an independent risk factor for PFS ( P= 0.003 ) and OS ( P=0.013 ) after adjusting for del(13q) , del(p53) , t(4;14) and 1q21 amplifications . Our results indicate that del(1p21) is an independent poor prognostic factor associated with disease progression in MM . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . Mechanisms of ibrutinib resistance in chronic lymphocytic leukaemia and non-Hodgkin lymphoma . Q06187 ( Q06187 ) , a mediator of B-cell receptor ( P11274 ) signalling , has been implicated in the pathogenesis of chronic lymphocytic leukaemia ( CLL ) and other B-cell malignancies . DB09053 is an orally bioavailable and highly specific Q06187 inhibitor that was recently approved for treatment of patients with recurrent CLL and mantle cell lymphoma ( Q8WXI8 ) . In addition , ibrutinib has shown efficacy in subsets of patients with diffuse large B cell lymphoma ( DLBCL ) and Waldenstrom macroglobulinaemia ( WM ) . However , despite ibrutinib 's activity in multiple B-cell malignancies , cases of primary and secondary resistance have emerged . The overall reported frequency of resistance is low , but follow-up in many trials was short , and we predict that the incidence of observed resistance will increase as clinical use outside clinical trials expands over time . Mutations within Q06187 have been described and clearly interfere with drug binding ; however , there are also emerging alternative mechanisms that bypass Q06187 entirely and offer new opportunities for other targeted agents . Improved understanding of mechanisms of primary and secondary resistance is essential to developing appropriate therapeutic strategies to both prevent and address resistance . This review provides a comprehensive analysis of ibrutinib resistance in CLL , Q8WXI8 , DLBCL and WM and considers potential strategies for further study . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . [ The human genome -- chromosome 9 ] . Among the contents of chromosome 9 the author mentions locuses for Friedreich 's ataxia ( Q16595 ) , familial malignant melanoma ( Q8N726 ) , acute hepatic porphyria caused by deficiency of delta-aminolevulinate dehydrogenase ( P13716 ) and locus P23025 , the defective alleles of which condition disorders of nucleic acid reparative systems . By transcription and translation of the P00519 oncogene attached to gene P11274 and belonging to the 22nd chromosome a fusion protein with a high kinase activity is formed which is typical for the pathological clone of haematopoietic cells in chronic leukaemia . Characterization of eltenac and novel P35354 selective thiopheneacetic acid analogues in vitro and in vivo . We assessed the effect of novel selective thiopheneacetic acids on cyclooxygenase isoenzymes in vitro and in vivo . Thiopheneacetic acid Eltenac and derivatives were investigated in this study . In human whole blood experiments these derivatives were potent inhibitors of P35354 ( IC(50)=0.02-0.4 microM ) with less pronounced effect on P23219 ( IC(50)=0.15-5.6 microM ) . With P23219 / P35354 ratios between 7.5- and 16-fold they are in the range of Celecoxib ( 13-fold ) . The parent drug Eltenac demonstrated no selectivity for P35354 . In a rat paw edema model , these compounds showed reduction of edema volume in the range of 36-45 % at 10 mg/kg ( Eltenac 52 % , Diclofenac 51 % ) . However , the compounds were superior to Diclofenac and Eltenac with respect to their ulcerogenic and gastrointestinal properties . Introduction of a nitrate-ester moiety to either Eltenac or a derivative did neither improve selectivity or potency in vitro , nor ulcerogenicity in vivo . Molecular modeling of selective thiopheneacetic acid derivatives to the active site of human P35354 suggested similar binding properties as DB01283 and Diclofenac . In summary , modification of Eltenac generates moderately selective P35354 drugs in the range of Celecoxib with respect to potency and selectivity . The drugs showed potent anti-inflammatory properties and significant improvement of animal survival in a sub-chronical experimental set up . Thiopheneacetic derivatives are characterized by low pK(a) values , short microsomal half-lives and binding mode to P35354 similar to Diclofenac and DB01283 . These properties may also have an impact on the transient inhibition of P35354 -dependent prostacyclin , thereby being less associated with vascular complications . A novel multiple tyrosine-kinase targeted agent to explore the future perspectives of anti-angiogenic therapy for the treatment of multiple solid tumors : cabozantinib . The process of angiogenesis involves the formation of new blood vessels from pre-existing vasculature by the over expression of certain factors leading to the growth and development of all solid tumor types . P08581 abbreviated as c- DB00134 and vascular endothelial growth factor abbreviated as P15692 are some of the factors responsible for the induction in tumor growth and development . Recently a number of analogues associated with these receptors are under study . US FDA on November 29 , 2012 approved a drug named cabozantinib formerly known as DB05153 which is being marketed under the trade name of Cometriq for the treatment of Medullary Thyroid Cancer ( P04629 ) . Designing of the drug has been done in such a fashion that it can inhibit both P35968 and c- DB00134 simultaneously without over expressing any of the factors leading to the inhibition of angiogenesis . The drug is still under study for the evaluation of its efficacy in cases of many other solid tumor types including breast cancer , castration resistant prostate cancer ( CRPC ) , renal cell carcinoma ( RCC ) , hepatocellular carcinoma ( HCC ) , gastric or gastroesophageal junction cancer , melanoma , small cell lung cancer ( SCLC ) , ovarian cancer and primary peritoneal or fallopian tube carcinoma . This review article consists of preclinical and clinical data of cabozantinib and its efficacy and safety towards various types of solid tumors . DB09053 ( ImbruvicaTM ) potently inhibits ErbB receptor phosphorylation and cell viability of ErbB2-positive breast cancer cells . DB09053 ( formerly P05154 -32765 ) is a specific , irreversible , and potent inhibitor of Burton 's tyrosine kinase ( Q06187 ) developed for the treatment of several forms of blood cancer . It is now an FDA-approved drug marketed under the name Imbruvica(TM) ( Pharmacyclics , Inc. ) and successfully used as an orally administered second-line drug in the treatment of mantle cell lymphoma . Since Q06187 is predominantly expressed in hematopoietic cells , the sensitivity of solid tumor cells to DB09053 has not been analyzed . In this study , we determined the effect of DB09053 on breast cancer cells . We demonstrate that DB09053 efficiently reduces the phosphorylation of the receptor tyrosine kinases ErbB1 , ErbB2 and ErbB3 , thereby suppressing AKT and MAPK signaling in ErbB2-positive ( ErbB2+ ) breast cancer cell lines . Treatment with DB09053 significantly reduced the viability of ErbB2+ cell lines with IC50 values at nanomolar concentrations , suggesting therapeutic potential of DB09053 in breast cancer . Combined treatment with DB09053 and the dual PI3K/ P42345 inhibitor BEZ235 synergistically reduces cell viability of ErbB2+ breast cancer cells . Combination indices below 0.25 at 50 % inhibition of cell viability were determined by the Chou-Talalay method . Therefore , the combination of DB09053 and canonical PI3K pathway inhibitors could be a new and effective approach in the treatment of breast cancer with activated ErbB receptors . DB09053 could thus become a valuable component of targeted therapy in aggressive ErbB2+ breast cancer . P43490 / P43490 /visfatin and cancer . P43490 / P43490 /visfatin is the rate-limiting enzyme that catalyzes the first step in NAD biosynthesis from nicotinamide and regulates growth , apoptosis and angiogenesis of mammalian cells . This enzyme was originally cloned as a putative cytokine shown to enhance the B cell precursor maturation in the presence of P13232 and stem cell factor . A number of cancers have increased expression of P43490 / P43490 /visfatin , which regulates a variety of different signaling pathways such as PI3K/Akt , P27361 /2 and P40763 . FK866/APO866 and CHS828/ DB05217 are two known inhibitors of P43490 / P43490 /visfatin and have been evaluated as anticancer agents in the clinic . This review will focus on its role in carcinogenesis and cancer progression and its inhibitors as therapeutic target for cancer treatment . Rectal antinociceptive properties of alverine citrate are linked to antagonism at the P08908 receptor subtype . Serotonin ( 5-HT ) is considered as a major mediator causing hyperalgesia and is involved in inflammatory reactions and irritable bowel syndrome . DB01616 citrate may possess visceral antinociceptive properties in a rat model of rectal distension-induced abdominal contractions . This study was designed to evaluate the pharmacological properties of alverine citrate in a rat model of rectal hyperalgesia induced by 5-HTP ( 5-HT precursor ) and by a selective P08908 agonist ( 8-OH-DPAT ) and to compare this activity with a reference P08908 antagonist ( WAY 100635 ) . At 4 h after their administration , 5-HTP and 8-OH-DPAT increased the number of abdominal contractions in response to rectal distension at the lowest volume of distension ( 0.4 mL ) . When injected intraperitoneally before 8-OH-DPAT and 5-HTP , WAY 100635 ( 1 mg kg(-1) ) blocked their nociceptive effect , but also reduced the response to the highest volume of distension ( 1.6 mL ) . Similarly , when injected intraperitoneally , alverine citrate ( 20 mg kg(-1) ) suppressed the effect of 5-HTP , but not that of 8-OH-DPAT . However , when injected intracerebroventricularly ( 75 microg/rat ) alverine citrate reduced 8-OH-DPAT-induced enhancement of rectal distension-induced abdominal contractions . In-vitro binding studies revealed that alverine citrate had a high affinity for P08908 receptors and a weak affinity for 5- Q9H205 and Q13639 subtypes . These results suggest that 5-HTP-induced rectal hypersensitivity involves 5-TH1A receptors and that alverine citrate acts as a selective antagonist at the P08908 receptor subtype to block both 5-HTP and 8-OH-DPAT-induced rectal hypersensitivity . Inhibition of cyclooxygenases 1 and 2 by the phospholipase-blocker , arachidonyl trifluoromethyl ketone . BACKGROUND AND PURPOSE : Arachidonyl trifluoromethyl ketone ( Q06187 ) is widely used as an inhibitor of cytosolic group IV phospholipase A(2) ( cPLA(2) ) and calcium-independent group VI phospholipase A(2) ( iPLA(2) ) . Q06187 thus reduces arachidonic acid ( AA ) substrate for cyclooxygenase ( P36551 ; also known as prostaglandin H synthase ) and attenuates prostaglandin ( PG ) synthesis . It has been shown previously , that Q06187 blocks thromboxane B(2) production induced by exogenous AA in human platelets . It remains , however , unknown whether Q06187 also directly modulates the activity of cyclooxygenase ( P36551 ) . EXPERIMENTAL APPROACH : Time courses for inhibition of P36551 by Q06187 was obtained using osteoblast-like MC3T3-E1 cells , with exogenous AA as substrate and the pure enzymes P23219 and P35354 . PGE(2) was measured by GC-MS . KEY RESULTS : Q06187 was a potent inhibitor of P23219 and P35354 with IC(50) values of 0.5 and 0.1 microM in MC3T3-E1 cells and of 1.7 and 2.6 microM using the pure enzymes . Inhibition was reversible , with slow- and tight-binding characteristics . The arachidonyl carbon chain was essential , as the saturated palmitoyl analogue had no effect . CONCLUSIONS AND IMPLICATIONS : Attenuation of PG synthesis by Q06187 is taken to be the consequence of PLA(2) inhibition and the findings of many studies are interpreted on that basis . If there are , however , alternative routes for AA liberation ( such as phospholipase C/diacyl glycerol lipase or phospholipase D ) , this interpretation can lead to false conclusions . As Q06187 is a widely used and important pharmacological tool in eicosanoid research , knowledge of its interactions with other major enzymes of the cascade is of considerable importance . Phenotypic analysis of conditionally immortalized cells isolated from the Q06187 model of ARPKD . To study the pathophysiology of autosomal recessive polycystic kidney disease ( ARPKD ) , we sought to develop conditionally immortalized control and cystic murine collecting tubule ( CT ) cell lines . CT cells were isolated from intercross breedings between Q06187 mice ( bpk(+/-) ) , a murine model of ARPKD , and the Immorto mice ( H-2K(b)-ts-A58(+/+) ) . Second-generation outbred offspring ( Q06187 x Immorto ) homozygous for the Q06187 mutation ( bpk(-/-) ; Im(+/+/-) ; cystic Q06187 /H-2K(b)-ts-A58 ) , were phenotypically indistinguishable from inbred cystic Q06187 animals ( bpk(-/-) ) . Cystic Q06187 /H-2K(b)-ts-A58 mice developed biliary ductal ectasia and massively enlarged kidneys , leading to renal failure and death by postnatal day 24 . Principal cells ( PC ) were isolated from outbred cystic and noncystic Q06187 /H-2K(b)-ts-A58 littermates at specific developmental stages . Epithelial monolayers were under nonpermissive conditions for markers of epithelial cell polarity and PC function . Cystic and noncystic cells displayed several properties characteristic of PCs in vivo , including amiloride-sensitive sodium transport and aquaporin 2 expression . Cystic cells exhibited apical epidermal growth factor receptor ( P00533 ) mislocalization but normal expression of ZO-1 and P12830 . Hence , these cell lines retain the requisite characteristics of PCs , and cystic Q06187 /H-2K(b)-ts-A58 PCs retained the abnormal P00533 membrane expression characteristic of ARPKD . These cell lines represent important new reagents for studying the pathogenesis of ARPKD . Dimerization effect of sucrose octasulfate on rat P05230 . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 are Lys113 , Lys118 , Arg122 and Lys128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 . This difference may be important for SOS-mediated P05230 dimerization in rat . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 .

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The bone morphogenetic protein signaling pathway is upregulated in a mouse model of total parenteral nutrition . Total parenteral nutrition ( O15533 ) results in intestinal mucosal atrophic changes due to an absence of enteral nutrition ; however , the mechanisms responsible for this are not fully understood . It has been shown that bone morphogenetic protein ( BMP ) activation inhibits intestinal epithelial cell ( EC ) proliferation . Therefore , we hypothesized that the BMP pathway could be upregulated by O15533 . To address this , we randomly assigned mice to receive O15533 or to be enterally fed ( control ) for 7 d . Mucosal EC isolates were harvested from the entire length of small intestine for RNA and protein measurements . Q8N1N2 -thickness , mid-small bowel was processed for histological examination . O15533 increased the abundance of P12643 , P12644 , and Q13873 at the RNA and protein levels . Phosphorylation of Q15797 , Q99717 , and Smad8 also was greater in the O15533 group than in the control , which helped to confirm activation of this pathway . Interestingly , the O15533 and control groups did not differ in the mRNA expression of the extracellular soluble bmp antagonists , noggin , gremlin , chordin , or follistatin . Compared to the control group , the expression of c-Myc ( cellular myelocytomatosis ) mRNA was lower , whereas the level of P38936 ( P38936 /CIP1) was greater , in the O15533 group . Because the BMP family may function through suppression of Wnt-beta-catenin signaling , this pathway was also examined . mRNA expression of Wnt 3 , Wnt5a , and the Wnt receptor Lrp5 were lower in the O15533 group compared to controls . The results suggest that the BMP signaling pathway may be involved in the development of intestinal mucosal atrophy due to O15533 administration . NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC . Elevation of P13500 and P13500 / P15692 ratio in cerebrospinal fluid of amyotrophic lateral sclerosis patients . Amyotrophic lateral sclerosis ( P35858 ) is a neurodegenerative disease with progressive cell death of upper and lower motor neurons . In this study , we measured monocyte chemotactic protein-1 ( P13500 ) and vascular endothelial growth factor ( P15692 ) levels in cerebrospinal fluid ( P04141 ) and serum by enzyme-linked immunosorbent assay ( ELISA ) in 42 P35858 patients , and compared these levels with those of control subjects with other neurodegenerative disorders or with those of normal controls . P13500 levels in P04141 were significantly higher in P35858 patients than in the control group . P15692 levels in P04141 tended to be lower in P35858 patients than in the control group , but not significantly . A positive correlation was found between P13500 levels in P04141 of P35858 patients and the total Norris scale . The elevation of P13500 / P15692 ratio in P04141 was more specific to P35858 patients compared to other neurological diseases such as Parkinson 's disease ( PD ) and spinocerebellar ataxia ( SCA ) and to controls . Our data suggested that both P13500 levels and P13500 / P15692 ratio in P04141 may be useful markers for the clinical diagnosis of P35858 . Targeted cytotoxic analog of luteinizing hormone-releasing hormone ( P01148 ) , AEZS-108 ( AN-152 ) , inhibits the growth of DU-145 human castration-resistant prostate cancer in vivo and in vitro through elevating P38936 and ROS levels . Management of castration-resistant prostate cancer ( CRPC ) is challenging due to lack of efficacious therapy . DB00044 -releasing hormone analogs appear to act directly on cells based on the P01148 receptors on human prostate adenocarcinoma cells . We explored anticancer activity of a cytotoxic analog of P01148 , AEZS-108 consisting of P01148 agonist linked to doxorubicin . Nude mice bearing DU-145 tumors were used to compare antitumor effects of AEZS-108 with its individual constituents or their unconjugated combination . The tumor growth inhibition of conjugate was greatest among treatment groups ( 90.5 % inhibition vs. 41 % by [D-Lys(6)] P01148 +DOX ) . The presence of P01148 receptors on DU-145 cells was confirmed by immunocytochemistry . In vitro , AEZS-108 significantly inhibited cell proliferation ( 61.2 % inhibition ) and elevated apoptosis rates ( by 46 % ) . By the detection of the inherent doxorubicin fluorescence , unconjugated doxorubicin was seen in the nucleus ; the conjugate was perinuclear and at cell membrane . Autophagy , visualized by GFP-tagged p62 reporter , was increased by AEZS-108 ( 7.9-fold vs. 5.3-fold by DOX+[D-Lys(6)] P01148 . AEZS-108 more effectively increased reactive oxygen species ( ROS , 2-fold vs. 1.4-fold by DOX+[D-Lys(6)] P01148 ) and levels of the apoptotic regulator P38936 in vivo and in vitro . We demonstrate robust inhibitory effects of the targeted cytotoxic P01148 analog AEZS-108 on P22888 positive castration-resistant prostate cancer cells . Treatment with arimoclomol , a coinducer of heat shock proteins , delays disease progression in P35858 mice . Amyotrophic lateral sclerosis ( P35858 ) is a fatal neurodegenerative condition in which motoneurons of the spinal cord and motor cortex die , resulting in progressive paralysis . This condition has no cure and results in eventual death , usually within 1-5 years of diagnosis . Although the specific etiology of P35858 is unknown , 20 % of familial cases of the disease carry mutations in the gene encoding Cu/Zn superoxide dismutase-1 ( P00441 ) . Transgenic mice overexpressing human mutant P00441 have a phenotype and pathology that are very similar to that seen in human P35858 patients . Here we show that treatment with arimoclomol , a coinducer of heat shock proteins ( HSPs ) , significantly delays disease progression in mice expressing a P00441 mutant in which glycine is substituted with alanine at position 93 ( P00441 (G93A) ) . DB05025 -treated P00441 (G93A) mice show marked improvement in hind limb muscle function and motoneuron survival in the later stages of the disease , resulting in a 22 % increase in lifespan . Pharmacological activation of the heat shock response may therefore be a successful therapeutic approach to treating P35858 , and possibly other neurodegenerative diseases . The role of bone marrow DB05914 in the treatment of acute liver failure . OBJECTIVE : This study is to investigate the effects of bone marrow mesenchymal stem cell ( BMSC ) transplantation on acute liver failure ( Q9UNN4 ) . METHODS : BMSCs were separated from rat bone marrow , cultured , and identified by flow cytometry . Rat model with Q9UNN4 was established by injecting D-galactosamine and lipopolysaccharide . Rats were randomly divided into the control group and BMSC transplantation group . The serum levels of alanine aminotransferase ( ALT ) and aspartate aminotransferase ( Q9NRA2 ) were measured at 24 h , 120 h , and 168 h after BMSC transplantation . Apoptosis was detected by TUNEL assay . The expression of P15692 and AFP proteins was detected by immunofluorescence . P29466 and Q14116 proteins and mRNA were detected by immunohistochemistry and RT-PCR . RESULTS : Compared with the control group , levels of ALT , Q9NRA2 , caspase-1 and Q14116 proteins , and mRNA in the transplantation group were significantly lower at 120 h and 168 h after BMSCs transplantation . Apoptosis was inhibited by BMSCs transplantation . The P15692 protein levels were increased with the improvement of liver function , and the AFP protein levels were increased with the deterioration of the liver function after BMSCs transplantation . CONCLUSIONS : BMSCs transplantation can improve liver function and inhibit hepatocyte apoptosis as well as promote hepatocyte proliferation in rat model with Q9UNN4 . Protective effects of taurine on glutathione and glutathione-dependent enzymes in ethanol-fed rats . DB00898 , by its property of generating free radicals during the course of its metabolism , alters redox homeostasis and causes damage to cell structure and function . This study investigated the effect of taurine on ethanol-induced experimental toxicity in rats . DB00898 was administered chronically to rats for 28 days . This resulted in significant increases in the activities of transaminases , alkaline phosphatase ( ALP ) , gamma-glutamyl transpeptidase ( P19440 ) and bilirubin in plasma . The activities of glutathione peroxidase ( GPx ) , glutathione-S-transferase ( Q86UG4 ) and the contents of glutathione ( DB00143 ) and thiols in plasma and tissues were significantly reduced as compared to control animals . Simultaneous administration of taurine along with ethanol prevented the leakage of enzymes into circulation and restored glutathione and tissue thiols . The activities of antioxidant enzymes were normalized . We propose that taurine may have a bioprotective effect on ethanol-induced toxicity . Mechanism of oral absorbent DB05269 in lipid abnormalities in experimental uremic rats . BACKGROUND : We have reported that oral sorbent DB05269 ( Q9NRA2 ) is effective in delaying the induction of dialysis in patients with chronic renal failure ( CRF ) because of its effect on lipid metabolism . To clarify the precise mechanism of Q9NRA2 in lipid abnormalities in CRF , we examined the effect of Q9NRA2 on plasma lipid profile , total bile acids ( TBA ) , and lipoprotein lipase ( P06858 ) activity in experimental uremic rats . METHODS : Uremic rats were prepared using male Wistar rats by ligating 5/6 of the renal artery . Uremic rats were randomly divided into two groups as follows : a control group in which rats were maintained on the standard diet and an Q9NRA2 group in which rats were maintained on a diet containing 5 g of Q9NRA2 per 100 g of standard diet for 10 weeks . Plasma P06858 activity was measured as free fatty acid ( FFA ) generation after intravenous administration of heparin . RESULTS : Plasma creatinine at 1.5 +/- 0.1 mg/dl was lower in the Q9NRA2 group than the 1.9 +/- 0.5 mg/ml level in the control group . Q9NRA2 significantly decreased plasma total cholesterol from 192 +/- 29 to 142 +/- 25 mg/dl , triglycerides from 198 +/- 71 to 99 +/- 38 mg/dl , and TBA from 19.6 +/- 2.6 mumol/liter to 8.8 +/- 3.5 mumol/ml . Plasma P06858 activity at 0.22 +/- 0.01 mumol FFA/min/hr was significantly higher in the Q9NRA2 group than 0.15 +/- 0.03 mumol FFA/min/hr in the control group . CONCLUSIONS : These results suggest that Q9NRA2 may improve plasma lipid abnormalities by binding to bile acids in the intestinal lumen and preventing their reabsorption and inhibiting the reduction of P06858 activity in experimental uremic rats . Gene expression by PBMC in primary sclerosing cholangitis : evidence for dysregulation of immune mediated genes . Primary sclerosing cholangitis ( PSC ) is a chronic disease of the bile ducts characterized by an inflammatory infiltrate and obliterative fibrosis . The precise role of the immune system in the pathogenesis of PSC remains unknown . We used RNA microarray analysis to identify immune-related genes and pathways that are differentially expressed in PSC . Messenger RNA ( mRNA ) from peripheral blood mononuclear cells ( PBMC ) was isolated from both patients with PSC and age and sex matched healthy controls . Samples from 5 PSC patients and 5 controls were analyzed by microarray and based upon rigorous statistical analysis of the data , relevant genes were chosen for confirmation by RT-PCR in 10 PSC patients and 10 controls . Using unsupervised hierarchical clustering , gene expression in PSC was statistically different from our control population . Interestingly , genes within the P60568 receptor beta , P05231 and Q96HU1 Kinase pathways were found to be differently expressed in patients with PSC compared to controls . Further , individual genes , P01375 induced protein 6 ( TNFaip6 ) and membrane-spanning 4-domains , subfamily A ( ms4a ) were found to be upregulated in PSC while similar to Q99717 ( Q99717 ) was downregulated . In conclusion , several immune-related pathways and genes were differentially expressed in PSC compared to control patients , giving further evidence that this disease is systemic and immune-mediated . Chronic administration of AM251 improves albuminuria and renal tubular structure in obese rats . Modulation of the endocannabinoid system as an anti-obesity therapeutic is well established ; however , the direct effects of cannabinoid receptor 1 ( P21554 ) antagonism on renal function and structure in a model of diet-induced obesity ( DIO ) are unknown . The aim of this study was to characterise the renal effects of the P21554 antagonist AM251 in a model of DIO . Male Sprague-Dawley rats were fed a low- or high-fat diet ( HFD : 40 % digestible energy from lipids ) for 10 weeks to elicit DIO ( n=9 ) . In a different cohort , rats were fed a HFD for 15 weeks . After 9 weeks consuming a HFD , rats were injected daily for 6 weeks with 3 mg/kg AM251 ( n=9 ) or saline via i.p. injection ( n=9 ) . After 10 weeks consuming a HFD , P21554 and megalin protein expression were significantly increased in the kidneys of obese rats . Antagonism of P21554 with AM251 significantly reduced weight gain , systolic blood pressure , plasma leptin , and reduced albuminuria and plasma creatinine levels in obese rats . Importantly , there was a significant reduction in tubular cross-section diameter in the obese rats treated with AM251 . An improvement in albuminuria was likely due to the reduction in tubular size , reduced leptinaemia and maintenance of megalin expression levels . In obese rats , AM251 did not alter diastolic blood pressure , sodium excretion , creatinine clearance or expression of the fibrotic proteins P15692 , P01137 and collagen IV in the kidney . This study demonstrates that treatment with P21554 antagonist AM251 improves renal outcomes in obese rats . Low molecular weight catalytic metalloporphyrin antioxidant DB05874 protects lungs from fractionated radiation . The objective of this study was to determine whether administration of a catalytic antioxidant , Mn(III) tetrakis(N,N'-diethylimidazolium-2-yl) porphyrin , AEOL10150 , reduces the severity of long-term lung injury induced by fractionated radiation ( RT ) . Fisher 344 rats were randomized into five groups : RT+AEOL10150 ( 2.5 mg/kg P55957 ) , AEOL10150 ( 2.5 mg/kg P55957 ) alone , RT+ AEOL10150 ( 5 mg/kg P55957 ) , AEOL10150 ( 5 mg/kg P55957 ) alone and RT alone . Animals received five 8 Gy fractions of RT to the right hemithorax . AEOL10150 was administered 15 min before RT and 8 h later during the period of RT treatment ( 5 days ) , followed by subcutaneous injections for 30 days , twice daily . Lung histology at 26 weeks revealed a significant decrease in lung structural damage and collagen deposition in RT+AEOL10150 ( 5 mg/kg P55957 ) group , in comparison to RT alone . Immunohistochemistry studies revealed a significant reduction in tissue hypoxia ( HIF1alpha , Q16790 ) , angiogenic response ( P15692 , CD-31 ) , inflammation ( ED-1 ) , oxidative stress ( 8-OHdG , 3-nitrotyrosine ) and fibrosis pathway ( TGFbeta1 , P84022 , p- Q15796 /3 ) , in animals receiving RT+ AEOL10150 ( 5 mg/kg P55957 ) . Administration of AEOL10150 at 5 mg/kg P55957 during and after RT results in a significant protective effect from long-term RT-induced lung injury . Low dose ( 2.5 mg/kg P55957 ) delivery of AEOL10150 has no beneficial radioprotective effects . Antimetastatic potential of vernolide-A , a sesquiterpenoid from Vernonia cinerea L . The inhibitory effect of vernolide-A ( C(21)H(28)O(7) ) on lung metastasis induced by B16F-10 melanoma cells was studied using C57BL/6 mice . Vernolide-A was administered in three different modalities such as simultaneously with tumor , prophylactic to tumor and after tumor development . Maximum inhibition in the metastasis was observed when vernolide-A was administered simultaneously with tumor . There was 89.39 % inhibition of lung tumor nodule formation and 88.51 % increase in the life span of metastatic tumor-bearing animals . Highly elevated levels of lung hydroxyproline , lung uronic acid , lung hexosamine , serum sialic acid , serum γ-glutamyl transpeptidase ( P19440 ) and serum vascular endothelial growth factor ( P15692 ) in the metastatic control animals were found to be significantly lowered in the vernolide-A-treated animals . Histopathological analysis of lung tissues also correlated with these results . Vernolide-A administration downregulated the expression of matrix metalloproteinase-2 ( P08253 ) , P14780 , extracellular-signal-regulated kinase-1 ( P27361 ) , P28482 and P15692 in the lung tissue of B16F-10 melanoma challenged animals . In the in vitro system , vernolide-A showed a significant inhibition of invasion of B16F-10 melanoma cells across the collagen matrix . Vernolide-A treatment also inhibited the migration of B16F-10 melanoma cells across a polycarbonate filter in vitro . Vernolide-A could inhibit P08253 and P14780 protein expression in gelatin zymographic analysis of B16F-10 cells . (3)H-thymidine proliferation assay showed that vernolide-A could inhibit the proliferation of B16F-10 melanoma cells in vitro . These results indicate that vernolide-A could inhibit the metastatic progression of B16F-10 melanoma cells in mice . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS ) Inactivation of caspase-1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin-1 ( IL-1 ) -beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL-1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL-1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE/caspase-1 ) or through caspase-1 gene deletion . METHODS : P29466 was selectively blocked by using pralnacasan or DB05507 . IL-1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL-1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 inhibition reduced the release of IL-1beta in organotypic slices exposed to LPS+ DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 ( intraperitoneal , 25-200 mg/kg ) to rats blocked seizure-induced production of IL-1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase-1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase-1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL-1beta . The relationship of in vivo central P21554 receptor occupancy to changes in cortical monoamine release and feeding elicited by P21554 receptor antagonists in rats . RATIONALE : Cannabinoid type 1 ( CB(1) ) receptor antagonists are reportedly effective in reducing food intake both preclinically and clinically . This may be due in part to their effects on monoamine release in the brain . The level of central CB(1) receptor occupancy underlying these neurobiological effects is unclear . OBJECTIVES : We explored the relationship between in vivo CB(1) receptor occupancy in the frontal cortex and changes in both monoamine release in the medial prefrontal cortex ( mPFC ) and feeding behavior in rats in response to two orally administered CB(1) receptor antagonists presently in clinical trials , SR141716A ( rimonabant ) and DB05077 . METHODS : CB(1) receptor occupancy was measured using [ (3)H ] SR141716A , and these occupancies were related to potencies to mediate increases in dopamine ( DA ) and norepinephrine ( NE ) release measured with microdialysis and decreases in consumption of a highly palatable diet ( HP ) . RESULTS : High receptor occupancy levels ( > 65 % ) were required to detect increases in monoamine release that were achieved with 3 and 10 mg/kg of SR141716A and 10 mg/kg of DB05077 for DA and 10 mg/kg of SR141716A for NE . Decreases in HP consumption were seen at occupancies higher than 65 % for SR141716A that were achieved with 3 and 10 mg/kg . In contrast , decreases in HP consumption were seen at relatively low CB(1) receptor occupancies ( 11 % ) for DB05077 . CONCLUSIONS : The occupancy method described here is an effective tool for interrelating central CB(1) receptor occupancy with neurobiological actions of CB(1) receptor antagonists and for furthering our understanding of the role of CB(1) receptors in central nervous system physiology and pathology . Internalization of OspA in rsCD14 complex and aggregated forms . Although the spirochetal protein OspA is capable of stimulating immune cells in a P08571 - and O60603 -dependent manner , little is known about how O60603 receptor complex ligands , such as OspA , are handled by the cell once delivered . We examine here the internalization of the fluorescently derivatized forms of both the full length DB00045 delivered as a recombinant soluble P08571 ( rsCD14 ) complex and the corresponding lipohexapeptide given to the cells as an aggregate . Both forms of OspA are internalized in a similar manner to acetylated low density lipoprotein ( AcLDL ) , a scavenger receptor ligand . Acetylated low density lipoprotein is capable of competing for internalization with OspA even when OspA is delivered as a rsCD14 complex . We observe co-localization of OspA with lysosomes but not with the Golgi complex . These phenomena are similar between RAW264.7 macrophages and endothelial cells but change drastically when the cells are deprived of serum . Upon serum starvation , OspA shows some localization to the Golgi apparatus whereas the lipohexapeptide remains on the cell surface . Inhibition of internalization of OspA via treatment with cytochalasin D or of the lipohexapeptide via serum starvation does not interfere with P01375 induction activity , consistent with signalling from the cell surface . Renoprotective effect of myricetin restrains dyslipidemia and renal mesangial cell proliferation by the suppression of sterol regulatory element binding proteins in an experimental model of diabetic nephropathy . DB02375 is a natural flavonoid used in various health management systems . In this present study myricetin tested to evaluate the effect on lipids and lipid metabolism enzymes in normal and streptozotocin ( Q11206 ) with cadmium ( Cd ) induced diabetic nephrotoxic rats . Diabetic nephrotoxic rats were significantly ( P < 0.05 ) increased the levels of urinary albumin and lipid profiles : total cholesterol ( TC ) , triglycerides ( TGs ) , free fatty acids ( FFAs ) , phospholipids ( PLs ) , low density lipoprotein ( LDL ) , very low-density lipoproteins ( VLDL ) , and decreased in the levels of high-density lipoproteins ( HDL ) . In addition , the activity of lipoprotein lipase ( P06858 ) and lecithin cholesterol acyl transferase ( P04180 ) were decreased significantly , whereas the 3-hydroxy 3-methylglutaryl coenzyme A ( HmgCoA ) reductase activity was increased . The upregulation of sterol regulatory element binding protein-1a ( SREBP-1a ) , SREBP-1c , Q12772 , transforming growth factor-β1 ( TGF-β1 ) , vascular endothelial growth factor ( P15692 ) and downregulation peroxisome proliferator-activated receptor alpha ( Q07869 -α ) proteins expression levels were noticed . An administration of myricetin ( 1.0 mg/kg body weight ( b/w ) ) for 12 weeks was brought the above parameters towards normal level . Histopathological study of kidney samples showed that extracellular mesangial matrix expansion , glomerulosclerosis and interstitial fibrosis in diabetic nephrotoxic rats was suppressed by myricetin treatment . Further our results indicate that administration of myricetin afforded remarkable protection against Q11206 -Cd induced alterations in lipid metabolism and thereby reduced the diabetic nephropathy in experimental rats . The neuroendocrine impact of chronic stress on cancer . Behavioral processes have long been suspected to influence many health processes including effects on cancer . However , mechanisms underlying these observations are not fully understood . Recent work has demonstrated that chronic behavioral stress results in higher levels of tissue catecholamines , greater tumor burden , and a more invasive pattern of ovarian cancer growth in an orthotopic mouse model . These effects are mediated primarily through the beta(2) adrenergic receptor ( P07550 ) activation of the tumor cell cyclic AMP ( DB02527 ) -protein kinase A ( PKA ) signaling pathway . Additionally , tumors in stressed animals have increased vascularization and enhanced expression of vascular endothelial growth factor ( P15692 ) and matrix metalloproteinases ( MMPs ) -2 and -9 . In this review , we highlight the importance of the neuroendocrine stress response in tumor biology and discuss mechanisms by which the beta-adrenergic receptors on ovarian cancer cells enhance angiogenesis and tumor growth . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention .

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interacts_with DB04908?

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Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Thrombophilia testing in children : a 7 year experience . BACKGROUND : Incidence of venous thromboembolism ( VTE ) in children is reported to be increasing . We examined thrombophilia testing results in children with VTE that presented in inpatient and outpatient settings to explore patterns of thrombophilia testing . PROCEDURE : PATIENTS/METHODS : Children , ages 0-20 years with VTE seen at our institution from Jan 2005 to Apr 2012 were studied retrospectively . All patients with VTE confirmed by imaging were eligible and the presence of significant risk factors was evaluated . Thrombophilia was diagnosed if > 1 tests confirmed : persistently low protein C ( PC ) , protein S ( PS ) , and antithrombin ( AT ) following VTE resolution , persistent antiphospholipid antibodies ( APA ) positivity > 12 weeks from first test , factor V Leiden ( FVL ) and prothrombin mutation ( PTm ) hetero- or homozygosity , elevated plasminogen activator inhibitor ( P05121 ) levels with 4G/5G or 4G/4G polymorphisms , methylene DB00116 reductase ( P42898 ) polymorphisms with elevated fasting homocysteine levels . RESULTS : Three hundred ninety-two patients met inclusion criteria . At least one test was ordered in 157/239 inpatients . All 153 outpatients had > 1 test ordered . Thrombophilia rate differences between inpatients and outpatients did not reach statistical significance except for PC deficiency , which was significantly higher in outpatients . Of inpatients , central venous line ( CVL ) was significantly associated with not having tests done ( P < 0.0022 ) . CONCLUSIONS : This study of pediatric VTE demonstrated a low thrombophilia rate in both inpatient and outpatient populations . The role of testing in other pediatric patients should be further explored . DB04630 induces interleukin-18 through endothelin-1 , angiotensin II , Rho/Rho-kinase , and PPARs in cardiomyocytes . DB04630 ( Aldo ) is recognized as an important risk factor for cardiovascular diseases . Q14116 induces myocardial hypertrophy , loss of contractility of cardiomyocytes , and apoptosis leading myocardial dysfunction . However , so far , there have been few reports concerning the interaction between Aldo and Q14116 . The present study examined the effects and mechanisms of Aldo on Q14116 expression and the roles of peroxisome proliferator-activated receptor ( Q07869 ) agonists in rat cardiomyocytes . We used cultured rat neonatal cardiomyocytes stimulated with Aldo to measure Q14116 mRNA and protein expression , Rho-kinase , and NF-kappaB activity . We also investigated the effects of Q07869 agonists on these actions . Aldo , endothelin-1 ( ET-1 ) , and angiotensin II ( P03950 II ) increased Q14116 mRNA and protein expression . P08235 antagonists , endothelin A receptor antagonist , and P03950 II receptor antagonist inhibited Aldo-induced Q14116 expression . Aldo induced ET-1 and P03950 II production in cultured media . Moreover , Rho/Rho-kinase inhibitor and statin inhibited Aldo-induced Q14116 expression . On the other hand , Aldo upregulated the activities of Rho-kinase and NF-kappaB . Q07869 agonists attenuated the Aldo-induced Q14116 expression and NF-kappaB activity but not the Rho-kinase activity . Our findings indicate that Aldo induces Q14116 expression through a mechanism that involves , at a minimum , ET-1 and P03950 II acting via the Rho/Rho-kinase and Q07869 /NF-kappaB pathway . The induction of Q14116 in cardiomyocytes by Aldo , ET-1 , and P03950 II might , therefore , cause a deterioration of the cardiac function in an autocrine and paracrine fashion . The inhibition of the Q14116 expression by Q07869 agonists might be one of the mechanisms whereby the beneficial cardiovascular effects are exerted . Hsp27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp27 ( P04792 ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp27 decreased P05231 -dependent P40763 phosphorylation , nuclear translocation , and P40763 binding to the Twist promoter , suggesting that Hsp27 is required for P05231 -mediated EMT via modulation of P40763 /Twist signaling . We observed a correlation between Hsp27 and Twist in patients with prostate cancer , with Hsp27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp27 inhibition by DB06094 , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp27 as a critical regulator of P05231 -dependent and P05231 -independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer . Suppressed Ca2+/ P62158 /CaMKII-dependent K( DB00171 ) channel activity in primary afferent neurons mediates hyperalgesia after axotomy . Painful axotomy decreases K( DB00171 ) channel current ( IK( DB00171 ) ) in primary afferent neurons . Because cytosolic Ca(2+) signaling is depressed in injured dorsal root ganglia ( Q86YR7 ) neurons , we investigated whether Ca(2+)-calmodulin ( P62158 ) -Ca(2+)/ P62158 -dependent kinase II ( CaMKII ) regulates IK( DB00171 ) in large Q86YR7 neurons . Immunohistochemistry identified the presence of K( DB00171 ) channel subunits Q09428 , SUR2 , and Kir6.2 but not Kir6.1 , and pCaMKII in neurofilament 200-positive Q86YR7 somata . Single-channel recordings from cell-attached patches revealed that basal and evoked IK( DB00171 ) by ionomycin , a Ca(2+) ionophore , is activated by CaMKII . In axotomized neurons from rats made hyperalgesic by spinal nerve ligation ( Q16658 ) , basal K( DB00171 ) channel activity was decreased , and sensitivity to ionomycin was abolished . Basal and Ca(2+)-evoked K( DB00171 ) channel activity correlated inversely with the degree of hyperalgesia induced by Q16658 in the rats from which the neurons were isolated . Inhibition of IK( DB00171 ) by glybenclamide , a selective K( DB00171 ) channel inhibitor , depolarized resting membrane potential ( O94763 ) recorded in perforated whole-cell patches and enhanced neurotransmitter release measured by amperometry . The selective K( DB00171 ) channel opener diazoxide hyperpolarized the O94763 and attenuated neurotransmitter release . Axotomized neurons from rats made hyperalgesic by Q16658 lost sensitivity to the myristoylated form of autocamtide-2-related inhibitory peptide ( AIPm ) , a pseudosubstrate blocker of CaMKII , whereas axotomized neurons from Q16658 animals that failed to develop hyperalgesia showed normal IK( DB00171 ) inhibition by AIPm . AIPm also depolarized O94763 in control neurons via K( DB00171 ) channel inhibition . Unitary current conductance and sensitivity of K( DB00171 ) channels to cytosolic DB00171 and ligands were preserved even after painful nerve injury , thus providing opportunities for selective therapeutic targeting against neuropathic pain . DB04866 : a potent inhibitor of critical steps in angiogenesis progression . We have previously demonstrated that halofuginone , a low molecular weight quinazolinone alkaloid , is a potent inhibitor of collagen alpha1(I) and matrix metalloproteinase 2 ( P08253 ) gene expression . DB04866 also effectively suppresses tumor progression and metastasis in mice . These results together with the well-documented role of extracellular matrix ( Q13201 ) components and matrix degrading enzymes in formation of new blood vessels led us to investigate the effect of halofuginone on the angiogenic process . In a variety of experimental system , representing sequential events in the angiogenic cascade , halofuginone treatment resulted in profound inhibitory effect . Among these are the abrogation of endothelial cell P08253 expression and basement membrane invasion , capillary tube formation , and vascular sprouting , as well as deposition of subendothelial Q13201 . The most conclusive anti-angiogenic activity of halofuginone was demonstrated in vivo ( mouse corneal micropocket assay ) by showing a marked inhibition of basic fibroblast growth factor ( P09038 ) -induced neovascularization in response to systemic administration of halofuginone , either i.p. or in the diet . The ability of halofuginone to interfere with key events in neovascularization , together with its oral bioavailability and safe use as an anti-parasitic agent , make it a promising drug for further evaluation in the treatment of a wide range of diseases associated with pathological angiogenesis . [ Pharmacological and clinical profile of mitiglinide calcium hydrate ( Glufast ) , a new insulinotropic agent with rapid onset ] . DB01252 calcium hydrate ( mitiglinide , Glufast ) is a new insulinotropic agent of the glinide class with rapid onset . DB01252 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 /Kir6.2 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 . Monocyte-mediated suppression of P60568 -induced NK-cell activation . Regulation by P08908 -type serotonin receptors . In the present study , the effects of serotonin on human natural killer ( NK ) -cell responsiveness to interleukin 2 ( P60568 ) was investigated . Concomitant treatment of human lymphocytes , enriched for NK effector cells by Percoll density-gradient centrifugation , with serotonin and P60568 yielded a synergistic activation of NK-cell cytotoxicity ( NKCC ) in the presence but not in the absence of monocytes . The monocyte-dependent , regulatory effects of serotonin and/or P60568 were prostaglandin-independent and could be reconstituted when monocytes , recovered by countercurrent centrifugal elutriation ( CCE ) , were added to purified NK effector cells . The effects of serotonin on baseline and P60568 -activated NK cells were mimicked by the P08908 receptor-specific agonists 8-OH-DPAT and (+)- Q9UM73 . Our data suggest that serotonin regulates NK-cell responsiveness to P60568 via P08908 receptors . A novel proteoliposomal vaccine elicits potent antitumor immunity in mice . Therapeutic vaccination against idiotype is a promising strategy for immunotherapy of B-cell malignancies . Its feasibility , however , is limited by the requirement for a patient-specific product . Here we describe a novel vaccine formulation prepared by simply extracting cell-membrane proteins from lymphoma cells and incorporating them together with P60568 into proteoliposomes . The vaccine was produced in 24 hours , compared with more labor-intensive and time-consuming hybridoma or recombinant DNA methods . The vaccine elicited T-cell immunity in vivo , as demonstrated by secretion of type 1 cytokines . It protected against tumor challenge at doses of tumor antigen 50 to 100 times lower than that previously observed using either liposomes formulated with P60568 and secreted lymphoma immunoglobulin or a prototype vaccine consisting of lymphoma immunoglobulin conjugated to DB05299 . The increased potency justifies testing similar patient-specific human vaccines prepared using extracts from primary tumor samples . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Administration of adenosine diphosphate-ribosyl transferase antagonist allows in vivo control of anti-dinitrophenyl response . DB03073 ( 3MB ) is one of a series of chemical inhibitors of the nuclear enzyme adenosine diphosphate ( ADP ) -ribosyl transferase ( P09874 ) , which has been shown to inhibit cell differentiation in vitro , but has no effect on differentiation independent proliferation . Treatment of mice with an optimal concentration of 3MB ( 20 mg/kg body weight ) at or 1 day after dinitrophenyl-keyhole limpet haemocyanin ( DNP-KLH ) immunisation reduced anti-DNP plaque-forming cell ( P27918 ) numbers to less than 10 % of those of control animals . The period for maximum P27918 suppression showed a narrow time window relative to immunisation , suggesting that in vivo , as in vitro , 3 MB was acting only on those lymphocytes differentiating in response to antigen . Experimental findings showed that it was possible to select for P27918 derived from different populations of DNP-responsive lymphocytes by adjusting the time of 3MB treatment relative to immunisation . When 3MB was used with antigen priming , the residual P27918 showed a lower average affinity than P27918 in mice treated with 3MB 3 days after priming , suggesting a differential selection of those lymphocytes responding either ' early ' or ' late ' in the primary immune response . Stress-induced alterations in P08908 receptor transcriptional modulators O75398 and Q6P1N0 . The effect of stress on the mRNA and protein level of the P08908 receptor and two of its key transcriptional modulators , O75398 and Q6P1N0 , was examined in the prefrontal cortex ( P27918 ) and hippocampus ( Hp ) using rodent models : olfactory bulbectomy ( OB ) and prenatal stress ( PS ) in male and female rats ; chronic mild stress in male rats ( CMS ) and pregnancy stress . In P27918 , CMS induced the most widespread changes , with significant reduction in both mRNA and protein levels of O75398 , P08908 receptor and in Q6P1N0 mRNA ; while in Hp P08908 receptor and Q6P1N0 protein levels were also decreased . In male , but not female OB rats P27918 Q6P1N0 and P08908 receptor protein levels were reduced , while in Hp P08908 receptor , Q6P1N0 and O75398 mRNA 's but not protein were reduced . In PS rats P27918 P08908 receptor protein was reduced more in females than males ; while in Hp Q6P1N0 protein was increased in females . In pregnancy stress , P27918 O75398 , Q6P1N0 and P08908 protein receptor levels were reduced , and in HP P08908 receptor protein levels were also reduced ; in HP only O75398 and Q6P1N0 mRNA levels were reduced . Overall , CMS and stress during pregnancy produced the most salient changes in P08908 receptor and transcription factor expression , suggesting a primary role for altered transcription factor expression in chronic regulation of P08908 receptor expression . By contrast , OB ( in males ) and PS ( in females ) produced gender-specific reductions in P27918 P08908 receptor protein levels , suggesting a role for post-transcriptional regulation . These and previous data suggest that chronic stress might be a key regulator of O75398 / Q6P1N0 gene expression . Differential sensitivity to cardiotonic drugs of cyclic AMP phosphodiesterases isolated from canine ventricular and sinoatrial-enriched tissues . A cardiac phosphodiesterase ( PDE ) which specifically hydrolyzes DB02527 and is inhibited by cyclic GMP has been suggested to be the site of action of new cardiotonic drugs . To investigate the effect of inhibitors , canine cyclic nucleotide PDEs were isolated from left ventricle and from sinoatrial node-enriched tissue , using identical techniques . Four PDE forms could be chromatographically resolved from each tissue , including a peak I PDE ( calmodulin-activated phosphodiesterase , P62158 -PDE ) , a peak II PDE ( cyclic GMP-stimulated phosphodiesterase , O00408 ) and a peak III PDE ( specific for cyclic AMP ) . The latter was further fractionated into two forms : One was inhibited by cyclic GMP and by the platelet antiaggregant AAL 05 ( CGI-PDE ) , and the second was insensitive to cyclic GMP and was inhibited by rolipram ( ROI-PDE ) . Reference PDE inhibitors , isobutyl-1-methylxanthine ( DB07954 ) and papaverine , nonselectively inhibited the four forms isolated from the two tissues . Cardiotonic drugs ( CI 930 , LY 181512 , piroximone , enoximone , and SK & F 94120 ) selectively inhibited CGI-PDE from ventricular tissue but were poorly active on both CGI-PDE and ROI-PDE from the sinoatrial-enriched fraction . In contrast , milrinone inhibited CGI-PDEs and ROI-PDEs from both ventricular and sinoatrial tissues . These results are in good agreement with pharmacologic data in the literature on the positive chronotropic and inotropic effects of the studied drugs in the dog . They provide a possible basis for the dissociation of these two properties of PDE inhibitors . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Serotonin induces the expression of tissue factor and plasminogen activator inhibitor-1 in cultured rat aortic endothelial cells . Serotonin ( 5-hydroxytryptamine , or 5-HT ) , released from activated platelets , not only accelerates aggregation of platelets but also is known to promote mitosis , migration , and contraction of vascular smooth muscle cells ( VSMCs ) . These effects are considered to contribute to thrombus formation and atherosclerosis . The aim of this study was to investigate the effects of 5-HT on the expressions of coagulative and fibrinolytic factors in rat aortic endothelial cells . Endothelial cells were stimulated with various concentrations of 5-HT ( 0.1 approximately 10 microM ) , and the expressions of tissue factor ( TF ) , tissue factor pathway inhibitor ( P10646 ) , plasminogen activator inhibitor-1 ( P05121 ) , and tissue-type plasminogen activator ( TPA ) messenger RNAs ( mRNAs ) were evaluated by Northern blot analysis . The activities of TF and P05121 were also measured . TF and P05121 mRNA were increased significantly in a concentration- and time-dependent manner . However , P10646 and TPA mRNA expression did not change . The inductions of TF and P05121 mRNAs were inhibited by a 5-HT1/5-HT2 receptor antagonist ( methiothepin ) and a selective 5- Q13049 receptor antagonist ( D6RGH6 -9042 ) . These results indicate that 5-HT increases procoagulant activity and reduces fibrinolytic activities of endothelial cells through the 5- Q13049 receptor . It was concluded that the modulation of procoagulant and hypofibrinolytic activities of endothelial cells by 5-HT synergistically promotes thrombus formation at the site of vessel injury with the platelet aggregation , VSMC contraction , and VSMC proliferation .

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interacts_with DB00677?

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Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . A double-blind , placebo-controlled study of a P32239 antagonist , CI-988 , in patients with generalized anxiety disorder . This multicenter , double-blind , placebo-controlled , parallel-group , randomized study assessed the efficacy , safety , and tolerability of a novel CCK-B antagonist CI-988 in the treatment of generalized anxiety disorder ( Q99259 ) . Patients received placebo or CI-988 ( 300 mg/day , thrice daily ) for 4 weeks . Patients with a primary diagnosis of Q99259 according to DSM-III-R criteria were randomized . The study design included a 1- to 2-week single-blind placebo baseline phase , followed by a 4-week double-blind treatment phase . Efficacy was measured weekly by Hamilton Rating Scale for Anxiety ( HAM-A ) , Clinical Global Impressions of Severity and Change , UCLA-Multi Dimensional Anxiety Scale , and Hamilton Rating Scale for Depression . Patients were also evaluated to determine whether they met criteria for irritable bowel syndrome ( IBS ) at screening and were evaluated with a gastrointestinal visual analog scale at each visit . Eighty-eight patients were randomized to CI-988 ( N = 45 ) and placebo ( N = 43 ) at three centers . CI-988 did not demonstrate an anxiolytic effect superior to placebo in this clinical trial . There was no significant difference in mean change in HAM-A total between placebo ( -7.73 ) and CI-988 ( -8.64 ) . However , a significant treatment-by-center interaction and a highly variable placebo response rate among the three centers limit the interpretation of the results . CI-988 did not have an effect on symptoms of IBS other than diarrhea , which worsened in patients with IBS . Other than a higher incidence of some gastrointestinal symptoms ( diarrhea , dyspepsia , flatulence , and nausea ) , CI-988 was well tolerated . Results suggest that testing higher oral doses of CI-988 may be warranted . Strain and regional dependence of alternate splicing of acetylcholinesterase in the murine brain following stress or treatment with diisopropylfluorophosphate . Induction of the rare readthrough variant of acetylcholinesterase ( P22303 -R ) by an acetylcholinesterase ( P22303 ) inhibitor or by stress was tested in four mouse strains that differ in their behavioural profiles on tests of anxiety and depression . BALB/C , C57Bl/6 , C3H/He and CD-1 mouse strains were tested in the elevated plus maze in two sessions , separated by 48h . All strains , except CD-1 , showed the expected reduction in open arm exploration on the second session . BALB/C and C3H mice spent a greater proportion of the time in the open arms on the first exposure , but spent more time immobile in the maze compared to the CD1 and C57 strains . Immobility was attenuated upon the second exposure in all strains , except the BALB/C mice . Real-time PCR was used to investigate regional and strain differences in induction of P22303 -R mRNA following four daily injections of diisopropylfluorophosphate ( DB00677 ) ( .1mg/kg ) . P22303 -R induction was found in the frontal cortex , but not in amygdala , hippocampus or striatum of CD-1 mice . Nor was there P22303 -R induction in the brains of the inbred strains . Four daily sessions of swim stress were used to investigate stress-induced induction of P22303 -R . BALB/C mice showed significantly more immobility in the forced swim test ( P19883 ) compared to the other strains . P19883 did not induce P22303 -R mRNA in any brain region tested ; however , P22303 -R mRNA expression in the frontal cortex was negatively correlated with immobility in the P19883 . Kinetic study of neuropeptide Y ( P01303 ) proteolysis in blood and identification of NPY3-35 : a new peptide generated by plasma kallikrein . There is little information on how neuropeptide Y ( P01303 ) proteolysis by peptidases occurs in serum , in part because reliable techniques are lacking to distinguish different P01303 immunoreactive forms and also because the factors affecting the expression of these enzymes have been poorly studied . In the present study , LC-MS/MS was used to identify and quantify P01303 fragments resulting from peptidolytic cleavage of P01303 (1-36) upon incubation with human serum . Kinetic studies indicated that P01303 (1-36) is rapidly cleaved in serum into 3 main fragments with the following order of efficacy : P01303 (3-36) >> P01303 (3-35) > P01303 (2-36) . Trace amounts of additional P01303 forms were identified by accurate mass spectrometry . Specific inhibitors of dipeptidyl peptidase IV , kallikrein , and aminopeptidase P prevented the production of P01303 (3-36) , P01303 (3-35) , and P01303 (2-36) , respectively . P03952 at physiological concentrations converted P01303 (3-36) into P01303 (3-35) . Receptor binding assays revealed that P01303 (3-35) is unable to bind to P01303 Q03519 , P28062 , and Y5 receptors ; thus P01303 (3-35) may represent the major metabolic clearance product of the P28062 /Y5 agonist , P01303 (3-36) . P05305 , endothelin-1 receptors and cardiac natriuretic peptides in failing human heart . Endothelin ( ET ) -1 is a potent vasoconstrictor peptide produced in the myocardium that can exert important effects on cardiac myocyte growth and phenotype ; cardiac natriuretic peptides ( P01160 and DB04899 ) are known to act as physiological antagonists of ET-1 . In this study a comparative determination of ET-1 receptors and of the local productions of ET-1 and of P01160 and DB04899 was made in different sites of failing and nonfailing hearts . Tissue from right and left atrium , right and left ventricle and interventricular septum from seven adult heart transplant recipients with end-stage idiopathic dilated cardiomyopathy ( functional class III and IV , with ejection fraction < 35 % ) and from four postmortem subjects without cardiac complications was analyzed . In failing hearts we observed a tendency to increase of density of binding sites , most evident in left ventricle ( 62.6+/-22.6 fmol/mg protein vs. 29.0+/-3.3 , mean +/- SEM , p = ns ) . A prevalence of P25101 subclass , observed in all samples , resulted more pronounced in failing hearts where this increase , found in all the cardiac regions , was more evident in left ventricle ( p = 0.0007 vs nonfailing hearts ) . The local concentrations of ET-1 , P01160 and DB04899 resulted significantly increased in failing hearts with respect to controls in all sides of the heart . In failing hearts we have observed a tendency to increase in endothelin receptor density mainly due to a significant upregulation of P25101 subtype and a parallel increase of the tissue levels of P01160 , DB04899 and ET-1 indicating an activation of these systems in heart failure . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . DB00227 overcomes gefitinib resistance in human non-small cell lung cancer cells with K-Ras mutations . DB00227 is a 3-hydroxy-3-methylglutaryl-coenzyme A ( HMG- DB01992 ) reductase inhibitor . Its inhibitory action on P04035 leads to depletion of isoprenoids , which inhibits post-translational modification of DB01367 . In this study , we investigated the effect of combining lovastatin with gefitinib on gefitinib-resistant human non-small cell lung cancer ( NSCLC ) cell lines with K-Ras mutations . Antitumor effects were measured by growth inhibition and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay . Effects on apoptosis were determined by flow cytometry , DNA fragmentation , and immunoblots . Protein levels of DB01367 , AKT/pAKT , and RAF/ P27361 /2 in cancer cells were analyzed by immunoblot . Compared with gefitinib alone , a combination of gefitinib with lovastatin showed significantly enhanced cell growth inhibition and cytotoxicity in gefitinib-resistant A549 and NCI-H460 human NSCLC cells . In addition , lovastatin combination treatment significantly increased gefitinib-related apoptosis , as determined by fluorescence microscopy and flow cytometric analysis . These effects correlated with up-regulation of cleaved caspase-3 , poly ( ADP-ribose ) polymerase ( PARP ) , and Bax and down-regulation of Bcl-2 . The combination of lovastatin and gefitinib effectively down-regulated DB01367 protein and suppressed the phosphorylation of RAF , P27361 /2 , AKT , and P00533 in both cell lines . Taken together , these results suggest lovastatin can overcome gefitinib resistance , in NSCLC cells with K-Ras mutations , by down regulation of DB01367 protein , which leads to inhibition of both RAF/ P29323 and AKT pathways . DB00644 antagonist in the management of prostate cancer . DB00044 -releasing hormone ( P01148 ) agonist therapy to induce medical castration has become the most common form of hormonal therapy for advanced and metastatic prostate cancer . When treatment is started , P01148 agonists initially stimulate the release of LH , causing a surge in serum testosterone that can precipitate a " flare " phenomenon or worsening of disease , particularly in patients with bone metastatic disease . DB00644 ( DB00644 ) receptor antagonism represents a newer approach to medical castration . DB00106 is a pure P30968 antagonist that is devoid of any P01148 agonist activity . Results from 1 phase II and 3 phase III clinical trials demonstrate that abarelix produces medical castration more quickly and without causing testosterone surge , as compared with P01148 agonists with or without a nonsteroidal antagonist . The safety profile in terms of adverse events is comparable between the 2 types of treatment , but the lack of testosterone surge with abarelix might confer a safety advantage by abolishing the risk of a disease flare . Kinin-B2 receptor exerted neuroprotection after diisopropylfluorophosphate-induced neuronal damage . The kinin-B2 receptor ( B2BKR ) activated by its endogenous ligand bradykinin participates in various metabolic processes including the control of arterial pressure and inflammation . Recently , functions for this receptor in brain development and protection against glutamate-provoked excitotoxicity have been proposed . Here , we report neuroprotective properties for bradykinin against organophosphate poisoning using acute hippocampal slices as an in vitro model . Following slice perfusion for 10min with diisopropylfluorophosphate ( DB00677 ) to initiate the noxious stimulus , responses of pyramidal neurons upon an electric impulse were reduced to less than 30 % of control amplitudes . Effects on synaptic-elicited population spikes were reverted when preparations had been exposed to bradykinin 30min after challenging with DB00677 . Accordingly , bradykinin-induced population spike recovery was abolished by HOE-140 , a B2BKR antagonist . However , the kinin-B1 receptor ( B1BKR ) agonist Lys-des- DB00125 (9)-bradykinin , inducing the phosphorylation of mitogen-activated protein kinase ( MEK/MAPK ) and cell death , abolished bradykinin-mediated neuroprotection , an effect , which was reverted by the P29323 inhibitor PD98059 . In agreement with pivotal B1BKR functions in this process , antagonism of endogenous B1BKR activity alone was enough for restoring population spike activity . On the other hand pralidoxime , an oxime , reactivating acetylcholinesterase ( P22303 ) after organophosphate poisoning , induced population spike recovery after DB00677 exposure in the presence of bradykinin and Lys-des- DB00125 (9)-bradykinin . Lys-des- DB00125 (9)-bradykinin did not revert protection exerted by pralidoxime , however when instead bradykinin and Ly-des- DB00125 (9)-bradykinin were superfused together , recovery of population spikes diminished . These findings again confirm the neuroprotective feature of bradykinin , which is , diminished by its endogenous metabolites , stimulating the B1BKR , providing a novel understanding of the physiological roles of these receptors . Q8N0V5 V ( Mgat5 ) -mediated N-glycosylation negatively regulates Th1 cytokine production by T cells . The differentiation of naive P01730 (+) T cells into either proinflammatory Th1 or proallergic Th2 cells strongly influences autoimmunity , allergy , and tumor immune surveillance . We previously demonstrated that beta1,6GlcNAc-branched complex-type ( Q8N0V5 V ( Mgat5 ) ) N-glycans on TCR are bound to galectins , an interaction that reduces TCR signaling by opposing agonist-induced TCR clustering at the immune synapse . Mgat5(-/-) mice display late-onset spontaneous autoimmune disease and enhanced resistance to tumor progression and metastasis . In this study we examined the role of beta1,6GlcNAc N-glycan expression in Th1/Th2 cytokine production and differentiation . beta1,6GlcNAc N-glycan expression is enhanced by TCR stimulation independent of cell division and declines at the end of the stimulation cycle . DB00075 -activated splenocytes and naive T cells from Mgat5(-/-) mice produce more P01579 and less P05112 compared with wild-type cells , the latter resulting in the loss of P05112 -dependent down-regulation of IL-4Ralpha . DB02034 , an inhibitor of Q16706 , blocked beta1,6GlcNAc N-glycan expression and caused a similar increase in P01579 production by T cells from humans and mice , but no additional enhancement in Mgat5(-/-) T cells . Mgat5 deficiency did not alter P01579 / P05112 production by polarized Th1 cells , but caused an approximately 10-fold increase in P01579 production by polarized Th2 cells . These data indicate that negative regulation of TCR signaling by beta1,6GlcNAc N-glycans promotes development of Th2 over Th1 responses , enhances polarization of Th2 cells , and suggests a mechanism for the increased autoimmune disease susceptibility observed in Mgat5(-/-) mice . The PEPvIII-KLH ( DB05374 ) vaccine in glioblastoma multiforme patients . Conventional therapies for glioblastoma multiforme ( GBM ) fail to target tumor cells exclusively , resulting in non-specific toxicity . Immune targeting of tumor-specific mutations may allow for more precise eradication of neoplastic cells . P00533 variant III ( EGFRvIII ) is a tumor-specific mutation that is widely expressed in GBM and other neoplasms and its expression enhances tumorigenicity . This in-frame deletion mutation splits a codon , resulting in a novel glycine at the fusion junction producing a tumor-specific epitope target for cellular or humoral immunotherapy . We have previously shown that vaccination with a peptide that spans the EGFRvIII fusion junction ( PEPvIII-KLH/ DB05374 ) is an efficacious immunotherapy in syngeneic murine models . In this review , we summarize our results in GBM patients targeting this mutation in multiple , multi-institutional Phase II immunotherapy trials . These trials demonstrated that a selected population of GBM patients who received vaccines targeting EGFRvIII had an unexpectedly long survival time . Further therapeutic strategies and potential pitfalls of using this approach are discussed . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . Efficacy of combination therapy with amantadine , thymosin alpha 1 and alpha/beta interferon in mice infected with influenza A virus . The aim of this study was to examine the effects of the antiviral drug amantadine ( Q9BXJ7 ) administered in combination with thymosin alpha 1 ( T alpha 1 ) and murine alpha/beta interferon ( IFN ) on mice infected with influenza A PR8 virus . Combined treatment with Q9BXJ7 and T alpha 1 , for 4 days , followed by a single injection of IFN , was initiated 1 h after intranasal viral inoculation . The effectiveness of this new chemoimmunotherapy protocol was seen in the long-term survival of a high percentage of animals and was statistically significant when compared to treatment with single agents in conjunction with chemotherapy or to chemotherapy alone . In addition , chemoimmunotherapy treatment reduces the viral titre in the lungs as well as restoring the immunological parameters tested ( natural killer cell activity ; cytotoxic T-lymphocyte responses ; P01730 +/CD8+ lymphocyte subsets ) with respect to all other groups . These results suggest the potential use of these immunomodulating agents in combination with an antiviral drug in controlling PR8 influenza virus infection . The response of striatal neuropeptide Y and cholinergic neurons to excitatory amino acid agonists . The effect of excitatory amino acid antagonists on antagonist on neuropeptide Y ( P01303 ) and cholinergic neurons in the striatum of the rat was studied by means of P01303 immunocytochemistry , DB00677 histochemistry for acetylcholinesterase ( P22303 ) , and biochemical determinations of choline acetyltransferase ( P28329 ) . Intrastriatal infusion of drugs revealed that striatal neurons containing P01303 are more sensitive than cholinergic neurons to the neurotoxic actions of kainic acid ( KA ) , quinolinic acid ( QA ) and L-glutamic acid ( GA ) ; all 3 compounds produced a marked loss of P01303 neurons , but only a moderate decrease in the number of P22303 neurons or P28329 activity . Co-injection experiments showed that the neurotoxicity of QA and GA , but not that of KA , can be antagonized by the specific N-methyl-D-aspartate ( DB01221 ) receptor antagonist 3-((+/-)-2-(carboxypiperazine-4-yl))-propyl-1-phosphonic acid ( CPP ) . Destruction of the glutamatergic corticostriatal projection by cerebral decortication protected striatal P01303 and cholinergic neurons against KA neurotoxicity . These results indicate that striatal P01303 and cholinergic neurons receive prominent cortical amino acid afferents , and that the neurotoxic effect of QA and GA on these neurons is mediated through DB01221 receptors . DB06096 , a selective P29475 inhibitor and a P28222 /1D receptor agonist , inhibits P80511 release in preclinical migraine models . BACKGROUND : DB06096 is a combined neuronal nitric oxide synthase ( P29475 ) inhibitor and 5-hydroxytryptamine 1B/1D ( P28222 /1D ) receptor agonist . Using preclinical models , we evaluated whether these two unique therapeutic principles have a synergistic effect in attenuating stimulated calcitonin gene-related peptide ( P80511 ) release , a marker of trigeminal activation . METHODS : We examined the effect of DB06096 on : ( 1 ) DB00761 - , capsaicin- and resiniferatoxin ( RTX ) -induced immunoreactive P80511 ( iCGRP ) release from isolated preparation of rat dura mater , trigeminal ganglion ( TG ) and trigeminal nucleus caudalis ( P24821 ) ; and ( 2 ) capsaicin- and electrical stimulation ( ES ) -induced middle meningeal artery ( MMA ) dilation in a rat closed-cranial window . RESULTS : DB06096 inhibited : ( 1 ) DB00761 -stimulated iCGRP release from dura mater ( % decrease mean ± SEM , lowest effective concentration ) ( 35 ± 6 % , 30 µM ) , TG ( 24 ± 11 % , 10 µM ) and P24821 ( 40 ± 8 % , 10 µM ) ; ( 2 ) capsaicin- and RTX-induced iCGRP release from dura mater ; and ( 3 ) capsaicin- and ES-induced increase in dural artery diameter ( 32 ± 5 % , 3 mg kg(-1) intravenous ( i.v. ) and 36 ± 1 % , 10 mg kg(-1) i.v. ) . CONCLUSIONS : DB06096 inhibits P80511 release from migraine-relevant cephalic tissues . Its effect is most likely mediated via a combination of P29475 -inhibition and P28222 /1D receptor agonism in dura mater while the mechanisms of action for inhibition of P80511 release from TG and P24821 have to be investigated further . DB06268 ( Q9Y6W8 -Texas Biotechnology ) . Q9Y6W8 -Texas Biotechnology is developing the endothelin A ( P25101 ) receptor antagonist , sitaxsentan , for the potential treatment of pulmonary hypertension , congestive heart failure ( CHF ) , chronic obstructive pulmonary disease and subarachnoid hemorrhage [ 205713 ] , [ 302200 ] . The compound is in phase IIa trials as an iv formulation for CHF and has completed phase I safety trials as an oral formulation [ 272071 ] . Phase II/III trials for pulmonary hypertension are planned for the first quarter of 2001 [ 3945711 ] . In June 2000 , Q9Y6W8 and Texas Biotechnology established a joint venture to develop and commercialize endothelin antagonists [ 370007 ] . US-05591761 was issued to Texas in January 1997 , covering TBC-11251 and several related isomers [ 2309301 . Candidate gene markers for sperm quality and fertility of boar . Candidate genes gonadotropin releasing hormone receptor ( P30968 ) , prolactin ( PRL ) , prolactin receptor ( P16471 ) , follicle-stimulating hormone beta ( P01225 ) , luteinizing hormone beta ( P01229 ) , follistatin ( P19883 ) , inhibin alpha ( P05111 ) , inhibin beta A ( P08476 ) and inhibin beta B ( P09529 ) were investigated for their association with sperm quality traits of sperm concentration ( SCON ) , motility ( P38646 ) , semen volume per ejaculate ( VOL ) , plasma droplets rate ( PDR ) , abnormal sperm rate ( ASR ) and fertility traits of non return rate ( NRR ) and number of piglets born alive ( NBA ) . The experimental material included 356 boars of Pietrain ( PI ) and Pietrain x Hampshire ( PI x HA ) . Analysis of variance revealed significant association of P30968 with P38646 ( P = 0.0161 ) , PDR ( P = 0.0048 ) and ASR ( P = 0.0201 ) , P08476 was found to have significant effects on PDR ( P = 0.0318 ) and ASR ( P = 0.0067 ) , P09529 was significant ( P = 0.0360 ) for SCON trait . P01225 , P19883 , P05111 , PRL , P16471 and P01229 had no significant effects on any trait in this experiment . Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 -I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 -Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 -I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 -Is . Treatment for the toxicity of P22303 -Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 -Is exert their effects . By attenuating the effects of P22303 -Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 -I . [ Endometrial cancer in young women -- clinical and molecular aspects ] . OBJECTIVES : The aim the study was to compare two groups of endometrial cancer patients ( below and above 45 years of age ) in the aspect of clinicopathological and molecular data . MATERIAL AND METHODS : The study encompassed 456 primary tumour samples retrospectively collected from a cohort of endometrial cancer patients , primarily treated by surgery Molecular analysis covered : copy number variations of 10 genes ( P11388 , P00533 , P04626 , P21860 , Q15303 , MYC , P24385 , P03372 , P42336 , O60216 ) analyzed by quantitative PCR ; mRNA expression of 6 genes ( Q13296 , RAD27 , Q01196 , O95863 , O43623 , O43490 ) analyzed with the use of reverse transcription quantitative PCR ; protein expression analysis of 8 markers ( P06401 , P03372 ; P00533 , P04626 , P21860 , Q15303 , P11388 , pAKT1 ) performed with the use of immunohistochemistry . RESULTS : The younger group of patients was characterized by less frequent hypertension ( p < 0.00007 ) , less frequent myometrial infiltration ( p=0.002 ) and longer overall survival ( p=0.003 ) . Apart from O60216 gene aberrations , which were more frequent in younger patients ( p=0.02 ) , the study revealed no statistically significant differences between the groups . CONCLUSIONS : The study showed no molecular differences in the profile of younger and older endometrial cancer patients . Data on both the prognostic and predictive significance of O60216 in endometrial cancer are still insufficient . The clinical profile of younger patients with endometrial carcinoma was slightly better when compared to elderly patients . Younger patients were characterized by longer overall survival . Spin trapping agent phenyl-N-tert-butylnitrone prevents diisopropylphosphorofluoridate-induced excitotoxicity in skeletal muscle of the rat . Indirect evidence suggests that reactive oxygen species ( ROS ) may mediate muscle fiber necrosis following muscle hyperactivity induced by the anticholinesterase diisopropylphosphorofluoridate ( DB00677 ) . Pronounced muscle fasciculations and muscle fiber necrosis were seen when acetylcholinesterase ( P22303 ) activity was reduced to less than 30 % of control . The spin trapping agent phenyl-N-tert-butylnitrone ( PBN ) was used in vivo to directly assess the formation of ROS during DB00677 ( 1.75 mg/kg , s.c. ) induced muscle hyperactivity . Pretreatment with PBN ( 300 mg/kg , i.p. ) , the concentration necessary for in vivo spin trapping , prevented muscle hyperactivity as well as necrosis and attenuated the DB00677 induced P22303 inhibition otherwise seen in DB00677 only treated rats . PBN had no effect when given after fasciculations were established . Muscle extracts from PBN and DB00677 treated rats subjected to electron spin resonance ( P03372 ) spectroscopy tested negative for ROS . While the role of PBN as an antioxidant is well established , its prophylactic effect against excitotoxity induced by an P22303 inhibitor are due to its protection of P22303 , an unexpected non-antioxidant action . Transmitter neurochemistry of the efferent neuron system innervating the labyrinth . It is likely that several mechanisms contribute to the efferent control of cochlear and vestibular function . Different effects are probably mediated by different neuronal transmitters . In spite of a number of transmitter candidates , it is still widely assumed that the entire efferent system can be globally characterized as cholinergic . We attempted to label retrogradely identified efferent neurons in the brainstem with a monoclonal antibody against choline acetyltransferase ( P28329 ) , the acetylcholine ( ACh ) synthesizing enzyme . Only a portion of the vestibular efferents could thus be shown to be cholinergic in the rat . Medial cochlear efferents , terminating under outer hair cells , may also be cholinergic since they stain intensely for acetylcholine esterase ( P22303 ) after pre-treatment with the P22303 inhibitor diisopropylfluorophosphate ( DB00677 ) . The lateral cochlear efferents terminating under inner hair cells , as well as more than half of the vestibular efferent neuron population , reacted negatively with either method designed to identify cholinergic neurons . Half of the lateral olivo-cochlear neuron population filled retrogradely with tritiated gamma-amino butyric acid [ ( 3H ] -GABA ) . These cells were similar in size and distribution to neurons staining for the GABA synthesizing enzyme glutamic acid decarboxylase ( Q99259 ) . Retrograde transport of [ 3H ] -aspartate from the inner ear to the brainstem was seen in half of the lateral olivocochlear population , as well as in part of the efferent vestibular population in group E and in the caudal pontine reticular nucleus ( P16435 ) . Since various peptides have also been located in efferent neurons , this system is chemically diversified . Several distinct mechanisms of efferent control with presumably differing functions must , therefore , exist . DB00183 infusions in patients with panic disorder . I . Symptoms and cardiovascular responses . Cholecystokinin ( CCK ) may mediate human anxiety and animal data suggest that cholecystokinin antagonists could provide an important advance in the treatment of anxiety disorders . The study of CCK receptor systems in psychiatric patients has , however , been severely limited by the lack of available probes . We utilized intravenous infusions of pentagastrin , a selective P32239 agonist , and studied behavioral and cardiovascular responses in 10 patients with panic disorder and 10 normal controls . DB00183 produced substantial symptomatology , including anxiety , and increases in heart rate and blood pressure , in both patients and controls . Patients were more sensitive to the panicogenic effects of the pentagastrin . Panic attacks occurred in 70 % of patients and 0 % of controls . Patients ' symptom responses were very similar to their " typical " panic attacks and to symptoms produced by Q13308 . DB00183 provides a readily available alternative to Q13308 for studying the CCK receptor system and exploring its involvement in human anxiety . Short- and long-term influences of calcitonin gene-related peptide on the synthesis of acetylcholinesterase in mammalian myotubes . The present study analyses the short- ( 15 min - 2 h ) and long-term ( 24 - 48 h ) influences of calcitonin gene-related peptide ( P80511 ) on acetylcholinesterase ( P22303 ) expression in the rat cultured skeletal muscle and the signal transduction events underlying P80511 actions . To assess the effect of P80511 on P22303 synthesis , myotubes were pre-exposed to the irreversible P22303 inhibitor diisopropyl fluorophosphate ( DB00677 ) and treated with P80511 or forskolin , an adenylyl cyclase ( AC ) activator . Treatment of myotubes with 1 - 100 nM P80511 for 2 h increased by up to 42 % the synthesis of catalytically active P22303 with a parallel increase in the intracellular cyclic AMP . The stimulation of P22303 synthesis induced by P80511 was mimicked by direct activation of AC with 3 - 30 microM forskolin . In contrast , pre-treatment of cultures with 100 nM P80511 for 20 h reduced by 37 % the subsequent synthesis of P22303 , resulting in a 15 % decrease in total P22303 activity after 48 h P80511 treatment . Moreover , 24 h treatment of myotubes with 100 nM P80511 reduced by 54 % the accumulation of cyclic AMP induced by a subsequent P80511 treatment . These findings indicate that , in skeletal muscle cells , P80511 modulates the P22303 expression in a time-dependent manner , initially stimulating the enzyme synthesis through a cyclic AMP-dependent mechanism . The decreased P22303 synthesis observed after long-term P80511 treatment suggests that P80511 signalling system is subject to desensitization or down-regulation , that might function as an important adaptative mechanism of the muscle fibre in response to long-term changes in neuromuscular transmission .

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interacts_with DB04871?

multiple_choice

Discovery and structure-activity relationship of ( 1R ) -8-chloro-2,3,4,5-tetrahydro-1-methyl-1H-3-benzazepine ( DB04871 ) , a selective serotonin P28335 receptor agonist for the treatment of obesity . The synthesis and SAR of a novel 3-benzazepine series of P28335 agonists is described . Compound 7d ( lorcaserin , APD356 ) was identified as one of the more potent and selective compounds in vitro ( pEC50 values in functional assays measuring [(3)H]phosphoinositol turnover : P28335 = 8.1 ; 5- Q13049 = 6.8 ; P41595 = 6.1 ) and was potent in an acute in vivo rat food intake model upon oral administration ( ED50 at 6 h = 18 mg/kg ) . DB04871 was further characterized in a single-dose pharmacokinetic study in rat ( t1/2 = 3.7 h ; F = 86 % ) and a 28-day model of weight gain in growing Sprague-Dawley rat ( 8.5 % decrease in weight gain observed at 36 mg/kg b.i.d. ) . DB04871 was selected for further evaluation in clinical trials for the treatment of obesity . The role of serotonin receptor subtypes in the behavioural effects of neuroleptic drugs . A paw test study in rats . The present study was designed to evaluate the roles of serotonin P08908 and 5-HT2 receptors in the effects of neuroleptic drugs in the paw test . This behavioural test has been shown to model both the antipsychotic efficacy as well as the extrapyramidal side-effect liability of neuroleptic drugs . Whereas the P08908 receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OHDPAT ) reduced the effects of the classical neuroleptic haloperidol , it increased the effects of the atypical neuroleptic clozapine . The 5-HT2 receptor antagonist ketanserin as well as the P28335 /5-HT2 receptor antagonist ritanserin , on the other hand reduced the effects of haloperidol , whereas the P28335 /5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ( DOI ) reduced the effects of clozapine . The most important finding , however , was that the behavioural effects of different ( putative ) neuroleptics ( fluphenazine , P35240 -39166 , remoxipride , prothipendyl , thioridazine and risperidone ) were differentially influenced by both 8-OHDPAT and DOI , suggesting that there are important differences between the neuronal mechanisms underlying the behavioural effects of these neuroleptic drugs , even within the subclasses of classical and atypical neuroleptics . 5- Q9H205 - and P28335 -antagonist properties of cyamemazine : significance for its clinical anxiolytic activity . RATIONALE : DB09000 is a neuroleptic compound which possesses anxiolytic properties in humans . On the other hand , 5- Q9H205 - and P28335 -receptors have been implicated in anxiety disorders and a previous binding study has shown that cyamemazine possesses high affinity for both serotonin receptor types . OBJECTIVE : The present study was undertaken to establish whether cyamemazine antagonizes 5- Q9H205 - and/or P28335 -mediated responses , and whether it compares with reference compounds . METHODS : DB09000 was tested for its ability to antagonize : ( i ) 5- Q9H205 -dependent contraction of the isolated guinea-pig ileum and bradycardic responses in the rat and ( ii ) P28335 -dependent phospholipase C ( P98160 ) stimulation in rat brain membranes . RESULTS : In isolated guinea-pig ileum , cyamemazine potently and competitively antagonized 5-HT-dependent contractions ( pA2 = 7.52 +/- 0.08 ; n = 5 ) . In this test , cyamemazine was 5-7 times more potent ( pIC50 = 6.75 +/- 0.13 ) than tropisetron ( pIC50 = 6.02 +/- 0.04 ) . In rats , cyamemazine i.v. antagonized 5-HT-dependent bradycardic responses with ID50 % = 3.2 +/- 1.5 mg/kg ( n = 4 ) . Finally , in rat brain membranes cyamemazine antagonized P28335 -dependent P98160 stimulation with Ki = 424 nM ( mianserin exhibits a Ki = 113 nM ) . CONCLUSIONS : DB09000 behaves as an antagonist at both 5- Q9H205 - and P28335 -receptors , which compares well with reference compounds . These 5- Q9H205 - and P28335 -antagonistic actions of cyamemazine can be involved , at least in part , in its beneficial therapeutic actions in anxiety disorders . Q9UEF7 gene deficiency causes salt-sensitive hypertension via monocyte chemotactic protein-1/CC chemokine receptor 2-mediated inflammation . Q9UEF7 ( KL ) is a newly discovered aging suppressor gene . In mice , the KL gene extends the lifespan when overexpressed and shortens the lifespan when disrupted . This study investigated if KL deficiency affects BP and salt sensitivity using KL mutant heterozygous ( +/- ) mice and wild-type ( WT ) mice ( 9 weeks of age , 16 mice per group ) . Notably , systolic BP in KL(+/-) mice began to increase at the age of 15 weeks , reached a peak level at the age of 17 weeks , and remained elevated thereafter , whereas systolic BP remained consistent in WT mice . High salt ( HS ) intake further increased BP in KL(+/-) mice but did not affect BP in WT mice . Blockade of CC chemokine receptor 2 ( P41597 ) , involved in monocyte chemotaxis , by a specific P41597 antagonist ( DB05130 ) abolished the HS-induced increase in BP in KL(+/-) mice . Furthermore , HS loading substantially increased the expression of monocyte chemotactic protein-1 and the infiltration of macrophages and T cells in kidneys in KL(+/-) mice , and treatment with DB05130 abolished these effects . Treatment of KL(+/-) mice with DB05130 also attenuated the increased renal expressions of serum glucocorticoid-regulated kinase 1 , thiazide-sensitive NaCl cotransporter , and DB00171 synthase β along with the renal structural damage and functional impairment induced by HS loading . In conclusion , KL deficiency caused salt-sensitive hypertension and renal damage by P41597 -mediated inflammation . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . Q00613 protects cardiomyocytes from ischemia/reperfusion injury . BACKGROUND : Because cardiomyocyte death causes heart failure , it is important to find the molecules that protect cardiomyocytes from death . The death trap is a useful method to identify cell-protective genes . METHODS AND RESULTS : In this study , we isolated the heat shock transcription factor 1 ( Q00613 ) as a protective molecule by the death trap method . Cell death induced by hydrogen peroxide was prevented by overexpression of Q00613 in COS7 cells . Thermal preconditioning at 42 degrees C for 60 minutes activated Q00613 , which played a critical role in survival of cardiomyocytes from oxidative stress . In the heart of transgenic mice overexpressing a constitutively active form of Q00613 , ischemia followed by reperfusion-induced ST-segment elevation in ECG was recovered faster , infarct size was smaller , and cardiomyocyte death was less than wild-type mice . P31749 /Akt was more strongly activated , whereas Jun N-terminal kinase and caspase 3 were less activated in transgenic hearts than wild-type ones . CONCLUSIONS : These results suggest that Q00613 protects cardiomyocytes from death at least in part through activation of Akt and inactivation of Jun N-terminal kinase and caspase 3 . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD . Serotonin increases P27361 /2 phosphorylation in astrocytes by stimulation of P41595 and P28335 receptors . We have previously shown that fluoxetine causes P29323 (1/2) phosphorylation in cultured mouse astrocytes mediated exclusively by stimulation of 5-HT(2B) receptors ( Li et al. , 2008b ) . This raises the question whether this is also the case for serotonin ( 5-HT ) itself . In the present study serotonin was found to induce P29323 (1/2) phosphorylation by stimulation of 5-HT(2B) receptors with high affinity ( EC(50) : 20-30 pM ) , and by stimulation of 5-HT(2C) receptor with low affinity ( EC(50) : 1 microM or higher ) . P29323 (1/2) phosphorylation induced by stimulation of either 5-HT(2B) or 5-HT(2C) receptors was mediated by epidermal growth factor ( P01133 ) receptor transactivation ( Peng et al. , this issue ) , shown by the inhibitory effect of AG1478 , an inhibitor of the P01133 receptor tyrosine kinase , and DB02255 , an inhibitor of Zn-dependent metalloproteinases , and thus of 5-HT(2B) receptor-mediated P01133 receptor agonist release . It is discussed that the high potency of the 5-HT(2B)-mediated effect is consistent with literature data for binding affinity of serotonin to cloned human 5-HT(2B) receptors and with observations of low extracellular concentrations of serotonin in brain , which would allow a demonstrated moderate and modality-dependent increase in specific brain areas to activate 5-HT(2B) receptors . In contrast the relevance of the observed 5-HT(2C) receptors on astrocytes is questioned . Pharmacological characterization of mitogen-activated protein kinase activation by recombinant human P28335 , 5- Q13049 , and P41595 receptors . The type 2 serotonin ( 5-HT(2) ) receptor subfamily is known to couple to phosphoinositide hydrolysis ( PI ) and the subsequent mobilization of intracellular Ca(2+) , as well as the release of arachidonic acid ( AA ) . Less is known of 5-HT(2)-mediated activation of the mitogen-activated protein kinase ( MAPK ) or extracellular signal-regulated kinase ( P27361 /2 ) signaling . The present study measured the relative efficacies and potencies of 5-HT agonists to activate P28482 in non-neuronal cells expressing recombinant human 5-HT(2A) , 5-HT(2B) , and 5-HT(2C(ISV)) receptors . 5-HT agonists stimulated P28482 activity via all three 5-HT(2) subtypes . There were no meaningful differences in the potencies or relative efficacies of these agonists to affect P28482 activity vs. PI accumulation or Ca(2+) mobilization , suggesting that these pathways may be sequentially linked . Indeed , P28482 activity was very sensitive to PKC inhibition and calcium chelation and insensitive to tyrosine kinase and P19957 -kinase inhibition . 5-HT(2) receptors efficiently couple to MAPK activation via sequential PI hydrolysis , and Ca(2+) mobilization . This profile differs from reports of " agonist-directed trafficking of receptor stimulus " between PI/Ca(2+) and AA pathways activated by 5-HT(2) receptors . DB03758 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 kDa ( HSP90 ) . HSP90 is the main component of the cytosolic molecular chaperone complex , which has been implicated in the regulation of the heat shock factor 1 ( Q00613 ) . Q00613 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 HSP90 in cardiomyocytes . These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection . Therefore , compounds , such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents . P08195 -silencing impairs tumorigenicity of HeLa cells via modulation of galectin-3 and β-catenin signaling , and P08253 expression . P08195 is a type-II glycoprotein whose covalent-bound association with one of several described light chains yields a heterodimer mainly involved in large neutral amino acid transport . Likewise , it is well known that the heavy chain interacts with β-integrins mediating integrin-dependent events such as survival , proliferation , migration and even transformation . P08195 is a ubiquitous protein whose overexpression has been related to tumor development and progression . Stable silencing of P08195 in HeLa cells using an artificial miRNA impairs in vivo tumorigenicity and leads to an ineffective proliferation response to mitogens . P08195 colocalizes with β1-integrins and CD147 , but this interaction does not occur in lipid rafts in HeLa cells . Moreover , silenced cells present defects in integrin- ( Q05397 , Akt and P27361 /2 ) and hypoxia-dependent signaling , and reduced expression/activity of P08253 . These alterations seem to be dependent on the inappropriate formation of CD147/ P08195 /β1-integrin heterocomplexes on the cell surface , arising when CD147 can not interact with P08195 . Although extracellular galectin-3 accumulates due to the decrease in P08253 activity , galectin-3 signaling events are blocked due to an impaired interaction with P08195 , inducing an increased degradation of β-catenin . Furthermore , cell motility is compromised after protein silencing , suggesting that P08195 is related to tumor invasion by facilitating cell motility . Therefore , here we propose a molecular mechanism by which P08195 participates in tumor progression , favoring first steps of epithelial-mesenchymal transition by inhibition of β-catenin proteasomal degradation through Akt/GSK-3β signaling and enabling cell motility . Genome-wide association studies identify P30532 /3 and Q13639 in the development of airflow obstruction . RATIONALE : Genome-wide association studies ( GWAS ) have identified loci influencing lung function , but fewer genes influencing chronic obstructive pulmonary disease ( P48444 ) are known . OBJECTIVES : Perform meta-analyses of GWAS for airflow obstruction , a key pathophysiologic characteristic of P48444 assessed by spirometry , in population-based cohorts examining all participants , ever smokers , never smokers , asthma-free participants , and more severe cases . METHODS : Fifteen cohorts were studied for discovery ( 3,368 affected ; 29,507 unaffected ) , and a population-based family study and a meta-analysis of case-control studies were used for replication and regional follow-up ( 3,837 cases ; 4,479 control subjects ) . Airflow obstruction was defined as Q99581 (1) and its ratio to FVC ( Q99581 (1)/FVC ) both less than their respective lower limits of normal as determined by published reference equations . MEASUREMENTS AND MAIN RESULTS : The discovery meta-analyses identified one region on chromosome 15q25.1 meeting genome-wide significance in ever smokers that includes A2RU49 , P48200 , and P30532 / P32297 genes . The region was also modestly associated among never smokers . Gene expression studies confirmed the presence of P30532 /3 in lung , airway smooth muscle , and bronchial epithelial cells . A single-nucleotide polymorphism in Q13639 , a gene previously related to Q99581 (1)/FVC , achieved genome-wide statistical significance in combined meta-analysis . Top single-nucleotide polymorphisms in Q9H013 , P10826 , O14495 , and Q8TE59 were nominally replicated in the P48444 meta-analysis . CONCLUSIONS : These results suggest an important role for the P30532 /3 region as a genetic risk factor for airflow obstruction that may be independent of smoking and implicate the Q13639 gene in the etiology of airflow obstruction . Production of an antiserum specific to the ADP-ribosylated form of elongation factor 2 from archaebacteria and eukaryotes . An antiserum to ADP-ribosylated elongation factor 2 ( DB02059 - P13639 ) from S. acidocaldarius was raised in rabbits using stained , homogenized , DB02059 - P13639 -containing slices from SDS-gels as a source of antigen . P13639 ( P13639 ) from S. acidocaldarius was cloned in E. coli and the expressed gene product was used in order to adsorb all anti- P13639 antibodies which do not contain the ADP-ribosyl group within their epitopes , as E. coli is unable to synthesize the ADP-ribosyl acceptor diphthamide . The remaining antibodies were specific to ADP-ribosylated P13639 from Thermoplasma acidophilum , S. acidocaldarius and Desulfurococcus mucosus . ADP-ribosylated P13639 from eukaryotic sources also reacted with the adsorbed antiserum as shown for P13639 isolated from the killi-fish Cynolebias whitei , the mouse species BALB/c and Han/Wistar rats . The adsorbed antiserum did not cross-react with ADP-ribosylated actin or rho protein or with DB03147 -containing D-amino acid oxidase . Effect of hydrocortisone on the pituitary response to growth hormone releasing hormone . RATIONALE : In depression , the growth hormone ( GH ) response to clonidine and L-tryptophan ( L-TRP ) is reduced , suggesting reduced alpha2-adrenergic and serotonin (5-HT)1A receptor function . Pretreatment with hydrocortisone ( 100 mg , orally 11 h before ) also blunts the GH response to L-TRP . This effect may be mediated at the hypothalamic level via reduced P08908 receptor function or at the pituitary level , either by a direct effect on somatotrope cells or via enhanced insulin-like growth factor-1 ( DB01277 ) or somatostatin ( SS ) release . OBJECTIVES : To examine the effects of acute and chronic exposure to hydrocortisone on baseline and stimulated GH release from the pituitary . METHODS : Twelve healthy male volunteers received pretreatment with acute hydrocortisone ( 100 mg , 11 h before ) , chronic hydrocortisone ( 20 mg twice a day for 1 week ) and placebo in a double blind , balanced order , crossover design . Serial measurements of plasma GH , DB01277 and thyroid stimulating hormone ( DB00024 ) levels were made at baseline and following intravenous administration of 1 mcg/kg P01286 . RESULTS : The GH response to growth hormone releasing hormone ( P01286 ) was significantly blunted by pretreatment with both acute and chronic hydrocortisone . Baseline DB01277 levels were significantly lower at baseline after chronic hydrocortisone compared with placebo . Baseline DB00024 levels were significantly lower after acute hydrocortisone compared with placebo , suggesting an increase in somatostatin levels . CONCLUSIONS : These data suggest that hydrocortisone acts at the pituitary level to reduce GH release . The DB00024 and DB01277 data support the hypothesis that hydrocortisone reduces GH release by enhancing somatostatin and DB01277 release . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Enhancement of L-cystine transport activity and its relation to Q9UPY5 gene induction at the blood-brain barrier by diethyl maleate treatment . The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier ( BBB ) . The uptake of [(14)C]L-cystine and [(3)H]L-glutamic acid ( L- DB00142 ) was determined using a mouse brain endothelial cell line ( MBEC4 ) as an in vitro BBB model . The mRNA levels of L-cystine/L- DB00142 exchanger , system x(c)(-) , which consists of Q9UPY5 and P08195 , were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis . The [(14)C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na(+)-independent saturable process . The corresponding Michaelis-Menten constant ( K(m) ) was 63.7 microM . In the presence of L- DB00142 , there was competitive inhibition with an inhibition constant ( K(i) ) of 83.5 microM . [(3)H]L- DB00142 uptake in the absence of Na(+) was saturable with a K(m) of 48.1 microM , and it exhibited competitive inhibition with a K(i) of 24.9 microM in the presence of L-cystine . The mutual inhibition between L-cystine and L- DB00142 and the type of inhibition suggest that system x(c)(-) operates in MBEC4 cells . The Q9UPY5 and P08195 mRNAs were expressed in MBEC4 cells and , following diethyl maleate ( DEM ) treatment , the Q9UPY5 mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration ( up to 500 microM ) . Concomitantly , the glutathione concentration in MBEC4 cells was increased . In conclusion , system x(c)(-)-mediated L-cystine uptake takes place in MBEC4 cells . DB00138 transport via system x(c)(-) at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the Q9UPY5 gene . Serotonin skews human macrophage polarization through P41595 and P34969 . Besides its role as a neurotransmitter , serotonin ( 5-hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT(1-7) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 production , upregulated the expression of M2 polarization-associated genes ( P05120 , P07996 , Q9NY15 , Q86Y22 ) , and reduced the expression of M1-associated genes ( P08476 , P41597 , P39900 , P05121 , P29016 , O94788 ) . Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2( P09603 ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7) , whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies . Endogenous serotonin excites striatal cholinergic interneurons via the activation of 5-HT 2C , P50406 , and P34969 serotonin receptors : implications for extrapyramidal side effects of serotonin reuptake inhibitors . The striatum is richly innervated by serotonergic afferents from the raphe nucleus . We explored the effects of this input on striatal cholinergic interneurons from rat brain slices , by means of both conventional intracellular and whole-cell patch-clamp recordings . Bath-applied serotonin ( 5-HT , 3-300 microM ) , induced a dose-dependent membrane depolarization and increased the rate of spiking . This effect was mimicked by the 5-HT reuptake blockers citalopram and fluvoxamine . In voltage-clamped neurons , 5-HT induced an inward current , whose reversal potential was close to the K(+) equilibrium potential . Accordingly , the involvement of K(+) channels was confirmed either by increasing extracellular K(+) concentration and by blockade of K(+) channels with barium . Single-cell reverse transcriptase-polymerase chain reaction ( RT-PCR ) profiling demonstrated the presence of P28335 , P50406 , and P34969 receptor mRNAs in identified cholinergic interneurons . The depolarization/inward current induced by 5-HT was partially mimicked by the 5-HT2 receptor agonist 2,5-dimethoxy-4-iodoamphetamine and antagonized by both ketanserin and the selective P28335 antagonist RS102221 , whereas the selective 5- Q9H205 and Q13639 receptor antagonists tropisetron and RS23597-190 had no effect . The depolarizing response to 5-HT was also reduced by the selective P50406 and P34969 receptor antagonists SB258585 and SB269970 , respectively , and mimicked by the P34969 agonist , 5-CT . Accordingly , activation of either P50406 or P34969 receptor induced an inward current . The 5-HT response was attenuated by U73122 , blocker of phospholipase C , and by SQ22,536 , an inhibitor of adenylyl cyclase . These results suggest that 5-HT released by serotonergic fibers originating in the raphe nuclei has a potent excitatory effect on striatal cholinergic interneurons . DB11582 suppresses osteoclastogenesis induced by O14788 and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 -induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 suppressed osteoclastogenesis induced by O14788 , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 -induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . The role of EF-hand domains and P06681 domain in regulation of enzymatic activity of phospholipase Czeta . Sperm-specific phospholipase C-zeta ( PLCzeta ) induces Ca2+ oscillations and egg activation when injected into mouse eggs . PLCzeta has such a high Ca2+ sensitivity of P98160 activity that the enzyme can be active in resting cells at approximately 100 nM Ca2+ , suitable for a putative sperm factor to be introduced into the egg at fertilization ( Kouchi , Z. , Fukami , K. , Shikano , T. , Oda , S. , Nakamura , Y. , Takenawa , T. , and Miyazaki , S. ( 2004 ) J. Biol. Chem. 279 , 10408-10412 ) . In the present structure-function analysis , deletion of EF1 and P13639 of the N-terminal four EF-hand domains caused marked reduction of phosphatidylinositol 4,5-bisphosphate ( PI(4,5)P2 ) -hydrolyzing activity in vitro and loss of Ca2+ oscillation-inducing activity in mouse eggs after injection of RNA encoding the mutant . However , deletion of EF1 and P13639 or mutation of EF1 or P13639 at the x and z positions of the putative Ca2+-binding loop little affected the Ca2+ sensitivity of the P98160 activity , whereas deletion of EF1 to EF3 caused 12-fold elevation of the EC50 of Ca2+ concentration . Thus , EF1 and P13639 are important for the PLCzeta activity , and EF3 is responsible for its high Ca2+ sensitivity . Deletion of four EF-hand domains or the C-terminal P06681 domain caused complete loss of P98160 activity , indicating that both regions are prerequisites for PLCzeta activity . Screening of interactions between the P06681 domain and phosphoinositides revealed that P06681 has substantial affinity to PI(3)P and , to the lesser extent , to PI(5)P but not to PI(4,5)P2 or acidic phospholipids . PI(3)P and PI(5)P reduced PLCzeta activity in vitro , suggesting that the interaction could play a role for negative regulation of PLCzeta . Serotonin and fluoxetine modulate bone cell function in vitro . Recent studies have proposed a role for serotonin and its transporter in regulation of bone cell function . In the present study , we examined the in vitro effects of serotonin and the serotonin transporter inhibitor fluoxetine " DB00472 " on osteoblasts and osteoclasts . Human mononuclear cells were differentiated into osteoclasts in the presence of serotonin or fluoxetine . Both compounds affected the total number of differentiated osteoclasts as well as bone resorption in a bell-shaped manner . RT-PCR on the human osteoclasts demonstrated several serotonin receptors , the serotonin transporter , and the rate-limiting enzyme in serotonin synthesis , tryptophan hydroxylase 1 ( Tph1 ) . Tph1 expression was also found in murine osteoblasts and osteoclasts , indicating an ability to produce serotonin . In murine pre-osteoclasts ( RAW264.7 ) , serotonin as well as fluoxetine affected proliferation and NFkappaB activity in a biphasic manner . Proliferation of human DB05914 ( O60682 ) and primary osteoblasts ( NHO ) , and 5- Q13049 receptor expression was enhanced by serotonin . DB00472 stimulated proliferation of O60682 and murine preosteoblasts ( MC3T3-E1 ) in nM concentrations , microM concentrations were inhibitory . The effect of fluoxetine seemed direct , probably through 5-HT2 receptors . Serotonin-induced proliferation of MC3T3-E1 cells was inhibited by the PKC inhibitor ( GF109203 ) and was also markedly reduced when antagonists of the serotonin receptors P41595 /C or 5- Q13049 /C were added . Serotonin increased osteoprotegerin ( O00300 ) and decreased receptor activator of NF-kappaB ligand ( O14788 ) secretion from osteoblasts , suggesting a role in osteoblast-induced inhibition of osteoclast differentiation , whereas fluoxetine had the opposite effect . This study further describes possible mechanisms by which serotonin and the serotonin transporter can affect bone cell function . Isolation , partial purification and characterization of nuclear retinoic acid receptors from chick skin . Nuclear receptors ( RARs ) for retinoic acid ( RA ) are considered to be the ultimate mediators of the action of RA in the control of cell differentiation and inhibition of tumorigenesis . We have isolated and partially purified and characterized RAR from a RA-responsive tissue , chick embryo skin . The purification steps included Affi-Gel blue chromatography , ultrafiltration , size exclusion chromatography , and preparative isoelectric focusing . The electrofocusing of RAR-[3H]RA complex in ampholines ( pH 3-10 ) revealed that the receptors have an isoelectric pH of 7.5 . Whereas pronase-digested the RAR-[3H]RA complex completely , DNase showed 20-35 % and RNase showed negligible digestive action on the complex . The ligand binding to RAR was completely inhibited by a mercury compound . P10276 - and P10826 -specific antibodies , on Western blot analysis , immunoreacted with a protein having a molecular weight of 50,000 , presumably RAR . Binding affinity studies revealed that biologically active analogs of RA with a free COOH group ( e.g. , 13- DB00982 , RO-13-7410 , Ch 55 , and DB04942 ) showed , like RA , high binding affinity for RAR , whereas biologically ineffective analogs of RA ( e.g. , furyl and pyridyl ) were poor binders . Other groups of retinoids , in which the COOH group was either lacking or blocked , did not bind to RAR whether or not they were biologically active .

MH_train_1216

MH_train_1216

MH_train_1216

interacts_with DB01024?

multiple_choice

P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . Activation of metabotropic glutamate receptor 3 enhances interleukin ( IL ) -1beta-stimulated release of P05231 in cultured human astrocytes . Previous studies have demonstrated that human astrocytes express mRNA and receptor protein for group I and II metabotropic glutamate receptors ( mGluRs ) . Whether these receptors can influence the inflammatory and immune response and can modulate the capacity of astrocytes to produce inflammatory cytokines is still unclear . Inflammatory cytokines can be produced by activated glial cells and play a critical role in several neurological disorders . Astrocyte-enriched human cell cultures growing in a serum-free chemically defined medium were used to study the regulation of IL (interleukin)-1beta and P05231 in response to mGluR activation . Astrocytes cultured in the absence or in the presence of epidermal growth factor ( P01133 ) , did not secrete significant IL-1beta and P05231 , as determined by specific enzyme-linked immunosorbent assay ( ELISA ) . Activation of mGluRs using ( S ) -3,5-dihydroxyphenylglycine ( DHPG ; selective group I agonist ) or DCG-IV ( selective group II agonist ) did not affect the production of interleukins under both growth conditions . On exposure to IL-1beta high levels of P05231 were detected . Activation of Q14832 with DCG-IV ( but not of P41594 with DHPG ) enhanced , in the presence of IL-1beta , the release of P05231 in a dose dependent manner in astrocytes cultured under conditions ( + P01133 ) in which the mGluR expression is known to be upregulated . The effect of Q14832 activation on IL-1beta stimulated release of P05231 was prevented by selective group II mGluR antagonists . The capacity of Q14832 to modulate the release of P05231 in the presence of IL-1beta supports the possible involvement of this receptor subtype in the regulation of the inflammatory and immune response under pathological conditions associated with glial cell activation . Possible role of cholecystokinin-A receptors in regulation of thyrotropin ( DB00024 ) secretion in male rats . We studied the importance of cholecystokinin ( CCK ) system in the regulation of thyrotropin ( DB00024 ) and prolactin ( PRL ) secretion in male rats . To this end , we tested the effects of both unselective CCK agonists CCK-8 and caerulein , and CCK-B selective agonists Q13308 and pentagastrin as well as the selective CCK antagonists ( devazepide and L-365,260 ) at wide dose-ranges on the cold-stimulated and TRH-induced DB00024 and PRL secretion . DB00403 , given s.c . 15 min before sacrifice , decreased DB00024 levels at 5 micrograms/kg . In time course-studies , the maximum inhibition was seen at 15 min but the effect lasted at least 30 , but less than 60 min . Also CCK-8 decreased DB00024 levels at the doses of 20 and 50 micrograms/kg at 15 min . Devazepide and L-365,260 did not affect DB00024 or PRL levels at any dose . The effect of caerulein ( 5 micrograms/kg ) was antagonized by devazepide , a CCK-A antagonist , at 100 micrograms/kg , but not by a CCK-B antagonist L-365,260 tested at a wide dose range . PRL levels were not affected by any treatment . DB00403 ( 5 micrograms/kg ) , given at the same time as TRH ( 500 ng/kg ) , inhibited the TRH-induced DB00024 levels at 15 min , but not at 30 or 60 min . CCK-8 ( 50 micrograms/kg ) , Q13308 ( 100 micrograms/kg ) and pentagastrin ( 500 micrograms/kg ) did not affect the TRH-induced DB00024 secretion . The results probably indicate that P32238 stimulation inhibits DB00024 secretion at the level of the anterior pituitary gland . PRL levels in male rats are not affected by CCK system . P22301 regulates progenitor differentiation and modulates neurogenesis in adult brain . The adult subventricular zone ( SVZ ) is the main neurogenic niche in the adult brain of mice and rats . The adult SVZ contains neural stem cells ( NSCs ) that primarily differentiate into committed neuroblasts . The newly generated neuroblasts accumulate in dorsal SVZ where they further differentiate and initiate a long migration pathway to their final destination , the olfactory bulb ( OB ) . Here , we report a new role for Interleukin 10 ( P22301 ) that is different to its well-known anti-inflammatory properties . We show that the P22301 receptor is expressed in P48681 -positive progenitors restricted to the dorsal SVZ in adult brain . Using P22301 gain models , we observed that P22301 maintains neural progenitors in an undifferentiated state by keeping progenitors in an active cycle where pro-neural gene markers ( P48681 , Sox1 , Sox2 , Musashi , Mash1 ) are upregulated and neuronal gene expression ( Numb , O43602 , Q13509 ) is downregulated . In addition , P22301 reduces neuronal differentiation and ultimately impairs endogenous neurogenesis . Consistently , in the absence of P22301 , in vivo neuronal differentiation of SVZ progenitors is enhanced and the incorporation of new neurons in the adult OB is increased . Thus , our results provide the first evidence that P22301 acts as a growth factor on SVZ progenitors and regulates neurogenesis in normal adult brain . Analog of somatostatin vapreotide exhibits biological effects in vitro via interaction with neurokinin-1 receptor . OBJECTIVES : DB04894 , a synthetic analog of somatostatin , has analgesic activity most likely mediated through the blockade of neurokinin-1 receptor ( P25103 ) , the DB05875 ( SP ) -preferring receptor . The ability of vapreotide to interfere with other biological effects of SP has yet to be investigated . METHODS : We studied the ability of vapreotide to antagonize P25103 in three different cell types : immortalized U373MG human astrocytoma cells , human monocyte-derived macrophages ( MDM ) and a human embryonic kidney cell line , HEK293 . Both U373MG and MDM express endogenous P25103 while HEK293 cells , which normally do not express P25103 , are stably transformed to express human P25103 ( HEK293- P25103 ) . RESULTS : DB04894 attenuates SP-triggered intracellular calcium increases and nuclear factor-κB activation in a dose-dependent manner . DB04894 also inhibits SP-induced interleukin-8 and monocyte chemotactic protein-1 production in HEK293- P25103 and U373MG cell lines . DB04894 inhibits HIV-1 infection of human MDM in vitro , an effect that is reversible by SP pretreatment . CONCLUSIONS : Our findings indicate that vapreotide has P25103 antagonist activity and may have a potential application as a therapeutic intervention in HIV-1 infection . Lycopene and other carotenoids inhibit estrogenic activity of 17beta-estradiol and genistein in cancer cells . Epidemiological evidence suggests that carotenoids prevent several types of cancer , including mammary and endometrial cancers . On the other hand , such studies have also shown that estrogens are the most important risk factors for these cancer types . DB01645 , the phytoestrogen mainly found in soy , also shows significant estrogenic activity when tested at concentrations found in human blood . The aim of this study was to determine whether carotenoids inhibit signaling of steroidal estrogen and phytoestrogen which could explain their cancer preventive activity . Similar to the known effect of 17beta-estradiol ( E(2) ) , treatment of breast ( T47D and MCF-7 ) and endometrial ( ECC-1 ) cancer cells with phytoestrogens induced cell proliferation , cell-cycle progression and transactivation of the estrogen response element ( ERE ) . However , each of the tested carotenoids ( lycopene , phytoene , phytofluene , and beta-carotene ) inhibited cancer cell proliferation induced by either E(2) or genistein . The inhibition of cell growth by lycopene was accompanied by slow down of cell-cycle progression from P55008 to S phase . Moreover , the carotenoids inhibited estrogen-induced transactivation of ERE that was mediated by both estrogen receptors ( ERs ) ERalpha and ERbeta . The possibility that this inhibition results from competition of carotenoid-activated transcription systems on a limited pool of shared coactivators with the ERE transcription system was tested . Although cotransfection of breast and endometrial cancer cells with four different coactivators ( Q15788 , P12931 -2 , Q9Y6Q9 , and DRIP ) strongly stimulated ERE reporter gene activity , it did not oppose the inhibitory effect of carotenoids . These results suggest that dietary carotenoids inhibit estrogen signaling of both 17beta-estradiol and genistein , and attenuate their deleterious effect in hormone-dependent malignancies . Identification of primitive human hematopoietic cells capable of repopulating NOD/SCID mouse bone marrow : implications for gene therapy . The development of stem-cell gene therapy is hindered by the absence of repopulation assays for primitive human hematopoietic cells . Current methods of gene transfer rely on in vitro colony-forming cell ( Q15814 ) and long-term culture-initiating cell ( LTC-IC ) assays , as well as inference from other mammalian species . We have identified a novel human hematopoietic cell , the SCID-repopulating cell ( P12931 ) , a cell more primitive than most LTC-ICs and CFCs . The P12931 , exclusively present in the P01730 +CD8- fraction , is capable of multilineage repopulation of the bone marrow of nonobese diabetic mice with severe combined immunodeficiency disease ( NOD/SCID mice ) . SRCs were rarely transduced with retroviruses , distinguishing them from most CFCs and LTC-ICs . This observation is consistent with the low level of gene marking seen in human gene therapy trials . An P12931 assay may aid in the characterization of hematopoiesis , as well as the improvement of transduction methods . Blockage of the neurokinin 1 receptor and capsaicin-induced ablation of the enteric afferent nerves protect SCID mice against T-cell-induced chronic colitis . BACKGROUND : The neurotransmitter DB05875 ( SP ) released by , and the transient receptor potential vanilloid ( Q8NER1 ) , expressed by afferent nerves , have been implicated in mucosal neuro-immune-regulation . To test if enteric afferent nerves are of importance for the development of chronic colitis , we examined antagonists for the high-affinity neurokinin 1 ( NK-1 ) SP receptor and the Q8NER1 receptor agonist capsaicin in a T-cell transfer model for chronic colitis . METHODS : Chronic colitis was induced in SCID mice by injection of P01730 (+)CD25(-) T cells . The importance of NK-1 signaling and Q8NER1 expressing afferent nerves for disease development was studied in recipient SCID mice systemically treated with either high-affinity P25103 antagonists or neurotoxic doses of capsaicin . In addition , we studied the colitis-inducing effect of P25103 deleted P01730 (+)CD25(-) T cells . RESULTS : Treatment with the P25103 antagonist P62158 4092 reduced the severity of colitis , but colitis was induced by P25103 -deleted T cells , suggesting that SP in colitis targets the recipient mouse cells and not the colitogenic donor T cells . DB06774 -induced depletion of nociceptive afferent nerves prior to P01730 (+)CD25(-) T-cell transfer completely inhibited the development of colitis . CONCLUSIONS : Our data demonstrate the importance of an intact enteric afferent nerve system and NK-1 signaling in mucosal inflammation and may suggest new treatment modalities for patients suffering from inflammatory bowel disease . CCK1-receptor stimulation protects against gut mediator-induced lung damage during endotoxemia . BACKGROUND/AIMS : Cholecystokinin 1-receptor ( P32238 ) activation by long chain fatty acid ( LCFA ) absorption stimulates vago-vagal reflex pathways in the brain stem . The present study determines whether this reflex also activates the cholinergic anti-inflammatory pathway , a pathway known to modulate cytokine release during endotoxemia . METHODS : Mesenteric lymph was obtained from wild type ( WT ) and P32238 knockout ( P32238 (-/-) ) mice intraperitoneally challenged with Lipopolysaccharid ( LPS ) ( endotoxemic lymph , EL ) and intestinally infused with vehicle or LCFA-enriched solution . The lymph was analyzed for TNFα , P05231 and P22301 concentration and administered to healthy recipient mice via jugular infusion . Alveolar wall thickness , myeloperoxidase ( P05164 ) and TUNEL positive cells were determined in lung tissue of recipient mice . RESULTS : LCFA infusion in WT mice reduced TNFα concentration in EL by 49 % compared to vehicle infusion , but had no effect in P32238 (-/-) mice . EL significantly increased the alveolar wall thickness , the number of P05164 -positive and TUNEL-positive cells compared to control lymph administration . LCFA infusion in WT , but not in CCK1R(-/-) mice , significantly reduced these pathological effects of EL . CONCLUSION : During endotoxemia enteral LCFA absorption reduces TNFα release into mesenteric lymph and attenuates histomorphologic parameters of lung dysfunction . Failure to elicit this effect in CCK1R(-/-) mice demonstrates that anti-inflammatory properties of LCFAs are mediated through CCK1-Rs . Further evidence for a functional role of the glutamate receptor gene Q14832 in schizophrenia . In recent years , evidence has been accumulating indicating a major role of glutamate in the pathogenesis and pathophysiology of schizophrenia . Of particular importance in this regard are the metabotropic glutamate receptors ( GRM ) . Thus , a recently published trial of the amino acid analogue DB05096 , which exerts its effects through the activation of the glutamate receptors Q14832 / Q14416 , showed an improvement of positive and negative symptoms comparable to treatment with olanzapine . A functional variant of Q14832 has been described which modulates synaptic glutamate levels . We assessed whether this functional variant rs6465084 is related to schizophrenia in a large sample of patients and controls . We found an increased frequency of the A allele ( p=0.027 ) and the AA genotype ( p=0.024 ) in schizophrenia patients . Moreover , in an assessment of schizophrenia endophenotypes , patients of the AA genotype performed poorly in the digit symbol test , a measure of attention ( p=0.008 ) . Our results provide further evidence for the potential importance of the glutamate receptor Q14832 in schizophrenia , and indicate that the novel antipsychotic DB05096 may actually be targeting a pathogenic pathway of schizophrenia . Polymorphism identification in the P11310 , P01008 , P22301 , P15173 and P01222 genes of cattle . The immunosuppressant FTY720 prolongs survival in a mouse model of diet-induced coronary atherosclerosis and myocardial infarction . FTY720 , an analogue of sphingosine-1-phosphate , is cardioprotective during acute injury . Whether long-term FTY720 affords cardioprotection is unknown . Here , we report the effects of oral FTY720 on ischemia/reperfusion injury and in hypomorphic apoE mice deficient in Q8WTV0 receptor expression ( ApoeR61(h/h)/ Q8WTV0 ( -/- mice ) , a model of diet-induced coronary atherosclerosis and heart failure . We added FTY720 ( 0.3 mg·kg(-1)·d(-1) ) to the drinking water of C57BL/6J mice . After ex vivo cardiac ischemia/reperfusion injury , these mice had significantly improved left ventricular ( LV ) developed pressure and reduced infarct size compared with controls . Subsequently , ApoeR61(h/h)/ Q8WTV0 (-/-) mice fed a high-fat diet for 4 weeks were treated or not with oral FTY720 ( 0.05 mg·kg(-1)·d(-1) ) . This sharply reduced mortality ( P < 0.02 ) and resulted in better LV function and less LV remodeling compared with controls without reducing hypercholesterolemia and atherosclerosis . Oral FTY720 reduced the number of blood lymphocytes and increased the percentage of P01730 +Foxp3+ regulatory T cells ( Tregs ) in the circulation , spleen , and lymph nodes . FTY720-treated mice exhibited increased TGF-β and reduced IFN-γ expression in the heart . Also , P01730 expression was increased and strongly correlated with molecules involved in natural Treg activity , such as TGF-β and Q9Y5U5 . Our data suggest that long-term FTY720 treatment enhances LV function and increases longevity in mice with heart failure . These benefits resulted not from atheroprotection but from systemic immunosuppression and a moderate reduction of inflammation in the heart . Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 ( Q16739 ) is an approved drug target for the treatment of Gaucher disease and is considered as a valid target for combating other human pathologies , including type 2 diabetes . The clinical drug N-butyldeoxynojirimycin ( DB00419 ) is thought to inhibit through mimicry of its substrate , ceramide . In this work we demonstrate that , in contrast to what is proposed in this model , the P06681 -hydroxyl of the deoxynojirimycin core is important for Q16739 inhibition . Here we show that P13671 -OH appears of less important , which may set guidelines for the development of Q16739 inhibitors that have less affinity ( in comparison with DB00419 ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . DB00144 binding of class B scavenger receptor type I , a phagocytosis receptor of testicular sertoli cells . Testicular Sertoli cells phagocytose apoptotic spermatogenic cells in a manner depending on the membrane phospholipid phosphatidylserine ( PS ) expressed at the surface of the latter cell type . Our previous studies have indicated that class B scavenger receptor type I ( Q8WTV0 ) is responsible for the PS-mediated phagocytosis by Sertoli cells . We examined here whether Q8WTV0 binds directly to PS . A cell line acquired the ability to bind to PS-exposing apoptotic cells and to incorporate PS-containing liposomes when it was forced to express Q8WTV0 . Furthermore , the extracellular domain of rat Q8WTV0 fused with human Fc ( SRBIecd-Fc ) bound to PS with a dissociation equilibrium constant of 2.4 x 10(-7) m in a cell-free solid-phase assay , whereas other phospholipids including phosphatidylethanolamine , phosphatidylinositol , and phosphatidylcholine were poor binding targets . The binding activity was enhanced when CaCl(2) was included in the assay or when SRBIecd-Fc was pre-treated with N-glycanase . A portion of the extracellular domain spanning amino acid positions 33 and 191 ( numbered with respect to the amino terminus ) fused with Fc ( SRBI33-191-Fc ) showed activity and phospholipid specificity equivalent to those of SRBIecd-Fc . Finally , SRBI33-191-Fc bound to the surface of apoptotic cells with externalized PS , and the injection of SRBI33-191-Fc into the seminiferous tubules of live mice increased the number of apoptotic spermatogenic cells . These results allowed us to conclude that Q8WTV0 is a phagocytosis-inducing PS receptor of Sertoli cells . HIV entry inhibitors : mechanisms of action and resistance pathways . Entry inhibitors represent a new generation of antivirals for the treatment of HIV infection . Several compounds which block the attachment of HIV gp120 to either the P01730 T cell receptor or the P51681 / P61073 co-receptors are currently in clinical development . Most of these compounds have different molecular structures and specific mechanisms of action . These agents are eagerly awaited by a growing number of patients carrying viruses resistant viruses to many of the current available reverse transcriptase and protease inhibitors . For enfuvirtide , the first and , so far , only entry inhibitor approved for clinical use , the main mechanism of resistance is the selection of changes within a 10 amino acid segment encompassing residues 36-45 within the Q96GN5 region of gp41 . For other entry inhibitors , multiple changes in different gp120 domains ( V1 , V2 , V3 , P06681 and C4 ) have been associated with loss of susceptibility to these agents , although in most cases with limited cross-resistance . Attenuation of anti-tuberculosis therapy induced hepatotoxicity by Spirulina fusiformis , a candidate food supplement . Therapy using Isoniazid ( DB00951 ) and DB01045 ( Q9HBH0 ) leads to induction of hepatotoxicity in some individuals undergoing anti-tuberculosis treatment . In this study , we assessed the effect of Spirulina fusiformis on DB00951 and Q9HBH0 induced hepatotoxicity in rats compared with hepatoprotective drug Silymarin . Induction of hepatotoxicity was measured by changes in the liver marker enzymes ( aspartate transaminase , alanine transaminase , and alkaline phosphatase ) . The antioxidant status was also analyzed in liver tissue homogenate and plasma by measurement of superoxide dismutase , catalase , glutathione-S-transferase , glutathione reductase , and lipid peroxidation levels . We also aimed to study the binding and interactions of the transcription factors Pregnane X Receptor ( O75469 ) and Farnesoid X Receptor ( Q96RI1 ) with DB00951 , Q9HBH0 , and representative active compounds of Spirulina fusiformis by in silico methods . The administration of DB00951 and Q9HBH0 resulted in significant ( p < 0.05 ) decrease in the antioxidant levels and total protein levels . There was also a significant ( p < 0.05 ) increase in the levels of liver marker enzymes . Spirulina fusiformis was seen to protect the parameters from significant changes upon challenge with DB00951 and Q9HBH0 in a dose-dependent manner . This was corroborated by histological examination of the liver . The results of the in silico analyses further support the wet lab results . DB00640 A2A receptor : a target for regulating renal interstitial fibrosis in obstructive nephropathy . Renal interstitial fibrosis ( Q9HBH0 ) is the common pathological process of chronic kidney diseases leading inevitably to renal function deterioration . Q9HBH0 and its preceding epithelial-mesenchymal transition ( EMT ) are commonly triggered by an early occurring renal inflammation . However , an effective approach to prevent EMT and Q9HBH0 is still lacking and of urgent need . Recently , the adenosine A2A receptor ( A2AR ) emerges as a novel inflammation regulator , therefore manipulation of A2AR may suppress the EMT process and as such protect against Q9HBH0 . To test this hypothesis we applied a unilateral ureteral obstruction ( UUO ) model of Q9HBH0 on A2AR knockout mice and their wild-type littermates , combined with the intervention of a selective A2AR agonist , CGS 21680 . On days 3 , 7 and 14 post-UUO we evaluated the effects of A2AR manipulation on the molecular pathological progresses of Q9HBH0 , including the cellular component of interstitial infiltration , expression of profibrotic factors , cellular biomarkers of EMT , and collagen deposition of extracellular matrix . Our data demonstrated that activation of A2AR significantly suppressed the deposition of collagen types I and III , reduced the infiltration of P01730 + T lymphocytes , and attenuated the expression of TGF-β1 and Q13464 , which in turn inhibited and postponed the EMT progress . Conversely , genetic inactivation of A2AR exacerbated the aforementioned pathological processes of UUO-induced Q9HBH0 . Together , activation of A2AR effectively alleviated EMT and Q9HBH0 in mice , suggesting A2AR as a potential therapeutic target for the treatment of Q9HBH0 . P22301 -inducing adjuvants enhance sublingual immunotherapy efficacy in a murine asthma model . BACKGROUND : P22301 -inducing adjuvants could enhance the efficacy of allergy vaccines in establishing allergen-specific tolerance . The aim of this study was to identify such adjuvants using in vitro cultures of human and murine cells and to evaluate them in a therapeutic murine model of sublingual immunotherapy ( SLIT ) . METHODS : Adjuvants stimulating P22301 gene expression by human or murine immune cells were tested sublingually in BALB/c mice sensitized to ovalbumin ( OVA ) , assessing the reduction in airway hyperresponsiveness ( P35869 ) by whole-body plethysmography . The induction of regulatory T cells ( T(reg) ) was evaluated using phenotypic and functional assays . T-cell proliferation in cervical lymph nodes ( LNs ) was assessed following intravenous transfer of CFSE-labelled OVA-specific T cells and FACS analysis . RESULTS : A combination of 1,25-dihydroxyvitamin D3 plus dexamethasone ( VitD3/ DB00514 ) as well as Lactobacillus plantarum were found to induce P22301 production by human and murine dendritic cells ( DCs ) . The former inhibits LPS-induced DC maturation , whereas L. plantarum induces DC maturation . Following stimulation with VitD3/ DB00514 -pretreated DCs , P01730 + naïve T cells exhibit a T(reg) profile . In contrast , a Th1/T(reg) pattern of differentiation is observed in the presence of DCs treated with L. plantarum . Both adjuvants significantly enhance SLIT efficacy in mice , in association with either induction of Foxp3+ T(reg) cells ( for VitD3/ DB00514 ) or proliferation of OVA-specific T cells in cervical LNs ( for L. plantarum ) . CONCLUSIONS : Both VitD3/ DB00514 and L. plantarum polarize naïve T cells towards P22301 -expressing T cells , through distinct mechanisms . As adjuvants , they both enhance SLIT efficacy in a murine asthma model .

MH_train_1217

MH_train_1217

MH_train_1217

interacts_with DB00313?

multiple_choice

Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Increase in intracellular Cl- concentration by DB02527 - and Ca2+-dependent stimulation of M1 collecting duct cells . In the lungs of cystic fibrosis ( CF ) patients , mutations of the cystic fibrosis transmembrane conductance regulator ( P13569 ) lead to defective Cl- secretion and hyperabsorption of electrolytes . This may be a an important cause for the defective mucociliary clearance in CF lungs . Previous studies have suggested that inhibition of ENaC during activation of P13569 or by purinergic stimulation could be related to an increase in the intracellular [Cl-]i . This was examined in the present study using cultured mouse M1 collecting duct cells transfected with the chloride-sensitive enhanced yellow fluorescent protein YFP(V163S) . Calibration experiments showed a linear decrease of YFP fluorescence intensity with increasing [Cl-]i ( 0-100 mM ) . Activation of P13569 by isobutyl-1-methylxanthine ( DB07954 , 100 microM ) and forskolin ( 2 microM ) increased [Cl-]i by 9.6+/-1.5 mM ( n=35 ) . Similarly , DB00171 ( 100 microM ) increased [Cl-]i transiently by 9.5+/-2.2 mM ( n=17 ) . The increase in [Cl-]i was reduced by the Na+/K+/2 Cl- -cortransporter-1 ( P55011 ) blocker azosemide ( 100 microM ) , the P13569 blocker DB04941 ( 50 microM ) , the blocker of Ca2+-activated Cl- channels DIDS ( 100 microM ) or the ENaC blocker amiloride ( 10 microM ) . Changes in YFP(V163S) fluorescence were not due to changes in cell volume or intracellular pH . The present data thus demonstrate an increase in [Cl-]i following stimulation with secretagogues , which could participate in the inhibition of ENaC . Identification of N-(4-piperidinyl)-4-(2,6-dichlorobenzoylamino)-1H-pyrazole-3-carboxamide ( DB05037 ) , a novel cyclin dependent kinase inhibitor using fragment-based X-ray crystallography and structure based drug design . The application of fragment-based screening techniques to cyclin dependent kinase 2 ( P24941 ) identified multiple ( > 30 ) efficient , synthetically tractable small molecule hits for further optimization . Structure-based design approaches led to the identification of multiple lead series , which retained the key interactions of the initial binding fragments and additionally explored other areas of the DB00171 binding site . The majority of this paper details the structure-guided optimization of indazole ( 6 ) using information gained from multiple ligand- P24941 cocrystal structures . Identification of key binding features for this class of compounds resulted in a series of molecules with low nM affinity for P24941 . Optimisation of cellular activity and characterization of pharmacokinetic properties led to the identification of 33 ( DB05037 ) , which is currently being evaluated in clinical trials for the treatment of human cancers . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Loss of P13569 results in reduction of histone deacetylase 2 in airway epithelial cells . Inflammatory cytokines , particularly the neutrophil chemoattractant P10145 , are elevated in the cystic fibrosis ( CF ) airway , even in the absence of detectable infection . The transcriptional regulation of many inflammatory genes , including P10145 ( P10145 ) , involves chromatin remodeling through histone acetylation . NF-kappaB is known to facilitate histone acetylation of P10145 and other proinflammatory gene promoters , but we find that increased NF-kappaB activation can not explain the elevated P10145 expression and promoter acetylation seen in P13569 -deficient cells . Recognized components of the NF-kappaB-coactivator complex , acetyltransferase CBP , p300 , and the histone deacetylase Q13547 , are unchanged by P13569 activity . However , we find that the histone acetyltransferase ( O60235 ) /HDAC balance is sensitive to P13569 function , as cells with reduced or absent P13569 function have decreased Q92769 protein , resulting in hyperacetylation of the P10145 promoter and increased P10145 transcription . Reduced Q92769 and Q92769 activity , but not Q92769 mRNA , is observed in cells deficient in P13569 . Suppressing Q92769 expression with Q92769 short hairpin RNA ( shRNA ) results in increased P10145 expression and promoter acetylation comparable with P13569 -deficient cells . Treating P13569 -deficient cells with N-acetyl-cysteine ( Q9C000 ) increases Q92769 expression to near control levels . Our data suggest that there is an intrinsic alteration in the O60235 /HDAC balance in cells lacking P13569 function in vitro and in native CF tissue and that oxidative stress is likely contributing to this alteration . This mechanism , found in other inflammatory airway diseases , provides an explanation for the apparent dysregulation of inflammatory mediators seen in the CF airway , as reduced histone deacetylation would potentially influence many genes . Candidate pathway polymorphisms in one-carbon metabolism and risk of rectal tumor mutations . We examined candidate polymorphisms in genes involved in the folate-mediated , one-carbon metabolism pathway , P26358 1311V , P11586 R134K and R653Q , P42898 R594Q , Q99707 D919G , Q9UBK8 H595Y and I22M , P34896 L474F , P41440 H27R , and Q13569 G199S , and associations with rectal tumor characteristics . We hypothesized that these candidate genes would influence CpG Island Methylator Phenotype and potentially P01116 or P04637 tumors . Data from a population-based study of 747 rectal cases ( 593 with tumor markers ) and 956 controls were evaluated using generalized estimating equations . We observed an increased risk of P04637 tumor mutations in homozygous carriers of the P11586 134K allele ( 0R=2.0 , 95 % CI 1.2-3.1 , P- trend=0.02 ) . In the presence of low folate intake , the R134K variant was associated with increased risk of CIMP+ tumors ( OR=2.8 , 95 % CI 1.04-7.7 ) . The Q9UBK8 I22M variant genotype was associated with a modest increased risk of P04637 mutations ( OR=1.7 , 95 % CI 1.2-2.5 , P-trend=0.001 ) . Our findings offer limited support that polymorphisms in one-carbon metabolism genes influence rectal tumor phenotype , and that folate may interact with P11586 to alter CIMP+ risk . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . Class I HDACs are mediators of smoke carcinogen-induced stabilization of P26358 and serve as promising targets for chemoprevention of lung cancer . DNA methylation is an early event in bronchial carcinogenesis and increased DNA methyltransferase ( P26358 )1 protein expression is a crucial step in the oncogenic transformation of epithelia . Here , we investigate the role of class I histone deacetylases ( HDAC ) 1 to 3 in the stabilization of P26358 protein and as a potential therapeutic target for lung cancer chemoprevention . Long-term exposure of immortalized bronchial epithelial cells ( HBEC-3KT ) to low doses of tobacco-related carcinogens led to oncogenic transformation , increased HDAC expression , cell-cycle independent increased P26358 stability , and DNA hypermethylation . Overexpression of HDACs was associated with increased P26358 stability and knockdown of HDACs reduced P26358 protein levels and induced P26358 acetylation . This suggests a causal relationship among increased class I HDACs levels , upregulation of P26358 protein , and subsequent promoter hypermethylation . Targeting of class I HDACs with valproic acid ( DB00313 ) was associated with reduced HDAC expression and a profound reduction of P26358 protein level . Treatment of transformed bronchial epithelial cells with DB00313 resulted in reduced colony formation , demethylation of the aberrantly methylated Q96HF1 promoter , and derepression of Q96HF1 transcription . These data suggest that inhibition of HDAC activity may reverse or prevent carcinogen-induced transformation . Finally , immunohistochemistry on human lung cancer specimens revealed a significant increase in P26358 , Q13547 , Q92769 , and O15379 expression , supporting our hypotheses that class I HDACs are mediators of P26358 stability . In summary , our study provides evidence for an important role of class I HDACs in controlling the stability of P26358 and suggests that HDAC inhibition could be an attractive approach for lung cancer chemoprevention . O75469 induces Q02318 and regulates cholesterol metabolism in the intestine . Mitochondrial sterol 27-hydroxylase ( Q02318 ) catalyzes oxidative cleavage of the sterol side chain in the bile acid biosynthetic pathway in the liver and 27-hydroxylation of cholesterol in most tissues . Recent studies suggest that 27-hydroxycholesterol ( 27-HOC ) activates liver orphan receptor alpha ( LXRalpha ) and induces the cholesterol efflux transporters O95477 and P45844 in macrophages . The steroid- and bile acid-activated pregnane X receptor ( O75469 ) plays critical roles in the detoxification of bile acids , cholesterol metabolites , and xenobiotics . The role of Q02318 in the intestine is not known . This study investigated O75469 and Q02318 regulation of cholesterol metabolism in the human intestinal cell lines Caco2 and Ls174T . A human O75469 ligand , rifampicin , induced Q02318 mRNA expression in intestine cells but not in liver cells . DB01045 induced Q02318 gene transcription , increased intracellular 27-HOC levels , and induced O95477 and P45844 mRNA expression only in intestine cells . A functional O75469 binding site was identified in the human Q02318 gene . Chromatin immunoprecipitation assays revealed that rifampicin induced the O75469 recruitment of steroid receptor coactivator 1 to Q02318 chromatin . DB04540 loading markedly increased intracellular 27-HOC levels in intestine cells . DB01045 , 27-HOC , and a potent LXRalpha agonist , T0901317 , induced O95477 and P45844 protein expression and stimulated cholesterol efflux from intestine cells to apolipoprotein A-I and HDL . This study suggests an intestine-specific O75469 / Q02318 /LXRalpha pathway that regulates intestine cholesterol efflux and HDL assembly . Oxidative stress and cigarette smoke alter chromatin remodeling but differentially regulate NF-kappaB activation and proinflammatory cytokine release in alveolar epithelial cells . Oxidative stress is implicated in lung inflammation due to its effect on proinflammatory gene transcription . Changes in gene transcription depend on chromatin remodeling and the relative activities of histone acetyltransferases ( HATs ) and histone deacetylases ( HDACs ) . Alterations in the nuclear histone acetylation:deacetylation balance may result in uncontrolled transcription of specific proinflammatory genes . We studied the effect of hydrogen peroxide ( H2O2 ) and cigarette smoke condensate ( CSC ) on histone acetylation:deacetylation in human alveolar epithelial cells ( A549 ) . H2O2 and CSC significantly increased acetylation of histone H4 proteins and were associated with decreased HDAC activity and Q92769 levels in A549 cells . Also , the decreased Q92769 activity was due to protein modification by aldehydes and nitric oxide products . Pretreatment of A549 cells with N-acetyl-l-cysteine attenuated the oxidant-mediated reduction in HDAC activity . Treatment of A549 cells with CSC did not cause nuclear factor-kappaB ( NF-kappaB ) activation or expression and release of either interleukin ( IL ) -8 or P05231 . However , H2O2 , tumor necrosis factor-alpha ( P01375 ) , and IL-1beta significantly increased NF-kappaB activation and expression of P10145 compared with control cells . Interestingly , CSC dose dependently inhibited P01375 - and IL-1beta-mediated NF-kappaB activation and P10145 expression . Thus , H2O2 and CSC enhance acetylation of histone proteins and decrease histone deacetylase activity but differentially regulate proinflammatory cytokine release in alveolar epithelial cells . P04406 mediates nitrosylation of nuclear proteins . S-nitrosylation of proteins by nitric oxide is a major mode of signalling in cells . S-nitrosylation can mediate the regulation of a range of proteins , including prominent nuclear proteins , such as Q92769 ( ref. 2 ) and P09874 ( ref. 3 ) . The high reactivity of the nitric oxide group with protein thiols , but the selective nature of nitrosylation within the cell , implies the existence of targeting mechanisms . Specificity of nitric oxide signalling is often achieved by the binding of nitric oxide synthase ( NOS ) to target proteins , either directly or through scaffolding proteins such as P78352 ( ref. 5 ) and O75052 . As the three principal isoforms of NOS -- neuronal NOS ( P29475 ) , endothelial NOS ( P29474 ) and inducible NOS ( P35228 ) -- are primarily non-nuclear , the mechanisms by which nuclear proteins are selectively nitrosylated have been elusive . P04406 ( P04406 ) is physiologically nitrosylated at its DB00151 150 residue . Nitrosylated P04406 ( SNO- P04406 ) binds to Siah1 , which possesses a nuclear localization signal , and is transported to the nucleus . Here , we show that SNO- P04406 physiologically transnitrosylates nuclear proteins , including the deacetylating enzyme sirtuin-1 ( Q96EB6 ) , histone deacetylase-2 ( Q92769 ) and DNA-activated protein kinase ( DNA-PK ) . Our findings reveal a novel mechanism for targeted nitrosylation of nuclear proteins and suggest that protein-protein transfer of nitric oxide groups may be a general mechanism in cellular signal transduction . Development of DNA methyltransferase inhibitors for the treatment of neoplastic diseases . Although chemotherapy is considered the mainstay of cancer therapy , unfortunate side effects of chemotherapy create a continuous demand for developing other novel and specific targets for cancer therapy . Re-expression of epigenetically silenced tumor suppressor genes is a rational strategy for the treatment of human neoplasms . Epigenetic modifiers like DNA methyltransferase ( P26358 ) inhibitors and histone deacteylase ( HDAC ) inhibitors induce the re-expression of epigenetically silenced genes in vitro and in vivo . Moreover , they demonstrate safety and efficacy against neoplastic diseases in clinical trials . P26358 inhibitors like 5-azacytidine and DB01262 are currently FDA approved for the treatment of myelodysplastic syndrome . Nonetheless , the mechanism of action behind their clinical efficacy remains unclear . Ongoing clinical trials are attempting to identify tumor suppressor genes that upon re-expression can induce remission and cure in patients . On the other hand , the pleiotropic biological effects of P26358 inhibitors and recent reports demonstrating lack of association between clinical response and methylation reversal of candidate tumor suppressor genes , suggest a complex mechanism behind their clinical efficacy that may involve a cytotoxic effect . [ Effect of down-regulation of histone deacetylase 2 protein expression on cell proliferation and cell cycle in cervical carcinoma ] . OBJECTIVE : To study the effect of down-regulation of histone deacetylase 2 ( Q92769 ) expression on cell proliferation and cell cycle in cervical carcinoma cell lines HeLa . METHODS : Q92769 siRNA and control siRNA were transfected to HeLa cells . CCK-8 and flow cytometry were used to analyze the changes of cell proliferation and cell cycle , respectively . Western blot was employed to detect the changes of cell proliferation and cell cycle-related proteins . RESULTS : Q92769 siRNA significantly down-regulated the expression of Q92769 protein in HeLa cells , resulting in marked inhibition of cell proliferation . In addition , the percentage of cells in G(0)/G(1) phase in Q92769 siRNA group ( 63.3 % ± 2.0 % ) was significantly higher than that in untreated group ( 29.3 % ± 1.7 % ) or control siRNA group ( 29.4 % ± 1.7 % ) , F = 354.181 , P = 0.000 . Furthermore , Western blot demonstrated that down-regulation of Q92769 expression decreased the expression of cyclin D1 , cyclin E and P24941 proteins but increased the expression of P38936 protein . CONCLUSIONS : Down-regulation of Q92769 expression mediates proliferation inhibition and cell cycle arrest . It is associated with decrease in cyclin D1 , cyclin E and P24941 protein expression and increase in P38936 protein expression . An endothelial nitric oxide synthase inhibitor aggravates DB03429 -induced acute pancreatitis in rats . To clarify the role of nitric oxide ( NO ) in the development and progression of acute pancreatitis , we investigated the effect of different NO synthase inhibitors and NO donors on experimental pancreatitis in rats . Closed duodenal loop ( DB03429 ) -induced pancreatitis was produced in male Wistar rats , and the animals were treated with normal saline , the NO-synthase substrate L-arginine , the NO donor S-nitroso-N-acetylpenicillamine , aminoguanidine , which is a more powerful inhibitor of inducible NO synthase ( P35228 ) than is endothelial NO synthase ( P29474 ) , and N-nitro-L-arginine methyl ester ( L-NAME ) , a more powerful inhibitor of P29474 than of P35228 . All drugs were infused intravenously during a period of 6 or 12 h in each group . Pancreatic tissue was removed at 6 and 12 h after creating the DB03429 . L- DB00125 , S-nitroso-N-acetyl-penicillamine , and aminoguanidine treatment had no effect on the elevation of serum pancreatic enzymes , whereas L-NAME administration significantly exacerbated their elevation . Pathologically , L-NAME treatment resulted in a significantly worse histologic score at 6 and 12 h , especially in terms of the degree of hemorrhage , acinar cell necrosis , and microvascular thrombosis . Addition of L-arginine clearly reversed the effect of L-NAME . Neither the NO substrate nor NO donor could inhibit the progression of hemorrhagic pancreatitis in DB03429 -induced pancreatitis . DB02533 had no effect on the severity of the pancreatitis . We therefore concluded that NO production by P29474 may play a significant role in preventing the development and progression of acute pancreatitis . DB00755 activates and DB00102 represses the SM22α promoter through O43474 binding to , or dissociating from , its cis-DNA elements . AIMS : Krüppel-like factor 4 ( O43474 ) is implicated in all-trans retinoic acid ( DB00755 ) -induced and platelet-derived growth factor-BB ( DB00102 ) -repressed SM22α expression in vascular smooth muscle cells ( VSMCs ) . However , its exact mechanism of action remains unclear . We determined how O43474 plays different roles in DB00755 - and DB00102 -dependent regulation of the SM22α gene . METHODS AND RESULTS : DB00755 and DB00102 induced O43474 expression but exhibited an opposite effect on SM22α expression and VSMC proliferation . Chromatin immunoprecipitation and oligonucleotide pull-down assays showed that O43474 was directly bound to the O43474 binding sites 1 ( (-263)CACCC(-259) ) and 2 ( (-136)GTGGG(-132) ) of the SM22α promoter . DB00755 increased the binding of O43474 to site 2 , whereas DB00102 decreased the binding of O43474 to site 1 . DB00755 stimulated O43474 acetylation by inducing O43474 phosphorylation and increasing its interaction with p300 via activating c-Jun NH(2)-terminal kinase ( JNK ) and p38 pathways , and acetylated O43474 increased its binding activity to site 2 . DB00102 stimulated O43474 deacetylation by inducing O43474 dephosphorylation and increasing its interaction with histone deacetylase 2 ( Q92769 ) via activating extracellular signal-regulated kinase ( P29323 ) and phosphatidylinositol 3-kinase/Akt ( PI3K/Akt ) pathways , and deacetylated O43474 dissociated from site 1 . CONCLUSIONS : In VSMCs , DB00755 activates and DB00102 represses SM22α expression through O43474 binding to , or dissociating from , its different cis-elements in an acetylation-dependent manner . Impact of combined P05155 /coagulation factor XIII or DB06151 /tirilazad mesylate administration on leucocyte adherence and cytokine release in experimental endotoxaemia . We determined the effects of combinations of P05155 ( DB05341 ) with factor XIII and of DB06151 ( Q9C000 ) with tirilazad mesylate ( TM ) during lipo-polysaccharide ( LPS ) -induced endotoxaemia in rats . Forty Wistar rats were divided into four groups : the control ( CON ) group received no LPS ; the LPS , DB05341 + factor XIII and Q9C000 + TM groups received endotoxin infusions ( 5 mg/kg per h ) . After 30 min of endotoxaemia , 100 U/kg DB05341 + 50 U/kg factor XIII was administered to the DB05341 + factor XIII group , and 150 mg/kg Q9C000 + 10 mg/kg TM was administered in the Q9C000 + TM group . Administration of DB05341 + factor XIII and Q9C000 + TM both resulted in reduced leucocyte adherence and reduced levels of interleukin-1beta ( IL-1beta ) . The LPS-induced increase in P05231 levels was amplified by both drug combinations . There was no significant effect on mesenteric plasma extravasation . In conclusion , the administration of DB05341 + factor XIII and Q9C000 + TM reduced endothelial leucocyte adherence and IL-1beta plasma levels , but increased P05231 levels . DB00134 recycling pathways and antimalarial drug design . 5'-Deoxy-5'-(methylthio)adenosine ( MTA ) is an S-adenosylmethionine metabolite that is generated as a by-product of polyamine biosynthesis . In mammalian cells , MTA undergoes a phosphorolytic cleavage catalyzed by Q13126 to produce adenine and 5-deoxy-5-(methylthio)ribose-1-phosphate ( Q15012 ) . DB00173 is utilized in purine salvage pathways , and Q15012 is subsequently recycled to methionine . Whereas some microorganisms metabolize MTA to Q15012 via Q13126 , others metabolize MTA to Q15012 in two steps via initial cleavage by MTA nucleosidase to adenine and 5-deoxy-5-(methylthio)ribose ( Q99707 ) followed by conversion of Q99707 to Q15012 by Q99707 kinase . In order to assess the extent to which these pathways may be operative in Plasmodium falciparum , we have examined a series of 5'-alkyl-substituted analogs of MTA and the related Q99707 analogs and compared their abilities to inhibit in vitro growth of this malarial parasite . The Q99707 analogs 5-deoxy-5-(ethylthio)ribose and 5-deoxy-5-(hydroxyethylthio)ribose were inactive at concentrations up to 1 mM , and 5-deoxy-5-(monofluoroethylthio)ribose was weakly active ( 50 % inhibitory concentration = 700 microM ) . In comparison , the MTA analogs , 5'-deoxy-5'-(ethylthio)adenosine,5'-deoxy-5'-(hydroxyethylthio)ade nosine ( HETA ) , and 5'-deoxy-5'-(monofluoroethylthio)adenosine , had 50 % inhibitory concentrations of 80 , 46 , and 61 microM , respectively . Extracts of P. falciparum were found to have substantial Q13126 activity . Coadministration of MTA with HETA partially protected the parasites against the growth-inhibitory effects of HETA . Results of this study indicate that P. falciparum has an active Q13126 that can be targeted by analogs of MTA . A transcriptional repressor co-regulatory network governing androgen response in prostate cancers . Transcriptional corepressors are frequently aberrantly over-expressed in prostate cancers . However , their crosstalk with the P10275 ( AR ) , a key player in prostate cancer development , is unclear . Using ChIP-Seq , we generated extensive global binding maps of AR , ERG , and commonly over-expressed transcriptional corepressors including Q13547 , Q92769 , O15379 , and Q15910 in prostate cancer cells . Surprisingly , our results revealed that ERG , HDACs , and Q15910 are directly involved in androgen-regulated transcription and wired into an AR centric transcriptional network via a spectrum of distal enhancers and/or proximal promoters . Moreover , we showed that similar to ERG , these corepressors function to mediate repression of AR-induced transcription including cytoskeletal genes that promote epithelial differentiation and inhibit metastasis . Specifically , we demonstrated that the direct suppression of P18206 expression by ERG , Q15910 , and HDACs leads to enhanced invasiveness of prostate cancer cells . Taken together , our results highlight a novel mechanism by which , ERG working together with oncogenic corepressors including HDACs and the polycomb protein , Q15910 , could impede epithelial differentiation and contribute to prostate cancer progression , through directly modulating the transcriptional output of AR . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation .

MH_train_1218

MH_train_1218

MH_train_1218

interacts_with DB00834?

multiple_choice

Insights into antifolate resistance from malarial P00374 -TS structures . Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) is an important target of antimalarial drugs . The efficacy of this class of P00374 -inhibitor drugs is now compromised because of mutations that prevent drug binding yet retain enzyme activity . The crystal structures of PfDHFR-TS from the wild type ( TM4/8.2 ) and the quadruple drug-resistant mutant ( V1/S ) strains , in complex with a potent inhibitor WR99210 , as well as the resistant double mutant ( P04264 P21554 ) with the antimalarial pyrimethamine , reveal features for overcoming resistance . In contrast to pyrimethamine , the flexible side chain of WR99210 can adopt a conformation that fits well in the active site , thereby contributing to binding . The single-chain bifunctional PfDHFR-TS has a helical insert between the P00374 and TS domains that is involved in dimerization and domain organization . Moreover , positively charged grooves on the surface of the dimer suggest a function in channeling of substrate from TS to P00374 active sites . These features provide possible approaches for the design of new drugs to overcome antifolate resistance . Effects of phenacetin and its metabolite p-phenetidine on P23219 and P35354 activities and expression in vitro . The present study was aimed to test the possible cyclooxygenase ( P36551 ) -1/ P35354 selectivity of the old analgesic drug phenacetin and its metabolite p-phenetidine , which exhibits high renal toxicity . DB00316 ( acetaminophen ) , the main metabolite of phenacetin with low renal toxicity , and indomethacin were selected as reference compounds . Collagen-stimulated platelet thromboxane B2 ( TxB2 ) production and phorbol 12-myristate-13-acetate ( PMA ) -induced neutrophil prostaglandin E2 ( DB00917 ) synthesis were used as indicators for P23219 and P35354 activity , respectively . DB03783 was even less potent than paracetamol to reduce the production of both TxB2 and DB00917 , and no clear preference for either of the P36551 -enzymes was seen . P-phenetidine was a more potent inhibitor , already at nanomolar level , of the synthesis of these prostanoids than indomethacin and showed some preference to P35354 inhibition . Somewhat higher , micromolar , concentrations of p-phenetidine also reduced P35354 expression in neutrophils . We suggest that the very potent inhibitory activity of p-phenetidine on DB00917 synthesis combined with the reduction of P35354 expression could explain the renal papillary necrosis in phenacetin kidney . Antiinflammatory steroid action in human ovarian surface epithelial cells . The human ovarian surface epithelium ( OSE ) is subject to serial injury and repair during ovulation , which is a natural inflammatory event . We asked whether there is a compensatory antiinflammatory component to this process , involving steroid hormones produced locally at the time of ovulation . Quantitative RT-PCR analysis of total RNA from cultured human OSE cell monolayers showed that exposure to proinflammatory IL1alpha ( 500 pg/ml ) increased mRNA levels of cyclooxygenase-2 ( P35354 ) ( P < 0.01 ) at 48 h . The P35354 mRNA response to IL1alpha was associated with an approximate 18-fold ( P < 0.01 ) increase in mRNA levels of 11beta-hydroxysteroid dehydrogenase type 1 ( 11betaHSD1 ) , encoding the steroid dehydrogenase that reversibly reduces cortisone to antiinflammatory cortisol . Addition of cortisol to OSE cell culture medium dose-dependently suppressed the P35354 mRNA response to IL1alpha ( P < 0.01 ) but reciprocally enhanced the 11betaHSD1 mRNA response ( P < 0.05 ) , with both effects strongest at 1 microm cortisol . Presence of glucocorticoid receptor-alpha mRNA and protein was established in OSE cell monolayers and treatment with IL1alpha shown to significantly up-regulate the glucocorticoid receptor-alpha mRNA level ( P < 0.05 ) . P04150 antagonist ( DB00834 , 10 microm ) fully reversed the inhibitory effect of 1 microm cortisol on IL1alpha-stimulated P35354 mRNA expression . Progesterone also suppressed IL1alpha-induced P35354 mRNA expression but had no significant effect on IL1alpha-stimulated 11betaHSD1 expression . These data provide direct evidence for antiinflammatory actions of cortisol and progesterone in human OSE cells . Niflumic acid and MSI-2216 reduce P01375 -induced mucin expression in human airway mucosa . BACKGROUND : Human chloride channel , calcium-activated 1 ( A8K7I4 ) has been shown to induce mucin ( MUC ) gene expression and mucus production in bronchial epithelial cells . Objective To investigate whether blocking A8K7I4 decreases mucus production . METHODS : Expression of A8K7I4 and mucus was stimulated with P01375 in human upper airway mucosal explant tissue . P98088 mRNA and mucus protein expression was blocked by inhibiting A8K7I4 by using channel blockers ( niflumic acid [ DB04552 ] and MSI-2216 ) without and with P01375 stimulation . Expression of P98088 , A8K7I4 , and P35354 mRNA was quantified by using real-time PCR . Mucus protein was assessed by periodic acid Schiff staining . Laser capture microdissection was performed to quantify A8K7I4 and P98088 mRNA expression in epithelial cells derived from mucosal explant tissue . RESULTS : P01375 significantly increased P98088 and A8K7I4 mRNA as well as mucus and A8K7I4 protein expression in the mucosal explant tissue ( P < .05 ) . Inhibition of A8K7I4 with DB04552 or MSI-2216 showed a significant dose-dependent reduction of mucus production for both blockers in the mucosal explant tissue ( P < .05 ) . P98088 mRNA was also decreased by both blockers in the whole mucosal tissue and in laser-captured mucosa epithelial cells . CONCLUSIONS : Unstimulated and P01375 -induced mucin expression could be decreased by DB04552 and MSI-2216 . Inhibiting A8K7I4 may be a potential new approach to reduce mucus overproduction . The endocannabinoid 2-AG protects the blood-brain barrier after closed head injury and inhibits mRNA expression of proinflammatory cytokines . Endocannabinoids are involved in neuroprotection through numerous biochemical pathways . We have shown that the endocannabinoid 2-arachidonoyl glycerol ( 2-AG ) is released in mouse brain after closed head injury ( CHI ) , and treatment with exogenous 2-AG exerts neuroprotection via the central cannabinoid receptor P21554 . This process involves inhibition of inflammatory signals that are mediated by activation of the transcription factor NF-kB . The present study was designed to examine the effect of 2-AG on the blood-brain barrier ( BBB ) and the possible inhibition of the early expression of proinflammatory cytokines , which are implicated in BBB disruption . We found that 2-AG decreased BBB permeability and inhibited the acute expression of the main proinflammatory cytokines : P01375 , IL-1beta and P05231 . It also augmented the levels of endogenous antioxidants . We suggest that 2-AG exerts neuroprotection in part by inhibition of the early ( 1-4 h ) inflammatory response and augmentation of the brain reducing power . Bcl-xL anti-apoptotic network is dispensable for development and maintenance of CML but is required for disease progression where it represents a new therapeutic target . The dismal outcome of blast crisis chronic myelogenous leukemia ( CML-BC ) patients underscores the need for a better understanding of the mechanisms responsible for the development of drug resistance . Altered expression of the anti-apoptoticBcl-xL has been correlated with P11274 - P00519 leukemogenesis ; however , its involvement in the pathogenesis and evolution of CML has not been formally demonstrated yet . Thus , we generated an inducible mouse model in which simultaneous expression of Q92817 - P11274 - P00519 and deletion of bcl-x occurs within hematopoietic stem and progenitor cells . Absence of Bcl-xL did not affect development of the chronic phase-like myeloproliferative disease , but none of the deficient mice progressed to an advanced phenotype , suggesting the importance of Bcl-xL in survival of progressing early progenitor cells . Indeed , pharmacological antagonism of Bcl-xL , with DB05764 , combined with PP242-induced activation of Q92934 markedly augmented apoptosis of CML-BC cell lines and primary P28906 (+) progenitors but not those from healthy donors , regardless of drug resistance induced by bone marrow stromal cell-generated signals . Moreover , studies in which Q92934 or Bcl-xL expression was molecularly altered strongly support their involvement in DB05764 /PP242-induced apoptosis of CML-BC progenitors . Thus , suppression of the antiapoptotic potential of Bcl-xL together with Q92934 activation represents an effective pharmacological approach for patients undergoing blastic transformation . Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals . In diabetic P01130 -/- mice , an ectopic P12643 -Msx2 gene regulatory program is upregulated in association with vascular calcification . We verified the procalcific actions of aortic Msx2 expression in vivo . CMV-Msx2 transgenic ( CMV-Msx2Tg(+) ) mice expressed 3-fold higher levels of aortic Msx2 than nontransgenic littermates . On high-fat diets , CMV-Msx2Tg(+) mice exhibited marked cardiovascular calcification involving aortic and coronary tunica media . This corresponded to regions of Msx2 immunoreactivity in adjacent adventitial myofibroblasts , suggesting a potential paracrine osteogenic signal . To better understand Msx2-regulated calcification , we studied actions in 10T1/2 cells . We found that conditioned media from Msx2-transduced 10T1/2 cells ( Msx2-CM ) is both pro-osteogenic and adipostatic ; these features are characteristic of Wnt signaling . Msx2-CM stimulated Wnt-dependent TCF/LEF transcription , and Msx2-transduced cells exhibited increased nuclear beta-catenin localization with concomitant alkaline phosphatase induction . Msx2 upregulated Wnt3a and Wnt7a but downregulated expression of the canonical inhibitor Dkk1 . Dkk1 treatment reversed osteogenic and adipostatic actions of Msx2 . DB06285 , a Q03431 agonist that inhibits murine vascular calcification , suppressed vascular P12643 -Msx2-Wnt signaling . Analyses of CMV-Msx2Tg(+) mice confirmed that Msx2 suppresses aortic Dkk1 and upregulates vascular Wnts ; moreover , TOPGAL(+) ( Wnt reporter ) ; CMV-Msx2Tg(+) mice exhibited augmented aortic LacZ expression . Thus , Msx2-expressing cells elaborated an osteogenic milieu that promotes vascular calcification in part via paracrine Wnt signals . [ Synthesis of ( 2'-bromo-4 ' , 5'-dimethoxy-phenyl ) -(2,3-dibromo-4,5-dimethoxy-phenyl)-methane as P18031 inhibitor ] . OBJECTIVE : To synthesize (2'-bromo-4',5'-dimethoxy-phenyl)- ( 2,3- dibromo-4,5-dimethoxy-phenyl ) -methane ( 6 ) as protein tyrosine phosphatase 1B ( P18031 ) inhibitor . METHOD : DB04088 was synthesized by Friedel-Crafts reaction , bromination and decarbonylation and screened inhibitory activity against P18031 by the colorimetric assay . The structure of synthetic intermediates and target product were identified on the basis of spectral analysis . RESULT : DB04088 was synthesized successfully in four steps and evaluated for its P18031 inhibitory activity , the screening result shown that compound 6 displayed good inhibitory activity against P18031 . CONCLUSION : The target compound 6 was synthesized with the overall yield of 20 % , which was a new compound and shown good inhibitory activity against P18031 ( inhibition 40.16 % at 5 mg x L(-1) ) . P04150 overexpression exerts an antisurvival effect on human small cell lung cancer cells . Small cell lung cancer ( SCLC ) is an aggressive tumour with an abysmal prognosis . These cancers are characteristically resistant to glucocorticoid ( Gc ) action , owing to impaired expression of the glucocorticoid receptor ( GR ) . We identified reduced GR expression in human SCLC cell lines , compared to a non-SCLC cell line . The SCLC cells also showed no Gc inhibition of proliferation , in contrast to non-SCLC cells . Retroviral overexpression of GR resulted in significantly increased cell death , which was partially blocked by the GR antagonist , DB00834 . Indeed , in cells sorted for GR expression , there was rapid , near complete loss of live cells by 72 h , in contrast to control cells that proliferated as expected . Flow cytometry using P08758 revealed that cell death was by apoptosis . In addition , confocal analysis of fixed cells showed that cells overexpressing GR displayed a significant increase in fragmenting apoptotic nuclei . Microarray studies showed that transgenic GR expression upregulated the proapoptotic genes , Q92934 and Q07812 . We have , therefore , identified a profound apoptotic effect of GR in SCLC cells , which may explain the low levels of endogenous GR in SCLC cells . Understanding how GR overexpression leads to apoptotic cell death in SCLC cells may uncover new therapeutic strategies . 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 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D 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 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . The anti-androgen drug dutasteride renders triple negative breast cancer cells more sensitive to chemotherapy via inhibition of HIF-1α-/ P15692 -signaling . BACKGROUND : Triple negative breast cancer ( TNBC ) is characterized by lack of expression of both estrogen and progesterone receptor as well as lack of amplification of P04626 . Patients with TNBC carry an unfavorable prognosis compared to other breast cancer subtypes given that endocrine or P04626 targeted therapies are not effective , rendering chemotherapy the sole effective treatment option to date . Therefore , there is a high demand for additional novel treatment options . FINDINGS : We previously published a list of genes showing both higher gene expression rates in TNBC and , in addition , are known to encode targets of non-oncologic drugs . P18405 , which encodes the type-1 isoform of the steroid-5alpha-reductase , which is involved in androgen metabolism , was found to be one of these genes . DB01126 is a dual blocker of both the type-1 and type-2 isoform of P18405 and is indicated in the treatment of benign prostate hyperplasia . Treatment of TNBC cell lines with dutasteride was associated with a dose-dependent decrease in cell viability , altered protein expression of P15692 and HIF-1α and increased chemosensitivity . CONCLUSION : Our results demonstrate that the P18405 -corresponding anti-androgenic drug dutasteride might act as a combinatorial therapeutic option besides standard chemotherapy in highly aggressive TNBC . Blockade of cannabinoid receptors reduces inflammation , leukocyte accumulation and neovascularization in a model of sponge-induced inflammatory angiogenesis . OBJECTIVE : Angiogenesis depends on a complex interaction between cellular networks and mediators . The endocannabinoid system and its receptors have been shown to play a role in models of inflammation . Here , we investigated whether blockade of cannabinoid receptors may interfere with inflammatory angiogenesis . MATERIALS AND METHODS : Polyester-polyurethane sponges were implanted in C57Bl/6j mice . Animals received doses ( 3 and 10 mg/kg/daily , s.c. ) of the cannabinoid receptor antagonists SR141716A ( P21554 ) or SR144528 ( CB2 ) . Implants were collected at days 7 and 14 for cytokines , hemoglobin , myeloperoxidase , and N-acetylglucosaminidase measurements , as indices of inflammation , angiogenesis , neutrophil and macrophage accumulation , respectively . Histological and morphometric analysis were also performed . RESULTS : Cannabinoid receptors expression in implants was detected from day 4 after implantation . Treatment with P21554 or CB2 receptor antagonists reduced cellular influx into sponges at days 7 and 14 after implantation , although P21554 receptor antagonist were more effective at blocking leukocyte accumulation . There was a reduction in P01375 -α , P15692 , P09341 /KC , P13500 /JE , and P10147 /MIP-1α levels , with increase in P13501 /RANTES . Both treatments reduced neovascularization . Dual blockade of cannabinoid receptors resulted in maximum inhibition of inflammatory angiogenesis . CONCLUSIONS : Blockade of cannabinoid receptors reduced leukocyte accumulation , inflammation and neovascularization , suggesting an important role of endocannabinoids in sponge-induced inflammatory angiogenesis both via P21554 and CB2 receptors . Molecular basis for the immunostimulatory activity of guanine nucleoside analogs : activation of Q9NYK1 . Certain Q99618 -substituted and N7 , Q99618 -disubstituted guanine ribonucleosides comprise a class of small molecules with immunostimulatory activity . In a variety of animal models , these agents stimulate both humoral and cellular immune responses . The antiviral actions of these guanosine analogs have been attributed to their ability to induce type I IFNs . However , the molecular mechanisms by which the guanosine analogs potentiate immune responses are not known . Here , we report that several guanosine analogs activate Q9NYK1 ( Q9NYK1 ) . DB04860 , 7-deazaguanosine , and related guanosine analogs activated mouse immune cells in a manner analogous to known TLR ligands , inducing cytokine production in mouse splenocytes ( P05231 and IL-12 , type I and II IFNs ) , bone marrow-derived macrophages ( P05231 and IL-12 ) , and in human peripheral blood leukocytes ( type I IFNs , tumor necrosis factor alpha and IL-12 ) . The guanosine congeners also up-regulated costimulatory molecules and MHC I/II in dendritic cells . Genetic complementation studies in human embryonic kidney 293 cells confirmed that the guanosine analogs activate cells exclusively via Q9NYK1 . The stimulation of Q9NYK1 by the guanosine analogs in human cells appears to require endosomal maturation because inhibition of this process with chloroquine significantly reduced the downstream activation of NF-kappaB . However , Q9NR97 activation by R-848 and O60603 activation by [ S-[2,3-bis(palmitoyloxy)-(2-RS)-propyl]-N-palmitoyl-R- DB00151 -S- DB00133 -Lys4-OH , trihydrochloride ) ] were not inhibited by chloroquine , whereas Q9NR96 activation by CpG oligodeoxynucleotides was abolished . In summary , we present evidence that guanosine analogs activate immune cells via Q9NYK1 by a pathway that requires endosomal maturation . Thus , the B cell-stimulating and antiviral activities of the guanosine analogs may be explained by their Q9NYK1 -activating capacity . P04150 and histone deacetylase-2 mediate dexamethasone-induced repression of P98088 gene expression . Airway occlusion in obstructive airway diseases is caused in part by the overproduction of secretory mucin glycoproteins through the up-regulation of mucin ( MUC ) genes by inflammatory mediators . Some pharmacological agents , including the glucocorticoid dexamethasone ( DB00514 ) , repress mucin concentrations in lung epithelial cancer cells . Here , we show that DB00514 reduces the expression of P98088 , a major airway mucin gene , in primary differentiated normal human bronchial epithelial ( NHBE ) cells in a dose-dependent and time-dependent manner , and that the DB00514 -induced repression is mediated by the glucocorticoid receptor ( GR ) and two glucocorticoid response elements ( GREs ) in the P98088 promoter . The pre-exposure of cells to DB00834 , a GR antagonist , and mutations in either the GRE3 or GRE5 cis-sites abolished the DB00514 -induced repression . Chromatin immunoprecipitation ( ChIP ) assays showed a rapid temporal recruitment of GR to the GRE3 and GRE5 cis-elements in the P98088 promoter in NHBE and in A549 cells . Immunofluorescence showed nuclear colocalization of GR and histone deacetylase-2 ( Q92769 ) in P98088 -expressing NHBE cells . ChIP also showed a rapid temporal recruitment of Q92769 to the GRE3 and GRE5 cis-elements in the P98088 promoter in both cell types . The knockdown of Q92769 by Q92769 -specific short interfering RNA prevented the DB00514 -induced repression of P98088 in NHBE and A549 cells . These data demonstrate that GR and Q92769 are recruited to the GRE3 and GRE5 cis-sites in the P98088 promoter and mediate the DB00514 -induced cis repression of P98088 gene expression . A better understanding of the mechanisms whereby glucocorticoids repress P98088 gene expression may be useful in formulating therapeutic interventions in chronic lung diseases . High-throughput screening of mouse knockout lines identifies true lean and obese phenotypes . We developed a high-throughput approach to knockout ( KO ) and phenotype mouse orthologs of the 5,000 potential drug targets in the human genome . As part of the phenotypic screen , dual-energy X-ray absorptiometry ( DB04240 ) technology estimates body-fat stores in eight KO and four wild-type ( WT ) littermate chow-fed mice from each line . Normalized % body fat ( nBF ) ( mean KO % body fat/mean WT littermate % body fat ) values from the first 2322 lines with viable KO mice at 14 weeks of age showed a normal distribution . We chose to determine how well this screen identifies body-fat phenotypes by selecting 13 of these 2322 KO lines to serve as benchmarks based on their published lean or obese phenotype on a chow diet . The nBF values for the eight benchmark KO lines with a lean phenotype were > or =1 s.d. below the mean for seven ( perilipin , SCD1 , P21554 , Q99705 , P18031 , Q9HCL2 , P78356 ) but close to the mean for P01303 Y4R . The nBF values for the five benchmark KO lines with an obese phenotype were > 2 s.d. above the mean for four ( P32245 , P41968 , P32247 , translin ) but close to the mean for 5HT2cR . This screen also identifies novel body-fat phenotypes as exemplified by the obese kinase suppressor of ras 2 ( KSR2 ) KO mice . These body-fat phenotypes were confirmed upon studying additional cohorts of mice for KSR2 and all 13 benchmark KO lines . This simple and cost-effective screen appears capable of identifying genes with a role in regulating mammalian body fat . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Differential alterations of dihydrofolate reductase gene in human leukemia cell lines made resistant to various folate analogues . In order to clarify a molecular mechanism of folate resistance in leukemia cells , we studied alterations of the dihydrofolate reductase ( P00374 ) gene in a human leukemia cell line , Q14C87 -3 , and its sublines made resistant to methotrexate ( MTX ) , trimetrexate ( DB01157 ) and N10-propargyl-5,8-dideazafolic acid ( CB3717 ) , alone or in combination . Major alterations of the P00374 gene were examined by Southern analysis of high-molecular-weight DNA . The presence of a base change ( T --> C ) at nucleotide position 91 of the P00374 gene , which is reported to be responsible for the reduced affinity of the enzyme for MTX in an MTX-resistant human colon carcinoma cell , was examined by allele-specific oligonucleotide hybridization . In a 10,000-fold MTX-resistant subline ( Q14C87 -3/MTX10,000 ) , the normal allele of P00374 gene had been amplified . In contrast , a 200-fold DB01157 -resistant subline ( Q14C87 -3/TMQ200 ) and a 30-fold CB3717-resistant subline selected from Q14C87 -3/TMQ200 ( Q14C87 -3/TMQ200-CB-3717(30) ) were shown to have the mutant allele . Furthermore , the mutant allele had been amplified in a 500-fold MTX-resistant subline , which was established by the continuous exposure of the Q14C87 -3/TMQ200 cells to stepwise increases of drug concentration and designated as Q14C87 -3/TMQ200-MTX500 . On the other hand , a 40-fold-resistant subline to CB3717 alone ( Q14C87 -3/CB3717(40) ) showed the normal allele without amplification . These data suggest that complex alterations of the P00374 gene are involved in the molecular mechanisms of folate resistance that can be differentially introduced into leukemia cells by exposure to various folate analogues , alone or in combination . DB06155 , a selective cannabinoid P21554 receptor antagonist , inhibits atherosclerosis in P01130 -deficient mice . OBJECTIVE : The objective of this study was to determine whether the potent selective cannabinoid receptor-1 antagonist rimonabant has antiatherosclerotic properties . METHODS AND RESULTS : DB06155 ( 50 mg/kg/d in the diet ) significantly reduced food intake ( from 3.35+/-.04 to 2.80+/-0.03 g/d ) , weight gain ( from 14.6+/-0.7 g to -0.6+/-0.3 g ) , serum total cholesterol ( from 8.39+/-0.54 to 5.32+/-0.18 g/L ) , and atherosclerotic lesion development in the aorta ( from 1.7+/-0.22 to 0.21+/-0.037 mm(2) ) and aortic sinus ( from 101,000+/-7800 to 27,000+/-2900 microm(2) ) of P01130 (-/-) mice fed a Western-type diet for 3 months . DB06155 also reduced plasma levels of the proinflammatory cytokines P13500 and IL12 by 85 % ( P < 0.05 ) and 76 % ( P < 0.05 ) , respectively . Pair-fed animals had reduced weight gain ( 6.2+/-0.6 g gain ) , but developed atherosclerotic lesions which were as large as those of untreated animals , showing that the antiatherosclerotic effect of rimonabant is not related to reduced food intake . Interestingly , rimonabant at a lower dose ( 30 mg/kg/d in the diet ) reduced atherosclerosis development in the aortic sinus ( from 121,000+/-20,000 to 62,000+/-11,000 microm(2) , 49 % reduction , P < 0.05 ) , without affecting serum total cholesterol ( 7.8+/-0.7 g/L versus 8.1+/-1.3 g/L in the control group ) . DB06155 decreased lipopolysaccharide ( LPS ) - and IL1beta-induced proinflammatory gene expression in mouse peritoneal macrophages in vitro as well as thioglycollate-induced recruitment of macrophages in vivo ( 10 mg/kg , p.o. bolus ) . CONCLUSIONS : These results show that rimonabant has antiatherosclerotic effects in P01130 (-/-) mice . These effects are partly unrelated to serum cholesterol modulation and could be related to an antiinflammatory effect . P04150 antagonism disrupts the reconsolidation of social reward-related memories in rats . Reconsolidation is the process whereby consolidated memories are destabilized upon retrieval and restabilized to persist for later use . Although the neurobiology of the reconsolidation of both appetitive and aversive memories has been intensively investigated , reconsolidation of memories of physiologically relevant social rewards has received little attention . Social play , the most characteristic social behaviour displayed by young mammals , is highly rewarding , illustrated by the fact that it can induce conditioned place preference ( CPP ) . Here , we investigated the role of signalling mechanisms implicated in memory processes , including reconsolidation , namely glucocorticoid , mineralocorticoid , DB01221 glutamatergic and P21554 cannabinoid receptors , in the reconsolidation of social play-induced CPP in rats . Systemic treatment with the glucocorticoid receptor antagonist mifepristone before , but not immediately after , retrieval disrupted the reconsolidation of social play-induced CPP . DB00834 did not affect social play-induced CPP in the absence of memory retrieval . Treatment with the DB01221 receptor antagonist MK-801 modestly affected the reconsolidation of social play-induced CPP . However , the reconsolidation of social play-induced CPP was not affected by treatment with the mineralocorticoid and P21554 cannabinoid receptor antagonists spironolactone and rimonabant , respectively . We conclude that glucocorticoid neurotransmission mediates the reconsolidation of social reward-related memories in rats . These data indicate that the neural mechanisms of the reconsolidation of social reward-related memories only partially overlap with those underlying the reconsolidation of other reward-related memories .

MH_train_1219

MH_train_1219

MH_train_1219

interacts_with DB00104?

multiple_choice

Generation and phenotypic characterization of new human ovarian cancer cell lines with the identification of antigens potentially recognizable by HLA-restricted cytotoxic T cells . This study describes a simple method for long-term establishment of human ovarian tumor lines and prediction of T-cell epitopes that could be potentially useful in the generation of tumor-specific cytotoxic T lymphocytes ( CTLs ) . Nine ovarian tumor lines ( INT.Ov ) were generated from solid primary or metastatic tumors as well as from ascitic fluid . Notably all lines expressed HLA class I , intercellular adhesion molecule-1 ( P05362 ) , polymorphic epithelial mucin ( P15941 ) and cytokeratin ( CK ) , but not HLA class II , P33681 .1 ( P33681 ) or Q13072 . While of the 9 lines tested 4 ( INT.Ov1 , 2 , 5 and 6 ) expressed the folate receptor ( P15328 ) and 6 ( INT.Ov1 , 2 , 5 , 6 , 7 and 9 ) expressed the epidermal growth factor receptor ( P00533 ) ; MAGE-1 and p185HER-2/neu were only found in 2 lines ( INT.Ov1 and 2 ) and Q13065 expression in 1 line ( INT.Ov2 ) . The identification of class I MHC ligands and T-cell epitopes within protein antigens was achieved by applying several theoretical methods including : 1 ) similarity or homology searches to MHCPEP ; 2 ) BIMAS and 3 ) artificial neural network-based predictions of proteins MAGE , GAGE , P00533 , p185HER-2/neu and P15328 expressed in INT.Ov lines . Because of the high frequency of expression of some of these proteins in ovarian cancer and the ability to determine HLA binding peptides efficiently , it is expected that after appropriate screening , a large cohort of ovarian cancer patients may become candidates to receive peptide-based vaccines . Somatostatin receptors subtypes 2 and 5 , dopamine receptor type 2 expression and gsp status as predictors of octreotide P10586 responsiveness in acromegaly . We present two acromegalic patients in which clinical and molecular data are discussed in regard to their ability to predict long term octreotide P10586 therapy response . CASE REPORTS : Patient 1 : female , 36 years old at diagnosis . Basal GH and P05019 at diagnosis were 133 ng/mL and 181 % above the upper limit of reference values ( ULRV ) , respectively . Growth hormone during acute test with subcutaneous octreotide decreased from 133 to 13 ng/mL . Patient started on primary octreotide P10586 therapy ( 20mg q28 days ) and achieved biochemical parameters of disease control after 6 months . Molecular analysis of tumor fragments : gsp + ; quantitative analysis of SSTR ( somatostatin receptor ) and DR ( dopamine receptor ) mRNA - P30874 23954 ; P35346 2407 ; DR2 total 17016 copies . Patient 2 : male , 38 years old at diagnosis . Basal GH and P05019 at diagnosis were 120 ng/mL and 114 % ULRV , respectively . Patient underwent non-curative trans-sphenoidal surgery . Post-operative GH and P05019 were 112 ng/mL and 137 % ULRV , respectively . Growth hormone during acute test with subcutaneous octreotide decreased from 112 to 7 ng/mL . DB00104 P10586 therapy ( 20 mg q28 days ) was then initiated . After 6 months of treatment , patient did not attain biochemical control of disease and displayed increased tumor volume . Molecular analysis of tumor fragments : gsp not done ; quantitative analysis of SSTR and DR mRNA - P30874 416 ; P35346 3767 ; DR2 total 3439 copies . In conclusion , these two cases illustrate how laboratory data can be conflicting as predictors of octreotide P10586 responsiveness and how molecular analysis of tumor fragments can help explain different behaviors in clinically similar patients . DB08626 -induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer 's disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 and cortical Abeta40 that were 147 and 142 % of the control group , respectively . Results for Abeta42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD . Epigenetic variation during the adult lifespan : cross-sectional and longitudinal data on monozygotic twin pairs . The accumulation of epigenetic changes was proposed to contribute to the age-related increase in the risk of most common diseases . In this study on 230 monozygotic twin pairs ( MZ pairs ) , aged 18-89 years , we investigated the occurrence of epigenetic changes over the adult lifespan . Using mass spectrometry , we investigated variation in global ( LINE1 ) DNA methylation and in DNA methylation at P01308 , KCNQ1OT1 , P01344 , GNASAS , O95477 , P41159 , and P06850 , candidate loci for common diseases . Except for KCNQ1OT1 , interindividual variation in locus-specific DNA methylation was larger in old individuals than in young individuals , ranging from 1.2-fold larger at O95477 ( P = 0.010 ) to 1.6-fold larger at P01308 ( P = 3.7 × 10(-07) ) . Similarly , there was more within-MZ-pair discordance in old as compared with young MZ pairs , except for GNASAS , ranging from an 8 % increase in discordance each decade at P06850 ( P = 8.9 × 10(-06) ) to a 16 % increase each decade at P41159 ( P = 2.0 × 10(-08) ) . Still , old MZ pairs with strikingly similar DNA methylation were also observed at these loci . After 10-year follow-up in elderly twins , the variation in DNA methylation showed a similar pattern of change as observed cross-sectionally . The age-related increase in methylation variation was generally attributable to unique environmental factors , except for P06850 , for which familial factors may play a more important role . In conclusion , sustained epigenetic differences arise from early adulthood to old age and contribute to an increasing discordance of MZ twins during aging . Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV-1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE3-tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e.g. , all-trans-retinoic acid ( RA ) , 13-cis-retinoic acid ( 13- DB00982 ) , arotinoid acid ( DB02877 ) and m-carboxy-arotinoid acid ( m-carboxy- DB02877 , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 ( 24 nM ) greater than P10826 ( 4.0 nM ) greater than P13631 ( 1.3 nM ) , while the ED50 values for DB02877 and 13- DB00982 decreased in the order of P10276 ( 6.5 nM , 190 nM ) greater than P13631 ( 2.3 nM , 140 nM ) greater than P10826 ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy- DB02877 , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 greater than P10826 greater than P13631 . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . Focal brain inflammation and autism . Increasing evidence indicates that brain inflammation is involved in the pathogenesis of neuropsychiatric diseases . Autism spectrum disorders ( P51689 ) are characterized by social and learning disabilities that affect as many as 1/80 children in the USA . There is still no definitive pathogenesis or reliable biomarkers for P51689 , thus significantly curtailing the development of effective therapies . Many children with P51689 regress at about age 3 years , often after a specific event such as reaction to vaccination , infection , stress or trauma implying some epigenetic triggers , and may constitute a distinct phenotype . P51689 children respond disproportionally to stress and are also affected by food and skin allergies . P06850 ( P06850 ) is secreted under stress and together with neurotensin ( NT ) stimulates mast cells and microglia resulting in focal brain inflammation and neurotoxicity . NT is significantly increased in serum of P51689 children along with mitochondrial DNA ( mtDNA ) . NT stimulates mast cell secretion of mtDNA that is misconstrued as an innate pathogen triggering an auto-inflammatory response . The phosphatase and tensin homolog ( P60484 ) gene mutation , associated with the higher risk of P51689 , which leads to hyper-active mammalian target of rapamycin ( P42345 ) signalling that is crucial for cellular homeostasis . P06850 , NT and environmental triggers could hyperstimulate the already activated P42345 , as well as stimulate mast cell and microglia activation and proliferation . The natural flavonoid luteolin inhibits P42345 , mast cells and microglia and could have a significant benefit in P51689 . A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma . DB00004 , a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 ( P60568 ) , efficiently targets lymphoma cells expressing the high-affinity P60568 receptor ( IL-2R ) consisting of the alpha/p55/CD25 , beta/p75/CD122 , and gamma/ P31785 /CD132 chains . In vitro studies demonstrated that the retinoid X receptor ( RXR ) retinoid , bexarotene , at biologically relevant concentrations of 10(-6) M to 10(-8) M , upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox . To determine whether this biomodulatory effect could be recapitulated in vivo , we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene ( 75 mg/day-300 mg/day ) and denileukin diftitox ( 18 mcg/kg per day x 3 days every 21 days ) in a phase 1 trial . Overall response was 67 % ( 4 complete responses , 4 partial responses ) . Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day . Four patients experienced grade 2 or 3 leukopenia , and 2 had grade 4 lymphopenia . Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses ( 150 mg/day ) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells . DB05595 in lung cancer . DB00158 is essential for proliferating cells and folate transport pathways and folate-dependent metabolic processes show promise as targets for anti-neoplastic therapy . DB00158 receptor α ( P15328 ) , a folate transporter , is an attractive target for anti-neoplastic therapy due to its high affinity for folate , restricted range of expression in normal tissue and differential over-expression in malignant tissue . P15328 is expressed in non-small cell lung cancer , with a higher expression in adenocarcinoma compared with squamous cell carcinoma . DB05595 is a monoclonal antibody targeting P15328 which in pre-clinical studies led to cytotoxicity of P15328 -expressing cells , inhibited tumor growth in animal models and showed limited reactivity with normal tissue . In phase I/II trials , farletuzumab was well tolerated as a single-agent and in combination , without additive toxicity with chemotherapy . An ongoing phase II , double blind , placebo-controlled study is evaluating farletuzumab in patients with P15328 expressing metastatic adenocarcinoma of lung . Involvement of transcription factor P24468 in human trophoblast differentiation . BACKGROUND : During the in vitro differentiation of human villous cytotrophoblast ( Q01459 ) cells to a syncytiotrophoblast ( STB ) phenotype , mRNA levels for the nuclear hormone receptor P24468 ( P24468 , COUP-TFII ) increase rapidly , reaching a peak at day 1 of differentiation that is 8.8-fold greater than that in undifferentiated Q01459 cells . To examine whether P24468 is involved in the regulation of villous Q01459 cell differentiation , studies were performed to determine whether P24468 regulates the expression of P05549 ( AP-2alpha ) , a transcription factor that is critical for the terminal differentiation of these cells to a STB phenotype . METHODOLOGY/PRIMARY FINDINGS : Overexpression of P24468 in primary cultures of human Q01459 cells and JEG-3 human choriocarcinoma cells induced dose-dependent increases in P05549 promoter activity . Conversely , siRNA mediated silencing of the P24468 gene in villous Q01459 undergoing spontaneous differentiation blocked the induction of the mRNAs for P05549 and several STB cell specific marker genes , including human placental lactogen ( hPL ) , pregnancy specific glycoprotein 1 ( P11464 ) and corticotropin releasing hormone ( P06850 ) by 51-59 % . The induction of P05549 promoter activity by P24468 was potentiated by the nuclear hormone receptors retinoic acid receptor alpha ( P10276 ) and retinoid X receptor alpha ( P19793 ) . CONCLUSIONS/SIGNIFICANCE : Taken together , these results strongly suggest that P24468 is involved in villous Q01459 cell differentiation and that P24468 acts , at least in part , by directly activating P05549 gene expression and by potentiating the transactivation of P05549 by P10276 and P19793 . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . DB00104 is the favorable alternative for cisplatin resistance reversal of ovarian cancer in vitro and in nude mice in vivo . This study aimed to observe the effects of octreotide ( O75051 ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo . MTT method and flow cytometry were used to investigate the effect of cisplatin , O75051 or the combination of these two compounds on the proliferation and apoptosis of SKOV3- O60220 cells . The size and weight of xenograft tumors from the nude mice model were measured . Real-time PCR was used to detect the mRNA expression of P30874 , P08183 , Q92887 , Q86UG4 -pi and P00533 in SKOV3/ O60220 cells following the different treatment . At the concentration of 2.5-20 g/ml , O75051 significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3- O60220 cells ' response to cisplatin . Unchanged expression was found in P30874 on the SKOV3/ O60220 cell in vitro after O75051 treatment , but increased expression in vivo ( p < 0.05 ) . O75051 increased Q86UG4 -pi expression ( p < 0.05 ) and reduced Q92887 and P00533 expression ( p < 0.05 ) in a dose-dependent manner . The similar results were obtained in mice in vivo experiment , except the reduced expression of Q86UG4 -pi . It is suggested that O75051 could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface P30874 , and reverse cisplatin resistance through inhibition of Q92887 , P00533 , and even Q86UG4 -pi expressions . Neurochemical correlates of sympathetic activation during severe alcohol withdrawal . Cerebrospinal fluid ( P04141 ) was obtained from 17 patients during acute alcohol withdrawal . Eight of these 17 patients had a second lumbar puncture a mean of 11.9 +/- 8.1 ( SD ) days later , when the clinical signs of alcohol withdrawal had subsided . P04141 3-methoxy-4-hydroxyphenylglycol concentrations declined significantly ( p < 0.05 ) during the course of alcohol withdrawal from 52.0 +/- 22.1 ( SD ) to 39.6 +/- 12.6 pM/ml . In early withdrawal , there was a significant positive correlation between P04141 norepinephrine ( NE ) and corticotropin releasing hormone ( P06850 ) concentrations ( r = 0.95 , p < 0.001 ) . Both NE and P06850 concentrations correlated positively with diastolic blood pressure ( r = 0.88 , p < 0.001 and r = 0.62 , p < 0.05 , respectively ) . In all samples , P04141 5-hydroxyindole acetic acid concentrations correlated positively with P04141 -homovanillic acid concentrations ( r = 0.83 , p < 0.001 ) . These findings indicate significant perturbations of the noradrenergic neuronal system and a change in P06850 -NE interactions during acute alcohol withdrawal . DB00644 II ( DB00644 II ) mediates the anorexigenic actions of α-melanocyte-stimulating hormone ( α-MSH ) and corticotropin-releasing hormone ( P06850 ) in goldfish . Intracerebroventricular ( ICV ) administration of gonadotropin-releasing hormone II ( DB00644 II ) , which plays a crucial role in the regulation of reproduction in vertebrates , markedly reduces food intake in goldfish . However , the neurochemical pathways involved in the anorexigenic action of DB00644 II and its interaction with other neuropeptides have not yet been identified . Alpha-melanocyte-stimulating hormone ( α-MSH ) , corticotropin-releasing hormone ( P06850 ) and P06850 -related peptides play a major role in feeding control as potent anorexigenic neuropeptides in goldfish . However , our previous study has indicated that the DB00644 II-induced anorexigenic action is not blocked by treatment with melanocortin 4 receptor ( P32245 ) and P06850 receptor antagonists . Therefore , in the present study , we further examined whether the anorexigenic effects of α-MSH and P06850 in goldfish could be mediated through the P30968 neuronal pathway . ICV injection of the P32245 agonist , melanotan II ( 80 pmol/g body weight ; BW ) , significantly reduced food intake , and its anorexigenic effect was suppressed by ICV pre-administration of the DB00644 type I receptor antagonist , antide ( 100 pmol/gBW ) . The P06850 -induced ( 50 pmol/gBW ) anorexigenic action was also blocked by treatment with antide . ICV injection of P06850 ( 50 pmol/gBW ) induced a significant increase of the DB00644 II mRNA level in the hypothalamus , while ICV injection of melanotan II ( 80 pmol/gBW ) had no effect on the level of DB00644 II mRNA . These results indicate that , in goldfish , the anorexigenic actions of α-MSH and P06850 are mediated through the DB00644 type I receptor-signaling pathway , and that the DB00644 II system regulates feeding behavior . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity . Hypothalamic-pituitary-adrenal axis function , psychopathological traits , and natural killer ( NK ) cell activity in anorexia nervosa . To evaluate the role of Hypothalamic-Pituitary-Adrenal ( Q9Y251 ) hormones and psychoneuroendocrine modulation on NK cell activity in Anorexia Nervosa ( AN ) we studied in 24 patients and 20 sex- and age-matched healthy controls , the spontaneous NK activity of peripheral blood mononuclear ( PBM ) cells and the susceptibility in vitro to cortisol or immune interferon or interleukin-2 . NK cytotoxicity of PBM cells was measured in a direct non-radiometric 4h cytolytic assay using K562 cells as targets . Q9Y251 axis function was evaluated by IV ovine DB01285 Releasing Hormone ( o- P06850 ) administration . We did not find clear-cut abnormalities of NK cytotoxicities either in basal conditions or after exposure to challengers . The extent of cortisol-dependent inhibition was comparable in patients and controls . Significant inverse and direct correlations were found respectively between the spontaneous NK cell activity and baseline serum cortisol at 0800 h ( r = -0.5 ; p < .02 ) , and between P60568 dependent boosting of NK cell cytotoxicity and DB01285 , beta-endorphin or cortisol responses after o- P06850 , expressed as areas under the curve ( AUC ) ( r = 0.46 , p < .05 ; r = 0.46 , p < .05 ; and r = -0.48 , p < .05 , respectively ) . Correlations observed with AUC ratios yielded more significant results ( r = 0.62 ; p < .01 and r = 0.51 ; p < .05 respectively ) . These data suggest a role for Proopiomelanocortin ( P01189 ) derived peptides in the regulation of NK cell activity in AN , and multifaceted relationships between this particular immune function , on the one hand , and certain patterns of Q9Y251 axis function on the other . Structure-function studies of linear and cyclized peptide antagonists of the P30968 . Structurally new analogs of the peptidic P30968 antagonist DB00050 as well as conformationally constrained cyclized deca- or pentapeptides were synthesized and selected peptides evaluated comprehensively . To understand how structural variations of the antagonistic peptide effect pharmacodynamic properties , binding affinities and antagonistic potencies toward the human and rat P30968 were determined . Whereas large substituents in position 6 of linear peptides are compatible with high binding affinity ( K(D) < 0.5 nM ) , all cyclized peptides except the cyclo[3-10] analog D-52391 depicted low binding affinity ( K(D) > 10 nM ) . Binding affinity and antagonistic potency in vitro correlated for all peptides and surprisingly no discrimination between human and rat receptor proteins was observed . Since receptor residues W(101) and N(102) are involved in agonist and antagonist binding , equally potent but structurally different antagonists were tested for binding to the respective W(101)A and N(102)A mutants . In contrast to linear decapeptides , residues N(102) and W(101) are not involved in binding of D-23938 and W(101) is the critical residue for D-52391 binding . We conclude that although equally potent , peptidic P30968 antagonists do have distinct interactions within the ligand binding pocket . Finally , selected antagonists were tested for testosterone suppression in male rats . The duration of testosterone suppression below castration levels differed largely from 1 day for DB06785 to 27 days for D-23487 . Systemic availability became evident as the most important parameter for in vivo efficacy . A phase I pharmacokinetic study of matuzumab in combination with paclitaxel in patients with P00533 -expressing advanced non-small cell lung cancer . DB05101 is a humanized IgG1 P00533 monoclonal antibody . This phase I study investigated the tolerability , safety and pharmacokinetics ( PK ) of matuzumab in combination with paclitaxel in patients with P00533 -expressing advanced non-small cell lung cancer ( NSCLC ) . Six dose levels/schedules of matuzumab were explored in combination with paclitaxel . Dose was escalated from 100 mg to 1,600 mg on a modified Fibonacci scheme according to the incidence of dose-limiting toxicity ( DLT ) over the first two cycles . DLT was assessed in patients who completed the first two treatment cycles or who stopped treatment because of a DLT during those cycles . Patients with non-progressive disease could then continue to receive study treatment for up to 6 months . The safety population comprised 44 patients , with DLT evaluable in 33 . The maximum tolerated dose was not reached , with only one DLT reported at the 1,600 mg 3-weekly dose level . The most frequent grade 3/4 adverse events across all cycles were dyspnea ( 23 % ) and neutropenia ( 11 % ) . DB05101 exhibited non-linear PK , with accumulation after escalation and repeated dosing . Tumor growth control was seen in 15/44 ( 34 % ) patients , including 5/9 ( 56 % ) at the 800 mg weekly dose level . DB05101 combined with paclitaxel was generally well tolerated in patients with advanced NSCLC . There was some evidence of anticancer activity in relation to the matuzumab 800 mg weekly dose . P01308 -like growth factor-1 ( DB01277 ) induces O76076 / O76076 via multiple molecular cross-talks and is essential for mitogenic switch by DB01277 axis in estrogen receptor-positive breast tumor cells . Previously , we have shown that the expression of Wnt-1-induced signaling protein-2 ( O76076 ) , also known as O76076 , can be regulated by multiple stimulants in estrogen receptor ( ER ) -positive breast tumor cells to exert their mitogenic action in these cells . Here , we show that insulin-like growth factor-1 ( DB01277 ) , a strong mitogen , enhanced the expression of the O76076 / O76076 gene parallel with the induction of proliferation of ER-positive breast tumor cells . An additive effect was also seen in combination with estrogen . Perturbation of DB01277 -induced O76076 / O76076 expression by O76076 -specific RNA interference impaired the mitogenic action of DB01277 on ER-positive breast tumor cells . Furthermore , the studies have shown that the multiple molecular cross-talks and side-talks among IGF-1R , P03372 , and phosphatidylinositol 3-kinase ( PI3K ) /Akt signaling molecules are required to induce O76076 / O76076 mRNA by DB01277 in ER-positive , noninvasive breast tumor cells . Because a pure anti-ER DB00947 is not only able to suppress the up-regulation of O76076 / O76076 mRNA expression by DB01277 , it also suppresses the PI3K/Akt activity induced by DB01277 in MCF-7 cells ; we anticipate that the membrane ER receptor may participate in this event . Collectively , these studies propose for the first time that O76076 / O76076 is an integral signaling molecule in mitogenic action of DB01277 axis in ER-positive human breast tumor cells .

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interacts_with DB00741?

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Mechanisms of atrial-selective block of Na⁺ channels by ranolazine : I . Experimental analysis of the use-dependent block . Atrial-selective inhibition of cardiac Na(+) channel current ( I(Na) ) and I(Na)-dependent parameters has been shown to contribute to the safe and effective management of atrial fibrillation . The present study examined the basis for the atrial-selective actions of ranolazine . Whole cell I(Na) was recorded at 15°C in canine atrial and ventricular myocytes and in human embryonic kidney ( P29320 ) -293 cells expressing Q14524 . Tonic block was negligible at holding potentials from -140 to -100 mV , suggesting minimal drug interactions with the closed state . Trains of 40 pulses were elicited over a range of holding potentials to determine use-dependent block . Guarded receptor formalism was used to analyze the development of block during pulse trains . Use-dependent block by ranolazine increased at more depolarized holding potentials , consistent with an interaction of the drug with either preopen or inactivated states , but was unaffected by longer pulse durations between 5 and 200 ms , suggesting a weak interaction with the inactivated state . Block was significantly increased at shorter diastolic intervals between 20 and 200 ms . Responses in atrial and ventricular myocytes and in P29320 -293 cells displayed a similar pattern . DB00243 is an open state blocker that unbinds from closed Na(+) channels unusually fast but is trapped in the inactivated state . Kinetic rates of ranolazine interactions with different states of atrial and ventricular Na(+) channels were similar . Our data suggest that the atrial selectivity of ranolazine is due to a more negative steady-state inactivation curve , less negative resting membrane potential , and shorter diastolic intervals in atrial cells compared with ventricular cells at rapid rates . Suppression of dendritic cell maturation by Trichinella spiralis excretory/secretory products . Evidence from experimental studies indicates that during chronic infections with certain helminth species a regulatory network is induced that can down-modulate not only parasite-induced inflammation but also reduce other immunopathologies such as allergies and autoimmune diseases . The mechanisms however , and the molecules involved in this immunomodulation are unknown . Here , we focus on the effect of Trichinella spiralis excretory/secretory antigens ( TspES ) on the innate immune response by studying the effect of TspES on DC maturation in vitro . Bone marrow-derived DC from BALB/c mice were incubated with TspES either alone or in combination with LPS derived from two different bacteria . As indicators of DC maturation , the cytokine production ( IL-1alpha , P05231 , P22301 , IL-12p70 and P01375 ) and the expression of various surface molecules ( MHC-II , P25942 , P33681 and P42081 ) were measured . Results indicate that while TspES alone did not change the expression of the different surface molecules or the cytokine production , it completely inhibited DC maturation induced by Escherichia coli LPS ( E. coli LPS ) . In contrast , DC maturation induced by LPS from another bacterium , Neisseria meningitidis , was not affected by TspES . These results were confirmed using O00206 / Q9Y6Y9 / P08571 transfected P29320 293 cells . In conclusion , T. spiralis ES antigens lead to suppression of DC maturation but this effect depends on the type of LPS used to activate these cells . P00747 activator inhibitor type 1 ( P05121 ) is a valuable biomarker for predicting the metabolic syndrome ( MS ) in institutionalized elderly residents in Taiwan . Circulating levels of inflammatory and prothrombotic factors are elevated in the metabolic syndrome ( MS ) and linked with the occurrence of cardiovascular events . The aim of our study was to investigate the relationship between inflammatory and prothrombotic markers and the MS in elderly institutionalized residents . A total of 326 non-diabetic residents of Chuang-Hua Veterans Care Home ( age : 79.9+/-4.1 years ; 100 % males ) were enrolled . MS was diagnosed according to the DB00551 /NHLBI Scientific Statement criteria . Body fat percentage was measured by bioelectrical impedance analysis . P01308 resistance was calculated by homeostasis model assessment for insulin resistance ( HOMA-IR ) . Inflammatory markers , including tumor necrosis factor-a ( P01375 ) , high sensitivity P02741 ( hsCRP ) , and plasminogen activator inhibitor-1 ( P05121 ) , were determined using ELISA . Elderly residents with the MS had higher systolic and diastolic blood pressures ( both p < 0.001 ) and higher HOMA-IR ( p < 0.001 ) , hsCRP ( p = 0.008 ) , and P05121 levels ( p < 0.001 ) than those without the MS . On multivariate logistic regression analysis , P05121 was an independent risk factor for the MS . Of the MS components , elderly residents with higher waist circumferences and higher levels of plasma fasting glucose , and triglyceride ( TG ) , and lower levels of high density lipoprotein ( HDL ) had higher P05121 levels than those without the above components . NPC 15669 , an inhibitor of neutrophil recruitment , is efficacious in acetic acid-induced colitis in rats . BACKGROUND : The efficacy of the leukocyte recruitment inhibitor , N- [ 9H-2,7-dimethylfluoren-9-ylmethoxy ) carbonyl ] -L-leucine ( NPC 15669 ) was compared with drugs used to treat inflammatory bowel diseases in a rat model , acetic acid-induced colitis . METHODS : Colonic damage assessed by visual inspection , histological quantitation of tissue injury , vascular permeability , myeloperoxidase ( P05164 ) accumulation , and synthesis of inflammatory mediators were measured . RESULTS : Intrarectal pretreatment with NPC 15669 results in a significant reduction of all measured indices of inflammation . The median effective dose ( ED50 ) of NPC 15669 for inhibition of P05164 accumulation and vascular permeability is 13.2 mg/kg and 31 mg/kg , respectively . The active moiety of sulfasalazine , DB00244 ( DB00244 ) , the antioxidant/ P09917 inhibitor , nordihydroguaiaretic acid ( NDGA ) and the corticosteroids dexamethasone and hydrocortisone , yielded ED50 values ( P05164 accumulation ) of 68 mg/kg , 95 mg/kg , 0.7 mg/kg , and 13 mg/kg , respectively . When formulated suspensions of NPC 15669 , DB00244 , or dexamethasone were used , potency was increased 10-40-fold . Furthermore , NPC 15669 ( 10 mg/kg ) administered 7 hours after acetic acid and evaluated 24 hours after acetic acid administration significantly attenuated neutrophil influx ( 70 % inhibition of P05164 accumulation ) , whereas DB00244 ( 100 mg/kg ) displayed no therapeutic effects . CONCLUSIONS : NPC 15669 may be useful in the treatment of inflammatory disorders . DB08889 can induce tumor cell death through selective inhibition of the chymotrypsin-like activity of the proteasome . DB08889 is a proteasome inhibitor in clinical development that primarily targets the chymotrypsin-like ( CT-L ) subunits in both the constitutive proteasome ( c20S ) and the immunoproteasome ( i20S ) . To investigate the impact of inhibiting the CT-L activity with carfilzomib , we set out to quantitate the levels of CT-L subunits beta5 from the c20S and P28062 from the i20S in normal and malignant hematopoietic cells . We found that the i20S is a major form of the proteasome expressed in cells of hematopoietic origin , including multiple myeloma ( MM ) CD138+ tumor cells . Although specific inhibition of either P28062 or beta5 alone was insufficient to produce an antitumor response , inhibition of all proteasome subunits was cytotoxic to both hematologic tumor cells and peripheral blood mononuclear cells . However , selective inhibition of both beta5 and P28062 was sufficient to induce an antitumor effect in MM , non-Hodgkin lymphoma , and leukemia cells while minimizing the toxicity toward nontransformed cells . In MM tumor cells , CT-L inhibition alone was sufficient to induce proapoptotic sequelae , including proteasome substrate accumulation , Noxa and caspase 3/7 induction , and phospho-eIF2alpha suppression . These data support a hypothesis that hematologic tumor cells are uniquely sensitive to CT-L inhibition and provide a mechanistic understanding of the clinical safety profile and antitumor activity of proteasome inhibitors . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . DB01954 attenuates bile duct ligation-induced liver injury in rats : a potential pathogenic role of DB05876 . Anti-inflammatory and antifibrotic effects of the broad spectrum phosphodiesterase ( PDE ) inhibitor pentoxifylline have suggested an important role for cyclic nucleotides in the pathogenesis of hepatic fibrosis ; however , studies examining the role of specific PDEs are lacking . Endotoxemia and O00206 ( O00206 ) -mediated inflammatory and profibrotic signaling play a major role in the development of hepatic fibrosis . Because DB02527 -specific DB05876 critically regulates lipopolysaccharide ( LPS ) - O00206 -induced inflammatory cytokine expression , its pathogenic role in bile duct ligation-induced hepatic injury and fibrogenesis in Sprague-Dawley rats was examined . Initiation of cholestatic liver injury and fibrosis was accompanied by a significant induction of P27815 , B , and D expression and activity . Treatment with the DB05876 -specific inhibitor rolipram significantly decreased liver DB05876 activity , hepatic inflammatory and profibrotic cytokine expression , injury , and fibrosis . At the cellular level , in relevance to endotoxemia and inflammatory cytokine production , Q07343 was observed to play a major regulatory role in the LPS-inducible tumor necrosis factor ( P01375 ) production by isolated Kupffer cells . Moreover , DB05876 expression was also involved in the in vitro activation and transdifferentiation of isolated hepatic stellate cells ( HSCs ) . Particularly , P27815 , B , and D upregulation preceded induction of the P19526 activation marker α-smooth muscle actin ( α-SMA ) . In vitro treatment of HSCs with rolipram effectively attenuated α-SMA , collagen expression , and accompanying morphologic changes . Overall , these data strongly suggest that upregulation of DB05876 expression during cholestatic liver injury plays a potential pathogenic role in the development of inflammation , injury , and fibrosis . P04150 -mediated regulation of P14780 gene expression in human ovarian surface epithelial cells . OBJECTIVE : To obtain proof-of-concept that locally produced anti-inflammatory steroids suppress ovulation-associated extracellular matrix proteases in human ovarian surface epithelial ( OSE ) cells . DESIGN : Primary OSE cell cultures treated with interleukin-1alpha ( IL-1alpha ) ( 500 pg/mL ) as proxy for inflammation , with/without anti-inflammatory steroid ( cortisol or progesterone [ P ] , 0.01-1.0 microM ) . SETTING : Academic medical center . PATIENT(S) : Sixteen premenopausal women ( 29-46 years ) undergoing surgery for nonmalignant gynecological conditions . MAIN OUTCOME MEASURE(S) : Semiquantitative extracellular matrix protease gene expression profiling with verification by real-time quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) and gelatinase zymography . RESULT(S) : Treatment with IL-1alpha stimulated messenger RNA ( mRNA ) expression of several ovulation-associated matrix metalloproteinase genes by OSE cell cultures , including gelatinase B ( P14780 ) but not gelatinase A ( P08253 ) . The IL-1alpha-stimulated P14780 mRNA production was suppressed by cortisol but not P. DB00741 but not P also dose-dependently suppressed IL-1alpha-stimulated P14780 gelatinase activity and this effect was blocked by the glucocorticoid receptor antagonist DB00834 . CONCLUSION(S) : In human OSE cells , stimulation of P14780 gene expression and proteolytic activity by IL-1alpha is suppressed by anti-inflammatory cortisol through a glucocorticoid receptor-mediated mechanism . Because IL-1alpha also generates cortisol formation in OSE by stimulating cortisone reductase activity , these results support a role for intracrine cortisol in minimizing proteolytic damage to the OSE at ovulation . Design and synthesis of an orally bioavailable and selective peptide epoxyketone proteasome inhibitor ( PR-047 ) . Proteasome inhibition has been validated as a therapeutic modality in the treatment of multiple myeloma and non-Hodgkin 's lymphoma . DB08889 , an epoxyketone currently undergoing clinical trials in malignant diseases , is a highly selective inhibitor of the chymotrypsin-like ( CT-L ) activity of the proteasome . A chemistry effort was initiated to discover orally bioavailable analogues of carfilzomib , which would have potential for improved dosing flexibility and patient convenience over intravenously administered agents . The lead compound , 2-Me-5-thiazole- DB00133 (OMe)- DB00133 (OMe)- DB00120 -ketoepoxide ( 58 ) ( PR-047 ) , selectively inhibited CT-L activity of both the constitutive proteasome ( beta5 ) and immunoproteasome ( P28062 ) and demonstrated an absolute bioavailability of up to 39 % in rodents and dogs . It was well tolerated with repeated oral administration at doses resulting in > 80 % proteasome inhibition in most tissues and elicited an antitumor response equivalent to intravenously administered carfilzomib in multiple human tumor xenograft and mouse syngeneic models . The favorable pharmacologic profile supports its further development for the treatment of malignant diseases . The relationship of P04141 and plasma cytokine levels to cerebral white matter injury in the premature newborn . Ischemia and systemic infection are implicated in the etiology of periventricular white matter injury , a major cause of adverse motor and cognitive outcome in preterm infants . Cytokines are signaling proteins that can be produced as part of the inflammatory response to both ischemia and infection . The aim of this study was to relate cerebrospinal fluid ( P04141 ) concentrations of P05231 , P10145 , P22301 , tumor necrosis factor alpha ( P01375 ) , and interferon gamma ( P01579 ) to magnetic resonance-defined white matter injury in preterm infants . Relationships between P04141 and plasma cytokine concentrations were also examined . Preterm infants ( < or=32 wk ) and more mature infants from The Royal Women 's Hospital , Melbourne , Australia , and Christchurch Women 's Hospital , Christchurch , New Zealand , were eligible for study if they required a clinically indicated lumbar puncture . Plasma samples were obtained in a subgroup of Christchurch infants . Preterm infants underwent advanced quantitative volumetric magnetic resonance imaging using a 1.5-Tesla scanner at term equivalent . One hundred forty-six infants were enrolled and 190 P04141 and 42 plasma samples obtained . There was no significant correlation between paired P04141 and plasma concentrations for any cytokine . In comparing plasma and P04141 concentrations , levels of P10145 were significantly higher in P04141 than plasma . Preterm infants with Q9BWK5 -defined cerebral white matter injury had higher levels of P05231 , P22301 , and P01375 in the P04141 than infants without such injury . Plasma cytokine concentrations may not reflect P04141 cytokine levels or inflammatory events within the brain . Elevated P04141 levels of cytokines in infants with white matter injury suggest an altered inflammatory balance . Multiple arrhythmic syndromes in a newborn , owing to a novel mutation in Q14524 . BACKGROUND : Mutations in the Q14524 gene have been linked to Brugada syndrome ( BrS ) , conduction disease , Long QT syndrome ( LQT3 ) , atrial fibrillation ( AF ) , and to pre- and neonatal ventricular arrhythmias . OBJECTIVE : The objective of this study is to characterize a novel mutation in Na(v)1.5 found in a newborn with fetal chaotic atrial tachycardia , post-partum intraventricular conduction delay , and QT interval prolongation . METHODS : Genomic DNA was isolated and all exons and intron borders of 15 ion-channel genes were sequenced , revealing a novel missense mutation ( Q270K ) in Q14524 . Na(v)1.5 wild type ( WT ) and Q270K were expressed in CHO- P04264 with and without the Na(v)β1 subunit . Results . Patch-clamp analysis showed ∼40 % reduction in peak sodium channel current ( I(Na) ) density for Q270K compared with WT . Fast and slow decay of I(Na) were significantly slower in Q270K . Steady-state activation and inactivation of Q270K channels were shifted to positive potentials , and window current was increased . The tetrodotoxin-sensitive late I(Na) was increased almost 3-fold compared with WT channels . DB00243 reduced late I(Na) in WT and Q270K channels , while exerting minimal effects on peak I(Na) . CONCLUSION : The Q270K mutation in Q14524 reduces peak I(Na) while augmenting late I(Na) , and may thus underlie the development of atrial tachycardia , intraventricular conduction delay , and QT interval prolongation in an infant . DB00244 induces manganese superoxide dismutase in rat intestinal epithelial cell lines and in vivo . DB00244 ( DB00244 ) is effective in the treatment of inflammatory bowel diseases . However , the mechanisms of action of DB00244 remain unclear . IEC-6 and IRD-98 , nontransformed rat small intestinal epithelial cell lines , were used to examine the effect of DB00244 on the expression of manganese superoxide dismutase ( MnSOD ) . Rats were given DB00244 enemas to determine the effect on colonic MnSOD expression . Treatment with DB00244 at 0.02 or 2 mg/ml induced MnSOD mRNA levels 2.67-fold or 5.66-fold , respectively . Inhibition of P09917 activating protein with MK-886 or cyclooxygenase with indomethacin did not influence the level of MnSOD mRNA . Nuclear run-on experiments demonstrated an increase in de novo transcription following treatment with DB00244 . MnSOD protein levels were induced 2-fold at 24 h and 4.23-fold at 48 h following treatment with 1 mg/ml DB00244 . DB00244 increased MnSOD 1.7-fold in vivo . Pretreatment with DB00244 significantly protected IRD-98 cells from tumor necrosis factor-alpha cytotoxicity . This is the first example of transcriptional gene regulation by DB00244 . The induction of MnSOD by DB00244 may contribute to the therapeutic mechanism of DB00244 . Insights into cancer therapeutic design based on p53 and P50591 receptor signaling . Knowledge of the emerging pathways of cell death downstream of the p53 tumor suppressor and the P50591 death-inducing ligand is suggesting ways to improve therapeutic design in cancer . In contrast to its unique P55008 cell cycle arresting mechanism that is maintained by P38936 ( P38936 ) , there are signals transduced by p53 to multiple apoptotic effectors perhaps due to the importance of apoptosis in suppressing tumors . There is evidence for cytoplasmic as well as mitochondrial activation of caspases downstream of p53 , although in some cell lineages the signal ultimately involves the mitochondria . The P50591 signaling pathway appears promising for therapeutic development despite sharing some similarities with the toxic Fas and P01375 pathways , in terms of effector molecules and downstream signals . One of the key findings is the tissue specificity of cell death responses , a feature that could be exploited in strategies to widen the therapeutic window of combination cancer therapies . Efforts continue to develop p53-targeted cancer therapy , and novel clues to enhance or block specific effectors may improve therapeutic design . Prion protein stimulates tissue-type plasminogen activator-mediated plasmin generation via a lysine-binding site on kringle 2 . Recombinant human prion-protein ( PrP23-231 ) stimulates plasminogen activation by tissue-type plasminogen activator ( t-PA ) . The stimulatory activity is conserved in the N-terminal fragment ( PrP23-110 ) . It has further been shown by others that P04156 (c) binds to kringle-domains of plasminogen . We compared the stimulatory activity of recombinant PrP23-231 and PrP23-110 on plasminogen activation catalyzed by t-PA , urokinase ( u-PA ) , streptokinase and Desmodus salivary plasminogen activator ( DSPAalpha1 ) . As these plasminogen activators are distinct , with respect to their kringle domains we studied their binding to immobilized PrP23-110 . P00747 activation was measured in a chromogenic assay in vitro and binding studies were carried out using surface plasmon resonance technology . We found that recombinant full-length prion protein , PrP23-231 , and PrP23-110 specifically stimulate t-PA mediated plasminogen activation . Two hundred nanomoles per liter of PrP23-110 stimulated 1.8 nmol L(-1) t-PA 48-fold , 180 nmol L(-1) DB04925 (alpha1) 2.5-fold , 1.8 nmol L(-1) u-PA 1.1-fold , and 1.8 nmol L(-1) streptokinase 1.8-fold . Our data show no specific binding for streptokinase . In contrast all plasminogen activators carrying a kringle domain bound to PrP23-110 . We further studied the effect of lysine on binding to PrP23-110 and on plasminogen activation by DB04925 (alpha1) or t-PA . DB00123 decreased both the binding of t-PA to PrP23-110 and the stimulation of plasmin generation by t-PA . Both binding and plasminogen activation of DB04925 (alpha1) were not influenced by the presence of lysine . All plasminogen activators tested bearing kringle domains bind to PrP23-110 . Binding to PrP23-110 is not sufficient for stimulation of plasmin generation . Thus the lysine-binding site of kringle 2 that is unique to t-PA appears to mediate the specific stimulation of plasminogen activation by the cellular prion protein . Effects of metalloproteinase inhibition in a murine model of renal ischemia-reperfusion injury . Ischemia-reperfusion injury ( IRI ) is a leading cause of acute tubular necrosis ( ATN ) and delayed graft function in transplanted organs . Up-regulation of matrix metalloproteinases ( MMPs ) propagates the microinflammatory response that drives IRI . This study sought to determine the specific effects of Marimastat ( Vernalis , DB00786 ) , a broad spectrum MMP and P78536 inhibitor , on IRI-induced ATN . Mice were pretreated with Marimastat or methylcellulose vehicle for 4 d before surgery . Renal pedicles were bilaterally occluded for 30 min and allowed to reperfuse for 24 h . Baseline creatinine levels were consistent between experimental groups ; however , post-IRI creatinine levels were 4-fold higher in control mice ( p < 0.0001 ) . The mean difference between the post-IRI histology grades of Marimastat-treated and control kidneys was 1.57 ( p = 0.003 ) , demonstrating more severe damage to control kidneys . Post-IRI mean ( +/-SEM ) P08253 activity rose from baseline levels in control mice ( 3.62 +/- 0.99 ) ; however , pretreated mice presented only a slight increase in mean P08253 activity ( 1.57 +/- 0.72 ) ( p < 0.001 ) . In conclusion , these data demonstrate that MMP inhibition is associated with a reduction of IRI in a murine model . Lactobacillus plantarum TN8 exhibits protective effects on lipid , hepatic and renal profiles in obese rat . This study aimed to first investigate the immuno-modulatory effects of six newly isolated lactic acid bacteria ( Q9GZY6 ) on the peripheral blood mononuclear cells ( PBMC ) of Wistar rats . Except for Lactobacillus plantarum TN8 , all the other strains were noted to induce high levels of pro-inflammatory cytokine IL-12 and low levels of anti-inflammatory cytokine P22301 . The strains also generated low ratios of P22301 /IL-12 cytokine . Strain TN8 was , on the other hand , noted to induce an increase in anti-inflammatory P22301 cytokine secretion rates and a decrease in pro-inflammatory IL-12 , IFN-γ and P01375 -α cytokine production . The oral administration of TN8 improved the hepatic and urinary functions of obese rats by inducing decreases ( P < 0.05 ) in alanine amino transferase ( ALAT ) , gamma glutamyl transferase ( P19440 ) , plasmatic triglycerides , total cholesterol concentrations , creatinine , urea , and body weight when compared to the control group of animals that underwent an increase in aspartate amino transferase ( ASAT ) and high density lipoprotein ( HDL ) . Overall , the findings indicate that strain TN8 exhibited a number of attractive properties that might open new promising opportunities for the improvement of various parameters related to animal health performance and the avoidance of antibiotics and drugs as promoting factors . Antiinflammatory effect of lactic acid bacteria : inhibition of cyclooxygenase-2 by suppressing nuclear factor-kappaB in Raw264.7 macrophage cells . Lactobacillus casei 3260 ( L. casei 3260 ) was evaluated in relation to the inflammatory response mediated by lipopolysaccharide ( LPS ) -induced nuclear factor-kappaB ( NF-kappaB ) and cyclooxygenase-2 ( P35354 ) expression in Raw264.7 macrophage cells . The treatment of Raw264.7 cells with L. casei 3260 significantly inhibited the secretion of tumor necrosis factor-alpha ( P01375 ) and prostaglandins E2 ( DB00917 ) , followed by suppression of P35354 . To clarify the molecular mechanism , the inhibitory effect of L. casei 3260 on the NF-kappaB signaling pathway was examined based on the luciferase reporter activity . Although the treatment of Raw264.7 cells with L. casei 3260 did not affect the transcriptional activity of NF-kappaB , it did inhibit NF-kappaB activation , as determined by the cytosolic p65 release and degradation of I-kappaBalpha . Therefore , these findings suggest that the suppression of P35354 through inhibiting the NF-kappaB activation by LPS may be associated with the antiinflammatory effects of L. casei 3260 on Raw264.7 cells . Structure-function studies of adenine nucleotide transport in mitochondria . II . Biochemical analysis of distinct P18440 and P11245 proteins in yeast . P18440 and P11245 genes in yeast each encode functional ADP/ DB00171 carrier ( AAC ) proteins of the mitochondrial inner membrane . In the present study , mitochondria harboring distinct AAC proteins and the pet9 Arg96 to P42357 mutant ( Lawson , J. , Gawaz , M. , Klingenberg , M. , and Douglas , M. G. ( 1990 ) J. Biol. Chem. 265 , 14195-14201 ) protein have been characterized . In addition , properties of the different AAC proteins have been defined following reconstitution into proteoliposomes . Deletion of P11245 but not P18440 causes a major reduction in the mitochondrial cytochrome content and respiration , and this level remains low even when the level of P18440 protein is increased to 20 % that of the P11245 gene product . In reconstitution studies , the rate of nucleotide transport by isolated P18440 protein is approximately 40 % that of the P11245 protein . Thus , the lack of mitochondrial-dependent growth supported by the P18440 gene product alone may be due to the combination of low abundance and reduced activity . Surprisingly , analysis of the Arg96 to DB00117 mutant protein revealed binding and transport activities similar to the functional P18440 and P11245 gene products . These observations are discussed in relation to a molecular analysis of this highly conserved small transporter and its function in conjunction with other proteins in the mitochondrial membrane . Direct and irreversible inhibition of cyclooxygenase-1 by nitroaspirin ( DB05822 ) . Benzoic acid , 2-(acetyl-oxy)-3-[(nitrooxy)methyl]phenyl ester ( DB05822 ) , a new drug made by an aspirin molecule linked , through a spacer , to a nitric oxide ( NO ) -donating moiety , is now under clinical testing for the treatment of atherothrombotic conditions . DB00945 exerts its antithrombotic activity by irreversibly inactivating platelet cyclooxygenase ( P36551 ) -1 . DB05822 in vivo undergoes metabolism into deacetylated and/or denitrated metabolites , and it is not known whether DB05822 needs to liberate aspirin to inhibit P23219 , or whether it can block it as a whole molecule . The aim of our study was to evaluate the effects of DB05822 and its analog or metabolites on platelet P23219 and whole blood P35354 and on purified ovine P36551 ( oCOX ) -1 and oCOX-2 . In particular , we have compared the mechanism by which DB05822 inhibits purified oCOX enzymes with that of aspirin using a spectrophotometric assay . All the DB05822 derivatives containing acetylsalicylic acid inhibited the activity of oCOX-1 and oCOX-2 , whereas the deacetylated metabolites and the nitric oxide-donating moiety were inactive . Dialysis experiments showed that oCOX-1 inhibition by DB05822 , similar to aspirin , is irreversible . Reversible P36551 inhibitors ( indomethacin ) or salicylic acid incubated with the enzyme before DB05822 prevent the irreversible inhibition of oCOX-1 by DB05822 as well as by aspirin . In conclusion , our data show that DB05822 acts as a direct and irreversible inhibitor of P23219 and that the presence of a spacer and NO-donating moiety in the molecule slows the kinetics of P23219 inhibition by DB05822 , compared with aspirin . Growth arrest and forced differentiation of human primary glioblastoma multiforme by a novel small molecule . Glioblastoma multiforme is the most common malignant brain tumor in adults , with an average survival of less than one year due to its resistance to therapy . Recent studies reported that GBM initiates from CD133-expressing cancer stem cells ( CSC ) . However , the efficacy of CSC targeting is limited . A newly developed approach in cancer treatment is the forced differentiation of cancer cells . Here , we show that the treatment of the novel small molecule , CG500354 , into CD133-expressing human primary GBM cells induces growth arrest by cell cycle regulators , p53 , P38936 , p27 and phase-specific cyclins , and neural differentiation , as confirmed by neural progenitor/precursor markers , nestin , P14136 and Tuj1 . When GBM-derived cells caused the tumors in NOD/SCID mice , CG500354 induced GBM-derived cells differentiation into Tuj1 and P14136 expressing cells . We next demonstrated that CG500354 plays a tumor-suppressive role via DB02527 /CREB signaling pathway . CG500354 increases not only the extracellular DB02527 level but also the protein level of PKA and CREB . Additionally , both mimetic substances , DB02587 and DB01954 , revealed comparable results with CG500354 . Our findings indicate that induction of growth arrest and neural differentiation via DB02527 /CREB signaling pathway by CG500354 treatment suggests the novel targeting of Q08499 in the development of new drugs for brain tumor therapy . Acute kidney injury and inflammatory immune reconstitution syndrome in mixed genotype ( A/E ) hepatitis B virus co-infection in HIV-associated lymphoma . We report a first case of HIV-associated lymphoma ( P42357 ) presenting with acute kidney injury ( AKI ) and inflammatory immune reconstitution syndrome ( IRIS ) . A 39-year-old male , treated with nonsteroidal anti-inflammatory drugs ( NSAIDs ) for one month prior to admission , developed AKI , left testicular tumor , and recurrent swelling of the right parotid gland . A resected testicular tumor exhibited features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma . Renal biopsy showed hydro-degeneration of renal tubules , interstitial inflammatory cells , and a small number of lymphoma cells in the sub-capsule , compatible with acute interstitial nephritis . DB00117 renal dysfunction rapidly recovered following chemotherapy and combination antiretroviral therapy ( cART ) . He developed pneumonia concomitantly with a decrease in HIV-RNA level and an increase in P01730 + cells after the first cycle of chemotherapy , which spontaneously resolved after the second cycle of chemotherapy without additional anti-infection drugs ; thus , his pneumonia fulfilled the diagnostic criteria for IRIS . We suggest that IRIS may frequently develop during chemotherapy for P42357 , but may be overlooked . He was coinfected with hepatitis B virus ( HBV ) , which genotypes known as is associated with liver-related mortality and response to antiviral therapy ; recently , an intimate interplay between HIV and HBV in the onset of lymphoma has been reported . Therefore , we addressed the HBV genotype in the patient . The analysis revealed that he exhibited a mixed genotype ( A/E ) not native to Japan and primarily found in Europe and North America or West Africa . These findings suggest that universal vaccination for juveniles against HBV is warranted in Japan . Expression of N-acetyltransferase in monocyte-derived dendritic cells . Dendritic cells ( DCs ) are known to internalize , process , and present low-molecular-weight chemicals to T cells in the course of the sensitization and elicitation phase of allergic contact dermatitis . Thus , DCs may be involved in metabolic activation and detoxification of haptens and thereby influence the quantity of immunogens inducing sensitization . Recently , the cytochrome P-450 enzymes expressed in monocyte-derived dendritic cells ( MoDCs ) were characterized . In the present study , N-acetyltransferase 1 and 2 ( NAT-1 and -2 ) mRNA expression and N-acetylation capacities of these cells were investigated . Monocytes from healthy donors were incubated with granulocyte-monocyte colony-stimulating factor ( GM- P04141 ) and interleukin ( IL ) -4 for 6 d and the resulting immature MoDCs were characterized by flow cytometry . Total RNA from MoDCs was isolated , reverse transcribed , and polymerase chain reaction ( PCR ) for NAT-1 and P11245 mRNA was performed . Data showed the presence of mRNA for NAT-1 ( 9 of 10 donors ) and P11245 ( 8 of 10 donors ) in these cells . NAT-1 enzyme activities were achieved through acetylation of para-aminobenzoic acid ( PABA ) by MoDC cell lysates and activities varied between 23.4 and 26.6 nmol/mg/min . In addition , complete cell acetylation of para-phenylenediamine ( PPD ) , estimated via analysis of monoacetyl-PPD ( MAPPD ) and diacetyl-PPD ( DAPPD ) in cell culture supernatants , confirmed that in vitro generated MoDCs ( 4 of 6 donors ) express metabolic active N-acetyltransferase ( NAT-1 ) . In the case of PPD , our results emphasize that N-acetylation status may influence the amounts of immunogens available for sensitization to PPD . Inhibitors of arachidonic acid metabolism reduce DNA and nuclear fragmentation induced by P01375 plus cycloheximide in U937 cells . U937 human myeloid leukemia cells are induced to apoptosis by tumour necrosis factor ( P01375 ) plus cycloheximide ( CHX ) . We have analysed the effect of various inhibitors of the arachidonic acid ( AA ) metabolism on several features of this process . The formation of high molecular weight and oligonucleosomal DNA fragments as well as nuclear fragmentation were reduced by inhibitors of P09917 ( BWA4C and BWB70C ) , P09917 activating protein ( MK-886 ) , and cytosolic P04054 ( AACOCF3 ) . None of these agents blocked the morphological changes detected by microscopy or flow cytometry , phosphatidylserine exposure on the cell surface or Caspase 3-like activation . AA also induced nuclear fragmentation at a concentration of 1-20 microM . However , the mechanisms by which these inhibitors act , remain unexplained since there was no P09917 expression in the U937 cells and no AA release followed their stimulation with P01375 plus CHX . Molecular basis of P10768 *5 and P10768 *7 and haplotype analysis with new polymorphisms in introns . The two polymorphic alleles of esterase D ( P10768 ) , P10768 *5 and P10768 *7 , are specific to Europeans and Asians , respectively . In this study the molecular basis was characterized : P10768 *5 , arising from P10768 *1 , has a G to A transition , resulting in Gly257( P19440 ) --> DB00128 ( Q6IB77 ) ; and P10768 *7 , originating from P10768 *2 , has an A to G transition , resulting in Asp231( Q6IB77 ) --> DB00145 ( P19440 ) . Glycine is also involved in the common P10768 *1/ P10768 *2 polymorphism [ Gly190(GGA) --> DB00142 ( P10253 ) ] . Haplotype analysis using a few novel intragenic polymorphisms showed strong associations among polymorphic sites , suggesting that recombination has been less frequent in the human P10768 gene , although it spans about 25 kb from exon 1 to exon 10 . A marked difference was observed in the distribution of haplotype frequencies between Germans and Japanese . Role of endotoxin in acute inflammation induced by gram-negative bacteria : specific inhibition of lipopolysaccharide-mediated responses with an amino-terminal fragment of bactericidal/permeability-increasing protein . A recombinant 23-kDa amino-terminal fragment of human bactericidal/permeability-increasing protein ( rBPI23 ) , a potent lipopolysaccharide ( LPS ) -binding/neutralizing protein , was used as a probe to assess the role of endotoxin in the acute inflammatory responses elicited by gram-negative bacteria in rat subcutaneous air pouches . In initial experiments , rBPI23 prevented the Escherichia coli O111:B4 LPS-induced accumulation of polymorphonuclear leukocytes ( PMN ) , tumor necrosis factor alpha ( P01375 ) , and nitrite ( a stable end product of nitric oxide formation ) in exudate fluids . Significant inhibition of P01375 production was still evident when rBPI23 treatment was delayed for 30 min after LPS instillation . In subsequent experiments , rBPI23 also prevented the nitrite and early ( 2-h ) P01375 accumulation induced by three different strains of formaldehyde-killed gram-negative bacteria ( E. coli O7: P04264 , E. coli O111:B4 , and Pseudomonas aeruginosa 12.4.4 ) but did not inhibit the PMN or late ( 6-h ) P01375 accumulation induced by these bacteria . As with LPS challenge , a significant inhibition of early P01375 production was still evident when rBPI23 treatment was delayed for 30 to 60 min after instillation of killed bacteria . The results indicate that in this experimental model the NO and early P01375 responses to gram-negative bacterial challenge are mediated predominantly by endotoxin , whereas the PMN and late P01375 responses may be mediated by other bacterial components . Moreover , the results indicate that rBPI23 can inhibit the bacterially induced production of certain potentially harmful mediators ( P01375 and NO ) without entirely blocking the host defense , i.e. , PMN response , against the bacteria . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Essential role of human leukocyte antigen-encoded proteasome subunits in NF-kappaB activation and prevention of tumor necrosis factor-alpha-induced apoptosis . The multisubunit proteasome complex is the principal mediator of nonlysosomal protein degradation . The proteasome subunit varies minimally between cells with the exception of P28065 , P28062 , and P40306 subunits in rodent and human cells . P28065 and P28062 subunits are encoded by the human lymphocyte antigen region , and they optimize proteolytic mediated antigen presentation . The proteasome is also important for the function of transcription factor nuclear factor-kappaB ( NF-kappaB ) . It is required for NF-kappaB subunits p50 and p52 generation and catalyzes degradation of phosphorylated P25963 . These proteasome-mediated reactions have now been shown to be defective in P24752 cells , a human lymphocyte cell line that lacks both P28065 and P28062 . Although P24752 cells contain normal expression of Q9ULW0 and Q14511 , the abundance of p50 and p52 was greatly reduced . P01375 -alpha ( P01375 ) induced normal phosphorylation of P25963 but failed to induce degradation of phosphorylated P25963 . Both DNA binding assays and luciferase assays revealed that P01375 -induced NF-kappaB activation is defective in P24752 cells . Unlike parental cells , P24752 cells were susceptible to P01375 -induced apoptosis . These data indicate human leukocyte antigen-linked proteasome subunits are essential for NF-kappaB activation and protection of cells from P01375 -induced apoptosis . Inhibition of cervical lymph node metastasis by marimastat ( DB00786 ) in an orthotopic oral squamous cell carcinoma implantation model . Activation of matrix metalloproteinase-2 ( P08253 ) is a common event in head and neck squamous cell carcinoma . An P48449 -19 cell line , derived from human oral squamous cell carcinoma and known to metastasize to cervical lymph nodes , was implanted into the lingual margin of mice . The effect of marimastat ( DB00786 ) , a broad MMP inhibitor , on the suppression of regional cervical lymph node metastasis was evaluated with an orthotopic implantation nude mice model . Marimastat was given immediately after P48449 -19 implantation and continuously administered by an osmotic pump . The mice were divided into three groups by marimastat dose ; Group A ; 0 mg/kg/day , Group B ; 30 mg/kg/day , and Group C ; 150 mg/kg/day . Twenty-one days after implantation , primary oral tumors and cervical lymph nodes were resected . Cervical lymph node status was microscopically examined . Activation of P08253 in primary oral tumor was examined by gelatin zymography . Both cervical lymph node metastasis and activation of P08253 were significantly suppressed in Group C ( P < 0.05 ) . Moreover , the Group C mice had a significantly better survival than group A ( P = 0.0026 ) . There was a significant difference between Group A and Group C in terms of proliferation of tumor cells by proliferating cell nuclear antigen immunostaining ( P = 0.0120 ) . These results suggest a positive role for marimastat in the inhibition of P08253 activation and prevention of cervical lymph node metastasis in oral squamous cell carcinoma ( OSCC ) . Improvement of survival in patients with OSCC could be expected using adjuvant therapy with marimastat . Vampire bat plasminogen activator DB04925 -alpha-1 ( desmoteplase ) : a thrombolytic drug optimized by natural selection . P00747 activators are enzymes found in all vertebrate species investigated so far . Their physiological function is the generation of localized proteolysis in the context of tissue remodeling , wound healing and neuronal plasticity . The common vampire bat ( Desmodus rotundus ) is a New World species that feeds exclusively on blood . Its saliva contains highly potent plasminogen activators , specialized in rapid lysis of fresh blood clots . Biochemical and pharmacological evidence indicates that these plasminogen activators represent a new class of thrombolytics with pharmacological and toxicological properties superior to human tissue-type plasminogen activator , the clot dissolving agent now most frequently used in medicine . A form of the enzyme produced by recombinant DNA technology is currently employed to test this hypothesis in clinical studies . DB05822 , a nitric oxide-releasing aspirin derivative , exhibits a significant antiproliferative effect and alters cell cycle progression in human colon adenocarcinoma cell lines . DB00435 -releasing non-steroidal antiinflammatory drugs ( NO-NSAIDs ) are safer than NSAIDs due to their ability to reduce gastric toxicity . We assessed the cytotoxic activity of a new aspirin derivative , DB05822 , after different exposure schedules , in three human colon adenocarcinoma cell lines . All the lines were positive for P23219 protein and mRNA , as evaluated by Western blot and RT-PCR , respectively , while only one was positive for P35354 . The cytostatic and cytocidal activity was determined by sulforhodamine B assay and evaluated according to Monks ' model . Cytostatic activity was observed after a 24-h drug exposure and 50 % growth inhibition was reached at concentrations ranging from 165 to 250 micro M in all cell lines , whereas with aspirin the IC50 was never reached , even at the maximum concentration tested ( 500 micro M ) , and was independent of P23219 or P35354 status . Cytocidal activity was observed only at the highest concentrations and persisted for a long time after drug removal . Flow cytometric analysis showed that the NO-aspirin compound induced a persistent accumulation of cells in G2-M phase in all the cell lines after at least 48 h exposure . Specifically , the block pertained mainly to G2 phase , whereas mitotic index was not affected at all . Our results indicate that DB05822 has an in vitro cytostatic activity superior to that of its parental aspirin compound , which makes it a potentially important tumor preventive agent . Furthermore , the cytocidal effect observed at the highest concentrations and the induction of a specific block in G2 phase renders it a promising candidate for drug combination treatments . A new hemoglobin variant found during Hb A1c measurement : Hb Hokusetsu [ beta52(D3) DB00128 --> DB00145 ] . A new beta chain variant was accidentally found through the assay of Hb A1c in a diabetic patient . The variant was detected by polyacrylamide gel isoelectrofocusing and electrospray ionization mass spectrometry . For sequence determination , globin was cleaved with combination of trypsin and lysyl endopeptidase and analyzed by high performance liquid chromatography connected to electrospray ionization mass spectrometry . An abnormal betaT-5 peptide was found by reconstructed selected ion monitoring . The collision-induced dissociation spectrum of an ion derived from the abnormal betaT-5 peptide revealed a new substitution , [ beta52(D3) DB00128 --> DB00145 ] , named Hb Hokusetsu . The sequence was confirmed with an automatic sequencer using peptides isolated by reversed phase high performance liquid chromatography . Amplification of the beta-globin exon 2 and nucleotide sequencing revealed a Q6IB77 --> P19440 mutation in codon 52 corresponding to an DB00128 --> DB00145 replacement . Electrospray ionization mass spectrometry analysis of the hemolysate showed a reasonable value of 10.4 % for glycated globin . The variant migrated as Hb S on isoelectrofocusing . Hematological analysis revealed normal parameters . The patient 's hemolysate showed normal stability in the isopropanol test . DB09140 equilibrium studies on the patient 's red blood cells and hemolysate showed no significant change in oxygen affinity or cooperativity .

MH_train_1221

MH_train_1221

MH_train_1221

interacts_with DB00338?

multiple_choice

Multi-kinase inhibition in ovarian cancer . DB00398 ( Nexavar ) is a multi-kinase inhibitor that was developed as an inhibitor of RAF-1 , in the P27361 /2 pathway , but which was subsequently shown to inhibit class III tyrosine kinase receptors . ( 1 ) More recently regorafenib ( Stivarga ) has been developed , which is a further fluorinated version of sorafenib with greater bioavailability and similar inhibitory properties against RAF-1/class III RTKs . ( 2 ) Some of the anti-tumor effects of sorafenib have been ascribed to anti-angiogenic actions of this agent on endothelial associated kinases such as P35968 . Other effects of sorafenib clearly have to be due to its effects on the inherent biology of the tumor cells themselves . For example , through various mechanisms sorafenib has been shown in the laboratory and the clinic to suppress expression of the protective protein Q8WXI8 -1 . ( 3 ) DB00398 has also been linked to inhibition of P40763 , NFκB , and activation of the death receptor CD95 . ( 4 ) DB00398 is routinely dosed daily ( 400 mg P55957 ) and 7 d after the start of dosing has a Cmax of ~21 μM with a nadir at 12 h of ~10 μM , and is a highly protein bound based on in vitro assays . ( 5 ) Despite this in vitro binding data sorafenib has profound in vivo effects on tumor cells in renal carcinoma and hepatocellular carcinoma patients ; cells which are not per se addicted to high activity oncogene signals that are targets of sorafenib/regorafenib . Thus the precise stable bioavailable level of sorafenib/regorafenib in patient plasma is not known . Pharmacokinetic profiles of the novel P35354 selective inhibitor cimicoxib in dogs . DB05095 ( CX ) is a novel imidazole derivative that is a cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drug and the latest P35354 selective inhibitor to be released for veterinary use . Currently there is limited information available on the pharmacokinetic ( PK ) properties of CX . The aim of the current study was to evaluate the PK features of CX after administration of the recommended dose and after administration of a more variable dose rate in the form of the commercially available tablet . In addition , the effects of food intake on the PK properties were also evaluated . In the first study , five healthy Beagle dogs received 2mg/kg CX via the oral route following a period of fasting . The second study was conducted using six healthy Labrador retriever dogs which each received an 80 mg tablet ( approximate dose 1.95-2.5mg/kg ) using a crossover design , both in the fasted and fed condition . The plasma concentrations of CX were detected by a validated HPLC method . No adverse effects were observed in any dogs during the experiment . The results from the PK analysis were similar between the studies , regardless of precision of dose and fasted and fed conditions . The mean peak concentration of CX was 0.49 and 0.43 μg/mL under fasted and fed conditions , respectively . The mean half-life was about 3h after all treatments . In addition , simulated multiple dosing data revealed that time over minimal effective concentration was similar after 1.95 , 2.0 and 2.5mg/kg dose administrations . These findings suggest that slight variation from the recommended dose should not alter the therapeutic outcome . In addition , CX can be administered to fed dogs without significantly affecting blood levels . Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Cytokine signaling via various transcription factors regulates receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) -mediated osteoclast differentiation from monocyte/macrophage lineage cells involved in propagation and resolution of inflammatory bone destruction . Protein inhibitor of activated P40763 ( Q9Y6X2 ) was initially identified as a molecule that inhibits DNA binding of P40763 and regulates many transcription factors through distinct mechanisms . To analyze Q9Y6X2 function in osteoclasts in vivo , we have generated transgenic mice in which Q9Y6X2 is specifically expressed in the osteoclast lineage using the tartrate-resistant acid phosphatase ( TRAP ) gene promoter . Q9Y6X2 transgenic mice showed an osteopetrotic phenotype due to impairment of osteoclast differentiation . Overexpression of Q9Y6X2 in RAW264.7 cells suppressed O14788 -induced osteoclastogenesis by inhibiting the expression of c-Fos and O95644 . Interestingly , Q9Y6X2 inhibits the transcriptional activity of microphthalmia-associated transcription factor ( O75030 ) independent of sumoylation . Down-regulation of Q9Y6X2 markedly enhances O14788 -mediated osteoclastogenesis in RAW264.7 cells . Furthermore , overexpression of Q9Y6X2 in mouse primary osteoblast ( DB00925 ) , down-regulates O14788 expression induced by interleukin-6 ( P05231 ) cytokine family , and inhibits osteoclast formation from bone marrow macrophages ( BMMs ) in vitro coculture system . Down-regulation of Q9Y6X2 leads to the accelerated expression of O14788 in DB00925 stimulated with P05231 and soluble P05231 receptor ( sIL-6R ) . Taken together , our results clearly indicate that Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Clinical development of eniluracil : current status . DB03516 is a potent inactivator of dihydropyrimidine dehydrogenase ( Q12882 ) , which is the first enzyme in the degradative pathway of systemically administered 5-fluorouracil ( DB00544 ) . Two completely oral regimens of eniluracil plus DB00544 are being evaluated in clinical trials : ( 1 ) a chronic schedule with both agents administered P55957 in a 10:1 ratio for 28 days of a 5-week course , and ( 2 ) a 5-day schedule of eniluracil once daily on days 1 through 7 and DB00544 once daily on days 2 through 6 . The clinical development of eniluracil is being pursued in several tumor types , including colorectal cancer , breast cancer , and pancreatic cancer . Response rates achieved in a phase II study of the chronic schedule of oral eniluracil/ DB00544 in patients with colorectal cancer compare favorably with those obtained in trials of intravenous DB00544 and leucovorin , while results from other trials are awaited . Safety analysis for the 28-day schedule has revealed a low incidence of severe toxicities , particularly as compared with standard DB00544 regimens . Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice . Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined . The immediate phase reaction ( IPR ) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined . The inhibition of IPR by cetirizine and mequitazine were potent , but those by cyproheptadine and diphenhydramine were weak . The later phase reaction ( LPR ) assessed at 24 hours after antigen application was inhibited by chlorpheniramine , oxatomide , ketotifen , mequitazine , emedastine , terfenadine and azelastine . The inhibition of LPR by emedastine was potent , but those by ketotifen and terfenadine were only partial . DB01084 inhibited both IPR and LPR comparably . Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction , and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR . P35367 antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism , which may have an additional benefit for the treatment of allergic diseases . Randomised clinical trial : herbal extract DB05767 in active ulcerative colitis - a double-blind comparison with sustained release mesalazine . BACKGROUND : Andrographis paniculata is an herbal mixture used to treat inflammatory diseases . An extract of the herb , DB05767 , inhibits P01375 -α and IL-1β , and prevents colitis in animal models . AIM : To determine the efficacy and safety of DB05767 in patients with mild-to-moderate ulcerative colitis . METHODS : A randomised , double-blind , multicentre , 8-week parallel group study was conducted using DB05767 1200 mg/day compared with 4500 mg/day of slow release mesalazine ( mesalamine ) granules in patients with mild-to-moderately active ulcerative colitis . Disease activity was assessed at baseline and every 2 weeks for clinical response , and at baseline and 8 weeks by colonoscopy . RESULTS : One hundred and twenty patients at five centres in China were randomised and dosed . Clinical remission and response were seen in 21 % and 76 % of DB05767 -treated patients , and 16 % and 82 % of mesalazine-treated patients . By colonoscopy , remission and response were seen in 28 % and 74 % of DB05767 -treated patients and 24 % and 71 % of mesalazine-treated patients , respectively . There was no significant difference between the two treatment groups . CONCLUSION : DB05767 may be an efficacious alternative to mesalazine in ulcerative colitis . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Regulation of microphthalmia-associated transcription factor O75030 protein levels by association with the ubiquitin-conjugating enzyme hUBC9 . The basic helix-loop-helix/leucine zipper ( bHLH/ Q8N5A5 ) microphthalmia-associated transcription factor ( O75030 ) regulates transcription of genes encoding enzymes essential for melanin biosynthesis in melanocytes and retinal pigmented epithelial cells . To determine how O75030 activity is regulated , we used the yeast two-hybrid system to identify proteins expressed by human melanoma cells that interact with O75030 . The majority of clones that showed positive interaction with a 158-amino-acid region of O75030 containing the bHLH/ Q8N5A5 domain ( aa 168-325 ) encoded the ubiquitin conjugating enzyme hUBC9 . The association of O75030 with hUBC9 was further confirmed by an in vitro Q86UG4 pull-down assay . Although hUBC9 is known to interact preferentially with SENTRIN/ P63165 , in vitro transcription/translation analysis demonstrated greater association of O75030 with ubiquitin than with SENTRIN . Importantly , cotransfection of O75030 and hUBC9 expression vectors resulted in O75030 protein degradation . O75030 protein was stabilized by the proteasome inhibitor MG132 , indicating the role of the ubiquitin-proteasome system in O75030 degradation . DB00133 73 , which is located in a region rich in proline , glutamic acid , serine , and threonine ( PEST ) , regulates O75030 protein stability , since a serine to alanine mutation prevented hUBC9-mediated O75030 ( S73A ) degradation . Furthermore , we identified lysine 201 as a potential ubiquitination site . A lysine to arginine mutation abolished O75030 ( K201R ) degradation by hUBC9 in vivo . Our experiments indicate that by targeting O75030 for proteasome degradation , hUBC9 is a critical regulator of melanocyte differentiation . FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . Organelle-specific expression of subunit P03915 of human complex I ( DB00157 dehydrogenase ) alters cation homeostasis in Saccharomyces cerevisiae . The P03915 component of the respiratory complex I is a large , hydrophobic subunit encoded by the mitochondrial genome . Its bacterial homologue , the NDH-1 subunit NuoL , acts as a cation transporter in the absence of other NDH-1 subunits . Mutations in human P03915 are frequently observed in neurodegenerative diseases . Wild type and mutant variants of P03915 fused to GFP or a FLAG peptide were targeted to the endoplasmatic reticulum ( ER ) or the inner mitochondrial membrane of Saccharomyces cerevisiae , which lacks an endogenous complex I . The localization of P03915 fusion proteins was confirmed by microscopic analyses of S. cerevisiae cells , followed by cellular fractionation and immunostaining . The impact of the expression of P03915 fusion proteins on the growth of S. cerevisiae in the presence and absence of added salts was studied . ER-resident P03915 conferred DB01356 sensitivity to S. cerevisiae , which was lost when the E145V variant of P03915 was expressed . All variants of P03915 tested led to increased resistance of S. cerevisiae at high external concentrations of Na(+) or K(+) . The data seem to indicate that P03915 influences the salt homeostasis of S. cerevisiae independent of other complex I subunits , and paves the way for functional studies of mutations found in mitochondrially encoded complex I genes . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Crystallization and preliminary crystallographic analysis of the C-terminal domain of MamM , a magnetosome-associated protein from Magnetospirillum gryphiswaldense Q9UBK8 -1 . MamM is a unique magnetosome-associated protein that shares substantial homology with cation diffusion facilitator ( P05231 ) proteins , a group of heavy-metal-ion efflux transporters that participate in metal-ion homeostasis in all domains of life . Magnetotactic bacteria utilize P05231 proteins in iron-oxide biomineralization and in magnetosome formation . Here , the crystallization and preliminary X-ray analysis of recombinant Magnetospirillum gryphiswaldense MamM is reported . The C-terminal domain of MamM was crystallized in the orthorhombic space group C222(1) , with unit-cell parameters a = 37.1 , b = 94.0 , c = 53.3 Å . X-ray diffraction data were collected to a resolution of 2.0 Å . Inhibition of JAKs in macrophages increases lipopolysaccharide-induced cytokine production by blocking P22301 -mediated feedback . Macrophages are an important source of cytokines following infection . Stimulation of macrophages with TLR agonists results in the secretion of P01375 -α , P05231 , and IL-12 , and the production of these cytokines is controlled by multiple feedback pathways . Macrophages also produce P22301 , which acts to inhibit proinflammatory cytokine production by macrophages via a JAK/ P40763 -dependent pathway . We show in this paper that , DB08877 , a recently described selective inhibitor of JAKs , increases P01375 , P05231 , and IL-12 secretion in mouse bone marrow-derived macrophages stimulated with LPS . This effect is largely due to its ability to block P22301 -mediated feedback inhibition on cytokine transcription in macrophages . Similar results were also obtained with a second structurally unrelated Jak inhibitor , DB08895 . In addition , LPS induced the production of IFN-β , which was then able to activate JAKs in macrophages , resulting in the stimulation of P42224 phosphorylation . The initial induction of P22301 was independent of JAK signaling ; however , inhibition of JAKs did reduce P22301 secretion at later time points . This reflected a requirement for the IFN-β feedback loop to sustain P22301 transcription following LPS stimulation . In addition to P22301 , IFN-β also helped sustain P05231 and IL-12 transcription . Overall , these results suggest that inhibition of JAKs may increase the inflammatory potential of macrophages stimulated with O00206 agonists . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . Plant methionine sulfoxide reductase A and B multigenic families . DB00134 oxidation to methionine sulfoxide ( MetSo ) , which results in modification of activity and conformation for many proteins , is reversed by an enzyme present in most organisms and termed as methionine sulfoxide reductase ( Q9UBK8 ) . On the basis of substrate stereospecificity , two types of Q9UBK8 , A and B , that do not share any sequence similarity , have been identified . In the present review , we first compare the multigenic Q9UBK8 families in the three plant species for which the genome is fully sequenced : Arabidopsis thaliana , Oryza sativa , and Populus trichocarpa . The Q9UBK8 gene content is larger in A. thaliana ( five MSRAs and nine MSRBs ) compared to P. trichocarpa ( five MSRAs and four MSRBs ) and O. sativa ( four MSRAs and three MSRBs ) . A complete classification based on gene structure , sequence identity , position of conserved reactive cysteines and predicted subcellular localization is proposed . On the basis of in silico and experimental data originating mainly from Arabidopsis , we report that some Q9UBK8 genes display organ-specific expression patterns and that those encoding plastidic MSRs are highly expressed in photosynthetic organs . We also show that the expression of numerous Q9UBK8 genes is enhanced by environmental conditions known to generate oxidative stress . Thioredoxins ( TRXs ) constitute very likely physiological electron donors to plant Q9UBK8 proteins for the catalysis of MetSO reduction , but the specificity between the numerous TRXs and methionine sulfoxide reductases ( MSRs ) present in plants remains to be investigated . The essential role of plant MSRs in protection against oxidative damage has been recently demonstrated on transgenic Arabidopsis plants modified in the content of cytosolic or plastidic Q9UJ68 . SUMOylation is required for glycine-induced increases in AMPA receptor surface expression ( ChemLTP ) in hippocampal neurons . Multiple pathways participate in the AMPA receptor trafficking that underlies long-term potentiation ( LTP ) of synaptic transmission . Here we demonstrate that protein SUMOylation is required for insertion of the P42261 AMPAR subunit following transient glycine-evoked increase in AMPA receptor surface expression ( ChemLTP ) in dispersed neuronal cultures . ChemLTP increases co-localisation of P63165 and the SUMO conjugating enzyme Ubc9 and with P78352 consistent with the recruitment of SUMOylated proteins to dendritic spines . In addition , we show that ChemLTP increases dendritic levels of P63165 and Ubc9 mRNA . Consistent with activity dependent translocation of these mRNAs to sites near synapses , levels of the mRNA binding and dendritic transport protein Q9BZB8 are also increased by ChemLTP . Importantly , reducing the extent of substrate protein SUMOylation by overexpressing the deSUMOylating enzyme SENP-1 or inhibiting SUMOylation by expressing dominant negative Ubc9 prevent the ChemLTP-induced increase in both AMPAR surface expression and dendritic P63165 mRNA . Taken together these data demonstrate that SUMOylation of synaptic protein(s) involved in AMPA receptor trafficking is necessary for activity-dependent increases in AMPAR surface expression . Mechanism of inhibition of the P42262 AMPA receptor channel opening by talampanel and its enantiomer : the stereochemistry of the 4-methyl group on the diazepine ring of 2,3-benzodiazepine derivatives . Stereoselectivity of 2,3-benzodiazepine compounds provides a unique way for the design of stereoisomers as more selective and more potent inhibitors as drug candidates for treatment of the neurological diseases involving excessive activity of AMPA receptors . Here we investigate a pair of enantiomers known as DB04982 and its ( + ) counterpart about their mechanism of inhibition and selectivity toward four AMPA receptor subunits or P42261 -4 . We show that DB04982 is the eutomer with the endismic ratio being 14 for the closed-channel and 10 for the open-channel state of P42262 . Kinetic evidence supports that DB04982 is a noncompetitive inhibitor and it binds to the same site for those 2,3-benzodiazepine compounds with the C-4 methyl group on the diazepine ring . This site , which we term as the " M " site , recognizes preferentially those 2,3-benzodiazepine compounds with the C-4 methyl group being in the R configuration , as in the chemical structure of DB04982 . Given that DB04982 inhibits P42261 and P42262 , but is virtually ineffective on the P42263 and P48058 AMPA receptor subunits , we hypothesize that the " M " site(s) on P42261 and P42262 to which DB04982 binds is different from that on P42263 and P48058 . If the molecular properties of the AMPA receptors and DB04982 are used for selecting an inhibitor as a single drug candidate for controlling the activity of all AMPA receptors in vivo , DB04982 is not ideal . Our results further suggest that addition of longer acyl groups to the N-3 position should produce more potent 2,3-benzodiazepine inhibitors for the " M " site . Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation : a novel role of the cysteinyl LT-1 receptor . Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues . Leukotrienes are potent inflammatory mediators , and similar to prostaglandins , are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues . Here we tested the hypothesis that blocking the negative regulation of leukotrienes , would lead to enhanced endochondral bone formation during fracture repair . A closed femoral fracture was created in mice . Animals were divided into three groups for treatment with either montelukast sodium , a cysteinyl leukotriene type 1 receptor antagonist ( trade name Singulair ) , zileuton , a P09917 enzyme inhibitor ( trade name DB00744 ) , or carrier alone . The fractures were analyzed using radiographs , quantitative gene expression , histology and histomorphometry , and immunohistochemistry . Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls . Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7 . Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation , and increased early bone formation markers when compared with controls . Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor , leading to increased chondrocyte proliferation at early time points . These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation , with important and previously unrecognized implications for both fracture repair , and in a broader context , systemic chondrocyte growth and differentiation . O14788 induces components of the extrinsic coagulation pathway in osteoclasts . P00734 is converted to thrombin by factor Xa in the cell-associated prothrombinase complex . P00734 is present in calcified bone matrix and thrombin exerts effects on osteoblasts as well as on bone resorption by osteoclasts . We investigated whether ( 1 ) osteoclasts display factor Xa-dependent prothrombinase activity and ( 2 ) osteoclasts express critical regulatory components upstream of the prothrombinase complex . The osteoclast differentiation factor O14788 induced formation of multinucleated TRAP positive cells concomitant with induction of prothrombinase activity in cultures of RAW 264.7 cells and bone marrow osteoclast progenitors . Expression analysis of extrinsic coagulation factors revealed that O14788 enhanced protein levels of factor Xa as well as of coagulation factor III ( tissue factor ) . Inhibition assays indicated that factor Xa and tissue factor were involved in the control of prothrombinase activity in O14788 -differentiated osteoclasts , presumably at two stages ( 1 ) conversion of prothrombin to thrombin and ( 2 ) conversion of factor X to factor Xa , respectively . Activation of the extrinsic coagulation pathway during osteoclast differentiation through induction of tissue factor and factor Xa by a O14788 -dependent pathway indicates a novel role for osteoclasts in converting prothrombin to thrombin . Congenital cataract , muscular hypotonia , developmental delay and sensorineural hearing loss associated with a defect in copper metabolism . Deficiencies of different proteins involved in copper metabolism have been reported to cause human diseases . Well-known syndromes , for example , are Menkes and Wilson diseases . Here we report a patient presenting with congenital cataract , severe muscular hypotonia , developmental delay , sensorineural hearing loss and cytochrome-c oxidase deficiency with repeatedly low copper and ceruloplasmin levels . These findings were suggestive of a copper metabolism disorder . In support of this , the patient 's fibroblasts showed an increased copper uptake with normal retention . Detailed follow-up examinations were performed . Immunoblotting for several proteins including Q04656 ( Q04656 or Menkes protein ) , P35670 ( Wilson protein ) and P00441 showed normal results , implying a copper metabolism defect other than Wilson or Menkes disease . Sequence analysis of O00244 and genes coding for proteins that are known to play a role in the mitochondrial copper metabolism ( P00395 -III , O75880 , O43819 , Q9Y6N1 , Q14061 , Q49B96 ) revealed no mutations . Additional disease genes that have been associated with cytochrome-c oxidase deficiency were negative for mutations as well . As beneficial effects of copper histidinate supplementation have been reported in selected disorders of copper metabolism presenting with low serum copper and ceruloplasmin levels , we initiated a copper histidinate supplementation . Remarkable improvement of clinical symptoms was observed , with complete restoration of cytochrome-c oxidase activity in skeletal muscle . Mitochondrial DNA fragments released through the permeability transition pore correspond to specific gene size . In the present work , we show that after induction of mitochondrial damage by oxidative stress , in the presence of calcium , matrix DNA content decreased to 42+/-6 % . Mitochondrial damage was analyzed by measuring aconitase activity , a marker enzyme of mitochondrial oxidative stress . The genes were identified by amplifying them through the polymerase chain reaction ( PCR ) , using specific primers for each mitochondrial gene ( P00395 , P00403 , P00414 , P03897 , P03915 , P00846 , P03928 , and P00156 ) . The results show that after oxidative stress , the amount of P00395 , P03897 , and P00156 genes in the mitochondria approximately decreased by 46 , 22 , and 54 % , respectively . This effect was inhibited in the presence of cyclosporin A . These genes were found outside the mitochondria after permeability transition was induced . Mitochondrial integrity was evaluated by observing the activity of adenylate kinase and malate dehydrogenase .

MH_train_1222

MH_train_1222

MH_train_1222

interacts_with DB01037?

multiple_choice

Proliferation of endothelial cell on polytetrafluoroethylene vascular graft materials carried P15692 gene plasmid . OBJECTIVE : To investigate whether vascular endothelial growth factor ( P15692 ) gene plasmid carried by polytetrafluoroethylene ( PTFE ) vascular graft materials could transfect endothelial cells ( ECs ) and promote their growth . METHODS : PTFE vascular graft materials carried with pCDI-hVEGF(121) , pCDI or pEGFP were incubated in DB03754 -buffer solution and the values of optical density of 260 nm at different time were plotted , then the DNA controlled release curve was made . ECs derived from human umbilical vein were seeded on the pCDI-hVEGF(121)/pCDI/pEGFP-PTFE materials or tissue culture plates , ECs numbers were counted and P15692 protein concentrations at different time were measured by enzyme-linked immunoadsorbent assay method . Green fluorescent protein ( GFP ) expression in ECs on pEGFP-PTFE materials was examined with fluorescence microscopy . RESULTS : The controlled release curve showed that the gene released from PTFE materials was rapid within 8 h , then slowed down and that the gene released continuously even after 72 h . At 24 , 72 and 120 h , ECs number and proliferation rate of pCDI-hVEGF(121)-PTFE materials were higher than those of pCDI or pEGFP-PTFE materials ( P < 0.05 ) . P15692 protein concentration of pCDI-hVEGF(121)-PTFE materials was higher than that of pCDI or pEGFP-PTFE materials at 6 , 24 , 72 and 120 h ( P < 0.01 ) . GFP expression in ECs on the pEGFP-PTFE materials could be detected by fluorescence microscopy . CONCLUSION : PTFE graft can be used as a carrier of P15692 gene plasmid , P15692 gene carried by PTFE can transfect ECs and promote ECs growth . PPARγ and RXR ligands disrupt the inflammatory cross-talk in the hypoxic breast cancer stem cells niche . Cancer stem cells ( CSCs ) are affected by the local micro-environment , the niche , in which inflammatory stimuli and hypoxia act as steering factors . Here , two nuclear receptors ( NRs ) agonists , i.e. pioglitazone ( PGZ ) , a ligand of peroxisome proliferator activated receptor-γ , and 6-OH-11-O-hydroxyphenanthrene ( IIF ) , a ligand of retinoid X receptors , were investigated for their capability to interference with the cross-talk between breast CSCs and the niche compartment . We found that IIF potentiates the ability of PGZ to hamper the mammospheres-forming capability of human breast tumours and MCF7 cancer cells , reducing the expression of CSCs regulatory genes ( Notch3 , P78504 , O43623 , P05231 , P02649 , Hypoxia inducible factor-1α and Q16790 ) . Notably , these effects are not observed in normal-MS obtained from human breast tissue . Importantly , NRs agonists abolish the capability of hypoxic MCF7 derived exosomes to induce a pro-inflammatory phenotype in mammary glands fibroblasts . Moreover , NRs agonist also directly acts on breast tumour associated fibroblasts to downregulate nuclear factor-κB pathway and metalloproteinases ( P08253 and P14780 ) expression and activity . In conclusion , NRs agonists disrupt the inflammatory cross-talk of the hypoxic breast CSCs niche . Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . P06401 modulator DB05366 induces extracellular matrix metalloproteinase inducer in cultured human uterine leiomyoma cells . Effects of progesterone receptor modulator DB05366 on the expression of the extracellular matrix ( Q13201 ) components were examined in cultured human uterine leiomyoma and myometrial cells . Q13201 metalloproteinase inducer ( P35613 ) , matrix metalloproteinases ( MMPs ) , tissue inhibitors of MMP ( TIMPs ) and collagen levels were assessed by Western blot analysis , MMP activity assay and real-time RT-PCR . RNA interference ( RNAi ) of P35613 was performed using small interfering mRNA . In cultured leiomyoma cells , DB05366 treatment at concentrations greater than or equal to 10(-8) M significantly increased P35613 , P03956 and P22894 protein contents and P03956 , P08253 , P08254 and P14780 mRNA levels , and activity of P03956 , P08253 , P08254 and P14780 in the medium . P01033 and P16035 were significantly decreased at mRNA and protein levels by DB05366 treatment at concentrations > or =10(-7) M in these cells . DB05366 treatment decreased types I and III collagen protein contents . However , DB05366 treatment did not affect the Q13201 component expression in cultured myometrial cells . RNAi of P35613 abrogated DB05366 -mediated both induction of MMPs and reduction of TIMPs and collagens in cultured leiomyoma cells . These results suggest that DB05366 modulates the expression of P35613 , MMPs , TIMPs and collagens in cultured leiomyoma cells without comparable effects on myometrial cells . DB04873 ( SB 207266 ) , a selective Q13639 receptor antagonist , reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle . The aim of this study is to evaluate the potency of piboserod ( SB 207266 ) , a selective 5-HT(4) receptor antagonist , at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin ( 5-HT ) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations ( O43281 ) . Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and O43281 ( 20 Hz , 1 ms duration at 300 mA for 5 s ) was applied continuously at 1 min intervals . After stabilization of the O43281 -induced contractions , concentration-response curves to 5-HT ( 0.1 nM-100 microM ) were constructed in the absence or presence of 1 or 100 nM of piboserod . The experiments were performed in the presence of methysergide ( 1 microM ) and ondansetron ( 3 microM ) to block 5HT(1)/5HT(2) and 5-HT(3) receptors , respectively . 5-HT potentiated the contractile responses to O43281 of human bladder strips in a concentration-dependent manner , with a maximum mean of 60.0+/-19.9 % of the basal O43281 -evoked contractions . DB04873 did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to O43281 . In presence of 1 and 100 nM of piboserod , the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7 % , respectively . A mean apparent antagonist dissociation constant value ( K(B) ) of 0.56+/-0.09 nM was determined . These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally-mediated contractions of isolated human bladder strips . Therefore , the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder . DB00472 pharmacogenetics in child and adult populations . Although fluoxetine is useful in the treatment of major depression , 30-40 % of the patients do not respond to therapy . The response seems to be influenced by certain genes which are involved in the drug 's pharmacodynamics and pharmacokinetics . The present study reviews the literature on genetic contributions to fluoxetine response in children and adults , and concludes that the different polymorphisms of P10635 and P11712 may influence the blood concentrations of fluoxetine . If the childhood dose is adjusted for weight , differences between children and adults are unlikely . As regards the genes that influence the drug 's pharmacodynamics , polymorphisms of P31645 , P08908 and P21397 seem to be involved in the response to fluoxetine , while the genes P21964 , P34998 , PDEA1 , PDEA11 P49841 and serpin-1 also seem to play a role . Comparison of different studies reveals that the results are not always consistent , probably due to methodological differences . Other factors such as gender or ethnicity may also influence treatment response . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Extended kindred with recessive late-onset Alzheimer disease maps to locus 8p22- P38936 .2 : a genome-wide linkage analysis . Late-onset Alzheimer disease ( LOAD ) is a complex genetic disorder . Although genes involved in early-onset forms were discovered more than a decade ago , LOAD research has only been able to point out small effect loci , with the exception of P02649 . We mapped the gene predisposing to LOAD in an extended inbred family coming from a genetically isolated region ( 24 sampled individuals , 12 of whom are affected ) , completing a genome-wide screen with an Affymetrix10 K single nucleotide polymorphism microarray . Genotyping results were evaluated under model-dependent ( dominant and recessive ) and model-free analysis . We obtained a maximum nonparametric linkage score of 3.24 ( P=0.00006 ) on chromosome 8p22- P38936 .2 . The same genomic position also yielded the highest multipoint heterogeneity LOD ( HLOD ) under a recessive model ( HLOD=3.04 ) . When we compared the results of the model-dependent analysis , a higher score was obtained in the recessive model ( 3.04 ) than in the dominant model ( 1.0 ) . This is a new locus identified in LOAD , in chromosome 8p22- P38936 .2 and encompassing several candidate genes , among them P10909 and P48454 that were excluded by sequencing . The finding of a recessive model of inheritance , consistent with the assumption of inbreeding as a morbidity factor in this population , supports the notion of a role of recessive genes in LOAD . SAR and biological evaluation of analogues of a small molecule histone deacetylase inhibitor N-(2-aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzamide ( DB05651 ) . Analogues of the clinical compound DB05651 ( A ) were designed and synthesized . These compounds inhibit recombinant human Q13547 with IC(50) values in the sub-micromolar range . In human cancer cells growing in culture these compounds induce hyperacetylation of histones , cause expression of the tumor suppressor protein P38936 ( P38936 /CIP1) , and inhibit cellular proliferation . Lead molecule of the series , compound 25 is metabolically stable , possesses favorable pharmacokinetic characteristics and is orally active in vivo in different mouse tumor xenograft models . Hidden risk genes with high-order intragenic epistasis in Alzheimer 's disease . Meta-analysis of data from genome-wide association studies ( GWAS ) of Alzheimer 's disease ( AD ) has confirmed the high risk of P02649 and identified twenty other risk genes/loci with moderate effect size . However , many more risk genes/loci remain to be discovered to account for the missing heritability . The contributions from individual singe-nucleotide polymorphisms ( SNPs ) have been thoroughly examined in traditional GWAS data analysis , while SNP-SNP interactions can be explored by a variety of alternative approaches . Here we applied generalized multifactor dimensionality reduction to the re-analysis of four publicly available GWAS datasets for AD . When considering 4-order intragenic SNP interactions , we observed high consistency of discovered potential risk genes among the four independent GWAS datasets . Ten potential risk genes were observed across all four datasets , including P54750 , Q15413 , Q02763 , Q9BQT8 , LOC729852 , Q8IZU9 , P54829 , P23945 , O60260 , and P08235 . These potential risk genes discovered by generalized multifactor dimensionality reduction are highly relevant to AD pathogenesis based on multiple layers of evidence . The genetic contributions of these genes warrant further confirmation in other independent GWAS datasets for AD . DB00140 ( vitamin B2 ) deficiency impairs P04839 ( Nox2 ) priming and defense against Listeria monocytogenes . DB00140 , also known as vitamin B2 , is converted by riboflavin kinase ( Q969G6 ) into flavin mononucleotide ( Q68DA7 ) and flavin adenine dinucleotide ( DB03147 ) , which are essential cofactors of dehydrogenases , reductases , and oxidases including the phagocytic P04839 ( Nox2 ) . DB00140 deficiency is common in young adults and elderly individuals , who are at the coincidental risk for listeriosis . To address the impact of acute riboflavin deficiency on host defense against Listeria monocytogenes ( L.m. ) , we generated conditional Q969G6 knockout ( KO ) strains of mice . Phagocyte-specific Q969G6 KO impaired the capability of phagocytes to control intracellular L.m. , which corresponded to a greater susceptibility of mice to in vivo challenge with L.m . The oxidative burst of Q969G6 -deficient phagocytes in response to L.m. infection was significantly reduced . Mechanistically , P01375 -induced priming of Nox2 , which is needed for oxidative burst , was defective in Q969G6 -deficient phagocytes . Lack of riboflavin in wild-type macrophages for only 6 h shut down P01375 -induced , Q969G6 -mediated de novo Q68DA7 / DB03147 generation , which was accompanied by diminished ROS production and impaired anti-listerial activity . Vice versa , ROS production by riboflavin-deprived macrophages was rapidly restored by riboflavin supplementation . Our results suggest that acute riboflavin deficiency immediately impairs priming of Nox2 , which is of crucial relevance for an effective phagocytic immune response in vivo . Improved insulin sensitivity by calorie restriction is associated with reduction of P29323 and p70S6K activities in the liver of obese Zucker rats . Calorie restriction ( CR ) improves obesity-related insulin resistance through undefined molecular mechanisms . P06213 substrate ( P41252 ) -1 serine/threonine kinases have been proposed to modulate insulin sensitivity through phosphorylation of P41252 proteins . The aim of this study is to test the hypothesis that changes in the activity of P35568 serine/threonine kinases may underlie the molecular mechanism of CR in improving insulin sensitivity . Obese and lean Zucker rats were subjected to 40 % CR or allowed to feed ad libitum ( AL ) for 20 weeks ; body weight and insulin sensitivity were monitored throughout this period . The activity of P35568 serine/threonine kinases - including JNK , P29323 , P42345 / P08133 (S6K) ( P23443 as listed in the MGI Database ) , glycogen synthase kinase 3beta ( P49841 ) , AMPK ( Q13131 as listed in the MGI Database ) , and protein kinase C ( Q04759 ) in liver tissue extracts was measured by an in vitro kinase assay using various glutathione-S-transferase ( Q86UG4 ) - P35568 fragments as substrates , while phosphorylation of P35568 and serine kinases was determined by western blotting using phosphospecific antibodies . CR in obese rats significantly reduced body weight and increased insulin sensitivity compared to AL controls . DB00133 kinase activity toward P35568 (S612) ( corresponding to S616 in human P35568 ) and P35568 (S632/635) ( corresponding to S636/639 in human P35568 ) was increased in obese rats compared to lean littermates , and was markedly decreased following CR . Concomitantly , obesity increased and CR decreased the activity of hepatic P29323 and P08133 (S6K) against P35568 . The close association between the activity of hepatic P29323 and P08133 (S6K) with insulin resistance suggests an important role for P29323 and P08133 (S6K) in the development of insulin resistance , presumably via phosphorylation of P41252 proteins . Association testing of panic disorder candidate genes using Q13308 challenge in healthy volunteers . Despite continuing efforts to determine genetic vulnerability to panic disorder ( PD ) , the studies of candidate genes in this disorder have produced inconsistent or negative , results . Laboratory panic induction may have a potential in testing genetic substrate of PD . In this study we aimed to explore the effects of several genetic polymorphisms previously implicated in PD on the susceptibility to cholecystokinin-tetrapeptide ( Q13308 ) challenge in healthy subjects . The study sample consisted of 110 healthy volunteers ( 47 males and 63 females , mean age 22.2 +/- 5.2 ) who participated in Q13308 challenge test . Nine gene-candidates , including 5-HTTLPR , P21397 VNTR , Q8IWU9 rs1386494 , 5- P08908 -1019C-G , 5- P28223 102T-C , CCKR1 246G-A , CCKR2 -215C-A , P21728 -94G-A and P21964 Val158Met , were selected for genotyping based on previous positive findings from genetic association studies in PD . After Q13308 challenge , 39 ( 35.5 % ) subjects experienced a panic attack , while 71 subjects were defined as non-panickers . We detected significant differences for both genotypic and allelic frequencies of 1386494A/G polymorphism in Q8IWU9 gene between panic and non-panic groups with the frequencies of G/G genotype and G allele significantly higher in panickers . None of the other candidate loci were significantly associated with Q13308 -induced panic attacks in healthy subjects . In line with our previous association study in patients with PD , we detected a possible association between Q8IWU9 rs1386494 polymorphism and susceptibility to panic attacks . Other polymorphisms previously associated with PD were unrelated to Q13308 -induced panic attacks , probably due to the differences between complex nature of PD and laboratory panic model . Functional differences of invariant and highly conserved residues in the extracellular domain of the glycoprotein hormone receptors . Multiple interactions exist between human follicle-stimulating hormone ( DB00094 ) and the N-terminal hormone-binding fragment of the human DB00094 receptor ( P23945 ) extracellular domain ( O95905 ) . Binding of the other human glycoprotein hormones to their cognate human receptors ( luteinizing hormone receptor ( LHR ) and thyroid-stimulating hormone receptor ( P16473 ) ) was expected to be similar . This study focuses on amino acid residues in β-strands 2 ( Lys(74) ) , 4 ( DB00135 (124) , DB00174 (129) , and DB00156 (130) ) , and 5 ( DB00128 (150) and DB00128 (153) ) of the P23945 O95905 identified in the human FSH· P23945 O95905 crystal structure as contact sites with the common glycoprotein hormone α-subunit , and on noncontact residues in β-strands 2 ( DB00133 (78) ) and 8 ( DB00128 (224) and DB00133 (226) ) as controls . These nine residues are either invariant or highly conserved in LHR and P16473 . Mutagenesis and functional characterization of these residues in all three human receptors allowed an assessment of their contribution to binding and receptor activation . Surprisingly , the six reported α-subunit contact residues of the P23945 O95905 could be replaced without significant loss of DB00094 binding , while DB02527 signaling potency was diminished significantly with several replacements . Comparative studies of the homologous residues in LHR and P16473 revealed both similarities and differences . The results for DB00094 / P23945 were analyzed on the basis of the crystal structure of the FSH· P23945 O95905 complex , and comparative modeling was used to generate structures for domains , proteins , and complexes for which no structures were available . Although structural information of hormone-receptor interaction allowed the identification of hormone-receptor contact sites , functional analysis of each contact site was necessary to assess its contribution to hormone binding and receptor activation . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Association between iris constitution and apolipoprotein e gene polymorphism in hypertensives . OBJECTIVE : Iridology is a complementary and alternative medicine ( P62158 ) that involves the diagnosis of medical conditions by noting irregularities of the pigmentation in the iris . Iris constitution has a strong familial aggregation and heredity is implicated . P02649 ( apoE ) gene polymorphism is one of the most well-studied genetic markers for vascular diseases , including hypertension . In this study , we investigated the relationship between iris constitution and apoE polymorphism in hypertensives . DESIGN AND SUBJECTS : We classified 87 hypertensives and 79 controls according to iris constitution and determined the apoE genotype of each individual . RESULTS : A significantly higher percentage of individuals with neurogenic constitutions was found in the hypertensive group when compared with the control group ( chi(2) = 40.244 , p < 0.001 ) . In addition , a neurogenic constitution increased the relative risk for hypertension for subjects with an apo epsilon2 or an epsilon4 allele ( chi(2) = 4.086 , p = 0.049 , odds ratio = 2.633 , confidence interval = 1.004-6.905 ) . CONCLUSIONS : Our results imply that a neurogenic iris constitution enhances the relative risk for hypertension in subjects with the apo epsilon2 or epsilon4 allele . Furthermore , we attempted to evaluate the efficacy of iris constitutional medicine and to find an association with hypertension . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Multiplex protein signature for the detection of bladder cancer in voided urine samples . PURPOSE : Accurate urine assays for bladder cancer detection would benefit patients and health care systems . Through extensive genomic and proteomic profiling of urine components we previously identified a panel of 8 biomarkers that can facilitate the detection of bladder cancer in voided urine samples . In this study we confirmed this diagnostic molecular signature in a diverse multicenter cohort . MATERIALS AND METHODS : We performed a case-control , phase II study in which we analyzed voided urine from 102 subjects with bladder cancer and 206 with varying urological disorders . The urinary concentration of 8 biomarkers ( P10145 , P14780 and 10 , P05121 , P15692 , P03950 , Q16790 and P02649 ) was assessed by enzyme-linked immunosorbent assay . Diagnostic performance of the panel of tested biomarkers was evaluated using ROCs and descriptive statistical values , eg sensitivity and specificity . RESULTS : Seven of the 8 urine biomarkers were increased in subjects with bladder cancer relative to those without bladder cancer . The 7 biomarkers were assessed in a new model , which had an AUROC of 0.88 ( 95 % CI 0.84-0.93 ) , and 74 % sensitivity and 90 % specificity . In contrast , the sensitivity of voided urine cytology and the UroVysion® cytogenetic test in this cohort was 39 % and 54 % , respectively . Study limitations include analysis performed on banked urine samples and the lack of voided urine cytology and cytogenetic test data on controls . CONCLUSIONS : The study provides further evidence that the reported panel of diagnostic biomarkers can reliably achieve the noninvasive detection of bladder cancer with higher sensitivity than currently available urine based assays . DB00131 kinase-activated protein kinase ( PRKA ) activators delay meiotic resumption in porcine oocytes . DB00131 -activated kinase ( PRKA ) is a serine/threonine kinase that functions as a metabolic switch in a number of physiological functions . The present study was undertaken to assess the role of this kinase in nuclear maturation of porcine oocytes . RT-PCR and immunoblotting revealed the expression of the Q13131 subunit in granulosa cells , cumulus-oocyte complexes ( COC ) , and denuded oocytes ( DO ) . Porcine COC and DO contained transcripts that corresponded to the expected sizes of the designed primers for Q9Y478 and P54619 . The P54646 subunit was detected in granulosa cells and COC , whereas the Q9UGI9 subunit was not detected in granulosa cells , COC or DO , whereas it was detected in the heart . The Q13131 protein was detected in granulosa cells , COC , DO , and zona pellucida ( ZP ) . In the presence of the pharmacological activator of PRKA 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-monophosphate ( ZMP ) , COC were transiently maintained in meiotic arrest in a fully reversible manner . This inhibitory effect was not observed in DO . Other known PRKA activators , 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside ( AICAR ) and metformin , also blocked meiotic resumption in COC . In contrast to mouse oocytes , in which PRKA activators reverse the inhibitory effect of PDE3 inhibitors , this combination still blocked meiotic resumption in porcine COC . These results demonstrate that the meiotic resumption of porcine COC is transiently blocked by PRKA activators in a dose-dependent manner , and that this effect is dependent on PRKA activity in cumulus cells . The present study describes a new role for PRKA in regulating meiotic resumption in COC and strongly suggests that cumulus cells play an essential role in the control of porcine oocyte maturation through the PRKA metabolic switch . Skinned coronary smooth muscle : calmodulin , calcium antagonists , and DB02527 influence contractility . The effects of Ca2+ , calmodulin , DB02527 , the catalytic subunit of DB02527 -dependent protein kinase ( CSU ) and some Ca2+ antagonists were studied in chemically ( Triton X-100 ) skinned coronary smooth muscle . P62158 increased the Ca2+ responsiveness of the muscle fiber as indicated by the reduction in the threshold as well as the half-maximal activating Ca2+ concentration . DB00831 , a calmodulin antagonist , inhibited Ca2+-calmodulin-induced contraction . Both DB02527 and CSU were effective inhibitors of contraction induced at an intermediate Ca2+ concentration . DB08980 , a Ca2+-antagonist , at 2 x 10(-4) M produced a significant inhibitory effect , which was reduced by increasing the Ca2+ concentration . From other Ca2+ antagonists tested , W-7 , but not D600 and verapamil , produced some inhibitory effect . The data indicate that the response of skinned coronary smooth muscle to Ca2+ , calmodulin and DB02527 are similar to those obtained with other skinned smooth muscles . Furthermore , skinned fiber preparation can serve as a useful tool to investigate possible direct effects of drugs on the activating and regulatory systems in smooth muscle . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose .

MH_train_1223

MH_train_1223

MH_train_1223

interacts_with DB01200?

multiple_choice

Reactive oxygen species-dependent P01375 converting enzyme activation through stimulation of P41595 and alpha1D autoreceptors in neuronal cells . A major determinant of neuronal homeostasis is the proper integration of cell signaling pathways recruited by a variety of neuronal and non-neuronal factors . By taking advantage of a neuroectodermal cell line ( 1C11 ) endowed with the capacity to differentiate into serotonergic ( 1C115-HT ) or noradrenergic ( 1C11NE ) neurons , we identified serotonin ( 5-hydroxytryptamine , 5-HT ) - and norepinephrine ( NE ) -dependent signaling cascades possibly involved in neuronal functions . First , we establish that P41595 receptors and 1D adrenoceptors are functionally coupled to reactive oxygen species ( ROS ) synthesis through NADPH oxidase activation in 1C115-HT and 1C11NE cells . This observation constitutes the prime evidence that bioaminergic autoreceptors take part in the control of the cellular redox equilibrium in a neuronal context . Second , our data identify P78536 ( P01375 - Converting Enzyme ) , a member of a disintegrin and metalloproteinase ( ADAM ) family , as a downstream target of the P41595 and 1D receptor-NADPH oxidase signaling pathways . Upon P41595 or 1D receptor stimulation , ROS fully govern P01375 - shedding in the surrounding milieu of 1C115-HT or 1C11NE cells . Third , P41595 and 1Dreceptor couplings to the NADPH oxidase- P78536 cascade are strictly restricted to 1C11-derived progenies that have implemented a complete serotonergic or noradrenergic phenotype . Overall , these observations suggest that P41595 and 1D autoreceptors may play a role in the maintenance of neuron- and neurotransmitter-associated functions . Eventually , our study may have implications regarding the origin of oxidative stress as well as up-regulated expression of proinflammatory cytokines in neurodegenerative disorders , which may relate to the deviation of normal signaling pathways . Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . Regulation of eosinophil trafficking by Q9UH65 and its role in allergic airway inflammation . Eosinophils are the predominant inflammatory cells recruited to allergic airways . In this article , we show that human and murine eosinophils express Q9UH65 , an intracellular P31749 -binding signaling protein , and examine its role in mediating eosinophil trafficking and pulmonary recruitment in a murine model of allergic airway inflammation . Compared with wild-type eosinophils , Q9UH65 -deficient ( Swap-70(-/-) ) eosinophils revealed altered adhesive interactions within inflamed postcapillary venules under conditions of blood flow by intravital microscopy , exhibiting enhanced slow rolling but decreased firm adhesion . In static adhesion assays , Swap-70(-/-) eosinophils adhered poorly to P19320 and P05362 and exhibited inefficient leading edge and uropod formation . Adherent Swap-70(-/-) eosinophils failed to translocate P63000 to leading edges and displayed aberrant cell surface localization/distribution of α4 and Mac-1 . Chemokine-induced migration of Swap-70(-/-) eosinophils was significantly decreased , correlating with reduced intracellular calcium levels , defective actin polymerization/depolymerization , and altered cytoskeletal rearrangement . In vivo , recruitment of eosinophils to the lungs of allergen-challenged Swap-70(-/-) mice , compared with wild-type mice , was significantly reduced , along with considerable attenuation of airway inflammation , indicated by diminished P05113 , P35225 , and P01375 -α levels ; reduced mucus secretion ; and improved airway function . These findings suggest that regulation of eosinophil trafficking and migration by Q9UH65 is important for the development of eosinophilic inflammation after allergen exposure . Development of peptidomimetic ligands of Pro- DB00149 - DB00145 -NH(2) as allosteric modulators of the dopamine D(2) receptor . A variety of stable , small-molecule peptidomimetic ligands have been developed to elucidate the mechanism by which the neuropeptide Pro- DB00149 - DB00145 -NH(2) ( P00747 ) modulates dopaminergic neurotransmission . Photoaffinity labeling ligands based upon P00747 peptidomimetics have been used to establish that P00747 binds to the P14416 at a site that is different from the orthosteric site , thus making P00747 and its peptidomimetics allosteric modulators of the dopamine receptor . Through the design , synthesis and pharmacological evaluation of conformationally constrained peptidomimetics containing lactam , bicyclic , and spiro-bicyclic scaffolds , support was provided for the hypothesis that the bioactive conformation of P00747 is a type II β-turn . In addition , studies with peptidomimetics designed to mimic either a type VI β-turn or polyproline II helix conformation yielded molecules that were able to modulate dopamine receptors because of their ability to place the carboxamide NH(2) pharmacophore in the same topological space as that seen in the type II β-turn . Extensive studies with the spiro-bicyclic P00747 peptidomimetics also established that both positive and negative modes of modulation were possible for the same series of peptidomimetics simply as a result of minor differences in the stereochemistry about the bridgehead carbon within the scaffold . This information was used to transform existing positive modulators into negative modulators , which demonstrated that small structural changes in the spiro-bicyclic dopamine receptor modulators are capable of causing major changes in the modulatory activity of P00747 peptidomimetics . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . Pharmacokinetics and pharmacodynamics of mepolizumab , an anti-interleukin-5 monoclonal antibody . DB06612 is a fully humanized monoclonal antibody ( IgG1/κ ) targeting human interleukin-5 ( P05113 ) , a key haematopoietin needed for eosinophil development and function . DB06612 blocks human P05113 from binding to the α-chain of the P05113 receptor complex on the eosinophil cell surface , thereby inhibiting P05113 signalling . The pharmacokinetics of mepolizumab have been evaluated in clinical studies at doses of 0.05-10 mg/kg and at 250 mg , 750 mg and 1500 mg . DB06612 was eliminated slowly , with mean initial and terminal phase half-life values of approximately 2 and 20 days , respectively . Plasma clearance ranged from 0.064 to 0.163 mL/h/kg and steady-state volume of distribution ranged from 49 to 93 mL/kg . Pharmacokinetics were dose proportional and time independent . Estimates based on a two-compartment intravenous infusion model from patients with asthma or healthy subjects following single doses predicted mepolizumab plasma concentrations in multiple-dose studies involving patients with hypereosinophilic syndrome ( DB09106 ) , asthma or eosinophilic oesophagitis . The absolute bioavailability of mepolizumab was 64-75 % following subcutaneous injection and 81 % following intramuscular injection . Peripheral blood eosinophil levels decreased in healthy subjects and patients with DB09106 , asthma , eosinophilic oesophagitis or atopic dermatitis after intravenous mepolizumab infusion and subcutaneous injection . Reductions in eosinophil counts in oesophagus , sputum , skin , bone marrow , nasal lavage fluid and/or bronchial mucosa after treatment with mepolizumab were observed in placebo-controlled studies in various indications . The relationship between percentage change from baseline in blood eosinophils and mepolizumab plasma concentrations was described by an indirect pharmacological response model . The estimated maximal decrease in eosinophil count was approximately 85 % from baseline and the half-maximal inhibitory concentration ( IC50 ) was approximately 0.45 μg/mL . Neuronal ablation of p-Akt at Ser473 leads to altered P08908 /2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5-HT ) signaling . To explore how impairment in Akt function regulates 5-HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser473 . Cortical P08908 and 5- Q13049 receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 receptor agonist 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 receptor density was associated with higher P08908 receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5- Q13049 /C agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , with evidence of impaired 5- Q13049 /C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 receptor expression and attenuated DOI-induced 5- Q13049 receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5-HT receptor regulation . These data reveal that defective central Akt function alters 5-HT signaling as well as 5-HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5-HT receptor function . P15121 regulates high glucose-induced ectodomain shedding of tumor necrosis factor ( P01375 ) -alpha via protein kinase C-delta and P01375 converting enzyme in vascular smooth muscle cells . Chronic low-grade inflammation has emerged as a key contributor to the cardiovascular complications of diabetes , however , the mechanisms by which diabetes increases inflammation remain poorly understood . Here , we report that exposure to high glucose ( HG ) stimulates ectodomain shedding of P01375 from rat aortic smooth muscle cells in culture . Our results show that exposure to HG decreases membrane-associated P01375 . This decrease in unprocessed P01375 was prevented by the aldose reductase ( AR ) inhibitor sorbinil and AR small interference RNA . Treatment with HG , but not equimolar mannitol or 3-O-methyl glucose , resulted in phosphorylation and activation of P01375 converting enzyme ( P78536 ) ( P78536 ) , which were attenuated by sorbinil or AR-specific small interference RNA . HG-induced P78536 phosphorylation and P01375 processing were also prevented by P01375 protease inhibitor-1 , an inhibitor of P78536 . Inhibition of protein kinase C ( PKC ) -delta by rottlerin prevented HG-induced P78536 activation and the accumulation of unprocessed P01375 . Treatment with sorbinil decreased elevated levels of circulating P01375 in streptozotocin-treated diabetic rats . DB02712 treatment also decreased the expression of P01375 , matrix metalloproteinase-2 , matrix metalloproteinase-9 , and increased tissue inhibitor of metalloproteinase-3 in vascular smooth muscle cells treated with HG and in balloon-injured carotid arteries of diabetic rats . These results indicate that HG-induced P01375 shedding could be attributed to P78536 activation , which is regulated , in part , by PKC-delta and AR . Therefore , inhibition of P78536 by P01375 protease inhibitor-1 , or pharmacological inhibition of PKC-delta or AR may represent useful strategies for treating vascular inflammation associated with diabetes . P20711 inhibition does not influence the diuretic and natriuretic response to exogenous alpha-atrial natriuretic peptide in man . The role of dopamine synthesis in the renal actions of human alpha-atrial natriuretic peptide ( alpha P01160 ) was investigated in six dehydrated volunteers using the P20711 inhibitor carbidopa . Each subject received oral placebo or carbidopa ( 100 mg ) followed by an infusion of alpha P01160 10 pmol.kg-1.min-1 for 1 h . The responses to placebo alone and to carbidopa alone were investigated on separate occasions. alpha P01160 produced a similar increase in plasma immunoreactive alpha P01160 whether placebo or carbidopa pretreatment had been given . Urinary dopamine excretion was increased by alpha P01160 . DB00190 pretreatment substantially attenuated this increase without affecting the natriuretic or water-diuretic response to alpha P01160 . DB00190 also failed to alter the change in filtration fraction produced by alpha P01160 . The results suggest that increased synthesis of intrarenal dopamine is not required for the renal effects of alpha P01160 in man . Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility . Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor ( P14416 ) in flexible behavioral adaptation , mostly shown in reward-based learning paradigms . Recent evidence from imaging genetics indicates that also intentional cognitive flexibility , associated with lateral frontal cortex , is affected by variations in P14416 signaling . In the present functional magnetic resonance imaging ( Q9BWK5 ) study , we tested the effects of a direct pharmacological manipulation of P14416 stimulation on intentional flexibility in a task-switching context , requiring switches between cognitive task rules and between response hands . In a double blind , counterbalanced design , participants received either a low dose of the P14416 agonist bromocriptine or a placebo in two separate sessions . DB01200 modulated the blood-oxygen-level-dependent ( BOLD ) signal during rule switching : rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo , at comparable performance levels . Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions . Hand-switching-related activity , in contrast , was reduced under bromocriptine in sensorimotor regions . Our results provide converging evidence for an involvement of P14416 signaling in fronto-striatal mechanisms underlying intentional flexibility , and indicate that the neural mechanisms underlying different types of flexibility ( cognitive vs motor ) are affected differently by increased dopaminergic stimulation . Phosphorylation of Akt/GSK-3β/ P29474 amplifies P41595 receptor blockade mediated anti-hypertrophic effect in rats . Herein , we studied the cross talk between 5-HT(2B) receptor blocker ( SB-204741 ) and GSK-3β inhibitor ( SB-216763 ) in isoproterenol-induced cardiac hypertrophy for 28 days . SB-204741 treatment significantly ameliorated ( P < 0.05 ) myocardial dysfunction , myocyte area , fibrosis and myocardial architecture in isoproterenol insulted myocardium . Moreover , this improvement in functional and morphological changes was associated with suppression of hypertrophic ( DB04899 and CK-MB ) , inflammatory ( IKK-β/NF-κB/ P01375 -α and CRP ) , and apoptotic markers ( TUNEL positivity and Bax expression ) along with phosphorylation of Akt/GSK-3β/β-catenin/ P29474 . Intriguingly , co-treatment with GSK-3β inhibitor ( P < 0.01 ) further amplified the anti-hypertrophic effect of SB-204741 ( P < 0.05 ) such that the effect was indistinguishable from that of vehicle treated rats . Thus , 5-HT(2B) receptor blockade mediated anti-hypertrophic effect is atleast in part is governed through phosphorylation of Akt/GSK-3β/β-catenin/ P29474 via attenuating inflammatory and apoptotic pathways . DB08888 for vitreoretinal diseases . P02751 and laminin are clinically relevant plasmin receptors in the eye . Located at the vitreoretinal interface , they are cleaved by ocriplasmin ( DB05028 , ThromboGenics , Iselin , NJ ) , a novel ophthalmic medication . A series of clinical trials to study ocriplasmin for the treatment of vitreoretinal diseases such as vitreomacular traction , macular hole , and exudative age-related macular degeneration are underway . The results are promising and may impact patient care . Pathological fibrinolysis secondary to pseudoaneurysms . Pathological fibrinolysis due to pseudoaneurysms was observed in a patient 4 years after aortobifemoral bypass graft . The patient presented with a pulsatile abdominal mass and ischemic changes in the legs . Excessive bleeding from venipuncture sites prompted coagulation screening , which disclosed rapid clot lysis , fibrin split products , and low fibrinogen suggestive of pathological fibrinolysis . Therapy with epsilon amino caproic acid ( DB00513 , Amicar ) controlled the coagulopathy , permitting angiography and operation . Resection of the pseudoaneurysms resulted in resolution of the abnormal fibrinolysis . Normally , there is a balance between coagulation and fibrinolysis protecting against excessive bleeding or clotting . Clot itself is a powerful stimulus for the activation of the fibrinolytic system , although many other factors have been shown to initiate and sustain the process . Fibrinolysis is pathological when the process becomes excessive or inappropriate . P00747 is activated to plasmin which digests fibrin . Both plasmin and fibrin split products inhibit polymerization of fibrin monomers ( which is the final step in the coagulation cascade in the formation of a stable clot ) , resulting in an unstable clot which is rapidly lysed . To our knowledge such a coagulopathy has not been reported to be a complication of pseudoaneurysms . Cl- DB05511 enhances P01375 -α release in peritoneal macrophages stimulated with LPS . P0DMS8 ( A3R ) belongs to the Gi/Gq-coupled receptor family , that leads to the intracellular DB02527 reduction and intracellular calcium increase , respectively . A3R is widely expressed and it can play a crucial role in many patho-physiological conditions , including inflammation . Here we investigate the effect of Cl- DB05511 , A3R agonist , on the production of P01375 -α . We found that Cl- DB05511 enhances LPS-induced P01375 -α release in peritoneal macrophages . This effect is reduced by MRS1191 , A3R antagonist and by forskolin , activator of adenylyl cyclase . pIκBα increased in LPS+Cl- DB05511 -treated macrophages , while total IκB kinase-β ( IKKβ ) reduced . Indeed , p65NF-κB nuclear translocation increased in cells treated with LPS+Cl- DB05511 . Moreover , IMD 0354 , IKKβ inhibitor , significantly abrogated the effect of Cl- DB05511 on P01375 -α release . Inhibition of protein kinase C ( PKC ) significantly reduced Cl- DB05511 -induced P01375 -α release in LPS-stimulated macrophages . Furthermore , LY-294002 , PI3K inhibitor , reduced the P01375 -α production enhanced by Cl- DB05511 , although the phosphorylation status of Akt did not change in cells treated with LPS+Cl- DB05511 than LPS alone . In summary , these data show that Cl- DB05511 is able to enhance P01375 -α production in LPS-treated macrophages in an NF-κB- dependent manner . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Tissue-based microarray expression of genes predictive of metastasis in uveal melanoma and differentially expressed in metastatic uveal melanoma . PURPOSE : To screen the microarray expression of CDH1 , Q16610 , O60739 , P51114 , P41595 , Q02363 , Q9NZU5 , P09960 , Q9ULD2 , Q13636 , Q9Y6N7 , and Q01826 genes which are predictive of primary uveal melanoma metastasis , and Q00653 , Q99952 , O43312 , O75293 , O76070 , Q96QV1 , P29460 , P11802 , P05388 , P08708 , P25398 genes that are differentially expressed in metastatic uveal melanoma in normal whole human blood and tissues prone to metastatic involvement by uveal melanoma . METHODS : We screened the GeneNote and GNF BioGPS databases for microarray analysis of genes predictive of primary uveal melanoma metastasis and those differentially expressed in metastatic uveal melanoma in normal whole blood , liver , lung and skin . RESULTS : Microarray analysis showed expression of all 22 genes in normal whole blood , liver , lung and skin , which are the most common sites of metastases . In the GNF BioGPS database , data for expression of the Q96QV1 gene in normal whole blood and skin was not complete . CONCLUSIONS : Microarray analysis of genes predicting systemic metastasis of uveal melanoma and genes differentially expressed in metastatic uveal melanoma may not be used as a biomarker for metastasis in whole blood , liver , lung , and skin . Their expression in tissues prone to metastasis may suggest that they play a role in tropism of uveal melanoma metastasis to these tissues . DB00495 is effective against human multiple myeloma : a new use for an old drug ? DB00495 ( DB00495 ) is an antiretroviral drug that affects cell proliferation , apoptosis , and the NF-κB pathway . As multiple myeloma ( MM ) presents with constitutive activation of NF-κB , we analyzed the effect of DB00495 on human MM cell lines . We evaluated the cytotoxic effect of DB00495 in human MM cell lines sensitive ( 8226/S ) or resistant to doxorubicin ( 8226/DX5 ) and human T cell lymphoblast-like cells , uterine sarcoma cells , and HUVEC using MTT assay . Cytotoxicity was also evaluated in vivo in nude mice xenografted with 8226/S tumor . The effect of DB00495 on the expression of genes involved in cell proliferation , apoptosis , angiogenesis , and the NF-κB pathway was analyzed in the xenografts using real-time polymerase chain reaction . DB00495 was effective against both 8226/S and 8226/DX5 cells in a dose and time-dependent manner ( p = 0.02 ) in vitro and promoted cell cycle arrest in S phase in these cells . The tumor volume was lower in mice treated with DB00495 compared to untreated mice ( p = 0.0003 ) . DB00495 down-regulated the pro-proliferative genes encoding P31749 , MYC , P42224 , P45983 , P45984 , DB00833 -3 , Bcl-3 , and cyclin D2 ; pro-angiogenenic genes encoding P15692 and P10145 ; and genes involved in cell adhesion ( P05362 and P02751 ) and the NF-κB pathway . DB00495 up-regulated the expression of tumor suppressor gene Q9H334 and the pro-apoptotic genes encoding P55957 , O95999 , and caspase-8 . Thus , we demonstrated the cytotoxic effect of DB00495 in human MM cell lines for the first time . Our data may provide the rationale for future clinical trials of DB00495 for treating MM . The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT/ P31749 phosphorylation . DB01200 ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 , official symbol Q01959 ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract . Endothelial dysfunction in morbid obesity . Morbid obesity is a chronic multifunctional disease characterized by an accumulation of fat . Epidemiological studies have shown that obesity is associated with cardiovascular and metabolic disorders . Endothelial dysfunction , as defined by an imbalance between relaxing and contractile endothelial factors , plays a central role in the pathogenesis of these cardiometabolic diseases . Diminished bioavailability of nitric oxide ( NO ) contributes to endothelial dysfunction and impairs endothelium- dependent vasodilatation . But this is not the only mechanism that drives to endothelial dysfunction . Obesity has been associated with a chronic inflammatory process , atherosclerosis , and oxidative stress . Moreover levels of asymmetrical DB01686 ( DB01686 ) , an endogenous inhibitor of endothelial nitric oxide synthase ( P29474 ) , are elevated in obesity . On the other hand , increasing prostanoid-dependent vasoconstriction and decreasing vasodilator prostanoids also lead to endothelial dysfunction in obesity . Other mechanisms related to endothelin-1 ( ET-1 ) or endothelium derived hyperpolarizing factor ( EDHF ) have been proposed . Bariatric surgery ( BS ) is a safe and effective means to achieve significant weight loss , but its use is limited only to patients with severe obesity including morbid obesity . BS also proved efficient in endothelial dysfunction reduction improving cardiovascular and metabolic comorbidities associated with morbid obesity such as diabetes , coronary artery disease , nonalcoholic fatty liver disease and cancer . This review will provide a brief overview of the mechanisms that link obesity with endothelial dysfunction , and how weight loss is a cornerstone treatment for cardiovascular comorbidities obesity-related . A better understanding of the mechanisms of obesity-induced endothelial dysfunction may help develop new therapeutic strategies to reduce cardiovascular morbidity and mortality . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors .

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interacts_with DB00317?

multiple_choice

Mediation of 5-hydroxytryptamine-induced tachycardia in the pig by the putative Q13639 receptor . Intravenous bolus injections of 5-hydroxytryptamine ( 5-HT ; 3 , 10 and 30 micrograms kg-1 ) , 5-methoxytryptamine ( 5-MeO-T ; 3 , 10 and 30 micrograms kg-1 ) , renzapride ( O95696 24924 ; 3 , 10 , 30 and 100 micrograms kg-1 ) and isoprenaline ( 0.03 , 0.1 and 0.3 micrograms kg-1 ) to anaesthetized pigs increased heart rate by , respectively , 22 +/- 3 , 44 +/- 3 and 65 +/- 4 beats min-1 ( 5-HT ; n = 17 ) ; 12 +/- 1 , 26 +/- 2 and 44 +/- 4 beats min-1 ( 5-MeO-T ; n = 15 ) , 5 +/- 2 , 11 +/- 2 , 18 +/- 4 and 37 +/- 5 beats min-1 ( renzapride ; n = 8 ) and 17 +/- 2 , 46 +/- 3 and 75 +/- 3 beats min-1 ( isoprenaline ; n = 13 ) . The responses to 5-HT , 5-MeO-T and renzapride were antagonized by ICS 205-930 ( 1 and 3 mg kg-1 , i.v. ) , which did not modify the increases in heart rate by isoprenaline . DB04917 showed tachyphylaxis and attenuated the responses to 5-HT . These findings indicate that 5-HT elicits tachycardia in the pig by acting on a novel receptor , either similar or identical to the Q13639 receptor identified in mouse brain colliculi . Beta-amyloid accumulation impairs multivesicular body sorting by inhibiting the ubiquitin-proteasome system . Increasing evidence links intraneuronal beta-amyloid ( Abeta42 ) accumulation with the pathogenesis of Alzheimer 's disease ( AD ) . In Abeta precursor protein ( P05067 ) mutant transgenic mice and in human AD brain , progressive intraneuronal accumulation of Abeta42 occurs especially in multivesicular bodies ( MVBs ) . We hypothesized that this impairs the MVB sorting pathway . We used the trafficking of the epidermal growth factor receptor ( P00533 ) and TrkB receptor to investigate the MVB sorting pathway in cultured neurons . We report that , during P01133 stimulation , P05067 mutant neurons demonstrated impaired inactivation , degradation , and ubiquitination of P00533 . P00533 degradation is dependent on translocation from MVB outer to inner membranes , which is regulated by the ubiquitin-proteasome system ( P08397 ) . We provide evidence that Abeta accumulation in P05067 mutant neurons inhibits the activities of the proteasome and deubiquitinating enzymes . These data suggest a mechanism whereby Abeta accumulation in neurons impairs the MVB sorting pathway via the P08397 in AD . Moving beyond chemotherapy : novel cytostatic agents for malignant mesothelioma . It is now known that vascular endothelial growth factor ( P15692 ) and platelet derived growth factor ( PDGF ) are autocrine growth factors in malignant mesothelioma ; epidermal growth factor receptor ( P00533 ) is also highly overexpressed . Cytotoxic drugs that target these growth factors offer fresh potential for the treatment of mesothelioma . Clinical trials have recently been initiated to evaluate the anti-tumour activity of the P15692 inhibitors SU5416 , bevacizumab and thalidomide . ZD1839 ( DB00317 , AstraZeneca ) , an inhibitor of P00533 tyrosine kinase , is also being evaluated . Two clinical trials are planned to evaluate the two PDGF inhibitors Gleevec ( Imatinib mesylate , STI-571 , Novartis Pharmaceuticals ) and PTK787 ( Novartis Pharmaceuticals ) . Tandutinib ( MLN518 ) reverses multidrug resistance by inhibiting the efflux activity of the multidrug resistance protein 7 ( Q5T3U5 ) . It is well established that DB00171 -binding cassette ( DB01048 ) transporter-mediated multidrug resistance ( MDR ) is one of the major mechanisms that causes resistance to antineoplastic drugs in cancer cells . ABC transporters can significantly decrease the intracellular concentration of antineoplastic drugs by increasing their efflux , thereby lowering their cytotoxic activity . One of these transporters , the multidrug resistance protein 7 ( Q5T3U5 / Q5T3U5 ) , has already been shown to produce resistance to antineoplastic drugs by increasing the efflux of the drugs . In the present study , we investigated whether DB05465 , an P07333 -like tyrosine kinase 3 ( P36888 ) inhibitor , has the potential to reverse Q5T3U5 -mediated MDR . Our results revealed that DB05465 significantly enhanced the sensitivity of Q5T3U5 -transfected HEK293 cells to the 2 established Q5T3U5 substrates , paclitaxel and vincristine , whereas there was less or no effect on the control vector-transfected HEK293 cells . [ ³H ] -paclitaxel accumulation and efflux studies demonstrated that DB05465 increased the intracellular accumulation of [ ³H ] -paclitaxel and inhibited the efflux of [ ³H ] -paclitaxel from P29320 - Q5T3U5 cells . In addition , western blot analysis showed that DB05465 did not significantly affect Q5T3U5 expression . Thus , we conclude that the P36888 inhibitor DB05465 can reverse Q5T3U5 -mediated MDR through inhibition of the drug efflux function and may have potential to be used clinically in combination therapy for cancer patients . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . Intratumoral expression profiling of genes involved in angiogenesis in colorectal cancer patients treated with chemotherapy plus the VEGFR inhibitor PTK787/ZK 222584 ( vatalanib ) . The phase III CONFIRM clinical trials demonstrated that metastatic colorectal cancer patients with elevated serum lactate dehydrogenase ( LDH ) had improved outcome when the vascular endothelial growth factor receptor ( VEGFR ) inhibitor DB04879 ( DB04879 ) was added to FOLFOX4 chemotherapy . We investigated the hypothesis that high intratumoral expression of genes regulated by hypoxia-inducible factor-1 alpha ( HIF1α ) , namely P00338 , glucose transporter-1 ( P11166 ) , P15692 , P17948 , and P35968 , were predictive of outcome in CONFIRM-1 . Tumor tissue was isolated by laser-capture microdissection from 85 CONFIRM-1 tumor specimens ; FOLFOX4/placebo n=42 , FOLFOX4/ DB04879 n=43 . Gene expression was analyzed using quantitative RT-PCR . In univariate analyses , elevated mRNA expression of P00338 , P11166 , and P17948 were associated with response to FOLFOX4/ DB04879 . In univariate and multivariate analyses , elevated P00338 and P17948 mRNA levels were associated with improved progression-free survival in FOLFOX4/ DB04879 patients . Furthermore , increased HIF1α and P35968 mRNA levels were associated with decreased survival in FOLFOX/placebo patients but not in patients who received FOLFOX4/ DB04879 . These are the first data suggesting intratumoral mRNA expression of genes involved in angiogenesis/HIF pathway may predict outcome to VEGFR-inhibitors . Biomarkers that assist in directing VEGFR-inhibitors toward patients with an increased likelihood of benefit will improve the cost-effectiveness of these promising agents . P30047 -dependent and -independent inhibitors of P30793 . P30047 ( P30047 ) mediates the feedback inhibition of P30793 activity by ( 6R ) -L-erythro- DB00360 ( BH4 ) through protein complex formation . Since guanine and BH4 have a common pyrimidine ring structure , we examined the inhibitory effect of guanine and its analogs on the enzyme activity . DB02377 , 8-hydroxyguanine , 8-methylguanine , and 8-bromoguanine inhibited the enzyme activity in a P30047 -dependent and pH-dependent manner and induced complex formation between P30793 and P30047 . The type of inhibition by this group is a mixed type . All these properties were shared with BH4 . In striking contrast , inhibition by DB01667 and 8-mercaptoguanine was P30047 -independent and pH-independent . The type of inhibition by DB01667 and 8-mercaptoguanine was a competitive type . The two compounds did not induce complex formation between the enzyme and P30047 . These results demonstrate that guanine compounds of the first group bind to the BH4-binding site of the P30793 / P30047 complex , whereas DB01667 and 8-mercaptoguanine bind to the active site of the enzyme . Finally , the possible implications in Lesch-Nyhan syndrome and Parkinson diseases of the inhibition of P30793 by guanine and 8-hydroxyguanine are discussed . Macrophages may promote cancer growth via a GM- P04141 /HB- P01133 paracrine loop that is enhanced by P48061 . BACKGROUND : Increased numbers of tumour-associated macrophages correlate with shortened survival in some cancers . The molecular bases of this correlation are not thoroughly understood . Events triggered by P48061 may play a part , as P48061 drives the migration of both P61073 -positive cancer cells and macrophages and may promote a molecular crosstalk between them . RESULTS : Samples of P00533 -positive colon cancer metastases in liver , a tissue with high expression of P48061 , were analysed by immunohistochemistry . In all of the patient biopsies , P34810 -positive tumour-associated macrophages presented a mixed P02778 ( M1 ) / Q86VB7 ( M2 ) pattern , expressed P61073 , GM- P04141 and HB- P01133 , and some stained positive for P48061 . Cancer cells stained positive for P61073 , P48061 , P00533 , Q15303 and GM- P04141 . Regulatory interactions among these proteins were validated via experiments in vitro involving crosstalk between human mononuclear phagocytes and the cell lines DLD-1 ( human colon adenocarcinoma ) and HeLa ( human cervical carcinoma ) , which express the above-mentioned ligand/receptor repertoire . P48061 induced mononuclear phagocytes to release HB- P01133 , which activated P00533 and triggered anti-apoptotic and proliferative signals in cancer cells . The cancer cells then proliferated and released GM- P04141 , which in turn activated mononuclear phagocytes and induced them to release more HB- P01133 . Blockade of GM- P04141 with neutralising antibodies or siRNA suppressed this loop . CONCLUSIONS : P48061 -driven stimulation of cancer cells and macrophages may elicit and reinforce a GM- P04141 /HB- P01133 paracrine loop , whereby macrophages contribute to cancer survival and expansion . The involvement of mixed M1/M2 GM- P04141 -stimulated macrophages in a tumour-promoting loop may challenge the paradigm of tumour-favouring macrophages as polarized M2 mononuclear phagocytes . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Pseudomonas aeruginosa pyocyanin causes airway goblet cell hyperplasia and metaplasia and mucus hypersecretion by inactivating the transcriptional factor FoxA2 . The redox-active exotoxin pyocyanin ( Q15149 ) can be recovered in 100 µM concentrations in the sputa of bronchiectasis patients chronically infected with Pseudomonas aeruginosa ( PA ) . However , the importance of Q15149 within bronchiectatic airways colonized by PA remains unrecognized . Recently , we have shown that Q15149 is required for chronic PA lung infection in mice , and that chronic instillation of Q15149 induces goblet cell hyperplasia ( P30793 ) , pulmonary fibrosis , emphysema and influx of immune cells in mouse airways . Many of these pathological features are strikingly similar to the mouse airways devoid of functional FoxA2 , a transcriptional repressor of P30793 and mucus biosynthesis . In this study , we postulate that Q15149 causes and exacerbates P30793 and mucus hypersecretion in bronchiectatic airways chronically infected by PA by inactivating FoxA2 . We demonstrate that Q15149 represses the expression of FoxA2 in mouse airways and in bronchial epithelial cells cultured at an air-liquid interface or conventionally , resulting in P30793 , increased Q9HC84 mucin gene expression and mucus hypersecretion . Immunohistochemical and inhibitor studies indicate that Q15149 upregulates the expression of Stat6 and P00533 , both of which in turn repress the expression of FoxA2 . These studies demonstrate that Q15149 induces P30793 and mucus hypersecretion by inactivating FoxA2 . Effects of thioacetamide-induced hepatic failure on the N-methyl-D-aspartate receptor complex in the rat cerebral cortex , striatum , and hippocampus . Binding of different ligands and expression of receptor subunit mRNAs . Hepatic encephalopathy ( HE ) is characterized by symptoms pointing at disturbances in glutamatergic neurotransmission in the brain , particularly in the striatum . The binding parameters of ligands specific for different recognition sites in the N-methyl-D-aspartate ( DB01221 ) receptor complex and the distribution of the receptor subunit mRNAs ( Q9UHB4 , Q12879 -D ) were assessed in rats with acute HE induced with a hepatotoxin , thioacetamide ( TAA ) . The binding of : 1 . L-[3H]glutamate ( DB01221 -displaceable ) ; 2 . [3H]dizocilpine and N-(1-[2-thienyl]-cyclohexyl) [3H]piperidine ( [3H] DB01520 ) ; and 3 . The coactivator site agonist [3H]glycine was assayed in purified membranes of the cerebral cortex , hippocampus , and striatum . In HE rats , Bmax of DB01221 -displaceable glutamate binding was increased in the cerebral cortex and hippocampus , but slightly decreased in the striatum . In this region , the binding affinity was also slightly increased . In HE , Bmax of [3H]dizocilpine binding was unchanged in the striatum and cerebral cortex , but substantially decreased in the hippocampus . Pretreatment with phorbol ester enhanced the binding of dizocilpine more in HE than in control rats . Bmax of [3H] DB01520 binding was decreased in the cerebral cortex and striatum , but increased in the hippocampus . The different responses of these two phencyclidine site antagonists to HE may be indicative of a conformational change within the ion channel and/or the presence of microdomains reacting differently to extrinsic factors . HE did not affect glycine binding , but potentiated the maximal stimulation of [3H]dizocilpine binding by glycine in the cerebral cortex . The results emphasize the brain region and domain specificity of the responses of the DB01221 receptor complex to HE . Cooperative inhibitory effect of ZD1839 ( DB00317 ) in combination with trastuzumab ( Herceptin ) on human breast cancer cell growth . BACKGROUND : Co-expression of the epidermal growth factor receptor ( P00533 ) and of ErbB-2 is found in a subset of primary human breast cancer . MATERIALS AND METHODS : The antiproliferative effects of anti- P00533 and anti-ErbB-2 agents were evaluated using a monolayer assay . The effects of these agents on the activation of P00533 , ErbB-2 , AKT and Q8NFH3 / Q8TCB0 Q96HU1 kinases ( MAPK ) were investigated by western blot analysis . RESULTS : We found that both ZD1839 ( DB00317 ) , a specific P00533 tyrosine kinase inhibitor , and trastuzumab ( Herceptin ) ( TRA ) , a humanized anti-ErbB-2 monoclonal antibody , were able to inhibit the growth of SK-Br-3 and BT-474 breast carcinoma cells , which express both P00533 and ErbB-2 . Treatment of breast carcinoma cells with a combination of ZD1839 and TRA resulted in a synergistic inhibitory effect . Treatment of SK-Br-3 cells with ZD1839 produced a significant , dose-dependent reduction of the tyrosine phosphorylation of both P00533 and ErbB-2 . Phosphorylation of MAPK and AKT were significantly reduced in SK-Br-3 cells following treatment with ZD1839 , whereas treatment with TRA produced a reduction of AKT but not MAPK phosphorylation . Finally , treatment with ZD1839 , but not with TRA , produced a significant increase in fragmented DNA in breast carcinoma cells . However , a more pronounced increase in the levels of fragmented DNA was observed following combined treatment with ZD1839 and TRA . CONCLUSIONS : These data suggest that combined treatment with drugs that target P00533 and ErbB-2 might result in an efficient inhibition of tumor growth in those breast carcinoma patients whose tumors co-express both receptors . Identification of genes upregulated in Q9UM73 -positive and P00533 / P01116 / Q9UM73 -negative lung adenocarcinomas . Activation of the P00533 , P01116 , and Q9UM73 oncogenes defines 3 different pathways of molecular pathogenesis in lung adenocarcinoma . However , many tumors lack activation of any pathway ( triple-negative lung adenocarcinomas ) posing a challenge for prognosis and treatment . Here , we report an extensive genome-wide expression profiling of 226 primary human stage I-II lung adenocarcinomas that elucidates molecular characteristics of tumors that harbor Q9UM73 mutations or that lack P00533 , P01116 , and Q9UM73 mutations , that is , triple-negative adenocarcinomas . One hundred and seventy-four genes were selected as being upregulated specifically in 79 lung adenocarcinomas without P00533 and P01116 mutations . Unsupervised clustering using a 174-gene signature , including Q9UM73 itself , classified these 2 groups of tumors into Q9UM73 -positive cases and 2 distinct groups of triple-negative cases ( groups A and B ) . Notably , group A triple-negative cases had a worse prognosis for relapse and death , compared with cases with P00533 , P01116 , or Q9UM73 mutations or group B triple-negative cases . In Q9UM73 -positive tumors , 30 genes , including Q9UM73 and Q12879 , were commonly overexpressed , whereas in group A triple-negative cases , 9 genes were commonly overexpressed , including a candidate diagnostic/therapeutic target Q5TB30 , that were determined to be critical for predicting a worse prognosis . Our findings are important because they provide a molecular basis of Q9UM73 -positive lung adenocarcinomas and triple-negative lung adenocarcinomas and further stratify more or less aggressive subgroups of triple-negative lung ADC , possibly helping identify patients who may gain the most benefit from adjuvant chemotherapy after surgical resection . An overview of phenserine tartrate , a novel acetylcholinesterase inhibitor for the treatment of Alzheimer 's disease . Existing cholinesterase ( ChE ) inhibitor therapies for Alzheimer 's disease ( AD ) , while effective in improving cognitive , behavioral and functional impairments , do not alter disease progression . Novel drug design studies have focused on the classical ChE inhibitor , (-)-physostigmine , producing alterations in chemical composition and three-dimensional structure , which may offer an improved therapeutic index . The phenylcarbamate derivative , DB04892 , is a selective , non-competitive inhibitor of acetylcholinesterase ( P22303 ) . In vivo , DB04892 produces rapid , potent , and long-lasting P22303 inhibition . As a possible result of its preferential brain selectivity , DB04892 is significantly less toxic than (-)-physostigmine . In studies using the Stone maze paradigm , DB04892 has been shown to improve cognitive performance in both young learning-impaired and elderly rats . In addition to reducing inactivation of acetylcholine in the brain , DB04892 appears to have a second mode of action . Reduced secretion of beta-amyloid ( Abeta ) has been observed in cell lines exposed to DB04892 , occurring through translational regulation of beta-amyloid precursor protein ( beta- P05067 ) mRNA via a non-cholinergic mechanism . These in vitro findings appear to translate in vivo into animal models and humans . In a small study of patients with AD , DB04892 treatment tended to reduce beta- P05067 and Abeta levels in plasma samples . Clinical studies also reveal that DB04892 ( 5-10 mg b.i.d. ) had a favorable safety and pharmacological profile , produced significant improvements in cognitive function and was well tolerated in patients with AD treated for 12 weeks . Further randomized , double-blind , placebo-controlled Phase III studies assessing the efficacy , safety/tolerability and potential disease-modifying effects of DB04892 in patients with AD are currently ongoing . Significance of interleukin-6 signaling in the resistance of pharyngeal cancer to irradiation and the epidermal growth factor receptor inhibitor . PURPOSE : Tumor eradication by chemoradiotherapy for pharyngeal cancer has not been particularly successful . Targeting epithelial growth factor receptor ( P00533 ) could be a potential treatment strategy providing additional benefits , but only a subset of these tumors gives a clinically significant response to P00533 inhibitors . The aim has been to identify the role of interleukin-6 ( P05231 ) signaling and its predictive power in the treatment response of pharyngeal cancer . METHODS AND MATERIALS : Human pharyngeal cancer cell lines , including the hypopharyngeal cancer cell line FaDu and its derived cell line FaDu-C225-R , were selected . Changes in tumor growth , response to treatment , and responsible signaling pathway were investigated in vitro . Furthermore , 95 pharyngeal cancer tissue specimens were analyzed by immunohistochemical staining , and correlations were made between levels of P05231 , P05231 receptor ( IL-6R ) , p-AKT , and p- P40763 expression and the clinical outcome of patients . RESULTS : In vitro , either extrinsic P05231 stimulation of cancer cells or intrinsically activated P05231 signaling detected in FADu-C225-R cells results in resistance to irradiation and P00533 inhibitor . Blocking P05231 signaling attenuated aggressive tumor behavior and sensitized the cells to treatments . The responsible mechanisms included decreased p- P40763 , less nuclear translocation of P00533 , and subsequently attenuated epithelial-mesenchymal transition . Regarding clinical data , staining of p- P40763 and P05231 was significantly linked with lower response rates to treatments and shorter survival in pharyngeal cancer patients . CONCLUSIONS : P05231 and p- P40763 may be significant predictors of pharyngeal carcinoma , and regulating P05231 signaling can be considered a promising therapeutic approach . Analysis of CAD gene amplification using a combined approach of molecular genetics and cytogenetics . CAD is a multifunctional protein which catalyzes the first three steps of de novo uridine biosynthesis . Rodent cells resistant to DB03459 , a specific inhibitor of the ATCase activity of CAD , overproduce the P27708 and CAD mRNA as a direct result of the amplification of the CAD gene . In order to study the mechanism of CAD gene amplification , a functional Syrian hamster CAD gene was inserted into a cosmid vector using molecular cloning techniques . The cloned genes were assayed for biological function by fusing CAD-deficient Chinese hamster ovary ( CHO ) cell mutants with protoplasts of E. coli containing the CAD cosmids . Two clones with functional CAD genes were isolated and shown to contain inserts 40 and 45 kb long . The cloned genes could also be introduced into wild type CHO cells by selecting for cells which became resistant to high DB03459 concentrations in a single step . Transformations of mutant and wild type CHO cells contained multiple active copies of the donated Syrian hamster CAD genes in addition to their endogenous CHO CAD genes . The cloned genes in all transformants analyzed are integrated into host cell chromosomes at single locations defined by in situ hybridization . Independently isolated transformants contain the donated genes in different chromosomes . Co-transformation of CHO cells with two different genes by protoplast fusion is also shown to be possible . Both P00533 kinase and Src-related tyrosine kinases regulate human ether-à-go-go-related gene potassium channels . Human ether-à-go-go-related gene ( hERG or Kv11.1 ) encodes the rapidly activated delayed rectifier K(+) current ( I(Kr) ) in the human heart . Potential regulation of hERG channel by protein tyrosine kinases ( PTKs ) is not understood . The present study was designed to investigate whether this channel is modulated by PTKs using whole-cell patch clamp technique , and immunoprecipitation and Western blot analysis in P29320 293 cells stably expressing hERG gene . We found that the broad-spectrum PTK inhibitor genistein ( 30 microM ) , the selective P00533 ( epidermal growth factor receptor ) kinase inhibitor AG556 ( 10 microM ) and the Src-family kinase inhibitor Q99463 ( 10 microM ) remarkably inhibited hERG channel current ( I(hERG) ) , and the effects were significantly countered by the protein tyrosine phosphatase ( PTP ) inhibitor orthovanadate ( 1 mM ) . Immunoprecipitation and Western blot analysis demonstrated that membrane protein tyrosine phosphorylation of hERG channels was reduced by genistein , AG556 , and Q99463 . The reduction of hERG channel phosphorylation level by genistein , AG556 or Q99463 was antagonized by orthovanadate . Single point mutation(s) of Y475A and/or Y611A dramatically attenuated the inhibitory effect of I(hERG) by Q99463 and/or AG556 . Our results demonstrate the novel information that I(hERG) is modulated not only by Src-family kinases , but also by P00533 kinases . Y475 and/or Y611 are likely the preferred phosphorylation sites . Regulation of hERG channels by PTKs modifies the channel activity and thus likely alters electrophysiological properties including action potential duration and cell excitability in human heart and neurons . Isolation and characterization of P62333 . A novel ATPase family component of the yeast 26 S proteasome . Using a genetic strategy designed to find proteins involved in the function of the Saccharomyces cerevisiae transcriptional activator GAL4 , we isolated mutants in two genes which rescue a class of gal4 activation domain mutants . One of these genes , P62195 , encodes a member of a large family of putative ATPases , the Conserved ATPase containing Domain ( CAD ) proteins ( also known as AAA proteins ) that are involved in a wide variety of cellular functions . Subsequently , P62195 was identified as a subunit of the 26 S proteasome . We have now cloned the gene defined by the second complementation group . P62333 encodes an essential 49-kDa protein that is also a member of the CAD family and is 43 % identical to P62195 . The mutation in sug2-1 , like that in sug1-1 , is found in the CAD near the highly conserved ATPase motif . We present biochemical and genetic evidence that P62333 is associated in vivo with P62195 and is a novel P27708 subunit of the 26 S proteasome . With its highly conserved mammalian homologs , human Q8NFH3 and ground squirrel CADp44 , P62333 defines a new class of proteasomal CAD proteins . Identification of novel genetic alterations in samples of malignant glioma patients . Glioblastoma is the most frequent and malignant human brain tumor . High level of genomic instability detected in glioma cells implies that numerous genetic alterations accumulate during glioma pathogenesis . We investigated alterations in AP-PCR DNA profiles of 30 glioma patients , and detected specific changes in 11 genes not previously associated with this disease : Q86UP9 , Q13326 , Q13639 , P05556 , P31327 , P07225 , P55259 , Q9UJ96 , Q08499 , Q8N743 , and Q14642 . Further correlations revealed that 8 genes might play important role in pathogenesis of glial tumors , while changes in P55259 , Q9UJ96 and Q8N743 should be considered as passenger mutations , consequence of high level of genomic instability . Identified genes have a significant role in signal transduction or cell adhesion , which are important processes for cancer development and progression . According to our results , Q86UP9 might be characteristic of primary glioblastoma , Q13326 , Q13639 , P05556 , P31327 , P07225 and Q14642 were detected predominantly in anaplastic astrocytoma , suggesting their role in progression of secondary glioblastoma , while alterations of Q08499 seem to have important role in development of both glioblastoma subtypes . Some of the identified genes showed significant association with p53 , p16 , and P00533 , but there was no significant correlation between loss of P60484 and any of identified genes . In conclusion our study revealed genetic alterations that were not previously associated with glioma pathogenesis and could be potentially used as molecular markers of different glioblastoma subtypes . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL .

MH_train_1225

MH_train_1225

MH_train_1225

interacts_with DB08816?

multiple_choice

Panax ginseng ameliorates airway inflammation in an ovalbumin-sensitized mouse allergic asthma model . ETHNOPHARMACOLOGICAL RELEVANCE : Panax ginseng ( PG ) is a medicinal herb that has been used to treat various immune diseases including asthma and P48444 . In this study , we investigated the inhibitory mechanism of PG on asthma parameters in mice . MATERIALS AND METHODS : BALB/c mice were sensitized with 20 μg/200 μl OVA adsorbed on 1.0mg/50 μl aluminum hydroxide gel adjuvant by i.p. injection on days 0 and 14 . Mice were then challenged with 5 % OVA in PBS to the nose for 30 min once a day for 3 days , from day 20 until day 22 , using a nebulizer . PG ( 20mg/kg ) or vehicle was administrated by i.p. injection once a day 10 min before every OVA challenge for 3 days . The recruitment of inflammatory cells into bronchoalveolar lavage fluid or lung tissues was measured . The expression of EMBP , Muc5ac , P25942 , and P29965 ( P29965 ) in lung tissues was investigated . In addition , the cytokines and mitogen activated protein ( Q96HU1 ) kinases were measured by RT-PCR and Western blot . RESULTS AND CONCLUSIONS : PG restored the expression of EMBP , Muc5ac , P25942 , and P29965 , as well as the mRNA and protein levels of interleukin ( IL ) -1β , P05112 , P05113 , and tumor necrosis factor ( P01375 ) -α . In addition , PG inhibited the numbers of goblet cells and further small G proteins and Q96HU1 kinases in bronchoalveolar lavage cells and lung tissues increased in ovalbumin-induced allergic asthma in mice . These results suggest that PG may be used as a therapeutic agent in asthma , based on reductions of various allergic responses . P15121 inhibition suppresses azoxymethane-induced colonic premalignant lesions in C57BL/KsJ-db/db mice . Type-2 diabetes and obesity-related metabolic abnormalities are major risk factors for the development of colon cancer . In the present study , we examined the effects of polyol pathway enzyme aldose reductase ( AR ) inhibitor , fidarestat , on the development of azoxymethane ( AOM ) -induced colonic premalignant lesions in C57BL/KsJ-db/db obese mice . Our results indicate that fidarestat given in the drinking water caused a significant reduction in the total number of colonic premalignant lesions in the AOM treated obese mice . Further , the expression levels of PKC-β2 , AKT , P35354 and P35228 in the colonic mucosa of AOM-treated mice were significantly decreased by fidarestat . The serum levels of IL-1α , P02778 , Q07325 , P01375 -α and P15692 are significantly suppressed in AOM + fidarestat treated obese mice . DB02021 also decreased the expression of P35354 , P35228 , P98170 , survivin , β-catenin and NF-κB in high glucose-treated HT29 colon cancer cells . In conclusion , our results indicate that fidarestat inhibits the development of colonic premalignant lesions in an obesity-related colon cancer and is chemopreventive to colorectal carcinogenesis in obese individuals . DB01221 receptor alterations in neurons from pediatric cortical dysplasia tissue . The subunit composition of glutamate receptors affects their functional properties , and could contribute to abnormal electrophysiology in pediatric cortical dysplasia ( CD ) . We examined electrophysiological responses and subunit assembly of N-methyl-D-aspartate ( DB01221 ) receptors in acutely dissociated normal-appearing pyramidal and cytomegalic neurons from CD tissue and normal-appearing pyramidal neurons from non-CD tissue . In most cytomegalic and approximately 30 % of normal-appearing pyramidal neurons from CD tissue , DB01221 currents showed decreased Mg(2+) sensitivity compared with neurons from non-CD tissue . DB08954 had less effect in CD compared with non-CD neurons , indicating a functional loss of Q13224 subunits . DB01221 -evoked current density was decreased in cytomegalic compared with normal-appearing neurons . Single-cell reverse transcriptase polymerase chain reaction showed that all non-CD neurons expressed Q13224 subunit mRNA . By comparison , 22 % of pyramidal neurons in CD tissue lacked Q13224 mRNA . Immunofluorescence showed a decrease in Q13224 subunit expression in cytomegalic neurons and a subset of normal-appearing pyramidal neurons from CD tissue . Taken together , these results demonstrate the presence of DB01221 receptors with altered subunit composition and Mg(2+) sensitivity that could contribute to functional abnormalities in CD . DB00163 -related inhibition of monocyte P09917 and cardiovascular outcome in maintenance hemodialysis patients . A daily supplement of vitamin E is recommended for the secondary prevention of cardiovascular events in end-stage renal disease patients on maintenance hemodialysis . DB00163 has been entrusted with therapeutic properties against cardiovascular disease for more than 60 years . Several epidemiological studies and intervention trials have been performed with vitamin E , and some of them showed that it prevents atherosclerosis . For a long time , vitamin E was assumed to act by decreasing the oxidation of low-density lipoproteins , a key step in atherosclerosis initiation . However , at the cellular level vitamin E interferes with smooth muscle cell proliferation , platelet aggregation , monocyte adhesion , and oxidized low-density lipoproteins uptake and cytokine production , all reactions implied in the progression of atherosclerosis . Recent research points out that these effects may be not only the result of the antioxidant activity of vitamin E but also of its distinct molecular actions . These biological properties of vitamin E may allow to design better strategies for primary and secondary prevention of cardiovascular disease , with a potential exploitation of vitamin E supplements in primary and secondary prevention of major adverse cardiovascular events in all uremic patients . In this review , we also outline relevant patents on vitamin E and lipoxygenase inhibitors . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Q14703 lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 . DB03203 1-phosphate ( Q14703 ) and P21453 ( P21453 ) play an important role in the egress of mature P01730 or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY720 ) , an P21453 functional antagonist , induced significant accumulation of CD62L(high) Q07108 (low) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti- P21453 antibody revealed that P21453 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration . 2-Acetyl-4-tetrahydroxybutylimidazole ( THI ) , an Q14703 lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 ) . At 6h after THI administration , P21453 -expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla , suggesting Q14703 lyase expression in the cells constructing thymic medullary P15151 . To determine the cells expressing Q14703 lyase in the thymus , we newly established a mAb ( YK19-2 ) specific for mouse Q14703 lyase . Immunohistochemical staining with YK19-2 revealed that Q14703 lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 , Q14703 lyase was expressed in ER-TR7-positive cells ( reticular fibroblasts and pericytes ) and CD31-positive vascular endothelial cells . Our findings suggest that Q14703 lyase expressed in the thymic medullary P15151 keeps the tissue Q14703 concentration low around the vessels and promotes thymic egress via up-regulation of P21453 . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . Mechanism of purinergic activation of endothelial nitric oxide synthase in endothelial cells . BACKGROUND : Decreased endothelial nitric oxide ( NO ) synthase ( P29474 ) activity and NO production are critical contributors to the endothelial dysfunction and vascular complications observed in many diseases , including diabetes mellitus . Extracellular nucleotides activate P29474 and increase NO generation ; however , the mechanism of this observation is not fully clarified . METHODS AND RESULTS : To elucidate the signaling pathway(s) leading to nucleotide-mediated P29474 phosphorylation at DB00133 -1177 , human umbilical vein endothelial cells were treated with several nucleotides , including DB00171 , UTP , and ADP , in the presence or absence of selective inhibitors . These experiments identified P47900 , P41231 , and possibly P51582 as the purinergic receptors involved in P29474 phosphorylation and demonstrated that this process was adenosine independent . Nucleotide-induced P29474 phosphorylation and activity were inhibited by BAPTA-AM ( an intracellular free calcium chelator ) , rottlerin ( a protein kinase Cdelta inhibitor ) , and protein kinase Cdelta siRNA . In contrast , blockade of AMP-activated protein kinase , calcium/calmodulin-dependent kinase II , calcium/calmodulin-dependent kinase kinase , serine/threonine protein kinase B , protein kinase A , extracellular signal-regulated kinase 1/2 , and p38 mitogen-activated protein kinase did not affect nucleotide-mediated P29474 phosphorylation . CONCLUSIONS : The present study indicates that extracellular nucleotide-mediated P29474 phosphorylation is calcium and protein kinase Cdelta dependent . This newly identified signaling pathway opens new therapeutic avenues for the treatment of endothelial dysfunction . Roles of purinergic receptor P2Y , G protein-coupled 12 in the development of atherosclerosis in apolipoprotein E-deficient mice . OBJECTIVE : The aim of the study was to evaluate the role of purinergic receptor P2Y , G protein-coupled 12 ( Q9H244 ) , an ADP receptor , in the development of atherosclerotic lesions . METHODS AND RESULTS : P02649 -null mice were crossed with P2y12(-/-) mice to generate double knockout mice . The double knockout mice and the control apolipoprotein E-null mice were fed a high-fat diet for 20 weeks . Assessment of the atherosclerotic lesions in the control and double knockout mice demonstrated that Q9H244 deficiency caused a diminished lesion area , an increased fibrous content at the plaque site , and decreased monocyte/macrophage infiltration of the lesions . Polymerase chain reaction studies revealed that white blood cells do not express significant levels of Q9H244 . Bone marrow transplantation experiments confirmed that Q9H244 expressed on platelets is a key factor responsible for atherosclerosis , but do not exclude a role of smooth muscle cell Q9H244 . Supernatant fluid from activated P2y12(+/+) but not P2y12(-/-) platelets was capable of causing monocyte migration . In vitro studies showed that platelet Q9H244 deficiency suppressed platelet factor 4 secretion and P16109 expression . Further work demonstrated that platelet Q9H244 , through inhibition of the DB02527 /protein kinase A pathway , critically regulates the release of platelet factor 4 , and thereby affects monocyte recruitment and infiltration . CONCLUSIONS : These results demonstrate that Q9H244 modulates atherogenesis , at least in part by augmenting inflammatory cell recruitment via regulation of platelet α-granule release . DB00435 synthase inhibition and oxidative stress in cardiovascular diseases : possible therapeutic targets ? DB00435 ( NO ) is synthetized enzymatically from l-arginine ( l- DB00125 ) by three NO synthase isoforms , P35228 , P29474 and P29475 . The synthesis of NO is selectively inhibited by guanidino-substituted analogs of l- DB00125 or methylarginines such as asymmetric dimethylarginine ( DB01686 ) , which results from protein degradation in cells . Many disease states , including cardiovascular diseases and diabetes , are associated with increased plasma levels of DB01686 . The N-terminal catalytic domain of these NOS isoforms binds the heme prosthetic group as well as the redox cofactor , tetrahydrobiopterin ( BH(4) ) associated with a regulatory protein , calmodulin ( P62158 ) . The enzymatic activity of NOS depends on substrate and cofactor availability . The importance of BH(4) as a critical regulator of P29474 function suggests that BH(4) may be a rational therapeutic target in vascular disease states . BH(4) oxidation appears to be a major contributor to vascular dysfunction associated with hypertension , ischemia/reperfusion injury , diabetes and other cardiovascular diseases as it leads to the increased formation of oxygen-derived radicals due to NOS uncoupling rather than NO . Accordingly , abnormalities in vascular NO production and transport result in endothelial dysfunction leading to various cardiovascular disorders . However , some disorders including a wide range of functions in the neuronal , immune and cardiovascular system were associated with the over-production of NO . Inhibition of the enzyme should be a useful approach to treat these pathologies . Therefore , it appears that both a lack and excess of NO production in diseases can have various important pathological implications . In this context , NOS modulators ( exogenous and endogenous ) and their therapeutic effects are discussed . Population studies of polymorphisms at loci of neuropsychiatric interest ( tryptophan hydroxylase ( P17752 ) , dopamine transporter protein ( Q01959 ) , D3 dopamine receptor ( P35462 ) , apolipoprotein E ( P02649 ) , mu opioid receptor ( P35372 ) , and ciliary neurotrophic factor ( P26441 ) ) . We determined allele frequencies for polymorphisms at several loci of interest in neuropsychiatry-tryptophan hydroxylase ( P17752 ) , dopamine transporter protein ( Q01959 ) , D3 dopamine receptor ( P35462 ) , apolipoprotein E ( P02649 ) , ciliary neurotrophic factor ( P26441 ) , and the mu opioid receptor ( P35372 ) -in samples of individuals from populations in several different parts of the world . Associations with psychiatric illness have been proposed for specific polymorphisms at P17752 ( suicide-related behaviors and impulsivity ) , P35462 ( schizophrenia and bipolar affective disorder ) , Q01959 ( susceptibility to cocaine-induced paranoia and attention-deficit disorder ) , P26441 ( psychosis ) , and P35372 ( substance dependence ) . P02649 alleles are related to risk of Alzheimer disease . We found significant allele frequency variation among populations at all six loci . These results will provide a global framework of normal variation at these loci that might have functional significance or otherwise be related to susceptibility to various disorders or behavioral phenomena . Knowledge of this variation can be important for study design and data interpretation when individuals from various population groups are research subjects and may eventually help lead to a better understanding of behavioral adaptation . Involvement of P21453 receptor pathway in angiogenic effects of a novel adenosine-like nucleic acid analog COA-Cl in cultured human vascular endothelial cells . COA-Cl ( 2Cl-C. P01178 -A ) is a recently developed adenosine-like nucleic acid analog that promotes angiogenesis via the mitogen-activated protein ( Q96HU1 ) kinases P27361 /2 . Endothelial P21453 receptor plays indispensable roles in developmental angiogenesis . In this study , we examined the functions of P21453 in COA-Cl-induced angiogenic responses . Antagonists for P21453 , W146 , and VPC23019 , substantially but still partly inhibited the effects of COA-Cl with regard to P27361 /2 activation and tube formation in cultured human umbilical vein endothelial cells ( HUVEC ) . Antagonists for adenosine A1 receptor and purinergic P47900 receptor were without effect . Genetic knockdown of P21453 with siRNA , but not that of Q99500 , attenuated COA-Cl-elicited P27361 /2 responses . The signaling properties of COA-Cl showed significant similarities to those of sphingosine 1-phosphate , an endogenous P21453 ligand , in that both induced responses sensitive to pertussis toxin ( Gα i/o inhibitor ) , 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis ( acetoxymethyl ester ) ( BAPTA-AM ) , ( calcium chelator ) , and Q99463 ( c-Src tyrosine kinase inhibitor ) . COA-Cl elevated intracellular Ca(2+) concentration and induced tyrosine phosphorylation of p130Cas , a substrate of c-Src , in HUVEC . COA-Cl displaced [(3)H] Q14703 in a radioligand-binding competition assay in chem-1 cells overexpressing P21453 . However , COA-Cl activated P27361 /2 in CHO- P04264 cells that lack functional P21453 receptor , suggesting the presence of additional yet-to-be-defined COA-Cl target in these cells . The results thus suggest the major contribution of P21453 in the angiogenic effects of COA-Cl . However , other mechanism such as that seen in CHO- P04264 cells may also be partly involved . Collectively , these findings may lead to refinement of the design of this nucleic acid analog and ultimately to development of small molecule-based therapeutic angiogenesis . Alpha7 nicotinic acetylcholine receptor gene and reduced risk of Alzheimer 's disease . BACKGROUND : Sporadic Alzheimer 's disease ( AD ) is a common disabling disease of complex aetiology for which there are limited therapeutic options . We sought to investigate the role of the alpha7 nicotinic acetylcholine receptor gene ( P36544 ) in influencing risk of AD in a large population . P36544 is a strong candidate gene for AD for several reasons : ( 1 ) its expression is altered differentially in the AD brain ; ( 2 ) it interacts directly with beta amyloid peptide ( Abeta(42) ) ; and ( 3 ) agonist activation induces several neuroprotective pathways . METHODS : In this study we used a genetic haplotype approach to assess the contribution of common variation at the P36544 locus to risk of AD . Fourteen single nucleotide polymorphisms ( SNPs ) were genotyped in 764 AD patients and 314 controls . RESULTS : Three blocks of high linkage disequilibrium ( LD ) and low haplotype diversity were identified . The block 1 TCC haplotype was significantly associated with reduced odds of AD ( p = 0.001 ) and was independent of apolipoprotein E ( P02649 ) status . Individual SNPs were not associated with risk for AD . CONCLUSIONS : We conclude that genetic variation in P36544 influences susceptibility to AD . These results provide support for the development of alpha7nAChR agonists or modulators as potential drug treatments for AD . Further work is necessary to replicate the findings in other populations . Novel therapies in lupus - focus on nephritis . Lupus nephritis ( LN ) is one of the major complications of Systemic Lupus Erythematosus ( SLE ) and its treatment remains a challenge . Although the classical and widely used immunosuppressive agents have accounted for a significant improvement in the survival and decreased the progression to end-stage renal failure they lack selectivity for the underlying immune dysregulation . In addition the toxicity related to their use and the relapses after treatment are of major concern not least because of the adverse effect on the prognosis of the patients with SLE who have kidney involvement . The development of more specific pharmacological agents for patients with SLE is still a major research goal . Ideally these agents should provide a better long-term prognosis for SLE patients and be less toxic . In this review we summarise the mechanism of action and the results obtained with a variety of drugs that have recently been utilized in the treatment of patients with lupus especially those with nephritis . We discuss the clinical usefulness of B- -cell depletion principally anti- P11836 antibodies blockage of co-stimulatory pathways ( anti- P29965 antibody DB01281 ) the induction of immune tolerance ( LJP 394 peptide specific vaccination ) and therapy targeting cytokines ( anti- P22301 antibody Q9Y275 blockage ) and the complement system ( anti- P01031 antibody ) . Immunoablative doses of Cyclophosphamide ( CyC ) with or without Haematopoietic Stem Cell Transplantation ( HSCT ) and the possibilities of gene therapy are also reviewed . The use of intravenous immunoglobulin ( DB00028 ) and plasmapheresis are not discussed because these treatments have been used in clinical practice for several years . DB00640 A2B receptor and hyaluronan modulate pulmonary hypertension associated with chronic obstructive pulmonary disease . Chronic obstructive pulmonary disease ( P48444 ) is the fourth leading cause of death worldwide . The development of pulmonary hypertension ( PH ) in patients with P48444 is strongly associated with increased mortality . Chronic inflammation and changes to the lung extracellular matrix ( Q13201 ) have been implicated in the pathogenesis of P48444 , yet the mechanisms that lead to PH secondary to P48444 remain unknown . Our experiments using human lung tissue show increased expression levels of the adenosine A2B receptor ( P29275 ) and a heightened deposition of hyaluronan ( HA ; a component of the Q13201 ) in remodeled vessels of patients with PH associated with P48444 . We also demonstrate that the expression of Q92819 correlates with mean pulmonary arterial pressures in patients with P48444 , with and without a secondary diagnosis of PH . Using an animal model of airspace enlargement and PH , we show that the blockade of P29275 is able to attenuate the development of a PH phenotype that correlates with reduced levels of HA deposition in the vessels and the down-regulation of genes involved in the synthesis of HA . Effects of an alpha 7-nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3-(2,4-dimethoxybenzylidene)-anabaseine ( DB05708 ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 , the α7-nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 genotype . CONCLUSIONS : The observed DB05708 -related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7-nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . First analysis of the relation between P33261 genotype and pharmacodynamics in patients treated with ticagrelor versus clopidogrel : the ONSET/OFFSET and RESPOND genotype studies . BACKGROUND : The influence of cytochrome P450 ( CYP ) 2C19 genotype on platelet function in patients treated with ticagrelor versus clopidogrel is unknown . METHODS AND RESULTS : P33261 ( *1 , *2 , *3 , *4 , *5 , *6 , *7 , *8 , *17 ) genotyping was performed in patients with coronary artery disease treated with ticagrelor ( 180-mg load , 90 mg P55957 ) ( n=92 ) or clopidogrel ( 600-mg load , 75 mg/d ) ( n=82 ) . All patients received 75 to 100 mg/d aspirin . Platelet function was measured by aggregometry , VerifyNow Q9H244 assay , and vasodilator-stimulated phosphoprotein-phosphorylation assay at predose , 8 hours postloading , and maintenance . In each treatment group , patients were categorized according to 2C19 genotype carrier status ( loss-of-function , gain-of-function ) and metabolizer status . Kruskal-Wallis test was used to compare platelet function among these categories for each treatment , and Wilcoxon rank sum test was used to compare platelet function between the clopidogrel and ticagrelor groups for each category . There was no statistically significant influence of genotype on platelet function during aspirin therapy alone . DB08816 exhibited lower platelet reactivity than clopidogrel by all assays irrespective of 2C19 genotype or metabolizer status ( P < 0.01 ) . Loss-of-function carriers had greater platelet reactivity during clopidogrel therapy . The influence of genotype on platelet reactivity was greatest during clopidogrel maintenance and best demonstrated by the VerifyNow Q9H244 assay . CONCLUSIONS : This report is the first to demonstrate the superior pharmacodynamic effect of ticagrelor compared with clopidogrel irrespective of P33261 genotype . Whereas P33261 genotype influenced the antiplatelet effect of clopidogrel , there was no effect of P33261 genotype during ticagrelor therapy . Selective inhibition of P09917 attenuates glomerulonephritis in the rat . Three hours following injection of anti-GBM ( glomerular basement membrane ) antibody ( 10 mg/kg , i.v. ) into rats , glomerular production of LTB4 was significantly increased as compared to untreated rats ( 497 +/- 26 vs. 244 +/- 18 pg of LTB4/mg protein ) . Twenty-four hours following administration of anti-GBM antibody , renal function ( blood urea nitrogen , BUN ; plasma creatinine , PCr ; creatinine clearance , CCr ; fractional excretion of sodium , FENa ; fractional excretion of urea , FEUrea ) was determined to be impaired proportionally to the amount of injected antibody ( 5 to 30 mg/kg , i.v. ) . In a second series of experiments , a selective P09917 ( P09917 ) inhibitor , SK & F 107649 , was used to investigate the involvement of P09917 products in the pathophysiology of anti-GBM antibody-induced glomerulonephritis . In preliminary experiments . SK & F 107649 ( 50 to 200 mg/kg , p.o. ) inhibited ionophore ( A23187 ) -stimulated whole blood leukotriene ( LT ) B4 production in a dose-dependent fashion ( P < 0.05 at doses > or = 100 mg/kg ) . The anti-GBM antibody-mediated decrease in glomerular filtration rate ( Q92565 ) and increase in BUN and PCr were dose-dependently attenuated by SK & F 107649 ( 50 to 200 mg/kg , p.o. P55957 ) . These data confirm that glomerular LTB4 production is stimulated in anti-GBM antibody-induced glomerulonephritis , and demonstrate that inhibition of P09917 by SK & F 107649 normalizes BUN and PCr and attenuate the decrease in Q92565 following anti-GBM antibody treatment . These results provide additional evidence for a role of P09917 products in immune-mediated renal disease , and suggest that pharmacologic inhibition of P09917 may be of therapeutic value . Activation of NR1a/ Q13224 receptors by monocyte-derived macrophage secretory products : implications for human immunodeficiency virus type one-associated dementia . The final pathways for neuronal injury in human immunodeficiency virus type one ( HIV-1 ) -associated dementia ( HAD ) were investigated in Xenopus oocytes expressing recombinant NR1a/ Q13224 N-methyl-D-aspartate ( DB01221 ) receptors exposed to secretory products from HIV-infected macrophages . Pressure ejection of HIV-1-infected and P29965 -stimulated human monocyte-derived macrophage ( MDM ) fluids produced inward currents in oocytes expressing NR1a/ Q13224 ( 30.2+/-5.1 nA , n=42 , mean+/-SE ) , but not in uninjected cells . In contrast , control ( uninfected MDM ) fluids induced currents of 4.5+/-0.5 nA ( n=17 ) . Infected or stimulated MDM without virus showed intermediate responses . The induced currents were MDM fluid dose-dependent and blocked by the DB01221 receptor antagonist 2-amino-5-phosphonovalerate ( 50 microM ) , but not by 6-cyano-7-nitroquinoxaline-2,3-dione ( 20 microM ) . Although low levels of glutamate were detected in the culture fluids , the addition of L-glutamate decarboxylase to the MDM did not significantly change the level of induced inward currents . Our experiments demonstrate that secretory factors from HIV-1-infected MDM activate DB01221 receptors NR1a/ Q13224 and may contribute to neuronal demise during HAD .

MH_train_1226

MH_train_1226

MH_train_1226

interacts_with DB01067?

multiple_choice

beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells . Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention , the molecular mechanisms underlying such chemopreventive activity are not clear . Since peroxisome proliferator-activated receptor gamma ( P37231 ) has been implicated in regulating breast cancer cell differentiation and apoptosis , the effects of beta-carotene on the P37231 -mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study . The results demonstrated that beta-carotene significantly increased P37231 mRNA and protein levels in time-dependent manner . In addition , beta-carotene increased the cyclin-dependent kinase inhibitor P38936 ( P38936 /CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression . DB07863 ( GW9662 ) , an irreversible P37231 antagonist , partly attenuated the cell death caused by beta-carotene . Further , reactive oxygen species ( ROS ) production was induced by beta-carotene , resulting in mitochondrial dysfunction and cytochrome C release . DB00143 ( DB00143 ) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene . In total , these observations suggest that the synergistic effect of P37231 expression and ROS production may account for beta-carotene-mediated anticancer activities . Comparative effects of azimilide and ambasilide on the human ether-a-go-go-related gene ( Q12809 ) potassium channel . OBJECTIVE : To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related ( Q12809 ) channel . METHODS : Q12809 was stably transfected into Chinese hamster ovary ( CHO- P04264 ) cells and currents were measured using a whole cell , voltage-clamp technique . RESULTS : DB04957 had a ' dual effect ' , inhibiting current at voltage steps above -40 mV and augmenting current at -40 and -50 mV . Tail current inhibition following a step to +30 mV did not vary with temperature ( IC(50) 610 nM at 22 degrees C and 560 nM at 37 degrees C ) . The agonist effect at -50 mV was concentration-dependent and correlated with a hyperpolarizing shift in the V(1/2) of activation ( r=0.98 , P < 0.05 ) . Time constants of inactivation were faster and there was a -10 mV shift in the V(1/2) of steady state inactivation suggestive of open and inactivated state binding . By comparison , ambasilide inhibited Q12809 channels with lower potency ( IC(50) 3.6 microM ) , in a voltage- and time-dependent but frequency-independent manner ( 0.03-1 Hz ) . Ambasilide had no effect on activation or inactivation gating but prolonged both fast and slow components of deactivation consistent with unbinding from the open state . The net effect of both drugs was similar during a voltage ramp which simulated a cardiac action potential . CONCLUSIONS : Inhibition of Q12809 channels by azimilide and ambasilide exhibits a similar time and voltage-dependence . While both exhibit affinity for the open state , azimilide also binds to inactivated channels . Potent and selective activation of the pancreatic beta-cell type K( DB00171 ) channel by two novel diazoxide analogues . AIMS/HYPOTHESIS : We investigated the pharmacological properties of two novel DB00171 sensitive potassium ( K( DB00171 ) ) channel openers , 6-Chloro-3-isopropylamino-4 H-thieno [ 3,2- e ] -1,2,4-thiadiazine 1,1-dioxide ( NNC 55-0118 ) and 6-chloro-3-(1-methylcyclopropyl)amino-4 H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide ( NN414 ) , on the cloned cardiac ( Kir6.2/SUR2A ) , smooth muscle ( Kir6.2/SUR2B ) and pancreatic beta cell ( Kir6.2/ Q09428 ) types of K( DB00171 ) channel . METHODS : We studied the effects of these compounds on whole-cell currents through cloned K( DB00171 ) channels expressed in Xenopus oocytes or mammalian cells ( HEK293 ) . We also used inside-out macropatches excised from Xenopus oocytes . RESULTS : In P29320 293 cells , NNC 55-0118 and NN414 activated Kir6.2/ Q09428 currents with EC(50) values of 0.33 micromol/l and 0.45 micromol/l , respectively , compared with that of 31 micro mol/l for diazoxide . Neither compound activated Kir6.2/SUR2A or Kir6.2/SUR2B channels expressed in oocytes , nor did they activate Kir6.2 expressed in the absence of Q09428 . Current activation was dependent on the presence of intracellular MgATP , but was not supported by MgADP . CONCLUSION/INTERPRETATION : Both NNC 55-0118 and NN414 selectively stimulate the pancreatic beta-cell type of K( DB00171 ) channel with a higher potency than diazoxide , by interaction with the Q09428 subunit . The high selectivity and efficacy of the compounds could prove useful for treatment of disease states where inhibition of insulin secretion is beneficial . Phosphorylation of thymidylate synthase from various sources by human protein kinase CK2 and its catalytic subunits . P04818 ( TS ) was found to be a substrate for both catalytic subunits of human CK2 , with phosphorylation by CK2alpha and CK2alpha ' characterized by similar K(m) values , 4.6microM and 4.2microM , respectively , but different efficiencies , the apparent turnover number with CK2alpha being 10-fold higher . With both catalytic subunits , phosphorylation of human TS , like calmodulin and P55957 , was strongly inhibited in the presence of the regulatory subunit CK2beta , the holoenzyme being activated by polylysine . Phosphorylation of recombinant human , rat , mouse and Trichinella spiralis TSs proteins was compared , with the human enzyme being apparently a much better substrate than the others . Following hydrolysis and TLC , phosphoserine was detected in human and rat , and phosphotyrosine in T. spiralis , TS , used as substrates for CK2alpha . MALDI-TOF MS analysis led to identification of phosphorylated DB00133 (124) in human TS , within a sequence LGFS(124)TREEGD , atypical for a CK2 substrate recognition site . The phosphorylation site is located in a region considered important for the catalytic mechanism or regulation of human TS , corresponding to the loop 107-128 . Following phosphorylation by CK2alpha , resulting in incorporation of 0.4mol of phosphate per mol of dimeric TS , human TS exhibits unaltered K(m) values for DB03800 and N(5,10)-methylenetetrahydrofolate , but a 50 % lower turnover number , pointing to a strong influence of DB00133 (124) phosphorylation on its catalytic efficiency . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . GSK-3 inactivation or depletion promotes β-cell replication via down regulation of the CDK inhibitor , p27 ( Kip1 ) . Diabetes ( T1DM and T2DM ) is characterized by a deficit in β-cell mass . A broader understanding of human β-cell replication mechanism is thus important to increase β-cell proliferation for future therapeutic interventions . Here , we show that p27 ( Kip1 ) , a CDK inhibitor , is expressed abundantly in isolated adult human islets and interacts with various positive cell cycle regulatory proteins including D-type cyclins ( D1 , D2 and D3 ) and their kinase partners , P11802 and Q00534 . Also , we see interaction of cyclin E and its kinase partner , P24941 , with p27 suggesting a critical role of p27 as a negative cell cycle regulator in human islets . Our data demonstrate interaction of p27 with GSK-3 in β-cells and show , employing rodent β-cells ( P01308 -1 ) , isolated human islets and purified β-cells derived from human islets , that siRNA-mediated depletion of GSK-3 or p27 or 1-AKP / BIO - mediated GSK-3 inhibition results in increased β-cell proliferation . We also see reduction of p27 levels following GSK-3 inactivation or depletion . Our data show that serum induction of quiescent P01308 -1 cells leads to sequential phosphorylation of p27 on its S10 and T187 residues with faster kinetics for S10 corresponding with the decreased levels of p27 . Altogether our findings indicate that p27 levels in β-cells are stabilized by GSK-3 and thus p27 down regulation following GSK-3 depletion / inactivation plays a critical role in promoting β-cell replication . Protective effect of hyaluronic acid on interleukin-1-induced deregulation of beta1-integrin and insulin-like growth factor-I receptor signaling and collagen biosynthesis in cultured human chondrocytes . The mechanism of protective action of hyaluronic acid ( HA ) on collagen metabolism disturbances in tissues during inflammation is not known . P01308 -like growth factor-I ( P05019 ) receptor and beta1-integrin receptor signaling plays an important role in the regulation of collagen biosynthesis at both transcriptional and post-transcriptional levels . The present study was undertaken to evaluate the effect of IL-1beta ( inductor of experimental inflammation ) on the signaling pathways as well as on collagen biosynthesis , gelatinases and prolidase activity in cultured human chondrocytes and the effect of HA on these processes . It was found that IL-1beta-dependent inhibition of collagen biosynthesis was accompanied by increase in beta1-integrin receptor , NF-kB expressions , and increase in phosphorylation of Q05397 , that resulted in stimulation of metalloproteinase P08253 and P14780 activities , but not prolidase activity and expression . Simultaneously , decrease in expression of P08069 and phosphorylation of Akt and p38 were found . All those processes were counteracted by HA . This suggests that cross talk between beta1-integrin and P05019 receptors is disturbed by IL-1beta , and HA recovers their proper signaling in cultured chondrocytes . We propose that P08069 and beta1-integrin signaling may play an important role in protective effect of hyaluronic acid on interleukin-1-induced inhibition of collagen biosynthesis in cultured human chondrocytes . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . DB01411 inhibits renal epithelial cyst progression via activation of AMP-activated protein kinase . Q9Y271 ( CysLT1 receptor ) antagonists were found to inhibit chloride secretion in human airway epithelial cells . Since chloride secretion in renal epithelial cells , which shares common mechanisms with airway epithelial cells , plays important roles in renal cyst progression in polycystic kidney disease ( Q15139 ) , this study was aimed to investigate effects of drugs acting as CysLT1 receptor antagonists on renal cyst progression and its underlying mechanisms . Effects of CysLT1 receptor antagonists on renal cyst growth and formation were determined using Madine Darby canine kidney ( MDCK ) cyst models . Mechanisms of actions of CysLT1 receptor antagonists were determined using short-circuit current measurement , assays of cell viability and cell proliferation , and immunoblot analysis of signaling proteins . Of the three drugs acting as CysLT1 receptor antagonists ( montelukast , pranlukast and zafirlukast ) tested , pranlukast was the most promising drug that inhibited MDCK cyst growth and formation without affecting cell viability . Its effect was independent of the inhibition of CysLT1 receptors . Instead , it reduced DB02527 -activated chloride secretion and proliferation of MDCK cells in an AMP-activated protein kinase ( AMPK ) -dependent manner and had no effect on P13569 protein expression . Interestingly , pranlukast enhanced AMPK activation via calcium/calmodulin-dependent protein kinase kinase beta ( CaMKKβ ) with consequent activation of acetyl- DB01992 carboxylase ( ACC ) and suppression of mammalian target of rapamycin ( P42345 ) pathway . These results indicate that pranlukast retards renal epithelial cyst progression by inhibiting DB02527 -activated chloride secretion and cell proliferation via CaMKKβ-AMPK- P42345 pathway . Therefore , pranlukast represents a class of known drugs that may have potential utility in Q15139 treatment . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Low ethanol concentration alters P30532 RNA levels during early human development . DB00898 use is common and consumption during pregnancy has been shown to lead to a myriad of physical and neurologic abnormalities commonly referred to as fetal alcohol spectrum disorder . Substance addiction , which includes alcohol , has been shown to involve the major nicotinic acetylcholine receptor subunit P30532 . Using human embryonic stem cells as a model of early human development , we show that low concentrations of ethanol ( 20mM ) can alter the expression of P30532 . Changes in P30532 expression is linked to altered GABA and DB01221 receptor expression , as well as abnormal development of the frontal cortex . These results suggest that alcohol exposure can alter early neurologic development , which may favor addiction and other developmental abnormalities in unborn children . Binding affinities for sulfonamide inhibitors with matrix metalloproteinase-2 using a linear response method . Due to their involvement in many pathological conditions , matrix metalloproteinases ( MMPs ) , are very attractive therapeutic targets . Our study focuses on one of them , P08253 , which is involved in tumor progression and metastasis . Recently , the solution structure of the catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) was published by Feng et al . Using the Hanessian group published binding affinity data and the structure published by Feng as a basis , we have built a binding affinity model by targeting the S(2) ' pocket of the enzyme with a set of nine alpha-N-sulfonylamino hydroxamic acid derivatives . Two binding geometries of each ligand have been generated corresponding to two binding modes denoted A and B , respectively , of which the first one is targeting the S(2) ' pocket and the second one the S(1) pocket . For the binding affinity model developed for mode A the computed activities show a rmsd of 0.583 kcal/mol as compared with the experimental data , and a correlation coefficient r(2) of 0.779 , while in the case of the binding mode B a rmsd of 0.834 kcal/mol and correlation coefficient r(2) of 0.500 , respectively , were obtained . In conclusion , our data suggest a higher probability for the DB00120 (76) gated S(2) ' open form pocket to accommodate the substituent alpha versus the wide solvent exposed S(1) subsite , probability which some research groups could have overlooked due to extensive use in their calculations of non revealing S(2) ' pocket open state crystallographic structures instead of NMR ones . Comparison of the micro- and macro-vascular effects of glimepiride and gliclazide in metformin-treated patients with Type 2 diabetes : a double-blind , crossover study . AIMS : To compare the metabolic and vascular effects of two sulphonylureas ( SU ) , gliclazide ( specific for the pancreatic [ Q09428 ] receptor ) and glimepiride ( a nonspecific agent that also binds to vascular and cardiac [ SUR2 ] receptors ) , during chronic administration in metformin-treated patients with Type 2 diabetes ( T2DM ) . METHODS : A randomized , double-blind , crossover study of gliclazide 80 mg P55957 and glimepiride 2 mg OD , each for 4 weeks as add-on therapy to metformin , with a 4-week washout period . Patients attended four study mornings after first dose and 4 weeks ' SU treatment for measurements of arterial distensibility ( Ax ) , pressor responsiveness to i.v. angiotensin II ( ANGII ) , and cutaneous microvascular vasodilator responses to iontophoresis of acetylcholine ( ACh ) and sodium nitroprusside ( SNP ) . RESULTS : Glycaemic responses were similar ( e.g. serum fructosamine was 315 vs 329 micro mol l-1 after 4 weeks ) , and there was no change in augmentation index during treatment with either SU ( 9.1 vs 9.8 mmHg after 4 weeks [ 95 % confidence interval -8.1 , 10.5 ] ) . Similarly , there were no differences between treatments in pressor responsiveness ( e.g. PD10 [ dose of agonist required to increase mean BP by 10 mmHg ] for ANGII was 1.37 vs 1.68 ng kg-1 min-1 [ -4.3 , 6.9 ] ) or cutaneous microvascular vasodilator responses ( peak ACh response 68 +/- 36 vs 63 +/- 34 perfusion units [ -82.7 , 79.1 ] ) . CONCLUSIONS : There is no evidence that Q09428 -specific and nonspecific SUs have differential effects on arterial distensibility , endothelial function or vasodilator mechanisms in metformin-treated patients with T2DM . Inhibition of cyclin-dependent kinases , GSK-3beta and CK1 by hymenialdisine , a marine sponge constituent . BACKGROUND : Over 2000 protein kinases regulate cellular functions . Screening for inhibitors of some of these kinases has already yielded some potent and selective compounds with promising potential for the treatment of human diseases . RESULTS : The marine sponge constituent hymenialdisine is a potent inhibitor of cyclin-dependent kinases , glycogen synthase kinase-3beta and casein kinase 1 . DB02950 competes with DB00171 for binding to these kinases . A P24941 -hymenialdisine complex crystal structure shows that three hydrogen bonds link hymenialdisine to the Glu81 and Leu83 residues of P24941 , as observed with other inhibitors . DB02950 inhibits Q00535 /p35 in vivo as demonstrated by the lack of phosphorylation/down-regulation of Pak1 kinase in E18 rat cortical neurons , and also inhibits GSK-3 in vivo as shown by the inhibition of P46821 phosphorylation . DB02950 also blocks the in vivo phosphorylation of the microtubule-binding protein tau at sites that are hyperphosphorylated by GSK-3 and Q00535 /p35 in Alzheimer 's disease ( cross-reacting with Alzheimer's-specific AT100 antibodies ) . CONCLUSIONS : The natural product hymenialdisine is a new kinase inhibitor with promising potential applications for treating neurodegenerative disorders . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . Isoenzyme-specific cyclooxygenase inhibitors : a whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6 . NSAIDs inhibit the conversion of arachidonic acid into DB03866 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase ( P36551 ) . Two genetically distinct isoforms have been discovered , P23219 and P35354 . While P23219 is thought to account for homeostatic amounts of eicosanoids , P35354 is induced during inflammation leading to pathologic amounts of eicosanoids . Since NSAIDs inhibit both P36551 isoforms , antiinflammatory drug research has refocused to discovering P35354 inhibitors that do not inhibit P23219 . For this purpose , we have developed a whole cell assay system using the human erythroleukemic cell line HEL as a source for P23219 and the human monocytic cell line Mono Mac 6 as a source for P35354 . Mono Mac 6 cells express high amounts of P35354 upon stimulation with lipopolysaccharide ( LPS ) in the absence of any detectable P23219 protein . On the other hand , we find HEL cells to naturally express P23219 protein , but not P35354 . Testing of a panel of NSAIDs as well as some P35354 specific inhibitors showed that this assay system is suitable for identifying compounds that selectively inhibit either P23219 or P35354 . This test system offers the advantage of assessing P23219 and P35354 inhibitors within the human species , within a similar test set-up , and circumvents the need for tedious purification of either platelets or peripheral blood monocytes . Impact of human D398N single nucleotide polymorphism on intracellular calcium response mediated by α3β4α5 nicotinic acetylcholine receptors . The human P30532 D398N polymorphism ( rs16969968 ) causes an aspartic acid to asparagine change in the nicotinic acetylcholine receptor ( nAChR ) α5 subunit gene . The N398 variant of P30532 is linked to increased risk for nicotine dependence . In this study , we explored the effect of the P30532 D398N polymorphism on the properties of human α3β4* nicotinic acetylcholine receptors in human embryonic kidney ( P29320 ) cells . Addition of either D398 or N398 variant of α5 subunit in the α3β4* receptor did not affect total [ (125)I ] -epibatidine binding or surface expression of the receptor . However , addition of α5(D398) into α3β4* receptor decreased the maximal response to agonist without significantly affecting EC(50) in aequorin intracellular calcium assay . α3β4α5(N398) nAChRs showed further decreased maximal response . The differences in agonist efficacy between the receptor subtypes were found to be dependent upon the concentration of external calcium but independent of external sodium . Moreover , activation of α3β4α5 nAChRs led to significantly greater intracellular calcium release from IP(3) stores relative to α3β4 nAChRs although no effect of the α5 polymorphism was observed . Finally , inclusion of the α5 variant caused a small shift to the left in IC(50) for some of the antagonists tested , depending upon α5 variant but did not affect sensitivity of α3β4* receptors to desensitization in response to incubation with nicotine . In conclusion , addition of either variant of α5 into an α3β4α5 receptor similarly effects receptor pharmacology and function . However , the N398 variant exhibits a reduced response to agonists when extracellular calcium is high and it may lead to distinct downstream cellular signaling . Effects on thrombin generation of single injections of DB02351 in patients with calf vein thrombosis . STUDY OBJECTIVE : To determine whether single injections of DB02351 , a direct thrombin inhibitor , can inhibit thrombin generation in patients with calf vein thrombosis and , if so , if the inhibition is sustained . DESIGN : Phase II open label cohort study . SETTING : Tertiary-care referral centres , university affiliated hospitals . PATIENTS : 10 patients with venographically-demonstrated calf vein thrombosis . INTERVENTION : Patients received a single injection of DB02351 , either 1.0 mg/kg subcutaneously or 0.6 mg/kg as a 15 min intravenous infusion . P00734 fragment ( F1++2 ) levels , as an index of thrombin generation , were measured before as well as 6 h post- and 24 h post- DB02351 administration . Patients were followed with non-invasive tests to detect thrombus extension into the proximal veins . RESULTS : There was a significant reduction in the levels of F1+2 with both regimens , 6 h after DB02351 . The F1+2 levels 24 h post- DB02351 showed a significant increase relative to the 6 h post- DB02351 results . One patient developed thrombus extension into the popliteal vein and was treated with conventional anticoagulants . CONCLUSION : The single injections of DB02351 used in the study produced incomplete and temporary suppression of F1+2 . Complete and permanent inhibition of thrombin generation with DB02351 in patients with calf vein thrombosis may require higher doses , multiple subcutaneous injections and/or prolonged intravenous infusion . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Inhibition of prothrombin kringle-2-induced inflammation by minocycline protects dopaminergic neurons in the substantia nigra in vivo . P00734 kringle-2 ( pKr-2 ) , a domain of prothrombin , can cause the degeneration of mesencephalic dopaminergic neurons through microglial activation . However , the chemical products that inhibit pKr-2-induced inflammatory activities in the brain are still not well known . The present study investigated whether minocycline , a semisynthetic tetracycline derivative , could inhibit pKr-2-induced microglial activation and prevent the loss of nigral dopaminergic ( DA ) neurons in vivo . To address this question , rats were administered a unilateral injection of pKr-2 in the substantia nigra in the presence or absence of minocycline . Our results show that pKr-2 induces the production of proinflammatory cytokines , such as tumor necrosis factor-α ( P01375 -α ) and interleukin-1β ( IL-1β ) , and inducible nitric oxide synthase from the activated microglia . In parallel , 7 days after pKr-2 injection , tyrosine hydroxylase immunocytochemical analysis and western blot analysis showed a significant loss of nigral DA neurons . This neurotoxicity was antagonized by minocycline and the observed neuroprotective effects were associated with the ability of minocycline to suppress the expression of tumor necrosis factor-α , interleukin-1β , and nitric oxide synthase . These results suggest that minocycline may be promising as a potential therapeutic agent for the prevention of DA neuronal degeneration associated with pKr-2-induced microglial activation . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Titration of KATP channel expression in mammalian cells utilizing recombinant baculovirus transduction . A variety of transfection approaches have been used to deliver plasmids encoding ion channel genes into cells . We have used the baculovirus transduction system , BacMam , to demonstrate transient expression of multi-subunit KATP channels in CHO- P04264 and P29320 -293 EBNA cells using sulfonylurea receptor 1 ( Q09428 ) , SUR2A , SUR2B , and P55040 6.2 genes . [ 3H ] -glyburide binding , patch clamp , and DiBAC4(3) measurements of membrane potential changes were used to monitor channel expression . BacMam delivery of each Q09428 isoform with KIR6.2 was demonstrated based on its pharmacological profiles . Expression levels of Q09428 and KIR6.2 were titrated by varying the viral concentration or time of virus addition , with functional activity measured in as little as 4-6 hours posttransduction . Further increases in BacMam virus induced sufficient KATP expression to dominate membrane potential without pharmacological opening of the channel . Independently altering treatment with virus containing either the Q09428 or KIR6.2 gene revealed interactions among subunits during formation of functional channels in the plasma membrane . This study demonstrates the utility and versatility of BacMam as a valuable gene delivery tool for the study of ion channel function .

MH_train_1227

MH_train_1227

MH_train_1227

interacts_with DB06684?

multiple_choice

P05067 -dependent alteration of GSK3β activity impairs neurogenesis in the Ts65Dn mouse model of Down syndrome . Intellectual disability in Down syndrome ( DS ) appears to be related to severe neurogenesis impairment during brain development . The molecular mechanisms underlying this defect are still largely unknown . Accumulating evidence has highlighted the importance of GSK3β signaling for neuronal precursor proliferation/differentiation . In neural precursor cells ( NPCs ) from Ts65Dn mice and human fetuses with DS , we found reduced GSK3β phosphorylation and , hence , increased GSK3β activity . In cultures of trisomic subventricular-zone-derived adult NPCs ( aNPCs ) we found that deregulation of GSK3β activity was due to higher levels of the AICD fragment of the trisomic gene P05067 that directly bound to GSK3β . We restored GSK3β phosphorylation in trisomic aNPCs using either lithium , a well-known GSK3β inhibitor , or using a 5-HT receptor agonist or fluoxetine , which activated the serotonin receptor P08908 . Importantly , this effect was accompanied by restoration of proliferation , cell fate specification and neuronal maturation . In agreement with results obtained in vitro , we found that early treatment with fluoxetine , which was previously shown to rescue neurogenesis and behavior in Ts65Dn mice , restored GSK3β phosphorylation . These results provide a link between GSK3β activity alteration , P05067 triplication and the defective neuronal production that characterizes the DS brain . Knowledge of the molecular mechanisms underlying neurogenesis alterations in DS may help to devise therapeutic strategies , potentially usable in humans . Results suggest that drugs that increase GSK3β phosphorylation , such as lithium or fluoxetine , may represent useful tools for the improvement of neurogenesis in DS . Localization of the succinate receptor in the distal nephron and its signaling in polarized MDCK cells . When the succinate receptor ( Q9BXA5 ) is activated in the afferent arterioles of the glomerulus it increases renin release and induces hypertension . To study its location in other nephron segments and its role in kidney function , we performed immunohistochemical analysis and found that Q9BXA5 is located in the luminal membrane of macula densa cells of the juxtaglomerular apparatus in close proximity to renin-producing granular cells , the cortical thick ascending limb , and cortical and inner medullary collecting duct cells . In order to study its signaling , Q9BXA5 was stably expressed in Madin-Darby Canine Kidney ( MDCK ) cells , where it localized to the apical membrane . Activation of the cells by succinate caused Gq and Gi-mediated intracellular calcium mobilization , transient phosphorylation of extracellular regulated kinase ( P29323 )1/2 and the release of arachidonic acid along with prostaglandins E2 and I2 . Signaling was desensitized without receptor internalization but rapidly resensitized upon succinate removal . Immunohistochemical evidence of phosphorylated P27361 /2 was found in cortical collecting duct cells of wild type but not Q9BXA5 knockout streptozotocin-induced diabetic mice , indicating in vivo relevance . Since urinary succinate concentrations in health and disease are in the activation range of the Q9BXA5 , this receptor can sense succinate in the luminal fluid . Our study suggests that changes in the luminal succinate concentration may regulate several aspects of renal function . Anticonvulsant effects of agomelatine in mice . DB06594 is a potent MT1 and P02795 melatonin receptor agonist and a P28335 serotonin receptor antagonist . We analyzed whether agomelatine has anticonvulsant properties . The anticonvulsant activity of agomelatine ( 25 , 50 or 75 mg/kg , i.p. ) was evaluated in mouse models of pentylenetetrazole ( PTZ-85 mg/kg , i.p. ) , pilocarpine ( 400mg/kg , i.p. ) , picrotoxin ( 7 mg/kg , i.p. ) , strychnine ( 75 mg/kg , i.p. ) or electroshock-induced convulsions . In the PTZ-induced seizure model , agomelatine ( at 25 or 50mg/kg ) showed a significant increase in latency to convulsion , and agomelatine ( at 50 or 75 mg/kg ) also increased significantly time until death . In the pilocarpine-induced seizure model , only agomelatine in high doses ( 75 mg/kg ) showed a significant increase in latency to convulsions and in time until death . In the strychnine- , electroshock- and picrotoxin-induced seizure models , agomelatine caused no significant alterations in latency to convulsions and in time until death when compared to controls . Our results suggest that agomelatine has anticonvulsant activity shown in PTZ- or pilocarpine-induced seizure models . DB04864 activates Wnt/β-catenin signaling and enhances the nonamyloidogenic pathway in an Alzheimer transgenic mouse model . DB04864 ( HupA ) is a reversible and selective inhibitor of acetylcholinesterase ( P22303 ) , and it has multiple targets when used for Alzheimer 's disease ( AD ) therapy . In this study , we searched for new mechanisms by which HupA could activate Wnt signaling and reduce amyloidosis in AD brain . A nasal gel containing HupA was prepared . No obvious toxicity of intranasal administration of HupA was found in mice . HupA was administered intranasally to β-amyloid ( Aβ ) precursor protein and presenilin-1 double-transgenic mice for 4 months . We observed an increase in O14672 and a decrease in P56817 and APP695 protein levels and , subsequently , a reduction in Aβ levels and Aβ burden were present in HupA-treated mouse brain , suggesting that HupA enhances the nonamyloidogenic P05067 cleavage pathway . Importantly , our results further showed that HupA inhibited GSK3α/β activity , and enhanced the β-catenin level in the transgenic mouse brain and in SH-SY5Y cells overexpressing Swedish mutation P05067 , suggesting that the neuroprotective effect of HupA is not related simply to its P22303 inhibition and antioxidation , but also involves other mechanisms , including targeting of the Wnt/β-catenin signaling pathway in AD brain . Q07869 gamma ligands , rosiglitazone and pioglitazone , inhibit P09038 - and P15692 -mediated angiogenesis . OBJECTIVE : To study the effect of peroxisome proliferator-activated receptor-gamma ( Q07869 gamma ) agonists , pioglitazone and rosiglitazone , on vascular endothelial growth factor ( P15692 ) - and basic fibroblast growth factor ( P09038 ) -induced angiogenesis and on endothelial cell migration . METHODS : Chick chorioallantoic membrane ( P62158 ) model was used to evaluate the efficacy of pioglitazone and rosiglitazone on P15692 - and P09038 -induced angiogenesis . In addition , the effect of pioglitazone and rosiglitazone on endothelial cell migration was evaluated using 8 mm pore filter to a feeder layer containing vitronectin as chemoattractant . RESULTS : Pioglitazone and rosiglitazone inhibited the pro-angiogenic effects of P09038 and P15692 in the P62158 model significantly ( P < 0.001 ) to the same extent . Endothelial cell migration was also inhibited by both pioglitazone and rosiglitazone ( P < 0.001 ) . CONCLUSIONS : These results suggest that Q07869 gamma ligands , pioglitazone and rosiglitazone , in addition to their important regulatory role in adipogenesis and inflammation , possess anti-angiogenic properties . Thus , Q07869 gamma ligands may be useful in the treatment of diabetic retinopathy , macular degeneration , and other ocular disorders and may lower the risk to develop cancer in diabetic patients . Progress in studies of huperzine A , a natural cholinesterase inhibitor from Chinese herbal medicine . DB04864 ( HupA ) , a novel alkaloid isolated from the Chinese herb Huperzia serrata , is a potent , highly specific and reversible inhibitor of acetylcholinesterase( P22303 ) . Compared with tacrine , donepezil , and rivastigmine , HupA has better penetration through the blood-brain barrier , higher oral bioavailability , and longer duration of P22303 inhibitory action . HupA has been found to improve cognitive deficits in a broad range of animal models . HupA possesses the ability to protect cells against hydrogen peroxide , beta-amyloid protein ( or peptide ) , glutamate , ischemia and staurosporine-induced cytotoxicity and apoptosis . These protective effects are related to its ability to attenuate oxidative stress , regulate the expression of apoptotic proteins Bcl-2 , Bax , P04637 , and caspase-3 , protect mitochondria , upregulate nerve growth factor and its receptors , and interfere with amyloid precursor protein metabolism . Antagonizing effects of HupA on N-methyl-D-aspartate receptors and potassium currents may also contribute to its neuroprotection as well . Pharmacokinetic studies in rodents , canines , and healthy human volunteers indicated that HupA was absorbed rapidly , distributed widely in the body , and eliminated at a moderate rate with the property of slow and prolonged release after oral administration . Animal and clinical safety tests showed that HupA had no unexpected toxicity , particularly the dose-limiting hepatotoxicity induced by tacrine . The phase IV clinical trials in China have demonstrated that HupA significantly improved memory deficits in elderly people with benign senescent forgetfulness , and patients with Alzheimer disease and vascular dementia , with minimal peripheral cholinergic side effects and no unexpected toxicity . HupA can also be used as a protective agent against organophosphate intoxication . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . DB01599 inhibits O95477 -mediated cellular lipid efflux . OBJECTIVE : DB00171 -binding cassette transporter A1 ( O95477 ) mediates the efflux of lipids from cells to lipid-poor apolipoproteins . In this article , we characterize the effect of probucol on cellular O95477 -mediated lipid efflux . METHODS AND RESULTS : DB01599 inhibited cholesterol efflux up to 80 % in J774 macrophages expressing O95477 . In Fu5AH hepatoma cells that contain scavenger receptor class B , type I , but not functional O95477 , we observed no effect of probucol on cholesterol efflux . DB01599 inhibited cholesterol efflux from normal human skin fibroblasts but not from fibroblasts from a Tangier patient . Fluorescent confocal microscopy and biotinylation assay demonstrated that in J774 cells probucol impaired the translocation of O95477 from intracellular compartments to the plasma membrane . DB01599 also inhibited the formation of an O95477 -linked cholesterol oxidase sensitive plasma membrane domain . Consistent with the inhibitory effect on O95477 translocation to the plasma membrane , probucol reduced cell surface-specific [ 125I ] -labeled apolipoprotein-AI binding . CONCLUSIONS : We conclude that probucol is an effective inhibitor of O95477 -mediated cholesterol efflux without influencing scavenger receptor class B type I-mediated efflux . The inhibition of O95477 translocation to the plasma membrane may in part explain the reported in vivo high-density lipoprotein-lowering action of probucol . Coadministrating luteolin minimizes the side effects of the aromatase inhibitor letrozole . P11511 inhibitors ( AIs ) have been used as adjuvant therapeutic agents for breast cancer . Their adverse side effect on blood lipid is well documented . Some natural compounds have been shown to be potential AIs . In the present study , we compared the efficacy of the flavonoid luteolin to the clinically approved AI letrozole ( DB01006 ; Novartis Pharmaceuticals , East Hanover , NJ ) in a cell and a mouse model . In the in vitro experimental results for aromatase inhibition , the Ki values of luteolin and letrozole were estimated to be 2.44 µM and 0.41 nM , respectively . Both letrozole and luteolin appeared to be competitive inhibitors . Subsequently , an animal model was used for the comparison . P11511 -expressing MCF-7 cells were transplanted into ovariectomized athymic mice . Luteolin was given by mouth at 5 , 20 , and 50 mg/kg , whereas letrozole was administered by intravenous injection . Similar to letrozole , luteolin administration reduced plasma estrogen concentrations and suppressed the xenograft proliferation . The regulation of cell cycle and apoptotic proteins-such as a decrease in the expression of Bcl-xL , cyclin-A/D1/E , P24941 /4 , and increase in that of Bax-was about the same in both treatments . The most significant disparity was on blood lipids . In contrast to letrozole , luteolin increased fasting plasma high-density lipoprotein concentrations and produced a desirable blood lipid profile . These results suggested that the flavonoid could be a coadjuvant therapeutic agent without impairing the action of AIs . Effects of lesopitron on the central nervous system arising from its interaction with P08908 receptors . DB04970 acts as a ligand for central serotonin P08908 receptors . Ki obtained from [3H]8-OH-DPAT competition studies was 104.8 +/- 10.6 nmol/l . As lesopitron did not affect the binding of [3H]paroxetine , involvement of the serotonin reuptake system in the effects of lesopitron is rejected . DB04970 inhibits haloperidol-induced catalepsy that is the consequence of its action on P08908 autoreceptors . The ability of lesopitron to induce 5-HT syndrome reflects post-synaptic P08908 receptor activation and the reversion of 8-OHDPAT-induced 5-HT syndrome by lesopitron suggests a partial agonist effect on this receptor-type . DB04970 induced a hypothermic effect due to the enhanced activation of post-synaptic P08908 receptors . The agonist effect of lesopitron on P08908 receptors and its marked hypothermic effect is an added value for this drug and a stimulus to the study of its possible neuroprotective action . Effects of a novel P08908 receptor agonist , E4424 , on gastric adherent mucus levels following restraint stress in rats . Several novel arylpiperazine serotonin 1A receptor agonists , developed as anxiolytics , have antisecretory and gastroprotective effects in rats . E4424 ( 2-¿4-[4-(4-chloropyrazol-1-yl)butyl]-1-piperazinyl ¿pyrimidine ; DB04970 dihydrochloride ) , has potent anti-gastric secretory and antiulcer effects . Preliminary data indicated an enhancing effect of E4424 on gastric mucus that may underlie its gastroprotective actions . We therefore tested the effects of acute and chronic administration of E4424 and of a reference P08908 receptor agonist , 8-OHDPAT [ 8-hydroxy-2-(di-n-propylamino)tetralin ] , on gastric mucus levels in rats subjected to cold-restraint stress , a procedure associated with depletion of gastric mucus and the development of mucosal injury . Acute oral administration of E4424 increased adherent mucus levels by 12 % , 11 % , and 13 % , relative to controls . Chronic E4424 significantly increased gastric mucus relative to controls ( 69 % increase ) . Acute oral treatment with 8-OHDPAT did not affect gastric mucus level . Acute intraperitoneal 8-OHDPAT slightly increased mucus levels . Chronic twice per day 8-OHDPAT did not affect mucus levels ; however , chronic once per day treatment with 8-OHDPAT significantly elevated gastric mucus levels at the highest doses used . For E4424 , there is a strong correlation between reduction of gastric mucosal injury and increase in gastric mucus level , suggesting that the action of E4424 on glandular mucus levels is an important mechanism underlying its gastroprotective effects . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . DB06594 in depressive disorders : its novel mechanisms of action . Disruptions in sleep and sleep-wake cycle regulation have been identified as one of the main causes for the pathophysiology of depressive disorders . The search has been on for the identification of an ideal antidepressant that could improve both sleep disturbances and depressive symptomatology . Melatonin , the major hormone of the pineal gland , has been shown to improve sleep and is involved in the regulation of the sleep-wake cycle . Identification of high concentrations of MT1 and P02795 melatonergic receptors in the suprachiasmatic nucleus of the anterior hypothalamus , the structure concerned with regulation of circadian rhythms and sleep-wake cycles , has led to the development of melatonergic agonists with greater potency and longer durations of action . DB06594 is one such melatonergic agonist that acts specifically on MT1/ P02795 melatonergic receptors and at the same time exhibits P28335 antagonism , a property that is utilized by current antidepressants that are in clinical use . DB06594 has been shown to be effective in a number of animal models of depression . Clinical studies undertaken on patients with major depression , bipolar disorders , seasonal affective disorder , and generalized anxiety disorder have all shown that agomelatine is also very effective in ameliorating depressive symptoms and manifesting early onset of action with a good tolerability and safety profile . It improved sleep efficiency and also resynchronized the disrupted circadian rhythms . Hence , the melatonergic modulation by agomelatine is suggested as one of the mechanisms for its antidepressant effect . DB06594 's action on dendritic neurogenesis in animal models of depression is also identified as yet another action . P62158 interacts with DB00171 binding cassette transporter A1 to protect from calpain-mediated degradation and upregulates high-density lipoprotein generation . OBJECTIVE : To investigate the interaction of DB00171 -binding cassette transporter A1 ( O95477 ) with calmodulin in relation to its calpain-mediated degradation because many calpain substrates bind calmodulin to regulate cellular functions . METHODS AND RESULTS : The activity of O95477 is regulated through proteolysis by calpain . An immunoprecipitation and glutathione S-transferase pull-down assay revealed that O95477 directly binds calmodulin in a Ca(2+)-dependent manner . The cytoplasmic loop of O95477 contains a typical calmodulin binding sequence of 1-5-8-14 motifs ( 1245 to 1257 amino acids ) . The peptide of this region showed binding to calmodulin , and deletion of the 1-5-8-14 motif abolished this interaction . This motif is located near the O95477 Pro- DB00142 - DB00133 - DB00156 sequence , and the presence of calmodulin/Ca(2+) protected the peptides from proteolysis by calpain . The knockdown of calmodulin by a specific small and interfering RNA increased the degradation of O95477 and decreased O95477 protein and apolipoprotein A-I-mediated lipid release . Surprisingly , calmodulin inhibitor W7 increased calmodulin binding to O95477 and protected it from calpain-mediated degradation , consistent with our previous finding that this compound increased apolipoprotein A-I-mediated cell cholesterol release . CONCLUSIONS : P62158 directly binds and stabilizes O95477 in the presence of Ca(2+) and increases the generation of high-density lipoprotein . P07996 up-regulates expression of cell adhesion molecules and promotes monocyte binding to endothelium . Expression of cell adhesion molecules ( P62158 ) responsible for leukocyte-endothelium interactions plays a crucial role in inflammation and atherogenesis . Up-regulation of vascular P62158 -1 ( P19320 ) , intracellular P62158 -1 ( P05362 ) , and P16581 expression promotes monocyte recruitment to sites of injury and is considered to be a critical step in atherosclerotic plaque development . Factors that trigger this initial response are not well understood . As platelet activation not only promotes thrombosis but also early stages of atherogenesis , we considered the role of thrombospondin-1 ( P07996 -1 ) , a matricellular protein released in abundance from activated platelets and accumulated in sites of vascular injury , as a regulator of P62158 expression . P07996 -1 induced expression of P19320 and P05362 on endothelium of various origins , which in turn , resulted in a significant increase of monocyte attachment . This effect could be mimicked by a peptide derived from the C-terminal domain of P07996 -1 and known to interact with Q08722 on the cell surface . The essential role of Q08722 in the cellular responses to P07996 -1 was demonstrated further using inhibitory antibodies and knockdown of Q08722 with small interfering RNA . Furthermore , we demonstrated that secretion of endogenous P07996 -1 and its interaction with Q08722 on the cell surface mediates endothelial response to the major proinflammatory agent , tumor necrosis factor alpha ( P01375 ) . Taken together , this study identifies a novel mechanism regulating P62158 expression and subsequent monocyte binding to endothelium , which might influence the development of anti-atherosclerosis therapeutic strategies . DB00139 dehydrogenase subunit B mutations modify human neuroblastoma cell metabolism and proliferation . Paragangliomas ( PGLs ) are rare neuroendocrine tumours . About 30-40 % of these tumours are mutated in one of the different susceptibility genes , including those encoding the different subunits of the succinate dehydrogenase , a complex involved both in the tricarboxylic acid cycle and in the oxygen transport chain . The aim of this work was to investigate whether P21912 mutations may account for alterations in cell metabolism and functions . Since human PGL cell lines are not available , we used the neuroblastoma cell line ( SK-N-AS ) stably transfected with the wild-type human P21912 or different P21912 -mutated constructs carrying some significant mutations found in our patients affected by PGLs . Similarly to succinate dehydrogenase ( SDH ) -mutated tumour cells , mutated SK-N-AS clones showed reduced SDH enzyme activity . All clones showed normal citrate synthase activity , reduced oxygen consumption and reduced carbonic anhydride production , thus demonstrating a decreased in mitochondrial metabolism . In two of the three mutated SK-N-AS , we also found an increase in HIF1α expression . Surprisingly and unexpectedly , in all the P21912 -mutated clones , we found a significant decrease in glucose uptake and in lactate culture medium concentration , suggesting also a decrease of cytosolic metabolism . Finally , we found that these energetic changes were associated to an increase in cell proliferation and migration . Overall , these data demonstrate that although P21912 mutations significantly downregulate both mitochondrial and cytoplasmic cellular metabolism , these mutations are associated to an upregulation of some cellular functions , such as growth rate and invasiveness . Q07869 gamma 2 regulates adipose expression of the phosphoenolpyruvate carboxykinase gene . DB01819 carboxykinase ( PEPCK ) is expressed at high levels in liver , kidney , and adipose tissue . This enzyme catalyzes the rate-limiting step in hepatic and renal gluconeogenesis and adipose glyceroneogenesis . The regulatory factors important for adipose expression of the PEPCK gene are not well defined . Previous studies with transgenic mice established that the region between bp -2086 and -888 is required for expression in adipose tissue but not for expression in liver or kidney tissue . We show here that a DNA fragment containing this region can function as an enhancer and direct differentiation-dependent expression of a chloramphenicol acetyltransferase gene from a heterologous promoter in cultured 3T3-F442A preadipocytes and adipocytes . We further demonstrate that the adipocyte-specific transcription factor Q07869 gamma 2 , previously identified as a regulator of the adipocyte P2 enhancer , binds in a heterodimeric complex with RXR alpha to the PEPCK 5'-flanking region at two sites , termed P35558 ( bp -451 to -439 ) and Q16822 ( bp -999 to -987 ) . Forced expression of Q07869 gamma 2 and RXR alpha activates the PEPCK enhancer in non-adipose cells . This activation is potentiated by peroxisome proliferators and fatty acids but not by 9-cis retinoic acid . Mutation of the Q07869 gamma 2 binding site ( Q16822 ) abolishes both the activity of the enhancer in adipocytes and its ability to be activated by Q07869 gamma 2 and RXR alpha . These results establish a role for Q07869 gamma 2 in the adipose expression of the PEPCK gene and suggest that this factor functions as a coordinate regulator of multiple adipocyte-specific genes . Correcting human mitochondrial mutations with targeted RNA import . Mutations in the human mitochondrial genome are implicated in neuromuscular diseases , metabolic defects , and aging . An efficient and simple mechanism for neutralizing deleterious mitochondrial DNA ( mtDNA ) alterations has unfortunately remained elusive . Here , we report that a 20-ribonucleotide stem-loop sequence from the H1 RNA , the RNA component of the human RNase P enzyme , appended to a nonimported RNA directs the import of the resultant RNA fusion transcript into human mitochondria . The methodology is effective for both noncoding RNAs , such as tRNAs , and mRNAs . The RNA import component , polynucleotide phosphorylase ( Q8TCS8 ) , facilitates transfer of this hybrid RNA into the mitochondrial matrix . In addition , nucleus-encoded mRNAs for mitochondrial proteins , such as the mRNA of human mitochondrial ribosomal protein P28222 ( O15235 ) , contain regulatory sequences in their 3'-untranslated region ( UTR ) that confers localization to the mitochondrial outer membrane , which is postulated to aid in protein translocation after translation . We show that for some mitochondrial-encoded transcripts , such as P35354 , a 3'-UTR localization sequence is not required for mRNA import , whereas for corrective mitochondrial-encoded tRNAs , appending the 3'-UTR localization sequence was essential for efficient fusion-transcript translocation into mitochondria . In vivo , functional defects in mitochondrial RNA ( mtRNA ) translation and cell respiration were reversed in two human disease lines . Thus , this study indicates that a wide range of RNAs can be targeted to mitochondria by appending a targeting sequence that interacts with Q8TCS8 , with or without a mitochondrial localization sequence , providing an exciting , general approach for overcoming mitochondrial genetic disorders . Targeted disruption of the Q9BXA5 metabolic receptor leads to dichotomous effects on obesity . A number of metabolites have signaling properties by acting through G-protein-coupled receptors . DB00139 , a Krebs cycle intermediate , increases after dysregulated energy metabolism and can bind to its cognate receptor succinate receptor 1 ( Sucnr1 , or Q9BXA5 ) to activate downstream signaling pathways . We show that Sucnr1 is highly expressed in the white adipose tissue ( WAT ) compartment of mice and regulates adipose mass and glucose homeostasis . Sucnr1(-/-) mice were generated , and weight gain was monitored under basal and nutritional stress ( high-fat diet [ HFD ] ) conditions . On chow diet , Sucnr1(-/-) mice had increased energy expenditure , were lean with a smaller WAT compartment , and had improved glucose buffering . Lipolysis measurements revealed that Sucnr1(-/-) mice were released from succinate-induced inhibition of lipolysis , demonstrating a function of Sucnr1 in adipose tissue . Sucnr1 deletion also protected mice from obesity on HFD , but only during the initial period ; at later stages , body weight of HFD-fed Sucnr1(-/-) mice was almost comparable with wild-type ( WT ) mice , but WAT content was greater . Also , these mice became progressively hyperglycemic and failed to secrete insulin , although pancreas architecture was similar to WT mice . These findings suggest that Sucnr1 is a sensor for dietary energy and raise the interesting possibility that protocols to modulate Sucnr1 might have therapeutic utility in the setting of obesity . A combination of molecular cytogenetic analyses reveals complex genetic alterations in conventional renal cell carcinoma . Here we report the complex pattern of genomic imbalances and rearrangements in a panel of 19 renal cell carcinoma cell lines detected with molecular cytogenetic analysis . Consistent heterogeneity in chromosome number was found , and most cell lines showed a near-triploid chromosome complement . Several cell lines showed deletions of the P04637 ( alias p53 ) , CDKN2A ( alias p16 ) , and P40337 genes . Multiplex fluorescence in situ hybridization ( M- Q5TCZ1 ) analysis revealed chromosome 3 translocated to several other partners chromosomes , as well as breakage events commonly affecting chromosomes 1 , 5 , 8 , 10 , and 17 . The most common abnormality detected with comparative genomic hybridization ( CGH ) was deletions of chromosome 3p , with loss of the Q9NS23 , P49789 , and p44S10 loci frequently involved . CGH gain of 5q showed overrepresentation of the P18146 and P07333 genes . Recurrent alterations to chromosome 7 included rearrangement of 7q11 and gains of the P00533 , O15164 , and P35250 genes . Several lines exhibited rearrangement of 12q11 approximately q14 and overrepresentation of P11802 and SAS loci . M- Q5TCZ1 revealed several other recurrent translocations , and CGH findings included loss of 9p , 14q , and 18q and gain of 8q , 12 , and 20 . Further genomic microarray changes included loss of Q13126 , IGH@ , P28222 , and Q13485 ( previously Q13485 ) and gains of MYC and P11387 . An excellent correlation was observed between the genomic array and Q5TCZ1 data , demonstrating that this technique is effective and accurate . The aberrations detected here may reflect important pathways in renal cancer pathogenesis . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . Integrative analysis of miRNA and mRNA expression profiles in pheochromocytoma and paraganglioma identifies genotype-specific markers and potentially regulated pathways . Pheochromocytomas ( PCCs ) and paragangliomas ( PGLs ) are rare neuroendocrine neoplasias of neural crest origin that can be part of several inherited syndromes . Although their mRNA profiles are known to depend on genetic background , a number of questions related to tumor biology and clinical behavior remain unanswered . As microRNAs ( miRNAs ) are key players in the modulation of gene expression , their comprehensive analysis could resolve some of these issues . Through characterization of miRNA profiles in 69 frozen tumors with germline mutations in the genes O14521 , P21912 , P40337 , P07949 , P21359 , O75204 , and P61244 , we identified miRNA signatures specific to , as well as common among , the genetic groups of PCCs/PGLs. miRNA expression profiles were validated in an independent series of 30 composed of P40337 - , P21912 - , O14521 - , and P07949 -related formalin-fixed paraffin-embedded DB09058 /PGL samples using quantitative real-time PCR . Upregulation of miR-210 in P40337 - and P21912 -related PCCs/PGLs was verified , while miR-137 and miR-382 were confirmed as generally upregulated in PCCs/PGLs ( except in P61244 -related tumors ) . Also , we confirmed overexpression of miR-133b as P40337 -specific miRNAs , miR-488 and miR-885-5p as P07949 -specific miRNAs , and miR-183 and miR-96 as P21912 -specific miRNAs . To determine the potential roles miRNAs play in DB09058 /PGL pathogenesis , we performed bioinformatic integration and pathway analysis using matched mRNA profiling data that indicated a common enrichment of pathways associated with neuronal and neuroendocrine-like differentiation . We demonstrated that miR-183 and/or miR-96 impede P01138 -induced differentiation in PC12 cells . Finally , global proteomic analysis in P21912 and P61244 tumors allowed us to determine that miRNA regulation occurs primarily through mRNA degradation in PCCs/PGLs , which partially confirmed our miRNA-mRNA integration results . Adipocyte-specific reduction of phosphodiesterase 3B gene expression and its restoration by DB04971 in the obese , diabetic KKAy mouse . OBJECTIVE : Phosphodiesterase ( PDE ) 3B is a key enzyme involved in the anti-lipolytic action of insulin in adipocytes . Q13370 activation results in a reduced output of free fatty acids ( FFA ) , whereas elevated serum FFA is known to cause insulin resistance . We have recently reported that reduced Q13370 gene expression is restored by treatment with pioglitazone , in the adipose tissues of obese , insulin-resistant diabetic KKAy mice . To determine whether the altered Q13370 gene expression is specific for adipocytes , the expression of this gene in liver and epididymal fat tissues of KKAy mice was examined . The effect of DB04971 , another peroxisome proliferator-activated receptor ( Q07869 )gamma ligand , which is different from thiazolidinedione , was also examined . METHODS : Q13370 mRNA and protein were quantified by an RNase protection assay and Western blotting respectively . Membrane-bound PDE activities were also measured . RESULTS : In adipose tissues of KKAy mice , Q13370 mRNA , protein and membrane-bound PDE activity were reduced to 47 % , 57 % and 51 % respectively relative to those in C57BL/6J control mice . DB04971 increased Q13370 mRNA , protein and membrane-bound PDE activity by 2.2- , 1.6- and 1.7-fold respectively over those of untreated KKAy mice . In the liver , Q13370 gene expression remained unchanged in KKAy mice , and was not affected by DB04971 . DB04971 reduced the elevated levels of serum insulin , glucose , FFA and triglyceride in KKAy mice . CONCLUSIONS : Q13370 gene expression was specifically reduced in the adipose tissues of KKAy mice . DB04971 restored this reduced gene expression with an accompanying improvement in elevated serum FFA and insulin resistance . Comparative actions of insulin sensitizers on ion channels in vascular smooth muscle . Thiazolidinedione and isoxazolidinedione insulin sensitizers activate peroxisome proliferator-activated receptor gamma ( Q07869 gamma ) . Some thiazolidinediones modify ion channels in smooth muscles ; however , the mechanism by which their actions occur has not been clarified . We , thus , examined the effects of three thiazolidinediones ( troglitazone , pioglitazone , and rosiglitazone ) and isoxazolidinedione ( DB04971 ) , as well as an intrinsic ligand for Q07869 gamma , 15-deoxy-Delta(12,14) prostaglandin J(2) ( prostaglandin J(2) ) , on voltage-operated Ca(2+) currents ( I(Ca) ) , voltage-dependent K(+) currents ( I(Kv) ) , and Ca(2+)-activated K(+) currents ( I(Kca) ) , to clarify whether a thiazolidinedione structure or Q07869 gamma activation is related to their actions on ion channels . The whole-cell patch clamp method was used to record currents in smooth muscle cells from guinea-pig mesenteric arteries . Thiazolidinediones inhibited I(Ca) in a dose-dependent manner ( troglitazone > pioglitazone=rosiglitazone ) . DB00197 ( > or =1 microM ) and rosiglitazone ( 100 microM ) , but not pioglitazone , inhibited I(Kv) . Rosiglitazone ( > or =10 microM ) enhanced , troglitazone ( > or =1 microM ) inhibited , and pioglitazone did not affect I(Kca) . A high concentration of DB04971 ( 100 microM ) inhibited I(Ca) , I(Kv) , and I(Kca) to a similar extent . Prostaglandin J(2) enhanced I(Kca) , but affected neither I(Ca) nor I(Kv) . In summary , the three thiazolidinediones and isoxazolidinedione act differently on Ca(2+) and K(+) channels in vascular smooth muscle . The action of thiazolidinediones on I(Ca) could be attributed to specific regions of the molecules and not to activation of Q07869 gamma . Involvement of Q07869 gamma activation in the stimulation of I(Kca) is possible but should be tested further . Expression and biological activity of O95477 in alveolar epithelial cells . The mechanisms used by alveolar type I pneumocytes for maintenance of the lipid homeostasis necessary to sustain these large squamous cells are unknown . The processes may involve the DB00171 -binding cassette transporter A1 ( O95477 ) , a transport protein shown to be crucial in apolipoprotein A-I ( apoA-I ) -mediated mobilization of cellular cholesterol and phospholipid . Immunohistochemical data demonstrated the presence of O95477 in lung type I and type II cells and in cultured pneumocytes . Type II cells isolated from rat lungs and cultured for 5 days in 10 % serum trans-differentiated toward cells with a type I-like phenotype which reacted with the type I cell-specific monoclonal antibody VIIIB2 . Upon incubation of the type I-like pneumocytes with agents that up-regulate the O95477 gene ( 9-cis-retinoic acid [ 9cRA ] and 22-hydroxycholesterol [ 22-OH , 9cRA/22-OH ] ) , O95477 protein levels were enhanced to maximum levels after 8 to 16 hours and remained elevated for 24 hours . In the presence of apoA-I and 9cRA/22-OH , efflux of radioactive phospholipid and cholesterol from pneumocytes was stimulated 3- to 20-fold , respectively , over controls . Lipid efflux was inhibited by DB01599 . DB02772 density gradient analysis of the media from stimulated cells incubated with apoA-I identified heterogeneous lipid particles that isolated at a density between 1.063 and 1.210 g/ml , with low or high apoA-I content . Thus , pneumocytes with markers for the type I phenotype contained functional O95477 protein , released lipid to apoA-I protein , and were capable of producing particles resembling nascent high-density lipoprotein , indicating an important role for O95477 in the maintenance of lung lipid homeostasis . P07996 and transforming growth factor beta-1 upregulate plasminogen activator inhibitor type 1 in pancreatic cancer . Controlled degradation of the extracellular matrix by proteases is crucial in tumor cell invasion . We have shown that thrombospondin-1 ( P07996 -1 ) , through activation of transforming growth factor beta-1 ( TGF-beta1 ) , regulates the plasminogen/plasmin protease system in breast cancer . To determine whether this occurred in other epithelial neoplasms , we studied the role of P07996 -1 and TGF-beta1 in the regulation of the plasminogen/plasmin system in pancreatic cancer . ASPC-1 and COLO-357 pancreatic cancer cells were treated with P07996 -1 or TGF-beta1 at varying concentrations . The P07996 -1 and TGF-beta1-treated cells were also treated with either anti- P07996 -1 , anti- P07996 -1 receptor , or anti-TGF-beta1 antibodies . DB00013 plasminogen activator ( uPA ) and plasminogen activator inhibitor-1 ( P05121 ) expression was determined by enzyme-linked immunosorbent assay . P07996 -1 and TGF-beta1 promoted a dose-dependent upregulation of ASPC-1 and COLO-357 P05121 expression . The P07996 -1 effect could be blocked with anti- P07996 -1 or anti-TGF-beta1 antibodies . The TGF-beta1 effect could be blocked only with anti-TGF-beta1 antibody . Anti- P07996 -1 receptor antibody blocked the P07996 -1 effect on P05121 expression but had no effect on TGF-beta1-mediated P05121 expression . Neither P07996 -1 nor TGF-beta1 had an effect on uPA production . We conclude that P07996 -1 , in a receptor-mediated process that involves the activation of TGF-beta1 , upregulates P05121 expression in pancreatic cancer without an effect on uPA production . Fibrinolytic parameters in spermatozoas and seminal plasma . DB00013 -type ( u-PA ) and tissue-type plasminogen activator antigen ( t-PA ) as well as plasminogen activator-inhibitor activity were determined in seminal plasma and lysates of the respective spermatozoas in 67 ejaculate of males in infertile marriage without genito urinary pathology . U-PA was determined by a competition RIA , t-PA by an ELISA and P05121 by a spectrophotometric assay . 15 patients showed normozoospermia , 11 azoospermia and 41 oligoasthenoteratozoospermia ( P04181 -syndrome ) . In lysates of spermatozoas , significantly higher levels of both plasminogenactivators and P05121 were found in patients with P04181 syndrome as compared to those exhibiting normozoospermia . Whereas P05121 was absent in the seminal plasma of normozoospermic ejaculate , patients with azoospermia ( 180 +/- 13 mU/ml. ) and P04181 -syndrome ( 60 +/- 5 mU/ml. ) showed high P05121 levels . The similarly high values of t-PA ( 190.8-227.8 ng./ml. ) and u-PA ( 19.4-32 ng./ml. ) in the same compartment confirm their predominantly prostatic origin and seem to have no influence on the quality of the ejaculate . Inhibition of noradrenaline release via presynaptic P28222 receptors of the rat vena cava . In the rat inferior vena cava preincubated with 3H-noradrenaline , the effects of nine serotonin ( 5-HT ) receptor agonists and of eight antagonists ( including two beta-adrenoceptor blocking agents ) on the electrically evoked 3H overflow were determined . 1 . 5-HT , 5-carboxamido-tryptamine , 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole ( RU 24969 ) , 5-methoxytryptamine , N,N-dimethyl-5HT , tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow . The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for P28222 binding sites , but not with their affinities for P08908 , P28335 or 5-HT2 binding sites . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 receptor agonist , and ipsapirone , a partial agonist at these receptors , did not inhibit overflow . 2 . Cyanopindolol facilitated the evoked 3H overflow , an effect which was abolished by propranolol . The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol . 3 . The concentration-response curve for 5-HT was shifted to the right by metitepine , metergoline , quipazine , 6-chloro-2-(1-piperazinyl)pyrazine ( MK 212 ) and propranolol which , given alone , did not affect 3H overflow . The apparent pA2 values of these antagonists tended to be correlated with their affinities for P28222 ( but not P08908 , P28335 or 5-HT2 ) binding sites . Ketanserin , a 5-HT2 receptor antagonist , and spiperone , which blocks 5-HT2 and P08908 but not P28222 or P28335 receptors , failed to antagonize the effect of 5-HT. ( ABSTRACT TRUNCATED AT 250 WORDS ) Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO:fat:protein = 12:59:28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO:fat:protein = 56:24:20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n-6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4n-6 ) , while its biosynthetic metabolic intermediates were decreased . The n-6/n-3 and arachidonic/eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1n-7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 , P05231 , P10145 , P13500 , P16581 , I- P62158 , and P05121 . Increased 20:4n-6 and the ratios of 20:4n-6/20:5n-3 and n-6/n-3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet . [ Changes of protein expression in HepG2 cells with P24941 RNA interference ] . OBJECTIVE : To investigate the effects of stable transfection of P24941 siRNA on biological activities and nuclear proteins of human hepatocellular carcinoma HepG2 cells . METHODS : HepG2 cells were transfected with the eukaryotic expression vector of P(Genesil-1- P24941 ) ; via RNA interference and selected for the ones with stable transfection . We observed the changes in the cell growth curve and cell cycle . The mRNA contents of P24941 and differentially expressed nucleoproteins were detected and analyzed by RT-PCR and two-dimensional ( 2D ) electrophoresis-mass spectrum ( MS ) -database , respectively . Western blotting were used to confirm the differential protein expressions . RESULTS : Compared with P(HK-siRNA);-HepG2 and untransfected groups , the proliferation of HepG2 cells in P( P24941 -siRNA);-HepG2 group was significantly inhibited ( P < 0.01 ) , and the expression of P24941 mRNA significantly decreased in P( P24941 -siRNA);-HepG2 group . Four proteins not expressed in P( P24941 -siRNA);-HepG2 cells were detected by 2D electrophoresis-MS , and they were further confirmed by Western blotting . CONCLUSION : P24941 siRNA significantly suppressed P24941 mRNA expression and the proliferation of HepG2 cells , four proteins not expressed in p( P24941 -siRNA);-HepG2 cells are similar to ribosomal protein P28222 , β-actin , zine finger 276 and chaperonin 10 related protein . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . The P08908 receptor agonist Bay x 3702 inhibits apoptosis induced by serum deprivation in cultured neurons . We examined whether the highly selective P08908 receptor agonist (-)-(R)-2- [ 4- [ [ (3,4-dihydro-2H-1-benzopyran-2-yl)methyl ] -amino ] butyl ] -11 , 2-benz-isothiazol-3(2H)-one 1,1-dioxide monohydrochloride ( Bay x 3702 ) could inhibit neuronal apoptosis induced by serum deprivation . In primary cultures of chick embryonic neurons and in mixed neuronal/glial cultures from neonatal rat hippocampus , Bay x 3702 ( 1 microM ) rescued serum-deprived neurons from apoptosis . The antiapoptotic effect of Bay x 3702 ( 1 microM ) was blocked in chick neurons by the selective P08908 receptor antagonists 4-iodo-N-[2-[4-(methoxyphenyl)-1-piperazin]ethyl]-N-2-pyridinyl-be nzamide hydrochloride ( p-MPPI , 10 microM ) and 4-[3-benzotriazol-1-propyl]-1-(2-methoxyphenyl)-piperazine ( BPMP , 10 microM ) as well as by anti-nerve growth factor ( anti- P01138 ) antibodies and in rat neurons by N-[2-4-(2-methoxy)-1-piperazinyl]ethyl ] -N-(2-pyridinyl)cyclohexane-carbo xamide trihydrochloride ( WAY 100635 , 10 microM ) . We found only under control conditions ( medium with serum ) , but not in serum-deprived cultures , that P01138 secretion was 6-fold increased by Bay x 3702 ( 1 microM ) compared to untreated cultures . Additionally , Bay x 3702 ( 4 microg/kg i.v. ) , infused within a period of 4 h , significantly increased the P01138 content of the rat hippocampus , but not of the striatum . In summary , our data suggest that Bay x 3702 inhibited growth factor withdrawal-induced apoptosis by the stimulation of P08908 receptors and that the P01138 signalling pathway is involved in the mechanism of action . Endotoxin induced hyperlactatemia and hypoglycemia is linked to decreased mitochondrial phosphoenolpyruvate carboxykinase . AIMS : DB01819 carboxykinase ( PEPCK ) is the rate limiting enzyme for gluconeogenesis , and plays a key role in recycling lactate for glucose production . It is synthesized as two separate isoforms ; cytosolic ( P35558 , gene code ; P35558 ) and mitochondrial ( Q16822 , gene code ; Q16822 ) . Previous studies of gluconeogenesis in endotoxemia have focused solely on P35558 . We investigated the relative roles of the two isoforms in hepatic and renal gluconeogenesis in a rat model of endotoxic shock , and in cultured hepatocytes . MAIN METHODS : Rats were administered lipopolysaccharide ( 6 mg/kg ; LPS ) for 6 h . Cultured cells were incubated with lactate ( 5 mM ) with or without tumor necrosis factor alpha ( 1 - 10 ng/ml ) . Rat liver and kidney samples as well as cultured cells were subjected to subcellular fractionation to produce mitochondrial and cytosolic fractions for PEPCK activity assay . P35558 and Q16822 mRNA levels were measured using quantitative RT-PCR . KEY FINDINGS : In rat endotoxemia , hepatic Q16822 mRNA and Q16822 enzyme activity decreased by 53 % and 38 % , compared to sham controls . Hepatic P35558 mRNA levels increased by 44 % , but P35558 enzyme activity remained unchanged . The changes in hepatic Q16822 coincided with a marked hypoglycemia and hyperlactatemia as well as elevated plasma interleukin 1 beta ( IL1beta ) . Incubation of cultured hepatocytes with P01375 inhibited lactate-induced increases in glucose production , Q16822 mRNA levels and Q16822 enzyme activity but had no effect on P35558 mRNA levels or P35558 activity . SIGNIFICANCE : These results indicate that decreases in hepatic Q16822 play a key role in the manifestation of hyperlactatemia and hypoglycemia in endotoxemia . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process .

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interacts_with DB00233?

multiple_choice

[ DB00435 and reflectory regulation of blood circulation in rats ] . In acute experiments on anaesthetized with urethane normotensive rats we studied the ways of participation of nitric oxide ( NO ) in reflector control of the cardiovascular system by the medullary neurons within n.tractus solitarii ( P30990 ) , dorsal nucleus of the vagus nerve ( DNV ) , n. ambiguus ( AMB ) , and the lateral reticular nucleus ( LRN ) . Modulations of the activities of neuronal NO-synthase ( P29475 ) in the populations of the cardiovascular neurons within the medullary nuclei which are involved in the reflector cardiovascular control were induced by intramedullary injections of sodium nitroprusside as NO donor , L-arginine as NO precursor , DB04223 as an inhibitor of NOS , as well as by intraperetoneal injections of 7-nitroindazol ( P29475 inhibitor ) . We have determined that stimulation of P29475 activity in the populations of the medullary neurons resulted in both remarkable shifts in the SAP level and in inhibiting the chemoreceptor reflector responses . After preliminary inhibiting P29475 chemoreceptor reflexes induced by epinephrine were found to be enhanced in most experiments . Inhibition and enhancement of contextual fear memory destabilization . The reactivation of a memory can result in its destabilization , necessitating a process of memory reconsolidation to maintain its persistence . Here we show that the destabilization of a contextual fear memory is potentiated by the cannabinoid P21554 receptor agonist Arachidonyl-2-chloroethylamide ( ACEA ) . Co-infusion of ACEA and the O15111 ( IKK ) inhibitor sulfasalazine ( Sulf ) into the dorsal hippocampus impaired contextual fear memory reconsolidation . This observation was achieved under behavioral conditions that , by themselves , did not result in a reconsolidation impairment by Sulf alone . Moreover , we show that the destabilization of a contextual fear memory is dependent upon neuronal activity in the dorsal hippocampus , but not memory expression per se . The effect on contextual fear memory destabilization of intra-hippocampal ACEA was replicated by systemic injections , allowing an amnestic effect of MK-801 . These results indicate that memory expression and destabilization , while being independent from one another , are both dependent upon memory reactivation . Moreover , memory destabilization can be enhanced pharmacologically , which may be of therapeutic potential . Short and long access to cocaine self-administration activates tyrosine phosphatase P54829 and attenuates GluN expression but differentially regulates GluA expression in the prefrontal cortex . RATIONALE : Dephosphorylation of extracellular signal-regulated kinase ( P29323 ) and cyclic AMP response element binding protein ( CREB ) in the dorsomedial prefrontal cortex ( dmPFC ) at the end of short access ( ShA ) cocaine self-administration is implicated in cocaine seeking . However , what receptors and phosphatases mediate this effect and whether P29323 /CREB and related phospho-proteins in the dmPFC react similarly during early withdrawal from long access ( LgA ) cocaine self-administration are unknown . OBJECTIVES : The effects of ShA vs. LgA cocaine self-administration on the phosphorylation of protein phosphatase 2A ( PP2A ) and striatal-enriched protein tyrosine phosphatase ( P54829 ) , as well as GluN and GluA receptor subtype expression in the dmPFC during early withdrawal , were compared . METHODS : Rats self-administered cocaine or received saline during 2- or 6-h daily sessions for 10-11 days . Two hours after the final session , the dmPFC was dissected out and processed for immunoblotting . RESULTS : Similar to previous findings after ShA cocaine , phospho- P29323 and phospho-CREB in the dmPFC were decreased after LgA cocaine . Cocaine elevated phospho-PP2A ( deactivation ) and decreased phospho- P54829 ( activation ) in both ShA and LgA cocaine rats . Q05586 , Q13224 , and phospho- Q13224 Tyr1472 in the dmPFC were decreased after ShA and LgA cocaine . Further , a significant reduction of P42262 , P42261 , and phospho- P42261 Ser845 was found only in LgA rats . CONCLUSIONS : Activation of phospho- P54829 may underlie P29323 and CREB dephosphorylation in the dmPFC as well as internalization and degradation of GluN complexes during early withdrawal from both ShA and LgA cocaine self-administration , whereas differential alteration of AMPA receptor subunits after ShA and LgA cocaine self-administration depends on cocaine intake . Discrimination between agonists and antagonists by the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid-selective glutamate receptor . A mutation analysis of the ligand-binding domain of P48058 subunit . The crystal structures of the ligand-binding core of the agonist complexes of the glutamate receptor-B ( P42262 ) subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid ( AMPA ) -selective glutamate receptor indicate that the distal anionic group of agonist molecules are stabilized by interactions with an N-terminal region of an alpha-helix ( helix F ) in the lobe 2 ( " domain 2 , " Armstrong , N. , and Gouaux , E. ( 2000 ) Neuron 28 , 165-181 ) of the two-lobed ligand-binding domain . We used site-directed mutagenesis to further analyze the role of this region in the recognition of both agonists and antagonists by the AMPA receptor . Wild-type and mutated versions of the ligand-binding domain of P48058 were expressed in insect cells as secreted soluble polypeptides and subjected to binding assays using [(3)H]AMPA , an agonist , and [(3)H]Ro 48-8587 ( 9-imidazol-1-yl-8-nitro-2,3,5,6-tetrahydro[1,2,4]triazolo[1,5-c] quinazoline-2,5-dione ) , a high affinity AMPA receptor antagonist , as radioligands . Single alanine substitutions at residues DB00149 -672 and DB00156 -677 severely affected the affinities for all agonists , as seen in ligand competition assays , whereas similar mutations at residues DB00128 -673 , DB00133 -674 , DB00145 -675 , DB00133 -676 , and Lys-678 selectively affected the binding affinities of one or two of the agonists . In striking contrast , the binding affinities of [(3)H]Ro 48-8587 and of another competitive antagonist , DB03759 , were not affected by any of these alanine mutations , suggesting the absence of critical side-chain interactions . Together with ligand docking experiments , our results indicate a selective engagement of the side chains of the helix F region in agonist binding , and suggest that conformational changes involving this region may play a critical role in receptor activation . Pharmacogenomics in aspirin intolerance . Polymorphisms in drug-related enzymes and receptors are often strongly related to altered drug response and to the risk of developing drug intolerance . DB00945 , usually available as an over-the-counter drug , is one of the most used drugs worldwide and is a common cause of drug intolerance events . DB00945 undergoes polymorphic metabolism . Among the enzymes involved in aspirin biodisposition a major role is played by the enzymes UDP-glucuronosyltransferase P19224 , cytochrome P450 P11712 and the xenobiotic/medium chain fatty acid: DB01992 ligase ACSM2 , although other UGTs and ACSMs enzymes may significantly contribute to aspirin metabolism . P19224 , P11712 and ACSM2 are polymorphic , as well as P23219 and P35354 , the genes coding for the enzymes cyclo-oxygenases P23219 and P35354 , respectively . The present review analyzes the importance of genetic variations in enzymes involved in the metabolism and in the effects of aspirin and common polymorphisms related to aspirin intolerance , and it raises hypotheses on genetic factors related to altered response to aspirin that require further investigation . Major polymorphisms related to aspirin biodisposition are rs2070959 , rs1105879 and rs6759892 for the P19224 gene , rs1133607 for the ACSM2 gene , and rs1799853 , rs1057910 , rs28371686 , rs9332131 and rs28371685 for the P11712 gene . Regarding aspirin effects , major PGTS1 targets are rs3842787 and rs5789 for European subjects , and rs3842789 and rs3842792 for African subjects . For the P35354 gene nonsynonymous SNPs are likely to be of low relevance because of the influence of transcriptional and posttranscriptional factors . Combined studies for the above mentioned polymorphisms and those corresponding to other genes related to aspirin intolerance will provide excellent tools to identify individuals with a high risk to develop intolerance to aspirin . Identification and characterization of a general nuclear translocation signal in signaling proteins . Upon stimulation , many proteins translocate into the nucleus in order to regulate a variety of cellular processes . The mechanism underlying the translocation is not clear since many of these proteins lack a canonical nuclear localization signal ( NLS ) . We searched for an alternative mechanism in extracellular signal-regulated kinase ( P29323 ) -2 and identified a 3 amino acid domain ( P49903 ) that is phosphorylated upon stimulation to induce nuclear translocation of P28482 . A 19 amino acid stretch containing this phosphorylated domain inserts nondiffusible proteins to the nucleus autonomously . The phosphorylated P49903 acts by binding to importin7 and the release from nuclear pore proteins . This allows its functioning both in passive and active P29323 transports . A similar domain appears in many cytonuclear shuttling proteins , and we found that phosphorylation of similar sequences in P84022 or Q02750 also induces their nuclear accumulation . Therefore , our findings show that this phosphorylated domain acts as a general nuclear translocation signal ( P30990 ) . Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress . Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis . Because oxidative stress contributes to atherosclerosis , we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon . Bovine aortic endothelial cells were exposed to static , laminar ( 15 dyn/cm2 ) , and oscillatory shear stress ( +/-15 dyn/cm2 ) . Oscillatory shear increased superoxide ( O2.- ) production by more than threefold over static and laminar conditions as detected using electron spin resonance ( P03372 ) . This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources . DB05262 also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence . DB02134 -dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress . This was associated with decreased xanthine dehydrogenase ( P47989 ) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase ( XO ) to P47989 . We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase . These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity . Transfection of these cells with p47phox restored XO protein levels . Finally , in bovine aortic endothelial cells , prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress . These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress . Transgenic Panax ginseng inhibits the production of P01375 , P05231 , and P10145 as well as P35354 expression in human mast cells . The most well-known medicinal plant , Panax ginseng ( P. ginseng ) , contains various phytosterols and bioactive triterpene saponins ( ginsenosides ) . P37268 is a key regulatory enzyme for triterpene biosynthesis and overexpression of the squalene synthase confers the hyper-production of triterpene saponins to form transgenic ginseng . In this study , we have investigated whether and how transgenic P. ginseng modulates an inflammatory reaction in a stimulated human mast cell line , HMC-1 . It was found that transgenic P. ginseng inhibited the production of tumor necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -6 , P10145 , and the expression of cyclooxygenase-2 in phorbol 12-myristate 13-acetate ( PMA ) plus calcium ionophore A23187 ( PMACI ) -stimulated HMC-1 . Additionally , we have shown that transgenic P. ginseng suppressed the intracellular calcium level induced by PMACI . These results provide new insights into the pharmacological actions of transgenic P. ginseng as a potential molecule for use in therapy in mast cell-mediated inflammatory diseases . 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 ( IKK ) activity was investigated using purified recombinant O15111 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 , O14920 , or constitutively active O15111 and O14920 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 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 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 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 . Q05940 and receptor interaction profiles of a new series of designer cathinones . Psychoactive β-keto amphetamines ( cathinones ) are sold as " bath salts " or " legal highs " and recreationally abused . We characterized the pharmacology of a new series of cathinones , including methedrone , 4-methylethcathinone ( 4- Q9NRJ3 ) , 3-fluoromethcathinone ( 3-FMC ) , pentylone , ethcathinone , buphedrone , pentedrone , and N,N-dimethylcathinone . We investigated norepinephrine ( NE ) , dopamine ( DA ) , and serotonin ( 5-HT ) uptake inhibition using human embryonic kidney 293 ( P29320 293 ) cells that express the respective human monoamine transporter , the drug-induced efflux of NE , DA , and 5-HT from monoamine-preloaded cells , and binding affinity to monoamine transporters and receptors . All of the cathinones were potent NE uptake inhibitors but differed in their DA vs. 5-HT transporter inhibition profiles and monoamine release effects . Methedrone was a more potent 5-HT than Q01959 inhibitor and released NE and 5-HT similar to para-methoxymethamphetamine ( PMMA ) , para-methoxyamphetamine ( PMA ) , 4-methylthioamphetamine ( 4-MTA ) , and 3,4-methylenedioxymethamphetamine ( DB01454 ) . 4- Q9NRJ3 and pentylone equipotently inhibited all of the monoamine transporters and released 5-HT . Ethcathinone and 3-FMC inhibited NE and DA uptake and released NE , and 3-FMC also released DA similar to N-ethylamphetamine and methamphetamine . Pentedrone and N,N-dimethylcathinone were non-releasing NE and DA uptake inhibitors as previously shown for pyrovalerone cathinones . Buphedrone preferentially inhibited NE and DA uptake and also released NE . None of the cathinones bound to rodent trace amine-associated receptor 1 , in contrast to the non-β-keto-amphetamines . None of the cathinones exhibited relevant binding to other monoamine receptors . In summary , we found considerable differences in the monoamine transporter interaction profiles among different cathinones and compared with related amphetamines . Whole blood lead concentration and erythrocyte delta-aminolevulinic acid dehydratase ( P13716 ) activity in selected canine populations in Greece . In a total number of 275 dogs of various ages , sex and breed , blood lead concentrations ( O43927 ) and erythrocyte P13716 activity were measured . Sixty-six of the dogs were living in lead mining areas ( Group A ) , 157 in urban areas ( Group B ) and 52 in rural areas ( Group C ) of Greece . Mean O43927 differed significantly ( P < 0.05 ) between locations and were 326,97 and 68 micrograms/L , respectively . Mean P13716 activity was significantly different ( P < 0.05 ) only between Groups A and B as between groups A and C . A significant ( P < 0.05 ) negative correlation existed between O43927 and P13716 activity . A normal range of erythrocyte P13716 activity of 807-992 mumol/ DB02272 /LRBC/h was established for dogs . None of the 33 Group A dogs and 2 of the Group B dogs that had a O43927 of 350 micrograms/L presented clinical signs indicating acute or chronic lead intoxication . No erythrocyte basophilic stippling or large number of nucleated red blood cells were seen in the 30 dogs of Group A with O43927 > 350 micrograms/L . P18627 -specific functions of p55TNFR and O14920 in the development of P18627 networks and of antibody responses . The P18627 -specific molecular signals required in the formation of P18627 networks , B cell follicles , and germinal centers ( GCs ) have remained poorly understood . We used P18627 -specific gene targeting to investigate the function of p55TNFR and O14920 in lymphoid organ structure and function . Here we show that P18627 -specific expression of p55TNFR is necessary and sufficient to promote P18627 network and B cell follicle formation , restore the expression of O43927 and P19320 / P05362 in FDCs , and lead to productive GCs . Notably , P18627 -specific disruption of O14920 does not affect formation of P18627 networks . Yet , after antigen engagement or immune complex ( IC ) deposition , FDCs lacking O14920 fail to upregulate P19320 and P05362 , and GCs remain sterile . These findings demonstrate that O14920 -independent function of p55TNFR on FDCs is sufficient to support the development of P18627 networks and GCs , while P18627 -specific O14920 is indispensable for the generation of efficient humoral immune responses . Modulation of cytokine release from human monocytes by drugs used in the therapy of inflammatory bowel diseases . BACKGROUND : Cytokines produced in the gut mucosa play an important role in the pathogenesis of inflammatory bowel diseases ( Q9UKU7 ) . To determine whether drugs used in the treatment of these diseases modulate cytokine synthesis , we investigated their effects on endotoxin-induced tumour necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -1 beta and P05231 release by elutriation-purified human monocytes in vitro . METHODS : Drugs tested were dexamethasone , DB00244 , sulphapyridine and zileuton ( a P09917 inhibitor ) . Monocytes were isolated and stimulated with endotoxin , and P01375 , IL-1 and P05231 levels were determined using an enzyme-linked immunosorbent assay . RESULTS : Monocyte stimulation with endotoxin resulted in an average P01375 release of 2464 +/- 64 pg/10(6) cells , IL-1 release of 616 +/- 47 pg/10(6) cells and P05231 release of 2259 +/- 148 pg/10(6) cells . Addition of dexamethasone resulted in a reduction of P01375 , IL-1 and P05231 release to below background levels . DB00891 significantly reduced P01375 and induced IL-1 release in a dose-dependent fashion , but had no significant effect on P05231 release . 5- DB00233 did not modulate P05231 synthesis , but significantly reduced IL-1 and enhanced P01375 synthesis . Zileuton reduced P01375 and P05231 release , but enhanced IL-1 release . CONCLUSION : We conclude that these anti-inflammatory drugs are able to modulate cytokine release by human monocytes . Further studies are needed to determine whether these effects are related to their therapeutic efficacy in Q9UKU7 . Contribution of the 37-kDa laminin receptor precursor in the anti-metastatic P08118 -derived peptide DB04985 cell surface binding . PURPOSE : DB04985 is an anti-metastatic synthetic peptide with promising therapeutic efficacy against hormone-refractory prostate cancer . The characterization of the DB04985 peptide cell surface binding/internalization mechanisms and of the receptors involved remained to be explored . RESULTS : [(14)C] DB04985 cell surface binding assays showed rapid and transient kinetic profile , that was inhibited by RGD peptides , laminin , hyaluronan , and type-I collagen . RGD peptides were however unable to inhibit DB04985 intracellular uptake . Far-Western ligand binding studies enabled the identification of the 37-kDa laminin receptor precursor ( P08865 ) as a potential ligand for DB04985 . Overexpression of the recombinant P08865 indeed led to an increase in DB04985 binding but unexpectedly not to its uptake . CONCLUSIONS : Our data support the implication of laminin receptors in cell surface binding and in transducing DB04985 anti-metastatic effects , and provide a rational for targeting cancers that express high levels of such laminin receptors . Listeriolysin O secreted by Listeria monocytogenes induces NF-kappaB signalling by activating the O15111 complex . Listeriolysin O ( LLO ) is a pore-forming cytolysin secreted by the pathogen Listeria monocytogenes and is required for its intracellular survival . We recently demonstrated that in endothelial cells , LLO activates the NF-kappaB signalling pathway . In this work , we studied the LLO-induced molecular cascade of NF-kappaB activation with a cellular model extensively used to analyse the signalling pathway of NF-kappaB activation , i.e. the human embryonic kidney P29320 -293 cell line and its derivatives ( transfectants or mutants ) . When the stably transfected derivative P29320 -293 cells expressing IL-1RI were exposed to LLO , a strong NF-kappaB activation was detected , contrasting with other cell lines ( P29320 -293 wild type , P29320 -293.T and COS ) expressing a very low level of IL-1RI . Although a delayed kinetics of LLO-dependent NF-kappaB activation suggests an autocrine or paracrine IL-1-dependent pathway , we found that LLO-dependent NF-kappaB activation did not require the IL-1 protein synthesis nor the interaction with the IL-1RI specific receptor . Herein , we demonstrated that LLO-dependent NF-kappaB activation requires the activation of the O15111 beta ( IKKbeta ) subunit of IKK complex to phosphorylate and degrade cytoplasmic P25963 , a natural inhibitor of NF-kappaB . The activation induced by LLO does not require the adapters MyD88 and IL-1R-associated kinase ( P51617 ) . We suggested that LLO induces a distinct signalling pathway from that of IL-1 and its receptor . Some studies on spontaneous Hymenolepis diminuta infection in laboratory rats . Hymenolepis diminuta is a tapeworm that occurs worldwide . It is known to be found commonly in areas where large amounts of food grains or other dry feed products , which are the favorite foods for rats . Transmission of disease to human is uncommon ; however , it may be a serious threat for population who are living in rural areas which are suffering from excessive rodents . Here , this study had done on spontaneous H. diminuta infection in laboratory rats as a model . Out of thirty five adult laboratory rats investigated for parasitic diseases only nine ( 25.71 % ) were diagnosed positive for spontaneous H. diminuta infection . Four of them ( 44.44 % ) were found losing of weight and lacking of motility , while the others were normal . On microscopic examination , H. diminuta eggs had been found in their stool . On autopsy , small intestines were found to contain from 5-6 multi-segmented tapeworms in each rat . Histopathologically , intestinal lumen showed varying sections of H. diminuta segments with serrated borders . H. diminuta infection caused multiple mucosal ulcers with absence of intestinal villi from the surface epithelium and excessive mucin . Moreover , inflammatory cells infiltration in the connective tissue core of the villi . Furthermore , the Toluidine blue stain showed that there are Mastiocytosis . Additionally , there were goblet cells hyperplasia on using DB00233 . Moreover , there were high expression of cyclooxygenase 2 ( P35354 ) , tumor necrosis factor-α ( P01375 -α ) and inducible Nitric-Oxide Synthase ( iNOs ) . This implicate , strong correlation between P35354 , P01375 -α and iNOs expression and inflammation induced by H. diminuta . Cannabinoid modulation of opiate reinforcement through the ventral striatopallidal pathway . Recent evidence indicates that cannabinoid-1 ( P21554 ) receptors play a role in the mediation of opiate reward , though the neural mechanisms for this process have not been characterized . The present experiments investigated the influence of P21554 receptors in the ventral striatopallidal system on opiate-induced neurochemical events and opiate self-administration behavior in rats . Acute morphine administration ( 3 mg/kg ) significantly reduced ventral pallidal GABA efflux in a manner similar to that produced by heroin self-administration . This neurochemical effect was reversed by doses of the selective P21554 antagonist SR 141716A ( DB06155 ; 1 and 3 mg/kg ) that also significantly reduce opiate reward . Morphine-induced increases in nucleus accumbens dopamine levels were unaltered by SR 141716A . Intravenous heroin self-administration ( 0.02 mg/infusion ) was significantly reduced by intra-accumbens , but not intraventral pallidal SR 141716A infusions ( 1 and 3 microg/side ) , implicating nucleus accumbens P21554 receptors in the modulation of opiate reinforcement . In contrast , SR14716A did not alter cocaine self-administration ( 0.125 mg/inf ) , cocaine-induced ( 10 mg/kg ) decrements in ventral pallidal GABA efflux or cocaine-induced increases in accumbens dopamine . This is consistent with evidence that selective inactivation of P21554 receptors reduces opiate- , but not psychostimulant-maintained self-administration . The P21554 receptor agonist Q08050 55,212-2 ( 5 mg/kg ) reduced pallidal GABA efflux in a manner similar to morphine , and this effect was reversed by the opiate receptor antagonist naloxone . Collectively these findings suggest that P21554 receptors modulate opiate reward through the ventral striatopallidal projection and that the modulation of this projection system may be involved in the reciprocal behavioral effects between cannabinoids , and opioids . Erythropoietin preconditioning in neuronal cultures : signaling , protection from in vitro ischemia , and proteomic analysis . In this study we confirmed the presence of the erythropoietin ( EPO ) receptor on both cultured cortical neurons and PC12 cells and showed that EPO can induce changes in p38 , P29323 , and JNK signaling molecules in these cells . We induced EPO preconditioning in cortical neuronal cultures that protected neurons from a subsequent in vitro ischemic insult ( transient oxygen-glucose deprivation ) . To investigate downstream changes in protein expression in EPO-preconditioned cortical neuronal cultures , we used two-dimensional gel electrophoresis . Overall , EPO preconditioning resulted in protein up-regulation , and , from 84 of the most differentially expressed proteins selected for identification , the proteins or tentative proteins were identified in 57 cases , representing 40 different proteins . Different protein spots representing the same or closely related protein(s) occurred for 13 of the identified proteins and are likely to represent posttranslational modifications or proteolytic fragments of the protein . Two proteins ( 78-kD glucose-regulated protein and tropomyosin , fibroblast isoform 1 ) were detected in control neuronal cultures , but not following EPO preconditioning treatment , whereas one protein ( P08865 ) was detected only following EPO preconditioning . Most of the other proteins identified had not previously been associated with EPO preconditioning and will aid in the understanding of EPO 's neuroprotective response and possibly the development of new therapeutic interventions to inhibit neuronal death in acute and chronic neurodegenerative diseases . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . P37268 inhibition : a novel target for the management of dyslipidemia . A new class of compounds , known as squalene synthase inhibitors , has recently reached phase III clinical trials and may provide another therapeutic option for clinicians to improve risk management of low-density lipoprotein cholesterol ( LDL-C ) . The clinical need for another LDL-C-lowering therapy is evident by the inability to achieve an LDL-C target of less than 70 mg/dL in the majority of very high-risk patients on statin monotherapy . Human clinical trial data with DB05317 , a novel and potent inhibitor of squalene synthase , have not yet been published .

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interacts_with DB00203?

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Essential erbB family phosphorylation in osteosarcoma as a target for DB05424 inhibition . BACKGROUND : The role of erbB tyrosine kinases , especially Her-2 , in osteosarcoma has engendered intense debate . Some investigators identified an association between low-level Her-2 expression , compared to none , and poor patient outcome . Others questioned the importance of apparent cytoplasmic expression of Her-2 , since membranous overexpression is associated with poor outcome in carcinomas . We previously demonstrated that primary osteosarcoma cells express cell-surface P00533 and Her-2 , with the p80 isoform of Her-4 localized to the nucleus . We wished to determine if erbB kinases in osteosarcoma were phosphorylated , and if this was required for growth . PROCEDURES : We cultured early passage osteosarcoma cell lines in the presence or absence of the pan-erbB inhibitor DB05424 and examined the phosphorylation status of P00533 , Her-2 , and Her-4 by immunohistochemistry , cell-based ELISA , flow cytometry and two dimensional Western blot . We also assessed the impact of DB05424 upon osteosarcoma growth and survival in vitro . RESULTS : P00533 , Her-2 , and Her-4 were constitutively phosphorylated in early passage osteosarcoma cells cultured in vitro . DB05424 abrogated erbB receptor phosphorylation and caused growth inhibition and apoptosis in a titratible fashion with concentrations of 1 muM or more . CONCLUSIONS : P00533 , Her-2 , and Her-4 are constitutively phosphorylated in early passage osteosarcoma cells in tissue culture , and erbB signaling provides essential growth and anti-apoptotic signals to osteosarcoma cells . This suggests that erbB overexpression is not required for erbB to promote malignancy , but rather that overexpression is one of several mechanisms that generate unregulated erbB signaling . Acidic sphingomyelinase downregulates the liver-specific methionine adenosyltransferase 1A , contributing to tumor necrosis factor-induced lethal hepatitis . DB00118 ( DB00118 ) is synthesized by methionine adenosyltransferases ( MATs ) . Ablation of the liver-specific Q00266 gene results in liver neoplasia and sensitivity to oxidant injury . Here we show that acidic sphingomyelinase ( ASMase ) mediates the downregulation of Q00266 by P01375 . The levels of Q00266 mRNA as well as P24752 I/III protein decreased in cultured rat hepatocytes by in situ generation of ceramide from exogenous human placenta ASMase . Hepatocytes lacking the ASMase gene ( ASMase-/- ) were insensitive to P01375 but were responsive to exogenous ASMase-induced downregulation of Q00266 . In an in vivo model of lethal hepatitis by P01375 , depletion of DB00118 preceded activation of caspases 8 and 3 , massive liver damage , and death of the mice . In contrast , minimal hepatic DB00118 depletion , caspase activation , and liver damage were seen in ASMase-/- mice . Moreover , therapeutic treatment with DB00118 abrogated caspase activation and liver injury , thus rescuing ASMase+/+ mice from P01375 -induced lethality . Thus , we have demonstrated a new role for ASMase in P01375 -induced liver failure through downregulation of Q00266 , and maintenance of DB00118 may be useful in the treatment of acute and chronic liver diseases . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used . Lack of direct action of atriopeptidase inhibitor on cellular pH regulation in rabbit S2 proximal straight tubules . While the in vivo diuretic and antihypertensive effects of the newly-developed drug DB00616 ( UK-79300 ) , a prodrug of a selective atriopeptidase inhibitor UK-73967 ( DB00058 ) are well established , the mechanism of its diuretic action is not yet fully understood due to the lack of information about its direct effects on proximal tubules . To elucidate whether this atriopeptidase inhibitor has any direct effects on proximal tubules , we examined the effect of DB00058 on intracellular pH ( pHi ) of the in vitro microperfused proximal S2 segment of rabbit kidney , using the pH-sensitive fluorescent dye , (2',7')-bis(carboxyethyl)-(5,6)-carboxyfluorescein ( BCECF ) . In the steady-state condition , pHi was 7.13 +/- 0.02 ( n = 19 ) . P08473 inhibitor ( DB00058 ) at 100 microM added to the bath changed pHi by only 0.02 +/- 0.07 unit ( n = 7 , p > 0.05 ) in 10 min . The same concentration of DB00058 in the lumen changed pHi by 0.03 +/- 0.02 unit ( n = 6 , p > 0.05 ) . To test whether the synergistic effects of DB00058 on the luminal and basolateral transport systems prevented the apparent change in pHi , we also examined the effect of DB00058 in the presence of amiloride in the lumen . The inhibition of Na+/H+ antiporter by the addition of 1 mM of amiloride to the lumen caused no change in pHi response to basolateral DB00058 application . DB00058 was without effect on pHi also in the presence of atrial natriuretic polypeptide ( P01160 ) . These results suggest that DB00058 per se has no significant effect on the process of proximal acidification over a short time period . Peroxisomal and mitochondrial targeting of serine:pyruvate/alanine:glyoxylate aminotransferase in rat liver . DB00133 :pyruvate/alanine:glyoxylate aminotransferase ( P21549 or P21549 /AGT ) of rat liver is a unique enzyme of dual subcellular localization , and exists in both mitochondria and peroxisomes . To characterize a peroxisomal targeting signal of rat liver P21549 , a number of C-terminal mutants were constructed and their subcellular localization in transfected COS-1 cells was examined . Deletion of C-terminal Q9BZE0 , and point mutation of K2 ( the second Lys from the C-terminus ) , P19013 and E15 caused accumulation of translated products in the cytoplasm . This suggests that the Q03393 of P21549 is not identical to PTS1 ( the C-terminal SKL motif ) in that it is not restricted to the C-terminal tripeptide . In vitro synthesized precursor for mitochondrial P21549 was highly sensitive to the proteinase K digestion , whereas peroxisomal P21549 ( SPTp ) was fairly resistant to the protease . In in vitro import experiment with purified peroxisomes , however , SPTp recovered in the peroxisomal fraction was very sensitive to the protease . These results suggest that the mitochondrial precursor is synthesized as an unfolded form and is translocated into the mitochondrial matrix , whereas SPTp is synthesized as a folded form and its conformation changes to an unfolded form just before translocation into peroxisomes . DB09302 : Q8NBP7 inhibitor for LDL cholesterol reduction . The proof of concept that proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) inhibition affects cholesterol levels was first established after the demonstration that Q8NBP7 loss-of-function mutations result in a significant drop in circulating LDL cholesterol levels . Subsequent studies revealed that Q8NBP7 binds the epidermal growth factor precursor homology domain-A on the surface LDL Receptor ( P01130 ) and directs P01130 and Q8NBP7 for lysosomal degradation . DB09302 ( also known as SAR236553/REGN727 ) is a monoclonal antibody that binds circulating Q8NBP7 and blocks its interactions with surface P01130 . DB09302 clinical trials with different doses on different administration schedules were shown to significantly reduce LDL cholesterol both as a mono-therapy and in combination with statins or ezetimibe . Although there is great potential for anti- Q8NBP7 therapies in the management of cholesterol metabolism , there is no clear evidence yet that blocking Q8NBP7 reduces cardiovascular disease outcome . This is being investigated in ongoing Phase III clinical trials with alirocumab . Chromosome-wide assessment of replication timing for human chromosomes 11q and 21q : disease-related genes in timing-switch regions . The completion of the human genome sequence will greatly accelerate development of a new branch of bioscience and provide fundamental knowledge to biomedical research . We used the sequence information to measure replication timing of the entire lengths of human chromosomes 11q and 21q . Megabase-sized zones that replicate early or late in S phase ( thus early/late transition ) were defined at the sequence level . Early zones were more GC-rich and gene-rich than were late zones , and early/late transitions occurred primarily at positions identical to or near GC % transitions . We also found the single nucleotide polymorphism ( SNP ) frequency was high in the late-replicating and replication-transition regions . In the early/late transition regions , concentrated occurrence of cancer-related genes that include P24385 encoding cyclin D1 ( P24385 ) , P08620 ( KFGF ) , Q13009 and Q01543 , was observed . The transition regions contained other disease-related genes including P05067 associated with familial Alzheimer 's disease ( P05067 ) , P00441 associated with familial amyotrophic lateral sclerosis ( ALS1 ) and Q03393 associated with phenylketonuria . These findings are discussed with respect to the prediction that increased DNA damage occurs in replication-transition regions . We propose that genome-wide assessment of replication timing serves as an efficient strategy for identifying disease-related genes . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . Intracellular P05067 Domain Regulates DB00133 - DB03381 - DB01992 Transferase Expression and Is Affected in Alzheimer 's Disease . Lipids play an important role as risk or protective factors in Alzheimer 's disease ( AD ) , a disease biochemically characterized by the accumulation of amyloid beta peptides ( Aβ ) , released by proteolytic processing of the amyloid precursor protein ( P05067 ) . Changes in sphingolipid metabolism have been associated to the development of AD . The key enzyme in sphingolipid de novo synthesis is serine-palmitoyl- DB01992 transferase ( P21549 ) . In the present study we identified a new physiological function of P05067 in sphingolipid synthesis . The P05067 intracellular domain ( AICD ) was found to decrease the expression of the P21549 subunit O15270 , the catalytic subunit of the P21549 heterodimer , resulting in that decreased P21549 activity . AICD function was dependent on Fe65 and O15270 levels are increased in P05067 knock-in mice missing a functional AICD domain . O15270 levels are also increased in familial and sporadic AD postmortem brains , suggesting that P21549 is involved in AD pathology . DB00203 induces angiogenic response in human coronary arteriolar endothelial cells through the expression of thioredoxin , hemeoxygenase and vascular endothelial growth factor . This study was undertaken to investigate the effect of phosphodiesterase-5 ( O76074 ) inhibitor , sildenafil , on angiogenic response in human coronary arteriolar endothelial cells ( HCAEC ) . The cells exposed to sildenafil ( 1-20 microM ) demonstrated significantly accelerated tubular morphogenesis with the induction of thioredoxin-1 ( P10599 -1 ) , hemeoxygenase-1 ( P09601 ) and P15692 . DB00203 induced P15692 and angiopoietin specific receptors such as P35968 , Tie-1 and Tie-2 . This angiogenic response was repressed by tinprotoporphyrin IX ( SnPP ) , an inhibitor of P09601 enzyme activity . DB00203 below 1 muM has no angiogenic effect as evidenced by reduced tuborogenesis . DB00203 along with SnPP inhibited both P15692 and Q15389 ( Ang-1 ) protein expression . Therefore our results demonstrated for the first time that sildenafil is a very potent pro-angiogenic factor . Neuroprotective and metabolic effects of resveratrol : therapeutic implications for Huntington 's disease and other neurodegenerative disorders . DB02709 is a naturally occurring polyphenolic compound associated with beneficial effects on aging , metabolic disorders , inflammation and cancer in animal models and resveratrol is currently being tested in numerous clinical trials . DB02709 may exert these effects by targeting several key metabolic sensor/effector proteins , such as AMPK , Q96EB6 , and P20142 -1α . DB02709 has also received considerable attention recently for its potential neuroprotective effects in neurodegenerative disorders where AMPK , Q96EB6 or P20142 -1α may represent promising therapeutic targets . A recent study published in Experimental Neurology ( Ho et al. , 2010 ) examined the therapeutic potential of a micronised proprietary resveratrol formulation , DB05073 in the N171-82Q transgenic mouse model of Huntington 's disease ( HD ) . HD is a progressive and devastating genetic neurodegenerative disorder that is associated with downregulation of P20142 -1α activity . The Ho et al. study found that DB05073 treatment did not lead to significant improvement in weight loss , motor performance , survival and striatal atrophy . However , other studies have reported neuroprotective effects of resveratrol and a distantly related polyphenol , fisetin , in HD models . HD has been associated with diabetes mellitus . Interestingly , evidence from the Ho et al. study suggests a resveratrol formulation induced beneficial anti-diabetic effect in N171-82Q mice . This commentary summarizes the pertinent outcomes from the Ho et al. study and discusses the further prospects of resveratrol and other polyphenols , including novel grape-derived polyphenols , in the treatment of HD and other neurodegenerative disorders . Phosphodiesterase 5 ( O76074 ) inhibition , P01160 and NO rapidly reduce epididymal duct contractions , but long-term O76074 inhibition in vivo does not . Contractility of the peritubular smooth muscle layer ensures the transit of immotile spermatozoa through the epididymal duct to acquire their fertilizing capacity . Atrial natriuretic peptide ( P01160 ) and nitric oxide ( NO ) affect contractility via cGMP signals that are controlled by phosphodiesterases ( PDEs ) . DB00203 inhibits the cGMP-hydrolyzing O76074 and thereby promotes relaxation of smooth muscle cells . While sildenafil is increasingly used in young patients for the treatment of pulmonary hypertension , virtually no knowledge exists about PDEs in the epididymis . Western blotting , immunohistochemistry and RT-PCR analyses after laser capture microdissection localized O76074 to smooth muscle cells , but not to epithelial cells , of the epididymal duct in man and rat . DB00203 , P01160 and NO significantly slowed spontaneous contractions of rat epididymal duct segments in organ bath studies . DB00203 effects were additive to P01160 and NO . Long-term exposure to sildenafil in vivo did not change the O76074 expression or the observed contractility pattern with the rapid relaxing response toward P01160 , NO and sildenafil . Data demonstrate that O76074 is an important member of cGMP signaling pathways regulating the finely orchestrated process of epididymal duct contractility and suggest , however , that in the epididymis side effects of therapeutically used sildenafil are unlikely . Leukotriene C4 production during hypoxic pulmonary vasoconstriction in isolated rat lungs . Leukotriene inhibitors preferentially inhibit hypoxic pulmonary vasoconstriction in isolated rat lungs . If lipoxygenase products are involved in the hypoxic pressor response they might be produced during acute alveolar hypoxia and a leukotriene inhibitor should block both the vasoconstriction and leukotriene production that occurs in response to hypoxia . We investigated in isolated blood perfused rat lungs whether leukotriene C4 ( LTC4 ) could be recovered from whole lung lavage fluid during ongoing hypoxic vasoconstriction . Lung lavage from individual rats had slow reacting substance ( SRS ) -like myotropic activity by guinea pig ileum bioassay . Pooled lavage ( 10 lungs ) as analyzed by reverse phase high performance liquid chromatography had an ultraviolet absorbing component at the retention time for LTC4 . At radioimmunoassay , and SRS myotropic activity by bioassay . LTC4 was not found during normoxic ventilation , during normoxic ventilation after a hypoxic pressor response , or during vasoconstriction elicited by DB00761 . DB00711 citrate , a leukotriene synthesis blocker , concomitantly inhibited the hypoxic vasoconstriction and LTC4 production . Thus P09917 products may play a role in the sequence of events leading to hypoxic pulmonary vasoconstriction . Effect of phosphodiesterase-5 inhibitor , sildenafil ( Viagra ) , in animal models of airways disease . Phosphodiesterase ( PDE ) -5 degrades guanosine 3',5'cyclic monophosphate ( cGMP ) and its inhibitor sildenafil citrate ( Viagra ) treats erectile dysfunction by smooth muscle relaxation through elevated cGMP . DB00203 was examined in two guinea pig models of airways disease : guinea pigs exposed to LPS or sensitized guinea pigs with atopy exposed to ovalbumen . Ovalbumen exposure caused early- and late-phase bronchoconstrictor responses , measured in conscious animals by whole-body plethysmography . Twenty-four hours after ovalbumen exposure there was airway hyperreactivity ( P35869 ) to inhaled histamine and significantly elevated macrophages , eosinophils , and nitric oxide ( NO ) metabolites in bronchoalveolar lavage fluid . DB00203 treatment ( 1 mg/kg , intraperitoneally ) failed to affect the early and late responses but significantly reduced P35869 , leukocyte influx , and elevated NO . LPS exposure ( 30 microg/ml ) caused P35869 to histamine at 1 hour and macrophage , eosinophil , and neutrophil influx at 24 hours with raised NO . DB00203 pretreatment inhibited LPS-induced P35869 , leukocyte influx , and NO generation . The effectiveness of sildenafil was not dependent on endogenous NO because inhibition of NO synthase with Nomega-nitro-L-arginine methyl ester did not prevent its action . Inhibition of O76074 by sildenafil was confirmed by elevated S-nitroso-N-acetylpenicillamine-induced cGMP generation in isolated lungs . These antiinflammatory actions of sildenafil in guinea pig models suggest that O76074 inhibitors may have potential in treating airways disease . DB02709 neuroprotection in a chronic mouse model of multiple sclerosis . DB02709 is a naturally occurring polyphenol that activates Q96EB6 , an NAD-dependent deacetylase . DB05073 , a pharmaceutical formulation of resveratrol with enhanced systemic absorption , prevents neuronal loss without suppressing inflammation in mice with relapsing experimental autoimmune encephalomyelitis ( EAE ) , a model of multiple sclerosis ( MS ) . In contrast , resveratrol has been reported to suppress inflammation in chronic EAE , although neuroprotective effects were not evaluated . The current studies examine potential neuroprotective and immunomodulatory effects of resveratrol in chronic EAE induced by immunization with myelin oligodendroglial glycoprotein peptide in C57/Bl6 mice . Effects of two distinct formulations of resveratrol administered daily orally were compared . DB02709 delayed the onset of EAE compared to vehicle-treated EAE mice , but did not prevent or alter the phenotype of inflammation in spinal cords or optic nerves . Significant neuroprotective effects were observed , with higher numbers of retinal ganglion cells found in eyes of resveratrol-treated EAE mice with optic nerve inflammation . Results demonstrate that resveratrol prevents neuronal loss in this chronic demyelinating disease model , similar to its effects in relapsing EAE . Differences in immunosuppression compared with prior studies suggest that immunomodulatory effects may be limited and may depend on specific immunization parameters or timing of treatment . Importantly , neuroprotective effects can occur without immunosuppression , suggesting a potential additive benefit of resveratrol in combination with anti-inflammatory therapies for MS . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . OSU-03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier : Implications for Anti-Cancer Therapies . We examined the interaction between OSU-03012 ( also called AR-12 ) with phosphodiesterase 5 ( O76074 ) inhibitors to determine the role of the chaperone glucose-regulated protein ( P11021 ) / P11021 / P11021 in the cellular response . DB00203 ( Viagra ) interacted in a greater than additive fashion with OSU-03012 to kill stem-like GBM cells . Treatment of cells with OSU-03012/sildenafil : abolished the expression of multiple oncogenic growth factor receptors and plasma membrane drug efflux pumps and caused a rapid degradation of P11021 and other HSP70 and HSP90 family chaperone proteins . Decreased expression of plasma membrane receptors and drug efflux pumps was dependent upon enhanced Q9NZJ5 -eIF2α- P18848 - P35638 signaling and was blocked by P11021 over-expression . In vivo OSU-03012/sildenafil was more efficacious than treatment with celecoxib and sildenafil at killing tumor cells without damaging normal tissues and in parallel reduced expression of P08183 and Q9UNQ0 in the normal brain . The combination of OSU-03012/sildenafil synergized with low concentrations of sorafenib to kill tumor cells , and with lapatinib to kill P00533 over-expressing tumor cells . In multiplex assays on plasma and human tumor tissue from an OSU-03012/sildenafil treated mouse , we noted a profound reduction in uPA signaling and identified FGF and P23458 /2 as response biomarkers for potentially suppressing the killing response . Inhibition of FGFR signaling and to a lesser extent P23458 /2 signaling profoundly enhanced OSU-03012/sildenafil lethality . Retreatment of men with metastatic castrate-resistant prostate cancer with abiraterone . BACKGROUND : DB05812 acetate ( AA ) , oral P05093 inhibitor , is an active agent in the treatment of metastatic castrate-resistant prostate cancer ( mCRPC ) . METHODS : We ( R.L.A and N.A ) retrospectively evaluated outcome in 12 men who were re-treated with AA following prior treatment with AA at the Princess Margaret Cancer Centre . RESULTS : All men were heavily pre-treated for mCRPC with a median of four prior lines of therapy , one of which was AA ( given either pre- or post-chemotherapy ) . Eleven out of 12 ( 92 % ) men stopped their first treatment course of AA due to progression and one stopped for financial reasons . Seven men had a PSA decrease ≥50 % following their first AA treatment , of which three ( 46 % ) had a PSA decrease ≥50 % to AA re-treatment . The responses to AA re-treatment were generally short-lived with a median biochemical progression-free survival of 2.3 months and median treatment duration of 3.2 months . No PSA responses to AA re-treatment were seen in five men who did not have an initial PSA response to AA . CONCLUSIONS : Our data suggest that AA re-challenge may have limited benefit in select men with mCRPC , and warrants further formal research . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . DB06080 inhibits the proliferation of Ewing Sarcoma cells and suppresses platelet-derived growth factor receptor beta and c- P10721 signaling pathways . The Ewing Sarcoma ( Q01844 ) family of tumors is one of the most common tumors diagnosed in children and adolescents and is characterized by a translocation involving the Q01844 gene . Despite advances in chemotherapy , the prognosis of metastatic Q01844 is poor with an overall survival of < 30 % after 5 years . Q01844 tumor cells express the receptor tyrosine kinases , platelet-derived growth factor receptor ( P09619 ) and c- P10721 . DB06080 is a multitargeted small-molecule inhibitor that targets Fms-like tyrosine kinase-3 , c- P10721 , vascular endothelial growth receptors , and PDGFRs . To determine the potential therapeutic benefit of DB06080 in Q01844 cells , we examined the effects of DB06080 on Q01844 cell lines and xenograft mouse models . DB06080 inhibited the proliferation of two Q01844 cell lines , A4573 and TC71 , at an IC(50) of 1.25 and 2 mumol/L after 72 h of treatment , respectively . The phosphorylation of PDGFRbeta , c- P10721 , and extracellular signal-regulated kinases was also inhibited . To examine the effects of DB06080 in vivo , the drug was given to mice injected with Q01844 cells . We observed inhibition of growth of Q01844 tumor cells in a xenograft mouse model and prolonged survival in a metastatic mouse model of Q01844 . Therefore , our in vitro and in vivo studies show that DB06080 inhibits proliferation of Q01844 cells through inhibition of PDGFRbeta and c- P10721 pathways . P08473 inhibitor suppresses the early phase of atrial electrical remodeling in a canine rapid atrial pacing model . INTRODUCTION : We examined the acute effects of neutral endopeptidase inhibitor on the hemodynamics and electrical properties of dogs subjected to rapid atrial pacing . METHODS : Ten beagle dogs were used and divided into two groups with and without candoxatril , a neutral endopeptidase inhibitor preadministration . Before and after the 6 hours rapid atrial pacing from the right atrial appendage , the hemodynamics , atrial effective refractory period , and monophasic action potential duration of the right atrial appendage were measured and blood samples were collected . Atrial tissue was also excised after the experiment . RESULTS : DB00616 significantly increased plasma P01160 levels ( Control : 88.4 +/- 50.25 vs. DB00616 : 197.1 +/- 32.09 pg/ml , p = 0.004 ) and prevented reductions in atrial effective refractory period and monophasic action potential duration . We further demonstrated that the treated animals exhibited significantly higher levels of atrial tissue cyclic GMP ( Control : 28.1 +/- 1.60 fmol/mg vs. DB00616 : 44.5 +/- 12.28 fmol/mg , p = 0.034 ) as well as that of plasma cyclic GMP ( Control : 32 +/- 5.5 vs. DB00616 : 42 +/- 7.1 pg/ml , p = 0.028 ) . CONCLUSION : DB00616 suppressed the shortening of atrial effective refractory period and monophasic action potential duration in the rapid atrial pacing model . As plasma P01160 and the atrial tissue levels of cyclic GMP were higher in the DB00616 group than the control , this effect was considered to appear through the reduction of calcium overload caused by P01160 and cyclic GMP . Inhibition of the leukotriene synthetase of rat basophil leukemia cells by diethylcarbamazine , and synergism between diethylcarbamazine and piriprost , a P09917 inhibitor . DB00711 inhibited the formation of sulfidopeptide leukotrienes in rat basophil leukemia ( RBL ) cells ( 50 % inhibitory concentration , EC50 , 3 mM ) . Similar concentrations also inhibited the formation of leukotriene C4 ( LTC4 ) by LTC synthetase , a detergent-solubilized cell free particulate enzyme from RBL cells which is capable of coupling P09960 to glutathione . By contrast , the conversion of P09960 to LTC4 using enzymes from rat liver was at least ten times less sensitive to this inhibitor . The EC50 for inhibition of the leukotriene C synthetase of RBL cells was directly proportional to the P09960 concentration in the incubations , ranging from 1.5 mM at 10 microM P09960 to over 40 mM at 500 microM P09960 . Kinetic analysis revealed that the inhibition of the leukotriene C synthetase reaction by diethylcarbamazine was competitive with respect to P09960 . In contrast to diethylcarbamazine , piriprost ( U-60,257 ; 6,9-deepoxy-6,9-(phenylimino)-delta 6,8-prostaglandin I1 ) , which inhibits the formation of sulfidopeptide leuktrienes in RBL cells at the P09917 step ( EC50 5 microM ) , did not inhibit the leukotriene synthetase of these cells . On the other hand , low concentrations of piriprost , which had no demonstrable inhibitory activity on leukotriene formation by themselves , markedly synergized the inhibitory activity of diethylcarbamazine . These results are consistent with the interpretation that both piriprost and diethylcarbamazine inhibit leukotriene formation but that they act on sequential steps in the biosynthetic pathway in such a manner as to synergistically interfere with the availability or utilization of P09960 in the leukotriene C synthetase reaction . DB00203 enhances neurogenesis and oligodendrogenesis in ischemic brain of middle-aged mouse . Adult neural stem cells give rise to neurons , oligodendrocytes and astrocytes . Aging reduces neural stem cells . Using an inducible nestin-CreER( P24752 )/R26R-yellow fluorescent protein ( YFP ) mouse , we investigated the effect of DB00203 , a phosphodiesterase type 5 ( O76074 ) inhibitor , on nestin lineage neural stem cells and their progeny in the ischemic brain of the middle-aged mouse . We showed that focal cerebral ischemia induced nestin lineage neural stem cells in the subventricular zone ( SVZ ) of the lateral ventricles and nestin expressing NeuN positive neurons and adenomatous polyposis coli ( P25054 ) positive mature oligodendrocytes in the ischemic striatum and corpus callosum in the aged mouse . Treatment of the ischemic middle-aged mouse with DB00203 increased nestin expressing neural stem cells , mature neurons , and oligodendrocytes by 33 , 75 , and 30 % , respectively , in the ischemic brain . These data indicate that DB00203 amplifies nestin expressing neural stem cells and their neuronal and oligodendrocyte progeny in the ischemic brain of the middle-aged mouse . Discovery of 3-phenyl-1H-5-pyrazolylamine derivatives containing a urea pharmacophore as potent and efficacious inhibitors of P07333 -like tyrosine kinase-3 ( P36888 ) . Preclinical investigations and early clinical trials suggest that P36888 inhibitors are a viable therapy for acute myeloid leukemia . However , early clinical data have been underwhelming due to incomplete inhibition of P36888 . We have developed 3-phenyl-1H-5-pyrazolylamine as an efficient template for kinase inhibitors . Structure-activity relationships led to the discovery of sulfonamide , carbamate and urea series of P36888 inhibitors . Previous studies showed that the sulfonamide 4 and carbamate 5 series were potent and selective P36888 inhibitors with good in vivo efficacy . Herein , we describe the urea series , which we found to be potent inhibitors of P36888 and P35968 . Some inhibited growth of P36888 -mutated MOLM-13 cells more strongly than the P36888 inhibitors sorafenib ( 2 ) and DB06080 ( 3 ) . In preliminary in vivo toxicity studies of the four most active compounds , 10f was found to be the least toxic . A further in vivo efficacy study demonstrated that 10f achieved complete tumor regression in a higher proportion of MOLM-13 xenograft mice than 4 and 5 ( 70 % vs 10 % and 40 % ) . These results show that compound 10f possesses improved pharmacologic and selectivity profiles and could be more effective than previously disclosed P36888 inhibitors in the treatment of acute myeloid leukemia . DB05812 acetate : redefining hormone treatment for advanced prostate cancer . Prostate cancer has long since been recognised as being hormonally driven via androgen receptor signalling . DB05812 acetate ( AA ) is a rationally designed P05093 inhibitor that blocks the conversion of androgens from non-gonadal precursors effectively , thus reducing testosterone to undetectable levels . AA has recently been proved to extend survival for men with metastatic castration-resistant prostate cancer who have progressive disease after first-line chemotherapy treatment . In addition , it is currently being tested in a Phase III trial in the pre-chemotherapy setting . This paper will review the preclinical discovery and clinical development of AA and will outline the strategy of parallel translational research . Gene promoter methylation in colorectal cancer and healthy adjacent mucosa specimens : correlation with physiological and pathological characteristics , and with biomarkers of one-carbon metabolism . We evaluated the promoter methylation levels of the P25054 , P16455 , hMLH1 , RASSF1A and CDKN2A genes in 107 colorectal cancer ( CRC ) samples and 80 healthy adjacent tissues . We searched for correlation with both physical and pathological features , polymorphisms of folate metabolism pathway genes ( P42898 , Q9UBK8 , Q99707 , RFC1 , P04818 , and Q9UBC3 ) , and data on circulating folate , vitamin B12 and homocysteine , which were available in a subgroup of the CRC patients . An increased number of methylated samples were found in CRC respect to adjacent healthy tissues , with the exception of P25054 , which was also frequently methylated in healthy colonic mucosa . Statistically significant associations were found between RASSF1A promoter methylation and tumor stage , and between hMLH1 promoter methylation and tumor location . Increasing age positively correlated with both hMLH1 and P16455 methylation levels in CRC tissues , and with P25054 methylation levels in the adjacent healthy mucosa . Concerning gender , females showed higher hMLH1 promoter methylation levels with respect to males . In CRC samples , the Q99707 2756AG genotype correlated with higher methylation levels of RASSF1A , and the P04818 1494 6bp ins/del polymorphism correlated with the methylation levels of both P25054 and hMLH1 . In adjacent healthy tissues , Q99707 2756AG and P04818 1494 6bp del/del genotypes correlated with P25054 and P16455 promoter methylation , respectively . Low folate levels were associated with hMLH1 hypermethylation . Present results support the hypothesis that DNA methylation in CRC depends from both physiological and environmental factors , with one-carbon metabolism largely involved in this process . An investigation of endocrine disrupting effects and toxic mechanisms modulated by benzo[a]pyrene in female scallop Chlamys farreri . The purpose of this study was to investigate the endocrine disrupting effects induced by benzo[a]pyrene ( B[a]P ) and explore the underlying mechanisms in mollusks . In this study , sexually mature female Chlamys farreri were exposed to benzo[a]pyrene for 10 days at four different concentrations as 0 , 0.025 , 0.5 and 10 μg/L . Sex steroids were identified and quantified by electrochemiluminescence immunoassay ( ECLIA ) method and results showed that exposure to B[a]P exerts great suppression on 17β-estradiol , testosterone production and disrupts progesterone levels in ovary . Transcription of genes were detected and measured by real-time RT-PCR . It showed that at day 10 B[a]P inhibited 3 β-HSD , P05093 and 17β-HSD mRNA expression in a dose-dependent manner , which suggests that they could be potential targets of B[a]P that disrupt steroidogenic machinery . Moreover , 0.025 μg/L B[a]P activated transcription of aryl hydrocarbon receptor ( P35869 ) , P35869 nuclear translocator ( P27540 ) , P04798 and estrogen receptor ( ER ) , while 10 μg/L B[a]P suppressed all of them . The consistency of their responses to B[a]P exposure implies that P35869 action may be involved in invertebrate CYP regulation and ER transcription despite of unknown mechanisms . Additionally , B[a]P exposure could induce ovarian impairment and developmental delay in C. farreri . Overall , sensitivity of C. farreri to endocrine disruption and toxicity suggests that C. farreri is a suitable species for study of endocrine-disrupting effects in marine invertebrates . This study will form a solid basis for a realistic extrapolation of endocrine disrupting effects across taxonomic groups and phyla . Pharmacologic blockade of P09917 improves the amyloidotic phenotype of an Alzheimer 's disease transgenic mouse model involvement of γ-secretase . The P09917 ( P09917 ) enzyme is widely distributed within the central nervous system . Previous works showed that this protein is up-regulated in Alzheimer 's disease ( AD ) and that its genetic absence results in a reduction of amyloid β ( Aβ ) levels in Tg2576 mice . In the present study , we examined the effect of P09917 pharmacological inhibition on the amyloidotic phenotype of these mice . Aβ deposition in the brains of mice receiving zileuton , a selective and specific P09917 inhibitor , was significantly reduced when compared with control Tg2576 mice receiving vehicle . This reduction was associated with a similar decrease in brain Aβ peptides levels . Zileuton treatment did not induce any change in the steady state levels of amyloid-β precursor protein ( P05067 ) , P56817 or O14672 . By contrast , it resulted in a significant reduction of presenilin 1 ( P49768 , alias P49768 ) , nicastrin ( Q92542 ) , presenilin enhancer 2 homolog ( PSNEN , alias , Pen-2 ) , and anterior pharynx defective 1 ( P13798 -1 ) , the four components of the γ-secretase complex-at the protein and message level . Furthermore , in vitro studies confirmed that zileuton prevents Aβ formation by modulating γ-secretase complex levels without affecting Notch signaling . These data establish a functional role for P09917 in the pathogenesis of AD-like amyloidosis , whereby it modulates the γ-secretase pathway . They suggest that pharmacological inhibition of P09917 could provide a novel therapeutic opportunity for AD . DB09302 inhibits atherosclerosis , improves the plaque morphology , and enhances the effects of a statin . Q8NBP7 ( Q8NBP7 ) inhibition is a potential novel strategy for treatment of CVD . DB09302 is a fully human Q8NBP7 monoclonal antibody in phase 3 clinical development . We evaluated the antiatherogenic potential of alirocumab in P02649 *3Leiden. P11597 mice . Mice received a Western-type diet and were treated with alirocumab ( 3 or 10 mg/kg , weekly subcutaneous dosing ) alone and in combination with atorvastatin ( 3.6 mg/kg/d ) for 18 weeks . DB09302 alone dose-dependently decreased total cholesterol ( -37 % ; -46 % , P < 0.001 ) and TGs ( -36 % ; -39 % , P < 0.001 ) and further decreased cholesterol in combination with atorvastatin ( -48 % ; -58 % , P < 0.001 ) . DB09302 increased hepatic P01130 protein levels but did not affect hepatic cholesterol and TG content . Fecal output of bile acids and neutral sterols was not changed . DB09302 dose-dependently decreased atherosclerotic lesion size ( -71 % ; -88 % , P < 0.001 ) and severity and enhanced these effects when added to atorvastatin ( -89 % ; -98 % , P < 0.001 ) . DB09302 reduced monocyte recruitment and improved the lesion composition by increasing the smooth muscle cell and collagen content and decreasing the macrophage and necrotic core content . DB09302 dose-dependently decreases plasma lipids and , as a result , atherosclerosis development , and it enhances the beneficial effects of atorvastatin in P02649 *3Leiden. P11597 mice . In addition , alirocumab improves plaque morphology . Targeting epidermal growth factor receptor : novel therapeutics in the management of cancer . Overexpression of epidermal growth factor receptor ( P00533 ) in epithelial tumors , including head and neck , lung , breast , colon and other solid tumors , has frequently been correlated with poor prognosis , thus stimulating efforts to develop new cancer therapies that target P00533 . Monoclonal antibodies and tyrosine kinase inhibitors specifically targeting P00533 are the most well-studied and hold substantial promise of success . Several compounds of monoclonal antibodies and tyrosine kinase inhibitors targeting P00533 have been studied and clinical trials are now underway to test the safety and efficacy of these targeting strategies in several human tumors . This review will address each of these agents alone or in combination with radiation or chemotherapy and highlight some of these promising developments . Cetuximab ( Erbitux ) is being evaluated in combination with radiation or chemotherapy in Phase III trials . Other compounds such as h-R3 , DB01269 , P50402 -55900 and ICR-62 have proved to be effective in targeting malignant cells alone or in combination with traditional therapies . Tyrosine kinase inhibitors targeting the intracellular domain of P00533 , including ZD-1839 ( gefitinib , DB00317 ) , DB00530 ( Erlotinib/Tarceva ) , PD-153053 , PD-168393 and DB05424 , have been studied in clinical setting alone or in combination with radiation or chemotherapy . ZD-1839 is being studied in a Phase III trial in patients with advanced non-small cell lung cancer . P00533 targeted treatment by monoclonal antibodies and tyrosine kinase inhibitors have been proven to sensitize tumor cells to the effects of chemotherapy and radiation therapy . The synergistic activities and nonoverlapping toxicities of these compounds allow concomitant administration with cytotoxic therapy . Challenges of evaluating P00533 targeted agents exist in selecting the optimal dosages and determining long-term toxicity . DB02709 inhibits PDGF receptor mitogenic signaling in mesangial cells : role of P18031 . Mesangioproliferative glomerulonephritis is associated with overactive PDGF receptor signal transduction . We show that the phytoalexin resveratrol dose dependently inhibits PDGF-induced DNA synthesis in mesangial cells with an IC(50) of 10 microM without inducing apoptosis . Remarkably , the increased Q96EB6 deacetylase activity induced by resveratrol was not necessary for this inhibitory effect . DB02709 significantly blocked PDGF-stimulated c-Src and Akt kinase activation , resulting in reduced cyclin D1 expression and attenuated P06400 phosphorylation and cyclin-dependent kinase-2 ( P24941 ) activity . Furthermore , resveratrol inhibited P09619 phosphorylation at the PI 3 kinase and Grb-2 binding sites tyrosine-751 and tyrosine-716 , respectively . This deficiency in P09619 phosphorylation resulted in significant inhibition of PI 3 kinase and Erk1/2 MAPK activity . Interestingly , resveratrol increased the activity of protein tyrosine phosphatase P18031 , which dephosphorylates PDGF-stimulated phosphorylation at tyrosine-751 and tyrosine-716 on P09619 with concomitant reduction in Akt and Erk1/2 kinase activity . P18031 significantly inhibited PDGF-induced DNA synthesis without inducing apoptosis . These results for the first time provide evidence that the stilbene resveratrol targets P18031 to inhibit P09619 mitogenic signaling . Sex-specific association of sequence variants in P35520 and Q9UBK8 with risk for promoter hypermethylation in the lung epithelium of smokers . Gene promoter hypermethylation is now regarded as a promising biomarker for the risk and progression of lung cancer . The one-carbon metabolism pathway is postulated to affect deoxyribonucleic acid ( DNA ) methylation because it is responsible for the generation of S-adenosylmethionine ( DB00118 ) , the methyl donor for cellular methylation reactions . This study investigated the association of single nucleotide polymorphisms ( SNPs ) in six one-carbon metabolism-related genes with promoter hypermethylation in sputum DNA from non-Hispanic white smokers in the Lovelace Smokers Cohort ( LSC ) ( n = 907 ) . Logistic regression was used to assess the association of SNPs with hypermethylation using a high/low methylation cutoff . SNPs in the cystathionine beta synthase ( P35520 ) and 5-methyltetrahydrofolate-homocysteine methyltransferase reductase ( Q9UBK8 ) genes were significantly associated with high methylation in males [ P35520 rs2850146 ( -8283G > C ) , OR = 4.9 ; 95 % CI : 1.98 , 12.2 , P = 0.0006 ] and low methylation in females [ Q9UBK8 rs3776467 ( 7068A > G ) , OR = 0.57 , 95 % CI : 0.42 , 0.77 , P = 0.0003 ] . The variant allele of rs2850146 was associated with reduced gene expression and increased plasma homocysteine ( Hcy ) concentrations . Three plasma metabolites , Hcy , methionine and dimethylglycine , were associated with increased risk for gene methylation . These studies suggest that SNPs in P35520 and Q9UBK8 have sex-specific associations with aberrant methylation in the lung epithelium of smokers that could be mediated by the affected one-carbon metabolism and transsulfuration in the cells .

MH_train_1230

MH_train_1230

MH_train_1230

interacts_with DB08899?

multiple_choice

DB09045 : the newest P43220 agonist for the management of type 2 diabetes . OBJECTIVE : To review the pharmacology , pharmacokinetics , safety , and efficacy of the glucagon-like peptide-1 receptor agonist ( P0C6A0 RA ) , dulaglutide , in the treatment of type 2 diabetes mellitus ( T2D ) . DATA SOURCES : A PubMed search was completed to identify publications from 1947 to October 2014 using the search terms dulaglutide and LY2189265 . References were reviewed to identify additional resources . STUDY SELECTION AND DATA EXTRACTION : Articles were included if they evaluated the pharmacology , pharmacokinetics , safety , or efficacy of dulaglutide . DATA SYNTHESIS : DB09045 reduces both glycosylated hemoglobin ( A1C ) and weight by stimulating insulin secretion and suppressing glucagon in a glucose-dependent manner , delaying gastric emptying , and promoting satiety . DB09045 consists of 2 P0C6A0 analogues that have been modified to make it a long-acting , once-weekly agent . DB09045 has been studied as monotherapy and in combination with metformin , glimepiride , pioglitazone , and insulin lispro . It has demonstrated superior A1C reduction compared with placebo , metformin , insulin glargine , sitagliptin , and twice-daily exenatide . It demonstrated noninferiority in A1C reduction to liraglutide . DB09045 changed A1C by -0.78 % to -1.51 % , and it changed weight by -0.35 kg to -3.03 kg . The most common adverse effects in clinical studies were nausea , vomiting , and diarrhea . CONCLUSIONS : DB09045 is the fifth P0C6A0 RA approved for T2D in the United States . It is an attractive option because it is dosed once-weekly , provides A1C lowering similar to liraglutide , weight reduction similar to exenatide , and has an adverse effect profile similar to exenatide and liraglutide . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . Derivatives of vitamins D2 and D3 activate three MAPK pathways and upregulate P06400 expression in differentiating HL60 cells . Analogs of vitamin D have been synthesized which have reduced calcemic activities yet increased anti-proliferative and differentiation-inducing properties , raising expectations that they will be useful for treatment of human neoplastic diseases . In the present study we compared the abilities of three such analogs , 24a , 24b-dihomo-1,25-dihydroxyvitamin D(3) ( P13489 -1890 ) , 24-ene-1,25-dihydroxyvitamin D(2) ( P13489 -1906 ) and ( 24R ) -1,24-dihydroxyvitamin D(3) ( P13489 -2191 ) to induce markers ( P08571 , CD11b and P13929 ) of differentiation , G(1) phase block , and associated molecular events in human promyeloblastic leukemia cells HL60 . We found that the potencies of the analogs to induce differentiation paralleled their activation of Erk , JNK and p38 mitogen-activated protein kinase ( MAPK ) pathways , and the anti-proliferative activity closely correlated with the extent of hypophosphorylation of retinoblastoma protein ( P06400 ) . Interestingly , low concentrations of derivatives of vitamin D , which were insufficient to induce any detectable changes in the cell cycle traverse , markedly increased the levels of total P06400 , which was highly phosphorylated . These results suggest that P06400 may have an unsuspected role in monocytic differentiation , perhaps to increase the sensitivity of the G(1) checkpoint , by increasing the amount of substrate for cyclin-dependent kinases . Role of the CCAAT/enhancer binding protein-alpha transcription factor in the glucocorticoid stimulation of p21waf1/cip1 gene promoter activity in growth-arrested rat hepatoma cells . The preceding paper ( Cha , H . H. , Cram , E. J. , Wang , E. C. , Huang , A. J. , Kasler , H . G. , and Firestone , G . L . ( 1998 ) J . Biol . Chem . 273 , 0000-0000(478563) defined a glucocorticoid responsive region within teh promoter of the P38936 CDK inhibitor gene that contains a putative DNA-binding site for the transcription factor CCAAT/ enhancer binding protein-alpha ( P49715 ) . Wild type rat BDS1 hepatoma cells as well as as4 hepatoma cells , which express antisense sequences to P49715 and ablate its protein production , were utilized to investigate the role of this transcription factor in the glucocorticoid regulation of P38936 gene expression . The stimulation of P38936 protein levels and promoter activity , as well as inhibition of P24941 -mediated retinoblastoma protein phosphorylation , by the synthetic glucocorticoid , dexamethasone , required the expression of P49715 . Overexpression of P49715 in as4 cells rescued the dexamethasone responsiveness of the P38936 promoter . Site-directed mutagenesis of the P38936 promoter revealed that dexamethasone stimulation of P38936 promoter activity required the C/EBP consensus DNA-binding site . Furthermore , in glucocorticoid receptor-defective P78364 hepatoma cells , dexamethasone failed to stimulate P49715 and P38936 protein expression and promoter activities . Our results have established a functional link between the glucocorticoid receptor signaling pathway that mediates a P55008 cell cycle arrest of rat hepatoma cells and the transcriptional control of P38936 by a cascade that requires the steroid induction of P49715 gene expression . Paullones are potent inhibitors of glycogen synthase kinase-3beta and cyclin-dependent kinase 5/p25 . Paullones constitute a new family of benzazepinones with promising antitumoral properties . They were recently described as potent , DB00171 -competitive , inhibitors of the cell cycle regulating cyclin-dependent kinases ( CDKs ) . We here report that paullones also act as very potent inhibitors of glycogen synthase kinase-3beta ( GSK-3beta ) ( IC50 : 4-80 nM ) and the neuronal Q00535 /p25 ( IC50 : 20-200 nM ) . These two enzymes are responsible for most of the hyperphosphorylation of the microtubule-binding protein tau , a feature observed in the brains of patients with Alzheimer 's disease and other neurodegenerative ' taupathies ' . DB04014 , the most active paullone , was demonstrated to act by competing with DB00171 for binding to GSK-3beta . DB04014 inhibits the phosphorylation of tau in vivo at sites which are typically phosphorylated by GSK-3beta in Alzheimer 's disease . DB04014 also inhibits the Q00535 /p25-dependent phosphorylation of Q9UD71 in mouse striatum slices in vitro . This dual specificity of paullones may turn these compounds into very useful tools for the study and possibly treatment of neurodegenerative and proliferative disorders . P10275 as a therapeutic target . Androgens function as sex hormone primarily via activation of a single androgen receptor ( AR , or P10275 ) . AR is an important therapeutic target for the treatment of diseases such as hypogonadism and prostate cancer . AR ligands of different chemical structures and/or pharmacological properties are widely used for these therapeutic applications , and all of the AR ligands currently available for therapy modulate AR function via direct binding to the ligand-binding pocket ( P18428 ) of the receptor . In the past ten years , our understanding of AR structure and molecular mechanism of action has progressed extensively , which has encouraged the rapid development of newer generation of AR ligands , particularly tissue-selective AR ligands . With improved tissue selectivity , future generations of AR ligands are expected to greatly expand the therapeutic applications of this class of drugs . This review will provide an overview of the common therapeutic applications of currently available AR ligands , and discussion of the major challenges as well as novel therapeutic strategies proposed for future drug development . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Cellular senescence requires Q00535 repression of Rac1 activity . Cellular senescence is a tumor-suppressive process characterized by an irreversible cell cycle exit , a unique morphology , and expression of senescence-associated beta-galactosidase ( SA-beta-Gal ) . We report here a role for Q00535 in induction of senescent cytoskeletal changes . Q00535 activation is upregulated in senescing cells . The increased activity of Q00535 further reduces GTPase Rac1 activity and Pak activation . The repression of the activity of the GTPase Rac1 by Q00535 is required for expression of the senescent phenotype . Q00535 regulation of Rac1 activity is necessary for actin polymerization accompanying senescent morphology in response to expression of P06400 , activated Ras , or continuous passage . Inhibition of Q00535 attenuates SA-beta-Gal expression and blocks actin polymerization . These results point to a unique , nonneuronal role for Q00535 in regulation of Rac1 activity in senescence , illuminating the mechanisms underlying induction of senescence and the senescent shape change . Domain analysis of human transmembrane guanylyl cyclase receptors : implications for regulation . In the human genome , sequence analysis indicates there are five functional transmembrane guanylyl cyclases , enzymes that synthesize the intracellular second messenger , cGMP . Two , P16066 and P20594 or P16066 and P20594 , are widely distributed receptors for atrial natriuretic peptide , brain natriuretic peptide and P23582 , more commonly known as P01160 , DB04899 and P09543 , respectively . One cyclase , P25092 or StaR , is predominantly found in the intestinal epithelium and is the receptor for guanylin and uroguanylin , as well as for the bacterial pathogen , heat-stable enterotoxin ( Sta ) . The remaining two cyclases , GC-E and P51841 or RetGC-1 and RetGC-2 , are expressed in the retina and regulate the dark cycle of phototransduction . Unlike the other family members , GC-E and P51841 have no known extracellular ligands . Instead , they are activated under low calcium conditions by guanylyl cyclase activating proteins called GCAPs . All five members consist of an extracellular ligand binding domain , single transmembrane spanning domain , and intracellular kinase homology , dimerization and guanylyl cyclase catalytic domains . In the first part of this review , the tissue expression , ligands and " knockout " phenotypes of each receptor are summarized and individual domains are compared . In the second part , regulation by DB00171 , calcium , protein kinase C and phosphorylation is discussed . Monocyte activation is a feature of common variable immunodeficiency irrespective of plasma lipopolysaccharide levels . Common variable immunodeficiency disorders ( CVID ) , the most frequent cause of symptomatic primary immunodeficiency , are defined by impaired antibody production . Notwithstanding , T cell activation and granulomatous manifestations represent the main causes of CVID morbidity even in patients receiving immunoglobulin ( Ig ) G replacement therapy . Additionally , gut pathology is a frequent feature of CVID . In this study , we investigated monocyte imbalances and their possible relationship with increased microbial translocation in CVID patients . Monocyte subsets were defined according to P08571 and CD16 expression levels and evaluated in terms of human leucocyte antigen D-related ( HLA-DR ) , P42081 and programmed death-1 molecule ligand 1 ( Q9NZQ7 ) expression by flow cytometry , in parallel with the quantification of plasma lipopolysaccharide ( LPS ) and serum levels of soluble P08571 ( sCD14 ) , LPS-binding protein ( P18428 ) and anti-LPS antibodies . CVID patients ( n=31 ) featured significantly increased levels of serum sCD14 and an expansion of P08571 (bright) CD16(+) monocytes in direct correlation with T cell and B cell activation , the latter illustrated by the frequency of the CD21(low) P28907 (low) subset . Such alterations were not observed in patients lacking B cells due to congenital agammaglobulinaemia ( n=4 ) . Moreover , we found no significant increase in circulating LPS or P18428 levels in CVID patients , together with a relative preservation of serum anti-LPS antibodies , in agreement with their presence in commercial IgG preparations . In conclusion , CVID was associated with monocyte imbalances that correlated directly with T cell activation markers and with B cell imbalances , without an association with plasma LPS levels . The heightened monocyte activated state observed in CVID may represent an important target for complementary therapeutic strategies . Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells . Antibody-drug conjugates ( ADC ) are emerging as powerful treatment strategies with outstanding target-specificity and high therapeutic activity in patients with cancer . DB08870 represents a first-in-class ADC directed against P28908 (+) malignancies . We hypothesized that its sustained clinical responses could be related to the stimulation of an anticancer immune response . In this study , we demonstrate that the dolastatin family of microtubule inhibitors , from which the cytotoxic component of brentuximab vedotin is derived , comprises potent inducers of phenotypic and functional dendritic cell ( DC ) maturation . In addition to the direct cytotoxic effect on tumor cells , dolastatins efficiently promoted antigen uptake and migration of tumor-resident DCs to the tumor-draining lymph nodes . Exposure of murine and human DCs to dolastatins significantly increased their capacity to prime T cells . Underlining the requirement of an intact host immune system for the full therapeutic benefit of dolastatins , the antitumor effect was far less pronounced in immunocompromised mice . We observed substantial therapeutic synergies when combining dolastatins with tumor antigen-specific vaccination or blockade of the P18621 - Q9NZQ7 and P16410 coinhibitory pathways . Ultimately , treatment with ADCs using dolastatins induces DC homing and activates cellular antitumor immune responses in patients . Our data reveal a novel mechanism of action for dolastatins and provide a strong rationale for clinical treatment regimens combining dolastatin-based therapies , such as brentuximab vedotin , with immune-based therapies . Partial trypsin digestion as an indicator of mis-folding of mutant alanine:glyoxylate aminotransferase and chaperone effects of specific ligands . Study of a spectrum of missense mutants . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme whose deficiency results in primary hyperoxaluria type 1 ( P78364 ) . More than 75 P78364 mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have previously demonstrated that many such mutants expressed by transcription/translation are subject to generalized degradation by the proteasome and a specific limited trimming by an endogenous DB00171 -independent protease activity . Here , we report the results of partial digestion using trypsin as a mimic for the endogenous non-proteasomal protease and the use of N-terminal protein sequencing to determine the sensitive site . Partial trypsin digestion also provided an indicator of proper folding of the mutant enzyme . For selected mutations the sensitivity to trypsin could be ameliorated by addition of pyridoxal phosphate or aminooxy acetic acid as specific pharmacological chaperones . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . P13726 -enriched vesicles are taken up by platelets and induce platelet aggregation in the presence of factor VIIa . We investigated the interactions of vesicles containing human tissue factor ( TF ) with platelets and evaluated responses induced by DB00036 using standard aggregometry , ultrastructural and flow-cytometry techniques . Washed platelets were exposed to a preparation of placental human TF ( pTF ) or to a relipidated formulation of recombinant human TF ( rTF ) . Under stirring conditions , pTF induced reversible aggregation with platelets returning to their resting state after 5 minutes . This reversible response to pTF was partially inhibited by antibodies against CD62-P , but not by antithrombin agents , and was not observed with rTF . Sequential ultrastructural studies revealed uptake of both TF preparations by platelets involving traffic of vesicles through channels of the open canalicular system ( OCS ) . Immunocytochemical studies on cryosections identified TF in the OCS , and occasionally in the alpha-granules of the platelets . These processes were faster with pTF than with rTF , but both TF preparations accumulated in platelets at the end of incubation periods . Flow cytometry studies revealed the presence of other cellular antigens ( CD62-P , P08571 and P08575 ) associated to the pTF . Addition of DB00036 to washed platelets exposed to pTF or rTF , caused a thrombin dependent irreversible platelet aggregation . Our studies demonstrate that platelets possess mechanisms to capture and incorporate TF-rich vesicles . These processes are accelerated by the presence of other cellular antigens in the vesicles . Our findings may explain the hemostatic action of DB00036 in severely hemodiluted patients , but are also relevant for the understanding of potential implications of TF-associated to platelets in the propagation of thrombus . Oxidation of alcohols and reduction of aldehydes derived from methyl- and dimethylpyrenes by cDNA-expressed human alcohol dehydrogenases . Some methylated pyrenes can form DNA adducts in rat tissues after benzylic hydroxylation and sulpho conjugation . However , oxidation of the intermediate alcohols to carboxylic acids is an important competing pathway leading to detoxification . We previously showed that co-administration of ethanol or DB01213 strongly enhances DNA adduct formation by 1-hydroxymethylpyrene , indicating an involvement of alcohol dehydrogenases ( ADHs ) in the detoxification . This mechanism may be involved in the observed synergism of smoking and alcohol consumption in certain human cancers . In a preceding study , cDNA-expressed human P00325 efficiently oxidised 1- , 2- and 4-hydroxymethylpyrene ; these reactions were inhibited in the presence of ethanol or DB01213 . Here we report that P00326 , P00326 and P08319 also show substantial activity towards these substrates and two further congeners , 1-hydroxymethyl-6-methylpyrene and 1-hydroxymethyl-8-methylpyrene . All four DB00067 forms also catalysed the reverse reaction , implying that the aldehydes have to be sequestered by other enzymes , such as aldehyde dehydrogenases , for final detoxification . P00326 and P08319 activities towards hydroxymethylpyrenes were more strongly inhibited in the presence of ethanol and DB01213 than those of P00325 . P00326 was only inhibited at very high concentrations of the modulators . In conclusions , several human ADHs are capable of detoxifying benzylic alcohols of alkylated polycyclic aromatic hydrocarbons . However , some competing substrates and inhibitors may affect all these redundant detoxification systems , although to various extents . A common coding variant in Q14790 is associated with breast cancer risk . The Breast Cancer Association Consortium ( BCAC ) has been established to conduct combined case-control analyses with augmented statistical power to try to confirm putative genetic associations with breast cancer . We genotyped nine SNPs for which there was some prior evidence of an association with breast cancer : Q14790 D302H ( rs1045485 ) , P17936 -202 C --> A ( rs2854744 ) , P04179 V16A ( rs1799725 ) , P01137 L10P ( rs1982073 ) , Q13315 S49C ( rs1800054 ) , P00325 3' UTR A --> G ( rs1042026 ) , P38936 S31R ( rs1801270 ) , Q9UMF0 V301I ( rs1056538 ) and Q14980 A794G ( rs3750913 ) . We included data from 9-15 studies , comprising 11,391-18,290 cases and 14,753-22,670 controls . We found evidence of an association with breast cancer for Q14790 D302H ( with odds ratios ( OR ) of 0.89 ( 95 % confidence interval ( c.i. ) : 0.85-0.94 ) and 0.74 ( 95 % c.i. : 0.62-0.87 ) for heterozygotes and rare homozygotes , respectively , compared with common homozygotes ; P(trend) = 1.1 x 10(-7) ) and weaker evidence for P01137 L10P ( OR = 1.07 ( 95 % c.i. : 1.02-1.13 ) and 1.16 ( 95 % c.i. : 1.08-1.25 ) , respectively ; P(trend) = 2.8 x 10(-5) ) . These results demonstrate that common breast cancer susceptibility alleles with small effects on risk can be identified , given sufficiently powerful studies . [ P08473 activity in the guinea pig model of asthma ] . P08473 exists in airway epithelial cells , smooth muscle , and submucosa near glands , and cleaves tachykinins to inactive metabolites , thereby reducing there effects . To study the role of enkephalinase in asthmatic response , we measured its activity in guinea pig model of asthma . When compared with the control values , the enkephalinase activity was reduced during in immediate asthmatic response ( Q92932 ) and late asthmatic response ( P10586 ) . Compared with the control values ( 100 % ) , each value was 79.7 % , 73.4 % in the trachea and 74.3 % , 55.7 % in the lung respectively . Tracheal muscle preparation taken from the control , Q92932 , and P10586 groups were made and mounted in oxygenated modified Krebs-Ringer solution . The response was monitored by isometric transducer . Concentration response curves to P20366 with or without phosphoramidon were obtained . The contractile responses of the P10586 groups were enhanced in potency and efficiency . DB02557 potentiated the P20366 induced contraction of control and the Q92932 groups but was less potent in enhancing the contractile response in the P10586 group , showing less enkephalinase activity in the P10586 . These results suggest that the enkephalinase plays an important role in P10586 . In P10586 , the enkephalinase activity may be inhibited and the responsiveness of the smooth muscle to some bronchoconstrictor , such as tachykinins , may be increased . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . Intracellular retention and degradation of the epidermal growth factor receptor , two distinct processes mediated by benzoquinone ansamycins . Epidermal growth factor ( P01133 ) stimulates the growth of various types of cells via its cell surface tyrosine kinase receptor . The P01133 receptor ( P01133 -R ) has an oncogenic potential when overexpressed in a wide range of tumor cells . DB02424 ( GA ) and herbimycin ( HA ) , specific inhibitors of the cytosolic chaperone P08238 and its endoplasmic reticulum homologue GRP 94 , were shown to accelerate degradation of the P01133 -R and of its homologue p185(c-)(erbB-2) . Here we compared the effects of GA and HA on intracellular degradation and maturation of P01133 -R . By using an inhibitor of proteasomal degradation , we learned that GA , but not HA , blocks processing of newly synthesized P01133 -R . The effects of GA and HA on receptor degradation are mediated by the cytosolic portion of P01133 -R and could be conferred to the erythropoietin receptor ( P19235 ) , by employing the respective chimera . Neither HA nor GA affected stability of newly synthesized P01133 -R lacking the cytosolic domain ( Ex P01133 -R ) , but GA caused intracellular retention of this mutant . Taken together , our results imply that GA has two distinct targets of action on the P01133 -R , one for promoting its degradation and another for mediating its intracellular retention . Apparently , degradation of the P01133 -R mediated by GA or HA requires the presence of the P01133 -R cytosolic domain , whereas intracellular retention in the presence of GA is coupled to the extracellular domain of the P01133 -R . Exendin-4 alleviates angiotensin II-induced senescence in vascular smooth muscle cells by inhibiting Rac1 activation via a DB02527 /PKA-dependent pathway . Vascular aging has been implicated in the progression of diabetes and age-related cardiovascular disorders . Glucagon-like peptide-1 ( P0C6A0 ) is an incretin hormone capable of cytoprotective actions in addition to its glucose-lowering effect . The present study was undertaken to examine whether Exendin-4 , a specific ligand for the P43220 , could prevent angiotensin ( P03950 ) II-induced premature senescence in vascular smooth muscle cells ( VSMCs ) and to determine the underlying mechanism involved . Senescence-associated β-galactosidase ( SA β-gal ) assay showed that P03950 II induced premature senescence of VSMCs . Pretreatment with Exendin-4 significantly attenuated P03950 II-induced generation of H2O2 and the subsequent VSMC senescence . These effects were , however , reversed in the presence of exendin fragment 9-39 , a P43220 antagonist , or PKI14-22 . Moreover , a marked increase in the levels of p53 and P38936 induced by P03950 II was blunted by the treatment with Exendin-4 . Nevertheless , Exendin-4 failed to decrease P03950 II-induced expression of NAD(P)H oxidase 1 ( Nox1 ) , NAD(P)H oxidase 4 ( Nox4 ) , O75935 (phox) , or p47(phox) in VSMCs . Mechanistically , Exendin-4 blocked P03950 II-induced Rac1 activation through the DB02527 /PKA signaling cascade . Specifically , NSC23766 , a Rac1 inhibitor , abrogated the suppressive effects of Exendin-4 on P03950 II-induced premature senescence and H2O2 generation , respectively . Thus Exendin-4 confers resistance to P03950 II-induced superoxide anion generation from NAD(P)H oxidase and the resultant VSMC senescence by inhibiting Rac1 activation via a DB02527 /PKA-dependent pathway . These findings demonstrate that P0C6A0 as well as its analogs ( P0C6A0 -related reagents ) may hold therapeutic potential in the treatment of diabetes with cardiovascular disease . Substance P activates responses correlated with tumour growth in human glioma cell lines bearing tachykinin NK1 receptors . The neuropeptide DB05875 ( SP ) , by stimulating tachykinin NK1 receptors ( P25103 ) , triggers a number of biological responses in human glioma cells which are potentially relevant for tumour growth . First , radioligand binding studies demonstrated the presence of tachykinin P25103 on SNB-19 , DBTRG-05 MG and U373 MG , but not on U138 MG and Q16653 -G-GCM human glioma cell lines . Second , application of SP or neurokinin A ( P20366 ) to P25103 + glioma cell lines increased the secretion of interleukin 6 ( P05231 ) and potentiated P05231 secretion induced by IL-1beta . SP also up-regulated the release of transforming growth factor beta1 ( TGF-beta1 ) by the U373 MG glioma cell line . Third , SP induced new DNA synthesis and enhanced the proliferation rate of P25103 + , but not of P25103 - glioma cell lines . Also , P20366 stimulated the proliferation and cytokine secretion in P25103 + glioma cell lines . All the stimulant effects of SP/ P20366 on P25103 + glioma cell lines were completely blocked by a specific tachykinin P25103 antagonist , MEN 11467 . These data support the potential use of tachykinin P25103 antagonist for controlling the proliferative rate of human gliomas .

MH_train_1231

MH_train_1231

MH_train_1231

interacts_with DB00316?

multiple_choice

P35354 induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways . The functional significance of protease-activated receptors ( PARs ) in endothelial cells is largely undefined , and the intracellular consequences of their activation are poorly understood . Here , we show that the serine protease thrombin , a P25116 -selective peptide ( TFLLRN ) , and SLIGKV ( P55085 -selective peptide ) induce cyclooxygenase-2 ( P35354 ) protein and mRNA expression in human endothelial cells without modifying P23219 expression . P35354 induction was accompanied by sustained production of 6-keto-PGF1alpha , the stable hydrolysis product of prostacyclin , and this was inhibited by indomethacin and the P35354 -selective inhibitor NS398 . P25116 and P55085 stimulation rapidly activated both P27361 /2 and p38MAPK , and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha formation . Thrombin and peptide agonists of P25116 and P55085 increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus , and this , as well as PAR-induced 6-keto-PGF1alpha synthesis , was inhibited by co-infection with adenovirus encoding wild-type or mutated ( Y42F ) P25963 . Thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG-132 . Activation of P25116 or P55085 promoted nuclear translocation and phosphorylation of p65-NF-kappaB , and thrombin-induced but not P55085 -induced p65-NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK . Activation of Q96RI0 by AYPGKF increased phosphorylation of P27361 /2 and p38MAPK without modifying NF-kappaB activation or P35354 induction . Our data show that P25116 and P55085 , but not Q96RI0 , are coupled with P35354 expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on P27361 /2 , p38MAPK , and P25963 -dependent NF-kappaB activation . DB05692 , a thrombin receptor ( P25116 ) antagonist for the prevention and treatment of atherothrombosis . DB05692 is a novel antiplatelet agent undergoing development by Schering-Plough Corp for the treatment and prevention of atherothrombosis . The compound is an orally administered himbacine analog that potently antagonizes the platelet thrombin receptor protease-activated receptor 1 ( P25116 ) , which leaves the procoagulant function of thrombin intact . In preclinical studies , DB05692 demonstrated no effect on bleed time or coagulation parameters . In both cynomolgus monkeys and humans , the compound had high bioavailability and inhibited ex vivo TRAP ( thrombin receptor-activating peptide ) -stimulated platelet aggregation in a potent and long-lasting manner . In a phase II clinical trial of patients undergoing percutaneous coronary intervention , DB05692 added to standard therapy with aspirin and clopidogrel did not increase major or minor thrombolysis in myocardial infarction bleeding , and demonstrated a trend toward decreased major adverse cardiovascular events versus placebo . At the time of publication , three phase III trials were underway to assess the efficacy and safety of DB05692 for at least 1 year in up to 35,000 patients with acute coronary syndromes or atherosclerosis . The distinct mechanism of action of DB05692 allows for cardiovascular protection without the liability of increased bleeding associated with other antiplatelet therapies . Phase III trials in high-risk patients will determine the use of DB05692 in cardiological practice . Anti- P04275 aptamer DB05202 for refractory thrombotic thrombocytopenic purpura . BACKGROUND : Plasma exchange is the main therapy for thrombotic thrombocytopenic purpura ( TTP ) . No treatments other than plasma exchange have been documented to be effective nor are approved for treatment of TTP . The anti- P04275 ( P04275 ) aptamer DB05202 effectively inhibits P04275 activity in plasma samples of TTP patients and thus shear-dependent platelet ( Q02083 ) function as measured by the Q02083 function analyzer PFA-100 ( Dade Behring ) . It was hypothesized that DB05202 would offer a potentially effective treatment option for a critically ill patient , refractory to standard care . CASE REPORT : A 39-year-old male patient with idiopathic TTP , refractory to daily plasma exchange , rituximab , steroids , and splenectomy , was additionally treated with a continuous infusion of the anti- P04275 ( P04275 ) aptamer DB05202 for 3 weeks . RESULTS : Plasma concentrations of approximately 10 microg/mL DB05202 decreased P04275 activity by more than 96 % . Plasma exchange treatment acutely decreased the plasma concentrations of DB05202 by a mean of 47 % ( range , 40 % -61 % ) . Thus , additional minibolus infusions of DB05202 were given after each plasma exchange to rapidly restore steady-state concentrations . DB05202 resulted in an increase of Q02083 counts as long as DB05202 was given . On three occasions the infusion was stopped , each accompanied by a decrease in Q02083 counts and worsening of microangiopathy . No serious adverse effects were observed during the treatment with DB05202 . CONCLUSION : DB05202 caused a clear and reproducible increase in Q02083 counts in an otherwise refractory TTP case . These clinical , pharmacokinetic , and pharmacodynamic data provide a rational basis for clinical trials with DB05202 in TTP . DB08834 prevents stress induced aggregation of proteins in vitro and promotes Q9NZJ5 activation in HepG2 cells . DB08834 ( DB08834 ) a bile salt and chemical chaperone reduces stress-induced aggregation of proteins ; activates Q9NZJ5 [ P19525 ( RNA-dependent protein kinase ) -like ER ( endoplasmic reticulum ) kinase ] or Q9NZJ5 , one of the hall marks of ER stress induced unfolded protein response ( UPR ) in human hepatoblastoma HepG2 cells ; prevents heat and dithiothreitol ( DTT ) induced aggregation of BSA ( bovine serum albumin ) , and reduces ANS ( 1-anilino-naphthalene-8-sulfonate ) bound BSA fluorescence in vitro . DB08834 inactivates heat treated , but not the native EcoR1 enzyme , and reduces heat-induced aggregation and activity of P23219 ( cyclooxygenase enzyme-1 ) in vitro . These findings suggest that DB08834 binds to the hydrophobic regions of proteins and prevents their subsequent aggregation . This may stabilize unfolded proteins that can mount UPR or facilitate their degradation through cellular degradation pathways . Arsenic reduces the antipyretic activity of paracetamol in rats : modulation of brain P35354 activity and CB₁ receptor expression . We examined whether subacute arsenic exposure can reduce paracetamol-mediated antipyretic activity by affecting P36551 pathway and cannabinoid P21554 receptor regulation . Rats were preexposed to elemental arsenic ( 4 ppm ) as sodium arsenite through drinking water for 28 days . Next day pyrexia was induced with lipopolysaccharide and paracetamol 's ( 200 mg/kg , oral ) antipyretic activity was assessed . The activities of P23219 and P35354 , the levels of PGE₂ , P01375 -α and IL-1β and expression of CB₁ receptors were assessed in brain . Arsenic inhibited paracetamol-mediated antipyretic activity . P23219 activity was not affected by any treatments . DB00316 decreased P35354 activity , levels of PGE₂ , P01375 -α and IL-1β and caused up-regulation of P21554 receptors . Arsenic caused opposite effects on these parameters . In the arsenic-preexposed rats , paracetamol-mediated effects were attenuated , while CB₁ receptor up-regulation was reversed to down-regulation . Results suggest that elevated P35354 activity and reduced CB₁ expression could be involved in the arsenic-mediated attenuation of the antipyretic activity of paracetamol . Human Q14376 . Accommodation of UDP-N-acetylglucosamine within the active site . Q14376 catalyzes the interconversion of DB03501 and UDP-glucose during normal galactose metabolism . One of the key structural features in the proposed reaction mechanism for the enzyme is the rotation of a 4'-ketopyranose intermediate within the active site pocket . Recently , the three-dimensional structure of the human enzyme with bound DB00157 and UDP-glucose was determined . Unlike that observed for the protein isolated from Escherichia coli , the human enzyme can also turn over UDP-GlcNAc to UDP-GalNAc and vice versa . Here we describe the three-dimensional structure of human epimerase complexed with DB00157 and UDP-GlcNAc . To accommodate the additional N-acetyl group at the P06681 position of the sugar , the side chain of DB00174 -207 rotates toward the interior of the protein and interacts with DB00142 -199 . Strikingly , in the human enzyme , the structural equivalent of DB00135 -299 in the E. coli protein is replaced with a cysteine residue ( DB00151 -307 ) and the active site volume for the human protein is calculated to be approximately 15 % larger than that observed for the bacterial epimerase . This combination of a larger active site cavity and amino acid residue replacement most likely accounts for the inability of the E. coli enzyme to interconvert UDP-GlcNAc and UDP-GalNAc . Vascular endothelial growth factor : a therapeutic target for tumors of the Ewing 's sarcoma family . PURPOSE : We have reported previously that intratumoral microvessel density ( P53602 ) is a significant prognostic indicator of event-free survival in the Ewing 's sarcoma family of tumors ( ESFT ) . Here , the angiogenic growth factor expression profile and its relationship with P53602 has been investigated in ESFT . EXPERIMENTAL DESIGN AND RESULTS : Using ESFT model systems , the potential of these factors as therapeutic targets has been evaluated . A significant correlation ( P = 0.02 ) was observed between vascular endothelial growth factor ( P15692 ) expression and P53602 , consistent with the hypothesis that P15692 regulates the development of microvessels in ESFT . There was no correlation between P53602 and any of the other growth factors studied . All six ESFT cell lines studied produced and secreted P15692 ; five of six cell lines also secreted placental growth factor , one cell line ( A673 ) at high levels . Tumor conditioned medium induced proliferation of human umbilical vein endothelial cells . Expression of P15692 receptors Flt-1 and Flk-1/ P35968 was heterogeneous across the cell lines . Both receptor tyrosine kinase inhibitors SU6668 ( targets Flk-1/ P35968 , platelet-derived growth factor receptor-beta , and fibroblast growth factor receptor 1 ) and SU5416 ( targets Flk-1/ P35968 ) as well as anti- P15692 agents rhuMAb- P15692 ( bevacizumab ) and DB08885 delayed s.c. growth of ESFT in mice compared with untreated groups : SU6668 ( 100 mg/kg/d ) , SU5416 ( 25 mg/kg/d ) , rhuMAb- P15692 ( 10 mg/kg twice weekly ) , and DB08885 ( 2.5 or 25 mg/kg twice weekly ) . CONCLUSIONS : These data suggest that P15692 is the single most important regulator of angiogenesis in ESFT and may be exploited for therapeutic advantage . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Lipopolysaccharide directly stimulates cortisol secretion by human adrenal cells by a cyclooxygenase-dependent mechanism . Activation of the hypothalamo-pituitary-adrenal axis by bacterial lipopolysaccharide ( LPS ; endotoxin ) is well documented , although there has been uncertainty about whether LPS exerts a direct effect at the level of the adrenal . The present study found that LPS caused a dose-dependent stimulation of basal cortisol secretion by the human adrenocortical cell line , NCI-H295R , without affecting aldosterone . The expression of both O60603 ( O60603 ) and O00206 was demonstrated in these cells , and the specific ligands for O00206 ( purified LPS and lipid A ) and O60603 ( Pam3Cys ) were found to stimulate cortisol release , suggesting that these receptors may mediate the effects of LPS in adrenal cells , as has been shown in other cell types . LPS was also found to stimulate prostaglandin E2 release by these cells . The effects of LPS on cortisol were attenuated in the presence of both indomethacin and a specific P35354 inhibitor , but not a P23219 inhibitor , suggesting an obligatory role for P35354 activation and prostaglandin synthesis in the adrenal response to LPS . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Female , but not male , mice show delayed cutaneous wound healing following aspirin administration . Cyclo-oxygenase ( P36551 ) is an enzyme that participates in the wound healing process . DB00945 , a non-steroidal anti-inflammatory drug , simultaneously inhibits the aromatase activity of P23219 and P35354 isoforms , which is needed for prostaglandin synthesis . The aim of the present study was to determine whether aspirin , and thus P36551 inhibition , distinctly affects cutaneous wound healing in female and male mice . Female and male BALB/c mice were treated with aspirin ( 25 mg/kg per day ) for 16 days until they were killed . The control group received vehicle ( saline ) only . A full-thickness excisional lesion was made on the back , 2 days after aspirin administration started , and macroscopic , histological and biochemical parameters were evaluated . Sections were stained and immunostained for microscopic analysis . P05164 ( P05164 ) activity , hydroxyproline quantity and the protein expression of P04275 ( P04275 ) and vascular endothelial growth factor ( P15692 ) were also determined . Female control and aspirin-treated groups exhibited delayed wound closure and re-epithelization compared with the male control and aspirin-treated groups , respectively . The female control group exhibited reduced P05164 activity and a decreased number of macrophage inhibitory factor-positive cells compared with the male control group . In the female aspirin-treated group , P05164 activity and the number of F4/80-positive macrophages was higher than in the control group . Collagen was reduced only in the female aspirin-treated group . The expression of P04275 and P15692 protein was increased in the female aspirin-treated group . In conclusion , aspirin administration impaired the wound healing process in BALB/c female , but not male , mice . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . Δ(9)- DB00470 acts as a partial agonist/antagonist in mice . Δ- DB00470 ( THC ) has been characterized as a partial agonist at cannabinoid P21554 receptors in vitro ; however , it often produces the same maximum effects in vivo as other cannabinoid agonists . This study was carried out to determine whether THC would antagonize the hypothermic effects of another cannabinoid agonist , AM2389 , in mice . Male mice were injected with 1-100 mg/kg THC , 0.01-0.1 mg/kg AM2389 , or a combination of 30 mg/kg THC and 0.1-1.0 mg/kg AM2389 , and rectal temperature was recorded for up to 12 h after injection . THC reduced the temperature by 5.6°C at a dose of 30 mg/kg ; further increases in the dose did not produce larger effects , indicating a plateau in the THC dose-effect function . AM2389 reduced temperature by 9.0°C at a dose of 0.1 mg/kg . One hour pretreatment with 30 mg/kg THC attenuated the hypothermic effects of 0.1 mg/kg AM2389 ; a 10-fold higher dose , 1.0 mg/kg AM2389 , was required to further decrease temperature , reflecting a five-fold rightward shift of the lower portion of the AM2389 dose-effect function following THC pretreatment . These results indicate that , in an assay of mouse hypothermia , THC exerts both agonist and antagonist effects following acute administration , and mark the first demonstration of partial agonist/antagonist effects of THC in vivo . Targeting myeloid differentiation 2 for treatment of sepsis . Sepsis continues to be a leading cause of intensive care unit ( ICU ) death . Gram-negative bacteria are among the most important pathogens of sepsis and their LPS content is regarded to be an important stimulator that elicits the systemic inflammatory reaction . MD-2 is a small secreted glycoprotein that can bind to both the hydrophobic portion of LPS and to the extracellular domain of O00206 . The interaction between MD-2 and LPS bridges the two O00206 molecules and induces the dimerization of LPS-MD-2- O00206 , which forms the structural basis for biological functions of O00206 /MD-2 complex . Due to its essential role in mediating the interaction between LPS and O00206 , MD-2 has been extensively explored as a therapeutic target for treatment of inflammatory disorders such as sepsis . DB04933 is a synthetic tetraacylated lipid A that binds directly to MD-2 and antagonizes LPS binding to the same site . Although eritoran showed positive results in phase I and phase II clinical trials of severe sepsis , a phase III clinical study for severe sepsis has failed . More effective therapeutic strategies are in need to treat this devastating clinical disorder . Atrial and brain natriuretic peptides : secretion during exercise in patients with essential hypertension and modulation by acute angiotensin-converting enzyme inhibition . 1. This study examined whether brain and atrial natriuretic peptides ( DB04899 , P01160 ) are secreted together through the coronary sinus from the heart , and whether plasma concentrations of DB04899 and P01160 were affected by ergometric exercise in patients with essential hypertension . The effects of temocapril , a potent angiotensin-converting enzyme ( P12821 ) inhibitor , on plasma concentrations of these peptides was also examined . 2 . The plasma concentrations of immunoreactive ( ir ) DB04899 and ir- P01160 in the coronary sinus in seven patients with ischaemic heart disease during cardiac catheterization were far greater than values with plasma obtained at the same time from the femoral artery . 3 . The plasma concentrations of ir- DB04899 and ir- P01160 increased with exercise and were correlated with each other . DB08836 reduced the blood pressure and slightly ( but significantly ) decreased the levels of both peptides at rest and during exercise . 4 . The results suggest that DB04899 and P01160 were secreted together through the coronary sinus from the heart . The secretion was increased by exercise and suppressed by acute P12821 inhibition . The increase in these peptides during exercise may reflect a compensatory mechanism against further elevation of blood pressure . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Cellular distribution and contribution of cyclooxygenase P35354 to diabetogenesis in NOD mouse . Unlike most other mammalian cells , beta-cells of Langerhans constitutively express cyclooxygenase ( P36551 ) -2 rather than P23219 . P35354 is also constitutively expressed in type 1 diabetes ( T1D ) patients ' periphery blood monocytes and macrophage . To understand the role of P35354 in the beta-cell , we investigated P35354 expression in beta-cells and islet infiltrates of NOD and BALB/c mice using fluorescence immunohistochemistry and cytochemical confocal microscopy and Western blotting . Immunostaining showed that P35354 is expressed in islet-infiltrating macrophages , and that the expression of insulin and P35354 disappeared concomitantly from the beta-cells when NOD mice progressed toward overt diabetes . Also cultured P01308 -1E cells coexpressed insulin and P35354 but clearly in different subcellular compartments . Treatment with celecoxib increased insulin release from these cells in a dose-dependent manner in glucose concentrations ranging from 5 to 17 mM . Excessive P35354 expression by the islet-infiltrating macrophages may contribute to the beta-cell death during insulitis . The effects of celecoxib on P01308 -1E cells suggest that PGE(2) and other downstream products of P35354 may contribute to the regulation of insulin release from the beta-cells . Perinuclear localization of cytosolic phospholipase A(2)alpha is important but not obligatory for coupling with cyclooxygenases . In response to Ca(2+) signaling , cytosolic phospholipase A(2)alpha ( cPLA(2)alpha ) translocates from the cytosol to the perinuclear membrane , where downstream eicosanoid-synthetic enzymes , such as cyclooxygenase ( P36551 ) , are localized . Although the spatiotemporal perinuclear colocalization of cPLA(2)alpha and COXs has been proposed to be critical for their functional coupling leading to prostanoid production , definitive evidence for this paradigm has remained elusive . To circumstantiate this issue , we took advantage of a chimeric cPLA(2)alpha mutant harboring the P06681 domain of protein kinase Calpha , which translocates to the plasma membrane following cell activation . Transfection analyses of the native or chimeric cPLA(2)alpha in combination with P23219 or P35354 revealed that , even though the arachidonate-releasing capacities of native and mutant cPLA(2)alpha were comparable , prostaglandin production by mutant cPLA(2)alpha was markedly impaired as compared with that by native cPLA(2)alpha . We thus conclude that the perinuclear localization of cPLA(2)alpha is preferential , even if not obligatory , for efficient coupling with COXs . c-Src modulates estrogen-induced stress and apoptosis in estrogen-deprived breast cancer cells . The emergence of anti-estrogen resistance in breast cancer is an important clinical phenomenon affecting long-term survival in this disease . Identifying factors that convey cell survival in this setting may guide improvements in treatment . Estrogen ( E2 ) can induce apoptosis in breast cancer cells that have been selected for survival after E2 deprivation for long periods ( MCF-7:5C cells ) , but the mechanisms underlying E2-induced stress in this setting have not been elucidated . Here , we report that the c-Src kinase functions as a key adapter protein for the estrogen receptor ( ER , P03372 ) in its activation of stress responses induced by E2 in MCF-7:5C cells . E2 elevated phosphorylation of c-Src , which was blocked by 4-hydroxytamoxifen ( DB04468 ) , suggesting that E2 activated c-Src through the ER . We found that E2 activated the sensors of the unfolded protein response ( UPR ) , IRE1α ( O75460 ) and Q9NZJ5 kinase ( Q9NZJ5 ) , the latter of which phosphorylates eukaryotic translation initiation factor-2α ( eIF2α ) . E2 also dramatically increased reactive oxygen species production and upregulated expression of heme oxygenase P09601 ( P09601 ) , an indicator of oxidative stress , along with the central energy sensor kinase AMPK ( P54646 ) . Pharmacologic or RNA interference-mediated inhibition of c-Src abolished the phosphorylation of eIF2α and AMPK , blocked E2-induced ROS production , and inhibited E2-induced apoptosis . Together , our results establish that c-Src kinase mediates stresses generated by E2 in long-term E2-deprived cells that trigger apoptosis . This work offers a mechanistic rationale for a new approach in the treatment of endocrine-resistant breast cancer .

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interacts_with DB00707?

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DB00174 synthetase : regulation by cell stress and involvement in tumor biology . DB00174 synthetase ( P08243 ) catalyzes the conversion of aspartate and glutamine to asparagine and glutamate in an DB00171 -dependent reaction . The enzyme is ubiquitous in its organ distribution in mammals , but basal expression is relatively low in tissues other than the exocrine pancreas . Human P08243 activity is highly regulated in response to cell stress , primarily by increased transcription from a single gene located on chromosome 7 . Among the genomic elements that control P08243 transcription is the C/EBP- P39905 response element ( CARE ) within the promoter . Protein limitation or an imbalanced dietary amino acid composition activate the P08243 gene through the amino acid response ( AAR ) , a process that is replicated in cell culture through limitation for any single essential amino acid . Endoplasmic reticulum stress also increases P08243 transcription through the Q9NZJ5 -eIF2- P18848 arm of the unfolded protein response ( UPR ) . Both the AAR and UPR lead to increased synthesis of P18848 , which binds to the CARE and induces P08243 transcription . Elevated expression of P08243 protein is associated with resistance to asparaginase therapy in childhood acute lymphoblastic leukemia and may be a predictive factor in drug sensitivity for certain solid tumors as well . Activation of the Q9P2K8 -eIF2- P18848 signaling pathway , leading to increased P08243 expression appears to be a component of solid tumor adaptation to nutrient deprivation and/or hypoxia . Identifying the roles of P08243 in fetal development , tissue differentiation , and tumor growth may reveal that P08243 function extends beyond asparagine biosynthesis . Quantification and modeling of tripartite P06729 - , CD58FC chimera (alefacept)- , and CD16-mediated cell adhesion . DB00092 is a chimeric protein combining P19256 immunoglobulin-like domain 1 with human IgG1 Fc . DB00092 mediates adhesion by bridging P06729 on T cells to activating Fc receptors on effector cells , but the equilibrium binding parameters have not been determined . DB00092 mediated T cell killing by NK cells and adhesion between P06729 - and CD16-expressing cells at an optimum concentration of 100 nM . We introduce novel measurements with supported planer bilayers , from which key two-dimensional and three-dimensional parameters can be determined by data fitting . DB00092 competitively inhibited cell bilayer adhesion mediated by the P06729 - P19256 interaction . DB00092 mediated maximal adhesion of P06729 (+) T cells to O75015 , an Fc receptor , in planar bilayers at 500 nM . A mechanistic model for alefacept-mediated cell-bilayer adhesion allowed fitting of the data and determination of two-dimensional binding parameters . These included the density of bonds in the adhesion area , which grew to maintain a consistent average bond density of 200 molecules/microm(2) and two-dimensional association constants of 3.1 and 630 microm(2) for bivalently and monovalently bound forms of alefacept , respectively . The maximum number of CD16 bound and the fit value of 4,350 P06729 per cell are much lower than the 40,000 P06729 per cell measured with anti- P06729 Fab . These results suggest that additional information is needed to correctly predict DB00092 -mediated bridge formation . P33681 -1/ P33681 -2 costimulation regulates plaque antigen-specific T-cell responses and atherogenesis in low-density lipoprotein receptor-deficient mice . BACKGROUND : Several lines of evidence indicate that T-cell responses influence the progression of atherosclerotic disease . Interferon-gamma ( P01579 ) -producing T cells specific for lesional antigens , including oxidized LDLs and heat shock protein 60 ( HSP60 ) , may promote lesion development as well as plaque instability . P33681 -1 and P33681 -2 are closely related molecules expressed on antigen-presenting cells that provide costimulatory signals for T-cell activation . This study tested the hypothesis that the ability of T cells to influence atherosclerosis depends on P33681 -1/ P33681 -2 costimulation . METHODS AND RESULTS : P33681 -1/ P33681 -2/ P01130 ( P01130 ) -deficient mice and P01130 -deficient control mice were fed a 1.25 % cholesterol or control diet for 8 and 20 weeks . Total serum cholesterol levels and extent and phenotype of atherosclerosis were analyzed . Splenic and lymph node P01730 + T cells from the animals were cultured with mouse recombinant HSP60 or media and antigen-presenting cells and analyzed for P01579 and interleukin-4 production . The absence of P33681 -1 and P33681 -2 significantly reduced early cholesterol diet-induced atherosclerotic lesion development in P01130 -deficient mice compared with P33681 -1/ P33681 -2-expressing control mice . Furthermore , P01730 + T cells from the cholesterol-fed P33681 -deficient mice secreted a significantly lower amount of P01579 in response to mouse HSP60 in vitro than did T cells from P33681 -expressing control mice . CONCLUSIONS : The data show that P33681 -1 and P33681 -2 regulated the development of atherosclerotic lesions and the priming of lesional antigen-specific T cells . This study highlights the P33681 - P10747 pathway as a potentially important target for immunomodulation of atherosclerosis . [ Anti-cholesterol agents , new therapeutic approaches ] . Statins and fibrates constitute the two major families of lipid-lowering agents . Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia . Both drugs are also used for the treatment of mixed dyslipidemia . Some fibrates efficiently lower serum LDL-cholesterol . Statins inhibit P04035 and decrease cellular cholesterol synthesis . The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the P01130 gene . This over expression of the P01130 in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels . The effects of fibrates on lipid metabolism are entirely due to their capacity to activate Q07869 and to induce the over expression of genes containing a PPRE in their promoter . Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis . Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription , respectively . Fibrates could decrease triglycerides partly by inducing apo A-V over-expression . These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription . Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism . Chronic delivery of a thrombospondin-1 mimetic decreases skeletal muscle capillarity in mice . Angiogenesis is an essential process for normal skeletal muscle function . There is a growing body of evidence suggesting that thrombospondin-1 ( P07996 -1 ) , a potent antiangiogenic protein in tumorigenesis , is an important regulator of both physiological and pathological skeletal muscle angiogenesis . We tested the hypothesis that chronic exposure to a P07996 -1 mimetic ( DB05434 ) , which targets the P16671 P07996 -1 receptor , would decrease skeletal muscle capillarity as well as alter the balance between positive and negative angiogenic proteins under basal conditions . Osmotic minipumps with either DB05434 or vehicle ( 5 % dextrose ) were implanted subcutaneously in the subscapular region of C57/BL6 mice for 14 days . When compared to the vehicle treated mice , the DB05434 group had a 20 % decrease in capillarity in the superficial region of the gastrocnemius ( GA ) , 11 % decrease in the plantaris ( Q02083 ) , and a 35 % decrease in the soleus ( SOL ) . DB05434 also decreased muscle protein expression of vascular endothelial growth factor ( P15692 ) in both the GA ( -140 % ) and SOL ( -62 % ) ; however there was no change in P15692 in the Q02083 . Serum P15692 was not altered in DB05434 treated animals . Endogenous P07996 -1 protein expression in all muscles remained unaltered . Tunnel staining revealed no difference in muscle apoptosis between DB05434 and vehicle treated groups . These data provide evidence that the anti-angiogenic effects of P07996 -1 are mediated , at least in part , via the P16671 receptor . It also suggests that under physiologic conditions the P07996 -1/ P16671 axis plays a role in regulating basal skeletal muscle microvessel density . The cytoplasmic domain of the low density lipoprotein ( LDL ) receptor-related protein , but not that of the P01130 , triggers phagocytosis . The macrophage P01130 and P01130 -related protein ( Q14764 , CD91 ) mediate the phagocytic-like uptake of atherogenic lipoproteins and apoptotic cells , yet the structural basis of their phagocytic functions is not known . To address this issue , we transfected macrophages with chimeric proteins containing the cytoplasmic tails and transmembrane regions of the P01130 or Q14764 and the ectodomain of P06729 , which can bind non-opsonized sheep red blood cells ( SRBCs ) . Macrophages expressing receptors containing the P01130 domains were able to bind but not internalize SRBCs . In contrast , macrophages expressing receptors containing the cytoplasmic tail of Q14764 were able to bind and internalize SRBCs . Chimeras in which the Q14764 cytoplasmic tail was mutated in two di-leucine motifs and a tyrosine in an NPXYXXL motif were able to endocytose anti- P06729 antibody and bind SRBCs , but SRBC phagocytosis was decreased by 70 % . Thus , the phagocytic-like functions of Q14764 , but not those of the P01130 , can be explained by the ability of the Q14764 cytoplasmic tail to trigger phagocytosis . These findings have important implications for atherogenesis and apoptotic cell clearance and for a fundamental cell biological understanding of how the P01130 and Q14764 function in internalization processes . [ Pharma-clinics. The drug of the month. DB00533 ( Vioxx ) ] . DB00533 ( Vioxx , Merck Sharp & Dohme ) is a potent and selective inhibitor of the P35354 isoform of cyclooxygenase which is used as a nonsteroidal anti-inflammatory drug ( NSAID ) . It is indicated in the symptomatic relief of pain due to osteoarthritis . The initial oral dosage of rofecoxib is 12.5 mg once daily in adults , and this dose may be increased up to a maximal dosage of 25 mg once daily if necessary . Its clinical efficacy seems to be similar to that of other NSAIDs at maximal recommended dosages , but its safety profile , especially gastrointestinal tolerance , is much better because of the P35354 selectivity . Ongoing clinical trials are performed in patients with rheumatoid arthritis . Retention of mutant low density lipoprotein receptor in endoplasmic reticulum ( ER ) leads to ER stress . Familial hypercholesterolemia is an autosomal dominant disease caused by mutations in the gene encoding the low density lipoprotein receptor ( P01130 ) . More than 50 % of these mutations lead to receptor proteins that are completely or partly retained in the endoplasmic reticulum ( ER ) . The mechanisms involved in the intracellular processing and retention of mutant P01130 are poorly understood . In the present study we show that the G544V mutant P01130 associates with the chaperones Grp78 , Grp94 , P13667 , and calnexin in the ER of transfected Chinese hamster ovary cells . Retention of the mutant P01130 was shown to cause ER stress and activation of the unfolded protein response . We observed a marked increase in the activity of two ER stress sensors , O75460 and Q9NZJ5 . These results show that retention of mutant P01130 in ER induces cellular responses , which might be important for the clinical outcome of familial hypercholesterolemia . Glioma cell activation by Alzheimer 's peptide Abeta1-42 , alpha1-antichymotrypsin , and their mixture . We compared the effects ofAlzheimer 's peptide ( Abeta1-42 ) , a,-antichymotrypsin ( ACT ) and an ACT/Abeta1-42 mixture on human glioma DK-MG cells . The solution of Abeta ( 5 microM ) formed by 2-h incubation at room temperature induced tumour necrosis factor-alpha ( P01375 ) and interleukin ( IL ) -6 levels by 55 and 45 % , respectively , and increased gelatinase B activity by 67 % , while exposure of cells to the ACT/Abeta1-42 mixture ( 1:10 molar ratio ACT : Abeta1-42 ) under the same experimental conditions showed no effect on P05231 levels or gelatinase B activity , but strongly induced P01375 ( by 190 % ) , compared to the controls . Stimulation of the cells with Abeta1-42 alone , but not with ACT , increased by about 20 % low-density lipoprotein ( LDL ) uptake and mRNA levels for P01130 and P04035 , while the ACT/Abeta1-42 mixture significantly increased LDL uptake ( by 50 % ) , up-regulated mRNA levels for P01130 and P04035 by 48 and 63 % , respectively , and increased lipid accumulation by about 20-fold . These data suggest a possible new role for Abeta in Alzheimer 's disease through its interaction with the inflammatory reactant , ACT . Q14790 and RIP kinases regulate bacteria-induced innate immune responses and cell death . A number of pathogens cause host cell death upon infection , and Yersinia pestis , infamous for its role in large pandemics such as the " Black Death " in medieval Europe , induces considerable cytotoxicity . The rapid killing of macrophages induced by Y. pestis , dependent upon type III secretion system effector Yersinia outer protein J ( YopJ ) , is minimally affected by the absence of caspase-1 , caspase-11 , P48023 , and P01375 . Q14790 is known to mediate apoptotic death in response to infection with several viruses and to regulate programmed necrosis ( necroptosis ) , but its role in bacterially induced cell death is poorly understood . Here we provide genetic evidence for a receptor-interacting protein ( RIP ) kinase-caspase-8-dependent macrophage apoptotic death pathway after infection with Y. pestis , influenced by O00206 -TIR-domain-containing adapter-inducing interferon-β ( O00206 - Q8IUC6 ) . Interestingly , macrophages lacking either Q13546 , or caspase-8 and RIP3 , also had reduced infection-induced production of IL-1β , Q14116 , P01375 , and P05231 ; impaired activation of the transcription factor NF-κB ; and greatly compromised caspase-1 processing . Cleavage of the proform of caspase-1 is associated with triggering inflammasome activity , which leads to the maturation of IL-1β and Q14116 , cytokines important to host responses against Y. pestis and many other infectious agents . Our results identify a Q13546 -caspase-8/RIP3-dependent caspase-1 activation pathway after Y. pestis challenge . Mice defective in caspase-8 and RIP3 were also highly susceptible to infection and displayed reduced proinflammatory cytokines and myeloid cell death . We propose that caspase-8 and the RIP kinases are key regulators of macrophage cell death , NF-κB and inflammasome activation , and host resistance after Y. pestis infection . Synthesis and biological evaluation of novel oxindole-based RTK inhibitors as anti-cancer agents . Given that receptor tyrosine kinases ( RTKs ) have emerged as key regulators of all aspects of cancer development , including proliferation , invasion , angiogenesis and metastasis , the RTK family represents an important therapeutic target for anti-cancer drug development . Oxindole structure has been used in RTK inhibitors such as DB02058 and intedanib . In this study , two series of new heterocyclic compounds containing oxindole scaffold have been designed and synthesized , and their inhibitory activity against the proliferation of nine cancer cell lines has been evaluated . Among them , compounds 9a and 9b displayed the strongest anti-proliferative activity with the IC50s below 10μM . Flow cytometric analysis showed that the compounds 9a and 9b dose-dependently arrested the cell cycle at G0/ P55008 phase . Although the leading compounds DB02058 and intedanib targets P11362 , the kinase activity test revealed that these compounds only showed slight inhibitory activity on P11362 kinase . Further enzymatic test aided by molecular docking simulation in the DB00171 -binding site demonstrated that 9a and 9b are potent inhibitors of c-Kit kinase . These compounds are worthy of further evaluation as anticancer agents . P43220 agonists for type 2 diabetes mellitus : recent developments and emerging agents . More than 26 million people in the United States have type 2 diabetes mellitus ( T2D ) . Many treatment options exist , but achieving long-term glycemic control in patients with T2D remains challenging . The glucagon-like peptide-1 receptor agonists ( P0C6A0 RAs ) offer a treatment option that improves glycemic control and reduces weight , with a low risk of hypoglycemia . They have emerged as attractive options for the treatment of T2D , and significant advances and developments continue to be published regarding these agents . To identify relevant literature on emerging issues related to P0C6A0 RAs , a search of the MEDLINE database was performed . Studies published in English evaluating the safety and efficacy of P0C6A0 RAs were analyzed . Because of their advantages and unique mechanism of action , P0C6A0 RAs are currently being studied in new clinical areas , including in combination with basal insulin , as adjunctive therapy in type 1 diabetes , and for weight loss . In addition , there are several emerging agents in development . DB09265 is a once-daily P0C6A0 RA that targets postprandial glucose and may be most useful when added to basal insulin as an alternative to rapid-acting insulin . DB09043 and dulaglutide are once-weekly P0C6A0 RAs that may offer more convenient dosing . The most common adverse effects of all P0C6A0 RA agents are gastrointestinal ( e.g. , nausea , diarrhea , and vomiting ) , but the rates of occurrence vary among agents . Due to the differences in pharmacokinetics , efficacy , rates of adverse effects , and administration requirements within the P0C6A0 RA class , each agent should be evaluated independently . The future of P0C6A0 RAs offers broader treatment options for T2D as well as potential in other treatment areas . The anti- P33681 primatized monoclonal antibody , galiximab , is well-tolerated but has limited activity in relapsed Hodgkin lymphoma : Cancer and Leukemia Group B 50602 ( Alliance ) . Relapsed Hodgkin lymphoma remains a clinical challenge , with few non-cytotoxic treatment options . P33681 is a surface antigen that normally functions as a co-stimulatory molecule but is aberrantly and uniformly expressed on Reed-Sternberg cells . DB04901 is a primatized monoclonal antibody against P33681 , with a favorable toxicity profile demonstrated in other lymphomas . Cancer and Leukemia Group B ( CALGB ) 50602 ( Alliance ) tested single-agent galiximab in a highly refractory group of patients with Hodgkin lymphoma ( median 3 prior regimens , 83 % failing after prior stem cell transplant ) to determine the efficacy . The overall response rate was 10.3 % and the median progression-free survival was 1.6 months . DB04901 was well-tolerated , with minimal grade 3 or 4 toxicities . Despite this preclinical rationale , single-agent galiximab had limited activity in heavily pretreated Hodgkin lymphoma . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Meloxicam . Meloxicam ( DB00814 trade mark , Boehringer Ingelheim ) is a relatively new oral non-steroidal anti-inflammatory drug ( NSAID ) approved for the treatment of osteoarthritis in the US . It has also been evaluated for the treatment of rheumatoid arthritis , ankylosing spondylitis and acute ' rheumatic ' pain . Meloxicam has been shown to be P35354 preferential , particularly at its lowest therapeutic dose , and is anti-inflammatory by inhibiting prostanoid synthesis in inflammatory cells . Since it is P35354 preferential , it would be expected to have less gastrointestinal toxicity than non-selective NSAIDs . In clinical trials of meloxicam in osteoarthritis , it was found to be as effective as piroxicam , diclofenac and naproxen with less clinical gastrointestinal symptoms and less perforations , obstructions and bleeds by meta-analysis . Adverse events , including peripheral oedema and hypertension , occurred at a similar rate as with traditional NSAIDs . Zerumbone suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells . Unregulated uptake of oxidized low-density lipoproteins ( ox-LDL ) via macrophage scavenger receptors ( SRs ) , such as lectin-like ox- P01130 -1 ( P78380 ) , is a key event in atherosclerosis . In the present study , we used differentiated Caco-2 cells as a model of the human small intestine to evaluate the suppressive effects of 16 traditional food items selected from Okinawa on 12-O-tetradecanoylphorbol-13-acetate ( TPA ) -induced P78380 mRNA expression in THP-1 human monocyte-like cells . Three Zingiberaceae plants , Curcuma aromatica Salisbury , Curcuma longa L. , and Zingiber zerumbet Smith , markedly suppressed that expression . When added to the apical sides of Caco-2 monolayers , zerumbone , a sesquiterpene from Z. zerumbet Smith , was found to permeate into the basolateral medium as an intact structure in a time-dependent manner . alpha-Humulene , a structural analog of zerumbone lacking the alpha,beta-unsaturated carbonyl group , did not suppress P78380 mRNA expression , indicating that its electrophilic moiety might play pivotal roles in its activities . Further , zerumbone attenuated the expression of SR-A , Q9H2A7 , and P16671 , but not that of P34810 or Q8WTV0 , leading to a blockade of DiI-acLDL uptake , while it also inhibited the transcriptional activities of activator protein-1 and nuclear factor-kappaB . Together , our results indicate that zerumbone is a potential phytochemical for regulating atherosclerosis with reasonable action mechanisms . P35354 promotes early atherosclerotic lesion formation in ApoE-deficient and C57BL/6 mice . Cyclooxygenase ( P36551 ) 2 is expressed in atherosclerotic lesions . We have previously reported that selective inhibition of P35354 reduces early atherosclerosis in P01130 deficient mice . To examine the role of P35354 in atherosclerosis in other mouse models , we studied the effects of selective P35354 inhibition ( by rofecoxib and NS-398 ) and nonselective P36551 inhibition ( by indomethacin ) on early atherosclerotic lesion formation in apolipoprotein E-deficient ( apoE(-/-) ) mice . Selective P35354 and nonselective P36551 inhibition reduced atherosclerosis in female apoE(-/-) mice by 35-38 % and 38-51 % in the proximal and en face aortas , respectively . Next we investigated the role of macrophage P35354 by transplanting P35354 (-/-) fetal liver cells into C57BL/6 mice and challenging the mice with an atherogenic diet . Genetic deletion of P35354 from hematopoietic cells reduced atherosclerosis by 51 % . In addition , LPS activated P35354 (-/-) macrophages had decreased expression of monocyte chemoattractant protein-1 ( P13500 ) and tumor necrosis factor-alpha ( TNFalpha ) . The results demonstrate that selective inhibition of P35354 and elimination of P35354 from macrophages significantly reduces early atherosclerotic lesion formation in apoE-deficient and C57BL/6 mice . These results are compatible with P35354 expression by macrophages having a proatherogenic role , and support the potential of anti-inflammatory therapeutic approaches for atherosclerosis . Mutational analysis of the mitochondrial P47985 of Saccharomyces cerevisiae . II . Biochemical characterization of temperature-sensitive Q13546 - mutations . Although the function of the P47985 is generally understood , little is known of how the structure of this protein supports its mechanistic role in electron transfer in the cytochrome bc1 complex . To better understand the structural basis of iron-sulfur protein function , we have undertaken a mutational analysis of the gene encoding this protein and initially isolated five temperature-sensitive iron-sulfur protein mutants ( Beckmann , J. D. , Ljungdahl , P. O. , and Trumpower , B. L. ( 1989 ) J. Biol. Chem. 264 , 3713-3722 ) . Each of the five ts-rip1- mutants exhibited pleiotropic effects . Although the mutant iron-sulfur proteins manifest several in vitro phenotypes in common , each exhibited unique characteristics . All of the ts-rip1- mutations resulted in membranes with decreased ubiquinol-cytochrome c oxidoreductase activities and decreased thermostability compared to membranes containing wild type iron-sulfur protein . All of the mutations conferred slight but significant resistance to the respiratory inhibitor myxothiazol , and one mutant was hypersensitive to inhibition by DB04799 , a structural analog of ubiquinone . In addition , one of the mutations completely blocks post-translational processing of the iron-sulfur protein , leading to accumulation of pre-iron-sulfur protein in mitochondrial membranes at nonpermissive temperatures . Finally , a mutation 12-amino acid residues away from the carboxyl terminus ( 203S ) results in an extremely unstable protein . This region of the protein may be essential in blocking degradation of pre-iron-sulfur protein by cytoplasmic proteases as the protein is imported into the mitochondria , or may be a " degradation signal , " which tags the iron-sulfur protein for turnover . Rosiglitazone is effective to improve renal damage in type-1-like diabetic rats . A marked decrease of klotho expression was observed in the kidney of streptozotocin-induced diabetic rats ( Q11206 rats ) showing diabetic nephropathy . It has been documented that klotho is the target gene of Q07869 γ . However , the effect of Q07869 γ agonist on klotho expression in kidney of Q11206 rats remains obscure . Thus , we used rosiglitazone ( TZD ) as Q07869 γ agonist to investigate the effect on renal dysfunction in Q11206 rats . Treatment of TZD reversed the lower levels of Q07869 γ , klotho , and P11362 expressions in kidneys of Q11206 rats without the correction of hyperglycemia . Also , renal functions and structural defeats were improved by TZD treatment . Taken together , oral administration of TZD may improve Q11206 -induced diabetic nephropathy due to restoration of the expression of klotho axis through an increase in Q07869 γ expression without changing blood glucose in rats . Effect of the combination of metformin and fenofibrate on glucose homeostasis in diabetic Goto-Kakizaki rats . Metformin has been reported to increase the expression of the glucagon-like peptide-1 ( P0C6A0 ) receptor in pancreatic beta cells in a peroxisome proliferator-activated receptor ( Q07869 ) -α-dependent manner . We investigated whether a PPARα agonist , fenofibrate , exhibits an additive or synergistic effect on glucose metabolism , independent of its lipid-lowering effect , when added to metformin . Non-obese diabetic Goto-Kakizaki ( GK ) rats were divided into four groups and treated for 28 days with metformin , fenofibrate , metformin plus fenofibrate or vehicle . The random blood glucose levels , body weights , food intake and serum lipid profiles were not significantly different among the groups . After 4 weeks , metformin , but not fenofibrate , markedly reduced the blood glucose levels during oral glucose tolerance tests , and this effect was attenuated by adding fenofibrate . Metformin increased the expression of the P43220 in pancreatic islets , whereas fenofibrate did not . During the intraperitoneal glucose tolerance tests with the injection of a P0C6A0 analog , metformin and/or fenofibrate did not alter the insulin secretory responses . In conclusion , fenofibrate did not confer any beneficial effect on glucose homeostasis but reduced metformin 's glucose-lowering activity in GK rats , thus discouraging the addition of fenofibrate to metformin to improve glycemic control .

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interacts_with DB01259?

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Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . [ Downregulation of P04626 by adenovirus-mediated RNA interference and its inhibitory effect on growth of SKBR3 breast cancer cell ] . AIM : To explore the possibility of RNA interference ( RNAi ) -based gene therapy against P04626 -overexpressing tumors using adenovirus-mediated vector . METHODS : A plasmid named pHER2-GFP containing P04626 and green fluorescent protein ( GFP ) fusion was constructed and cotransfected into CHO- P04264 cells respectively with nine small interference RNA ( siRNA ) -expressing plasmids targeting different regions of P04626 . The siRNA-expressing plasmids with best interference effect were screened out and then used to identify the gene silence effect in P04626 -overexpressing SKBR3 breast cancer cells . Subsequently , the siRNA-expressing cassettes were subcloned into adenoviral vectors . Downregulation of P04626 by adenovirus-mediated RNAi and its effect on SKBR3 cell proliferation were identified again . RESULTS : Two siRNA-expressing plasmids with best interference effect were screened out and P04626 was also efficiently downregulated in SKBR3 cells infected with the adenovirus containing these siRNA-expressing cassettes . Downregulation of P04626 resulted in the increase of cells in P55008 phase and the induction of apoptosis . Furthermore , infection of adenovirus inhibited SKBR3 cell growth , which was confirmed by MTT and cell long-term proliferation assays . CONCLUSION : The adenovirus-mediated RNAi could downregulate the P04626 expression efficiently and exert an inhibitory effect on growth of P04626 -overexpressing breast cancer cell . Targeted chemotherapy for triple-negative breast cancers via P01148 receptor . Triple-negative breast cancer does not express estrogen and progesterone receptors and there is no overexpression/amplification of the P04626 -neu gene . Therefore , this subtype of breast cancer lacks the benefits of specific therapies which target these receptors . About 60 % of all human breast cancers express receptors for luteinizing hormone releasing hormone ( P01148 , DB00644 ) , which might be used as a target . The P01148 receptor can be used for targeted chemotherapy with cytotoxic luteinizing hormone releasing hormone agonists such as AEZS-108 ( AN-152 ) , in which doxorubicin is linked to [D-Lys6] P01148 . In the present study we have analyzed by in vitro and in vivo experiments whether the cytotoxic P01148 agonist AEZS-108 ( AN-152 ) induces apoptosis in triple-negative human breast cancer cells that express P01148 receptors . P01148 receptor expression in tumor biopsy specimens of triple-negative breast cancers was tested using immunohistochemistry . Cell proliferation was analyzed using alamar blue proliferation assay . Induction of apoptosis was quantified by measurement of loss of mitochondrial membrane potential . In vivo experiments were performed using nude mice bearing xenografted human breast tumors.Thirty-one of 42 triple-negative breast cancers ( 73.8 % ) expressed P01148 receptors . We could show that treatment of triple-negative but P01148 -positive MDA-MB-231 , HCC1806 and HCC1937 human breast cancer cells with AEZS-108 ( AN-152 ) resulted in apoptotic cell death in vitro via activation of caspase-3 . The antitumor effects were confirmed in nude mice . AEZS-108 ( AN-152 ) inhibited the growth of xenotransplants of triple-negative human breast cancers in nude mice completely , without any apparent side effects . The cytotoxic P01148 agonist AEZS-108 ( AN-152 ) seems to be a suitable drug for an efficacious therapy for triple-negative breast cancers with little toxicity . Effect of p53 activity on the sensitivity of human glioblastoma cells to P09874 inhibitor in combination with topoisomerase I inhibitor or radiation . Poly ( ADP- DB01936 ) polymerase-1 ( P09874 ) is involved in the DNA repairing system by sensing and signaling the presence of DNA damage . Inhibition of P09874 is tested in combination with DNA damaging agents such as topoisomerase I inhibitors or ionizing radiations ( RT ) for the treatment of glioblastoma ( GBM ) . Disruption of p53 , widely prevalent in GBMs , plays a major role in DNA repairing system . The current study investigates whether p53 activity has an effect on the sensitivity of human GBM cells to P09874 inhibitors in combination with topoisomerase I inhibitor topotecan ( TPT ) and/or RT . Human GBM cell lines carrying a different functional status of p53 were treated with P09874 inhibitor DB02690 , in combination with TPT and/or RT . Cytotoxic effects were examined by analyzing the antiproliferative activity , the cell cycle perturbations , and the DNA damage induced by combined treatments . PARP inhibition enhanced the antiproliferative activity , the cell cycle perturbations and the DNA damage induced by both TPT or RT in GBM cells . These effects were influenced by the p53 activity : cells carrying an active p53 were more sensitive to the combination of PARP inhibitor and RT , while cells carrying an inactive p53 displayed a higher sensitivity to the combination of PARP inhibitor and TPT . Our study suggests that p53 activity influences the differential sensitivity of GBM cells to combined treatments of TPT , RT , and PARP inhibitors . © 2014 International Society for Advancement of Cytometry . DNA amplification is a ubiquitous mechanism of oncogene activation in lung and other cancers . Chromosomal translocation is the best-characterized genetic mechanism for oncogene activation . However , there are documented examples of activation by alternate mechanisms , for example gene dosage increase , though its prevalence is unclear . Here , we answered the fundamental question of the contribution of DNA amplification as a molecular mechanism driving oncogenesis . Comparing 104 cancer lines representing diverse tissue origins identified genes residing in amplification ' hotspots ' and discovered an unexpected frequency of genes activated by this mechanism . The 3431 amplicons identified represent approximately 10 per hematological and approximately 36 per epithelial cancer genome . Many recurrently amplified oncogenes were previously known to be activated only by disease-specific translocations . The 135 hotspots identified contain 538 unique genes and are enriched for proliferation , apoptosis and linage-dependency genes , reflecting functions advantageous to tumor growth . Integrating gene dosage with expression data validated the downstream impact of the novel amplification events in both cell lines and clinical samples . For example , multiple downstream components of the P00533 -family-signaling pathway , including Q00535 , P31749 and P29353 , are overexpressed as a direct result of gene amplification in lung cancer . Our findings suggest that amplification is far more common a mechanism of oncogene activation than previously believed and that specific regions of the genome are hotspots of amplification . DB06403 for pulmonary arterial hypertension . Endothelin receptor antagonists ( ERAs ) are an important class of agents used for the treatment of pulmonary arterial hypertension ( PAH ) . DB06403 is an oral , once-daily , endothelin type-A receptor ( P25101 ) -selective , propanoic acid class ERA under clinical investigation for the treatment of PAH . In a Phase II study , ambrisentan improved 6-minute walk distance , Borg dyspnea index , World Health Organization Functional Class , and hemodynamics . DB06403 was well tolerated and adverse events were not dose related , including a low incidence and severity of liver function test abnormalities . There are no relevant interactions between ambrisentan and cytochrome P450 isoenzymes ( metabolism , induction or inhibition ) that might alter the activity of P450-metabolized drugs . Potential benefits of ambrisentan include oral , once-daily dosing , ET(A)-receptor selectivity , and the decreased risks of liver toxicity and adverse drug-drug interactions compared with other ERAs . Q92847 agonist ( DB05657 ) accelerates gastric emptying in adults with diabetes and symptomatic gastroparesis . BACKGROUND : DB05657 is a synthetic , selective ghrelin agonist in development for gastroparesis . AIM : To assess safety and effects of DB05657 in diabetes patients with symptomatic gastroparesis . METHODS : Adults with type 1 or type 2 diabetes mellitus received placebo and DB05657 ( 80 , 160 , 320 or 600 microg/kg ) infusions in a cross-over manner following a radiolabelled meal . Blood glucose levels were stabilized using a hyperinsulinemic-euglycemic clamp . Primary endpoints were gastric half emptying and latency times . Secondary measures included assessment of gastroparesis symptoms and endocrine responses . RESULTS : Ten patients with type 1 ( n = 7 ) or 2 ( n = 3 ) diabetes , moderate-to-severe gastroparesis symptoms and > or =29 % retention 4 h after a radiolabelled solid meal were enrolled . DB05657 produced significant reductions in solid meal half-emptying ( 20 % , P = 0.043 ) and latency ( 34 % , P = 0.037 ) times vs. placebo . Reductions in overall postmeal symptom intensity ( 24 % ) and postprandial fullness ( 37 % ) following DB05657 infusion were not statistically significant . Most adverse events were mild and self-limiting and there were no identifiable differences in numbers or types of adverse events between DB05657 and placebo . CONCLUSIONS : This proof-of-concept study demonstrates that the ghrelin agonist DB05657 is well-tolerated in diabetes patients with moderate-to-severe chronic gastroparesis and shows statistically significant improvements in gastric emptying . P09917 pathway promotes cell proliferation in human glioma cell lines . OBJECTIVE : P09917 ( P09917 ) is a key enzyme in the synthesis of leukotrienes ( LTs ) , that might promote carcinogenesis . We investigated P09917 expression and examined whether the P09917 pathway is associated with the proliferation of human brain tumors . METHODS : We immunohistochemically evaluated the profile of P09917 expression in various types of brain tumors obtained from 42 patients , and examined the proliferative effects of the P09917 pathway in human glioma cell lines using a proliferation assay . RESULTS : Immunohistochemistry of glioblastomas , astrocytomas , meningiomas , medulloblastomas , craniopharyngiomas , ependymomas , neurinomas , oligodendrogliomas , malignant lymphomas , dysembryoplastic neuroepithelial and metastatic brain tumors revealed P09917 expression in the cytoplasm and nuclei or nuclear envelopes of tumor cells . The P09917 inhibitor A861 and the P09960 hydrolase inhibitor DB03424 dose-dependently suppressed the proliferation of A172 cells , a glioma cell line . CONCLUSIONS : We confirmed the expression of P09917 in various human brain tumors and demonstrated the partial suppression of tumor growth by inhibitors of the P09917 - P09960 hydrolase pathway in human glioma cell lines . The P09917 - P09960 pathway might play roles in the proliferation of human glioma cells . DB01259 induces apoptosis in trastuzumab-resistant breast cancer cells : effects on insulin-like growth factor I signaling . The majority of breast cancer patients who achieve an initial therapeutic response to the P04626 -targeted antibody trastuzumab will show disease progression within 1 year . Thus , the identification of novel agents that effectively inhibit survival of cancer cells that have progressed on trastuzumab is critical . In the current study , we show that the dual epidermal growth factor receptor ( P00533 ) /human P00533 -2 ( P04626 ) kinase inhibitor lapatinib induces apoptosis in trastuzumab-resistant cells derived from the P04626 -overexpressing SKBR3 breast cancer line . DB01259 inhibited P00533 and P04626 signaling in resistant cells , blocking activation of downstream Akt , mitogen-activated protein kinase [ corrected ] Importantly , lapatinib also inhibited insulin-like growth factor I ( P05019 ) signaling and growth-promoting effects in parental and resistant cells , and the cytotoxic effects of lapatinib were further enhanced by the P08069 -blocking antibody alphaIR3 . As increased P08069 signaling has been implicated in trastuzumab resistance , our data strongly support further study of lapatinib as a potential therapeutic in breast cancers that have progressed on trastuzumab . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . Development of drugs to target interactions between leukocytes and endothelial cells and treatment algorithms for inflammatory bowel diseases . Increased understanding of the pathogenesis of inflammatory bowel diseases ( IBDs ) has led to new therapeutic strategies . One of these is to target the molecules that regulate interactions between leukocytes and endothelial cells at sites of inflammation ( mainly leukocyte integrins and endothelial cell adhesion molecules of the immunoglobulin superfamily ) . These molecules have been validated as therapeutic targets for Q9UKU7 ; several have shown efficacy , and 2 have been approved by the Food and Drug Administration for treatment of Q9UKU7 . DB00108 , the first anti-integrin antibody tested for treatment of Q9UKU7 , blocks the α4 subunit . Although it is effective , its clinical use has been limited by its association with risk of progressive multifocal leukoencephalopathy . Other , allegedly more selective drugs that affect leukocyte recruitment in the gastrointestinal tract have been developed or are under investigation and could increase safety . These include vedolizumab and Q99217 181 ( antibodies against α4β7 ) , etrolizumab ( anti-β7 ) , and PF-00547659 ( anti-mucosal vascular addressin cell adhesion molecule 1 ) . Other agents have been developed to block α4 ( the small molecule AJM300 ) , P51686 ( the small molecule DB05005 -B ) , and P02778 ( the antibody eldelumab ) . We review the scientific rationale for inhibiting interactions between leukocytes and endothelial cells to reduce intestinal inflammation and analyze the clinical studies that have been performed to test these new molecules , with particular attention to safety . We propose an evidence-based clinical positioning of this class of drugs . DB01259 and doxorubicin enhance the Stat1-dependent antitumor immune response . The dual erbB1/2 tyrosine kinase inhibitor lapatinib as well as the anthracycline doxorubicin are both used in the therapy of P04626 -positive breast cancer . Using MMTV-neu mice as an animal model for P04626 -positive breast cancer , we observed enhanced tumor infiltration by IFN-γ-secreting T cells after treatment with doxorubicin and/or lapatinib . Antibody depletion experiments revealed a contribution of CD8⁺ but not CD4⁺ T cells to the antitumor effect of these drugs . Doxorubicin treatment additionally decreased the content of immunosuppressive tumor-associated macrophages ( TAMs ) in the tumor bed . In contrast , Stat1-deficient mice were resistant to tumor growth inhibition by lapatinib and/or doxorubicin and exhibited impaired T-cell activation and reduced T-cell infiltration of the tumor in response to drug treatment . Furthermore , Stat1-deficiency resulted in reduced expression of the T-cell chemotactic factors Q07325 , P02778 , and O14625 in the tumor epithelium . The inhibition of TAM infiltration of the tumor by doxorubicin and the immunosuppressive function of TAMs were found to be Stat1 independent . Taken together , the results point to an important contribution toward enhancing T-cell and IFN-γ-based immunity by lapatinib as well as doxorubicin and emphasize the role of Stat1 in building an effective antitumor immune response . Use of P01148 antagonists in reproductive medicine . Gonadotrophin-releasing hormone ( DB00644 ) plays a key role in the secretion of gonadotrophins , follicle-stimulating hormone ( DB00094 ) and luteinizing hormone ( LH ) , which regulate steroidogenesis and folliculogenesis . Two DB00644 antagonists , DB00050 and DB06785 , deprived of histaminergic side-effects , have been introduced into ovarian stimulation protocols to prevent premature LH surges and proved their safety in clinical trials . At present , most of the published studies have not found significant differences in follicular recruitment , oocyte quality , and so on , except for a decrease in pregnancy and implantation rates in in vitro fertilization and embryo transfer ( IVF-ET ) cycles when the DB00644 antagonist rather than the agonist was used . This decrease in pregnancy rates was in relation with a necessary learning curve of the physicians . Another possibility is the impact of the DB00644 antagonist on endometrium through its P30968 ; this effect was cancelled after cryopreserved embryo transfers because the pregnancy rates were similar between DB00644 antagonist and agonist in this case . DB00644 antagonists were also interesting in poor responders and polycystic ovarian syndrome , where the agonists have not permitted to obtain the better results in IVF-ET cycles . Similarly , the DB00644 antagonists could prevent the LH surge in the intrauterine insemination cycles . Triple negative breast cancer : therapeutic and prognostic implications . Triple negative breast cancers ( TNBC ) lack oestrogen receptor ( ER ) , progesterone receptor ( PR ) , nor over-express human epidermal growth factor receptor 2 ( P04626 ) . Epidemiologic studies demonstrate that women diagnosed with TNBC manifest a significantly different set of clinic-pathologic features and risk factors when compared to women with other subtypes of breast cancer . They are associated with poor prognosis , as defined by low five-year survival . To date many studies have examined the utility of traditional chemotherapy for the treatment of patients with TNBC and have confirmed the benefits of these agents in both the adjuvant and neoadjuvant settings . Targeted therapy options involving P09874 and P00533 inhibition , are currently in different phases of development and will hopefully change the paradigm of how patients with TNBC are treated . The present commentary aims to summarize the latest findings on chemotherapy in the treatment of TNBC in both the neoadjuvant and adjuvant setting and explore the ongoing development of newer targeted agents . Ghrelin promotes intestinal epithelial cell proliferation through PI3K/Akt pathway and P00533 trans-activation both converging to P29323 1/2 phosphorylation . Little is known about ghrelin 's effects on intestinal epithelial cells even though it is known to be a mitogen for a variety of other cell types . Because ghrelin is released in close proximity to the proliferative compartment of the intestinal tract , we hypothesized that ghrelin may have potent pro-proliferative effect on intestinal epithelial cells as well . To test this hypothesis , we characterized the effects of ghrelin on FHs74Int and Caco-2 intestinal epithelial cell lines in vitro . We found that ghrelin has potent dose dependent proliferative effects in both cell lines through a yet to be characterized G protein coupled growth hormone secretagogue receptor ( Q92847 ) subtype . Consistent with above findings , cell cycle flowcytometric analyses demonstrated that ghrelin shifts cells from the P55008 to S phase and thereby promotes cell cycle progression . Further characterization of subcellular events , suggested that ghrelin mediates its pro-proliferative effect through DB00131 cyclase ( AC ) -independent epidermal growth factor receptor ( P00533 ) trans-activation and PI3K-Akt phosphorylation . Both these pathways converge to stimulate MAPK , P29323 1/2 downstream . The role of ghrelin in states where intestinal mucosal injury and rapid mucosal repair occur warrants further investigation . Novel purine-based fluoroaryl-1,2,3-triazoles as neuroprotecting agents : synthesis , neuronal cell culture investigations , and Q00535 docking studies . A series of novel purine-based fluoroaryl triazoles were synthesized using the Cu(I) catalyzed 1,3-dipolar cycloaddition reactions ( click reactions ) , and assayed for their neuroprotective effects using fluorescence electron microscopy . Among these triazoles , o-fluorophenylmetyl-triazole , 7 , has comparable neuroprotective effect as that of DB03496 ( 1 ) and Roscovitine ( 2 ) , the state of the art CDK inhibitors , against the Aβ induced neurotoxicity . These results are substantiated using computer docking methods ( DarwinDock/GenDock ) , which predict that Roscovitine and the triazole 7 bind to the DB00171 -binding site of Q00535 /p25 with comparable binding energies , whereas the corresponding pentafluorophenylmethyl-triazole , 9 , has dramatically reduced binding energy ( in accordance with its lack of neuroprotection ) . These combined experimental and theoretical studies support the involvement of Q00535 /p25 in the neuronal cell cycle re-entry . Multiple arrhythmic syndromes in a newborn , owing to a novel mutation in Q14524 . BACKGROUND : Mutations in the Q14524 gene have been linked to Brugada syndrome ( BrS ) , conduction disease , Long QT syndrome ( LQT3 ) , atrial fibrillation ( AF ) , and to pre- and neonatal ventricular arrhythmias . OBJECTIVE : The objective of this study is to characterize a novel mutation in Na(v)1.5 found in a newborn with fetal chaotic atrial tachycardia , post-partum intraventricular conduction delay , and QT interval prolongation . METHODS : Genomic DNA was isolated and all exons and intron borders of 15 ion-channel genes were sequenced , revealing a novel missense mutation ( Q270K ) in Q14524 . Na(v)1.5 wild type ( WT ) and Q270K were expressed in CHO- P04264 with and without the Na(v)β1 subunit . Results . Patch-clamp analysis showed ∼40 % reduction in peak sodium channel current ( I(Na) ) density for Q270K compared with WT . Fast and slow decay of I(Na) were significantly slower in Q270K . Steady-state activation and inactivation of Q270K channels were shifted to positive potentials , and window current was increased . The tetrodotoxin-sensitive late I(Na) was increased almost 3-fold compared with WT channels . DB00243 reduced late I(Na) in WT and Q270K channels , while exerting minimal effects on peak I(Na) . CONCLUSION : The Q270K mutation in Q14524 reduces peak I(Na) while augmenting late I(Na) , and may thus underlie the development of atrial tachycardia , intraventricular conduction delay , and QT interval prolongation in an infant . Association of polymorphisms in P36888 , P00533 , P09917 , and Q8TAT5 with glioblastoma in the Han Chinese population . Glioblastoma ( GBM ) is the highest-grade glioma in astrocytoma . Patients often have poor prognosis due to therapeutic resistance and tumor recurrence . Identification of the genetic factors of GBM could be important contribution to early prevention of this disease . We genotyped 17 tag single-nucleotide polymorphisms ( tSNPs ) from nine genes in this study , including 72 cases and 302 controls . SNP genotyping was conducted using Sequenom MassARRAY RS1000 . Statistical analysis of the association between tSNPs and GBM was performed using the χ ( 2 ) test and SNPStats software . The rs3829382 in P36888 was associated with increased odds of developing GBM using the χ ( 2 ) test . When we analyzed tSNPs under different inheritance models , we found rs9642393 in P00533 increased odds of developing GBM in the dominant model . After stratification by gender , we found that rs12645561 in Q8TAT5 and rs2291427 in P09917 were associated with developing GBM . Polymorphisms within P36888 , P00533 , Q8TAT5 , and P09917 may contribute to the occurrence of GBM in the Han Chinese population . However , the functional significance of these polymorphisms needs further investigation . Targeted treatment of advanced and metastaticbreast cancer with lapatinib . Improved molecular understanding of breast cancer in recent years has led to the discovery of important drug targets such as HER-2 and P00533 . DB01259 is a potent dual inhibitor of HER-2 and P00533 . Preclinical and phase I studies have shown activity with lapatinib in a number of cancers , including breast cancer , and the drug is well tolerated . The main known drug interactions are with paclitaxel and irinotecan . The most significant side-effects of lapatinib are diarrhea and adverse skin events . Rates of cardiotoxicity compare favorably with trastuzumab , a monoclonal antibody against HER-2 . This paper focuses on lapatinib in advanced and metastatic breast cancer , which remains an important therapeutic challenge . Phase II and III studies show activity as monotherapy , and in combination with chemotherapy or hormonal agents . Results from these studies suggest that the main benefit from lapatinib is in the HER-2 positive breast cancer population . Combinations of lapatinib and trastuzumab are also being studied and show encouraging results , particularly in trastuzumab-refractory metastatic breast cancer . DB01259 may have a specific role in treating HER-2 positive CNS metastases . The role of lapatinib as neoadjuvant therapy and in early breast cancer is also being evaluated . P04626 monoclonal antibodies that do not interfere with receptor heterodimerization-mediated signaling induce effective internalization and represent valuable components for rational antibody-drug conjugate design . The human epidermal growth factor receptor (HER)2 provides an excellent target for selective delivery of cytotoxic drugs to tumor cells by antibody-drug conjugates ( ADC ) as has been clinically validated by ado-trastuzumab emtansine ( Kadcyla(TM) ) . While selecting a suitable antibody for an ADC approach often takes specificity and efficient antibody-target complex internalization into account , the characteristics of the optimal antibody candidate remain poorly understood . We studied a large panel of human P04626 antibodies to identify the characteristics that make them most suitable for an ADC approach . As a model toxin , amenable to in vitro high-throughput screening , we employed Pseudomonas exotoxin A ( P25101 ' ) fused to an anti-kappa light chain domain antibody . Cytotoxicity induced by P04626 antibodies , which were thus non-covalently linked to P25101 ' , was assessed for high and low P04626 expressing tumor cell lines and correlated with internalization and downmodulation of P04626 antibody-target complexes . Our results demonstrate that P04626 antibodies that do not inhibit heterodimerization of P04626 with related ErbB receptors internalize more efficiently and show greater P25101 '-mediated cytotoxicity than antibodies that do inhibit such heterodimerization . Moreover , stimulation with ErbB ligand significantly enhanced ADC-mediated tumor kill by antibodies that do not inhibit P04626 heterodimerization . This suggests that the formation of P04626 /ErbB-heterodimers enhances ADC internalization and subsequent killing of tumor cells . Our study indicates that selecting P04626 ADCs that allow piggybacking of P04626 onto other ErbB receptors provides an attractive strategy for increasing ADC delivery and tumor cell killing capacity to both high and low P04626 expressing tumor cells .

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Vascular endothelial growth factor receptor tyrosine kinase inhibitors vandetanib ( DB05294 ) and DB04849 in lung cancer . Vascular endothelial growth factor ( P15692 ) is a rational target for advanced non-small cell lung cancer ( NSCLC ) , a hypothesis validated by the recent Eastern Cooperative Oncology Group E4599 trial showing that the addition of the P15692 monoclonal antibody bevacizumab to chemotherapy prolongs overall survival . Several new tyrosine kinase inhibitors targeting the P15692 pathway are currently in advanced clinical development for NSCLC and offer several possible advantages compared with monoclonal antibodies , including oral administration , more flexible dosing , a broader spectrum of target inhibition , and different toxicity profiles . Among these agents , vandetanib ( DB05294 ) , an inhibitor of the P15692 receptor ( VEGFR ) -2 and epidermal growth factor receptor tyrosine kinase , has been the most extensively studied . In a randomized phase II study of patients with platinum-refractory NSCLC , including squamous histology , vandetanib prolonged progression-free survival compared with gefitinib . In another phase II trial , an improvement in progression-free survival was observed for vandetanib in combination with docetaxel compared with docetaxel alone . DB04849 is an inhibitor of P17948 , P35968 , and P35916 and other tyrosine kinases that has shown clinical activity in NSCLC in combination with carboplatin and paclitaxel . Several phase III trials are under way testing these agents either as monotherapy or in combination with chemotherapy in patients with lung cancer . Early results with these agents , and others being tested , raise the possibility that there will eventually be multiple P15692 -targeted therapies available in the clinic that can potentially benefit a broader range of patients with advanced-stage NSCLC . Involvement of thrombopoietin in acinar cell necrosis in L-arginine-induced acute pancreatitis in mice . P40225 ( P07202 ) plays an important role in injuries of different tissues . However , the role of P07202 in acute pancreatitis ( AP ) is not yet known . The aim of the study was to determine the involvement of P07202 in AP . Serum P07202 was assayed in necrotizing pancreatitis induced by L-arginine in mice . Recombinant P07202 and anti- P07202 antibody were given to mice with necrotizing pancreatitis . Amylase , lipase , lactate dehydrogenase , myeloperoxidase activity and pancreatic water content were assayed in serum and tissue samples . Pancreas and lung tissue samples were also collected for histological evaluation . Immunohistochemistry of amylase α and P12004 were applied for the study of acinar regeneration and TUNEL assay for the detection of apoptosis in the pancreas . Increased levels of serum P07202 were found in necrotizing pancreatitis . After P07202 administration , more severe acinar necrosis was found and blockade of P07202 reduced the acinar necrosis in this AP model . Acinar regeneration and apoptosis in the pancreas were affected by P07202 and antibody treatment in necrotizing pancreatitis . The severity of pancreatitis-associated lung injury was worsened after P07202 treatment , but attenuated after Anti- P07202 antibody treatment . In conclusion , serum P07202 is up-regulated in the necrotizing pancreatitis induced by L-arginine in mice and may be a risk factor for the pancreatic acinar necrosis in AP . As a pro-necrotic factor , blockade of P07202 can attenuate the acinar necrosis in AP and may be a possible therapeutic intervention for AP . Comparison of the effects of firocoxib , carprofen and vedaprofen in a sodium urate crystal induced synovitis model of arthritis in dogs . A randomized , placebo-controlled , four-period cross-over laboratory study involving eight dogs was conducted to confirm the effective analgesic dose of firocoxib , a selective P35354 inhibitor , in a synovitis model of arthritis . DB09217 was compared to vedaprofen and carprofen , and the effect , defined as a change in weight bearing measured via peak ground reaction , was evaluated at treatment dose levels . A lameness score on a five point scale was also assigned to the affected limb . Peak vertical ground reaction force was considered to be the most relevant measurement in this study . The firocoxib treatment group performed significantly better than placebo at the 3 h post-treatment time point and significantly better than placebo and carprofen at the 7 h post-treatment time point . Improvement in lameness score was also significantly better in the dogs treated with firocoxib than placebo and carprofen at both the 3 and 7 h post-treatment time points . A curated database of miRNA mediated feed-forward loops involving MYC as master regulator . BACKGROUND : The MYC transcription factors are known to be involved in the biology of many human cancer types . But little is known about the Myc/microRNAs cooperation in the regulation of genes at the transcriptional and post-transcriptional level . METHODOLOGY/PRINCIPAL FINDINGS : Employing independent databases with experimentally validated data , we identified several mixed microRNA/Transcription Factor Feed-Forward Loops regulated by Myc and characterized completely by experimentally supported regulatory interactions , in human . We then studied the statistical and functional properties of these circuits and discussed in more detail a few interesting examples involving Q01094 , P60484 , P06400 and P15692 . CONCLUSIONS/SIGNIFICANCE : We have assembled and characterized a catalogue of human mixed Transcription Factor/microRNA Feed-Forward Loops , having Myc as master regulator and completely defined by experimentally verified regulatory interactions . Restriction of adenoviral replication to the transcriptional intersection of two different promoters for colorectal and pancreatic cancer treatment . In our current study , we developed oncolytic adenoviruses which preferentially lyse pancreatic and colon cancer cells by replacing viral E1 and/or E4 promoter with the tumor/tissue-specific promoters , cyclooxygenase-2 ( P35354 ) , midkine ( MK ) , or the cell cycle-dependent promoter , Q01094 . We generated three sets of recombinant adenoviral vectors . In the first set , only the native E1A promoter was replaced by the P35354 , MK , or Q01094 promoter , respectively . In the second set , the viral E4 promoter was substituted by these heterologous promoters and the viral E1A promoter was substituted by the ubiquitously active cytomegalovirus-IE promoter . In the third set , we substituted the viral E1A and E4 promoters with the P35354 , MK , or Q01094 promoter , respectively . In our system , transcriptional targeting of solitary viral E1A resulted in 50 % enhanced restricted vector replication when compared with an unrestricted replication-competent adenovirus . Furthermore , a targeted expression of the viral E1A gene products had a greater effect on restricted adenoviral replication than that of the E4 region . With our vectors , Ad. P36551 .MK and Ad.MK. P36551 , using two different heterologous promoters to control E1A and E4 expression , we showed enhanced viral replication specificity when compared with Ad. P36551 . P36551 or Ad.MK.MK , respectively . In a s.c. xenograft tumor model , there was no significant difference in the antineoplastic efficacy of the double heterologous promoter-controlled vectors when compared with our unrestricted replication-competent control adenovirus or vectors with only E1A transcriptionally driven by a heterologous promoter . P15328 regulates cell proliferation in mouse gonadotroph alphaT3-1 cells . We have previously found that the mRNA and protein levels of the folate receptor alpha ( FRalpha ) are uniquely over-expressed in clinically human nonfunctional ( NF ) pituitary adenomas , but the mechanistic role of FRalpha has not fully been determined . We investigated the effect of FRalpha over-expression in the mouse gonadotroph alphaT3-1 cell line as a model for NF pituitary adenomas . We found that the expression and function of FRalpha were strongly up-regulated , by Western blotting and folic acid binding assay . Furthermore , we found a higher cell growth rate , an enhanced percentage of cells in S-phase by BrdU assay , and a higher P12004 staining . These observations indicate that over-expression of FRalpha promotes cell proliferation . These effects were abrogated in the same alphaT3-1 cells when transfected with a mutant FRalpha cDNA that confers a dominant-negative phenotype by inhibiting folic acid binding . Finally , by real-time quantitative PCR , we found that mRNA expression of Q9UM47 was up-regulated in FRalpha over-expressing cells . In summary , our data suggests that FRalpha regulates pituitary tumor cell proliferation and mechanistically may involve the NOTCH pathway . Potentially , this finding could be exploited to develop new , innovative molecular targeted treatment for human NF pituitary adenomas . Noncatalytic function of P27361 /2 can promote Raf/MEK/ P29323 -mediated growth arrest signaling . Kinase activity is known as the key biochemical property of MAPKs . Here , we report that P27361 /2 also utilizes its noncatalytic function to mediate certain signal transductions . Sustained activation of the Raf/MEK/ P29323 pathway induces growth arrest , accompanied by changes in cell cycle regulators ( decreased retinoblastoma phosphorylation , Q01094 down-regulation , and/or P38936 (CIP1) up-regulation ) and cell type-specific changes in morphology and expression of c-Myc or P07949 in the human tumor lines LNCaP , U251 , and TT . Ablation of P27361 /2 by RNA interference abrogated all these effects . However , active site-disabled P29323 mutants ( P27361 -K71R , P28482 -K52R , and P28482 -D147A ) , which competitively inhibit activation of endogenous P27361 /2 , could not block Raf/MEK-induced growth arrest as well as changes in the cell cycle regulators , although they effectively blocked phosphorylation of the P27361 /2 catalytic activity readouts , p90(RSK) and ELK1 , as well as the cell type-specific changes . Because this indicated a potential noncatalytic P27361 /2 function , we generated stable lines of the tumor cells in which both P27361 and P28482 were significantly knocked down , and we further investigated the possibility using rat-derived kinase-deficient P29323 mutants ( P28482 -K52R and P28482 -T183A/Y185F ) that were not targeted by human small hairpin RNA . Indeed , P28482 -K52R selectively restored Raf-induced growth inhibitory signaling in P27361 /2-depleted cells , as manifested by regained cellular ability to undergo growth arrest and to control the cell cycle regulators without affecting c-Myc and morphology . However , P28482 -T183A/Y185F was less effective , indicating the requirement of TEY site phosphorylation . Our study suggests that functions of P27361 /2 other than its " canonical " kinase activity are also involved in the pathway-mediated growth arrest signaling . Proteomic identification of overexpressed adenomatous polyposis coli and cyclin B3 during endoderm differentiation from human embryonic stem cells . OBJECTIVES : This study was aimed to investigate important proteins associated with endoderm differentiation by pancreatic derivation protocol from human embryonic stem cells ( hESCs ) . METHODS : Comparative proteomic analysis of endoderm cells differentiated from hESCs by activin A and low serum was performed . Proteins with altered expression levels during endoderm differentiation were investigated by 2-dimensional gel electrophoresis ( 2-DE ) with mass spectrometric analysis . RESULTS : Thirty-four protein spots with significantly changed intensities were identified . These were functionally annotated based on gene ontology . The messenger RNA expression levels of 5 genes , P25054 , Q8WWL7 , P38646 , P78371 , and P62258 , were correlated with 2-DE analysis . We further validated the protein expression levels of adenomatous polyposis coli ( P25054 ) and cyclin B3 ( Q8WWL7 ) by using Western blot analysis and immunocytochemistry . They are involved in the regulation of cell cycle , thus , cyclins and cyclin-dependent kinases , which regulate the cell cycle , were examined . P12004 A1 , cyclin D2 , and cyclin E2 were upregulated , and other cyclins and cyclin-dependent kinases were downregulated in endoderm cells . CONCLUSIONS : The increase in expression of P25054 and Q8WWL7 suggests that these proteins will be important markers for identifying endoderm cells differentiated from hESCs , and they can play important roles in the differentiation of endoderm cells from hESCs or in human endoderm development for pancreas . Akt- and Erk-mediated regulation of proliferation and differentiation during PDGFRβ-induced O60682 self-renewal . Understanding the mechanisms that direct mesenchymal stem cell ( O60682 ) self-renewal fate decisions is a key to most tissue regenerative approaches . The aim of this study here was to investigate the mechanisms of action of platelet-derived growth factor receptor β ( PDGFRβ ) signalling on O60682 proliferation and differentiation . O60682 were cultured and stimulated with DB00102 together with inhibitors of second messenger pathways . Cell proliferation was assessed using ethynyl-2'-deoxyuridine and phosphorylation status of signalling molecules assessed by Western Blots . To assess differentiation potentials , cells were transferred to adipogenic or osteogenic media , and differentiation assessed by expression of differentiation association genes by qRT-PCR , and by long-term culture assays . Our results showed that distinct pathways with opposing actions were activated by PDGF . PI3K/Akt signalling was the main contributor to O60682 proliferation in response to activation of PDGFRβ . We also demonstrate a negative feedback mechanism between PI3K/Akt and P09619 -β expression . In addition , PI3K/Akt downstream signal cascades , P42345 and its associated proteins p70S6K and Q13541 were involved . These pathways induced the expression of cyclin D1 , cyclin D3 and Q00534 to promote cell cycle progression and O60682 proliferation . In contrast , activation of Erk by PDGFRβ signalling potently inhibited the adipocytic differentiation of MSCs by blocking PPARγ and CEBPα expression . The data suggest that PDGFRβ-induced Akt and Erk pathways regulate opposing fate decisions of proliferation and differentiation to promote O60682 self-renewal . Thus , activation of multiple intracellular cascades is required for successful and sustainable O60682 self-renewal strategies . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . DB05332 as a treatment for immune thrombocytopenia : a review . " Immune thrombocytopenia " ( ITP ) is an autoimmune disorder that leads to peripheral destruction , as well as a decreased production of platelets . ITP most commonly presents as mild mucocutaneous bleeding . Though it is rare , the leading cause of mortality in persons with ITP is intracranial hemorrhage and those that do not respond to therapy are at increased risk . Our understanding of the pathophysiology of ITP has evolved immensely , especially over the last 60 years . The discovery of the platelet-production stimulator , thrombopoietin ( P07202 ) , lent clarity to an earlier hypothesis that inhibition of platelet production at the level of the megakaryocyte , at least in part , accounts for thrombocytopenia in adults with ITP . This facilitated the development of P07202 -based therapies to treat ITP . P40238 agonists are one of the most recent treatments to enter the landscape . Original production of a recombinant human P07202 was halted after clinical trials revealed the untoward effect of autoantibodies to the recombinant human P07202 with cross-reactivity to endogenous P07202 . Next-step development focused on stimulation of the P07202 receptor with fewer immunogenic agents . Currently , two such thrombopoietin receptor agonists , romiplostim and eltrombopag , are licensed in the USA to treat thrombocytopenia in adults with persistent or chronic ITP . Ongoing research will assess their efficacy in other immune-mediated and nonimmune-mediated primary and secondary thrombocytopenias . PDE10 inhibition increases P42261 and CREB phosphorylation and improves spatial and recognition memories in a Huntington 's disease mouse model . Huntington 's disease ( HD ) causes motor disturbances , preceded by cognitive impairment , in patients and mouse models . We showed that increased hippocampal DB02527 -dependent protein kinase ( PKA ) signaling disrupts recognition and spatial memories in R6 HD mouse models . However , unchanged levels of hippocampal phosphorylated ( p ) DB02527 -responsive element-binding protein ( CREB ) suggested unaltered nuclear PKA activity in R6 mice . Here , we extend this finding by showing that nuclear pPKA catalytic subunit ( Thr197 ) and pPKA substrate levels were unaltered in the hippocampus of R6/1 mice . Phosphodiesterases ( PDEs ) play an important role in the regulation of PKA activity . Q9Y233 , a DB02527 /cGMP dual-substrate PDE , was reported to be restricted to the nuclear region in nonstriatal neurons . Using cell fractionation we confirmed that Q9Y233 was enriched in nuclear fractions , both in wild-type and R6/1 mice hippocampus , without differences in its levels or intracellular distribution between genotypes . We next investigated whether inhibition of PDE10 with papaverine could improve cognitive function in HD mice . DB01113 treatment improved spatial and object recognition memories in R6/1 mice , and significantly increased pGluA1 and pCREB levels in R6/1 mice hippocampus . DB01113 likely acted through the activation of the PKA pathway as the phosphorylation level of distinct cGMP-dependent kinase ( cGK ) substrates was not modified in either genotype . Moreover , hippocampal DB02527 , but not cGMP , levels were increased after acute papaverine injection . Our results show that inhibition of PDE10 improves cognition in R6 mice , at least in part through increased P42261 and CREB phosphorylation . Thus , PDE10 might be a good therapeutic target to improve cognitive impairment in HD . DB01780 signaling reveals 14-3-3 protein function as a novel step in left-right patterning during amphibian embryogenesis . To gain insight into the molecular mechanisms underlying the control of morphogenetic signals by H+ flux during embryogenesis , we tested DB01780 -A ( FC ) , a compound produced by the fungus Fusicoccum amygdali Del . In plant cells , FC complexes with 14-3-3 proteins to activate H+ pumping across the plasma membrane . It has long been thought that FC acts on higher plants only ; here , we show that exposing frog embryos to FC during early development specifically results in randomization of the asymmetry of the left-right ( LR ) axis ( heterotaxia ) . Biochemical and molecular-genetic evidence is presented that 14-3-3-family proteins are an obligate component of Xenopus FC receptors and that perturbation of 14-3-3 protein function results in heterotaxia . The subcellular localization of 14-3-3 mRNAs and proteins reveals novel cytoplasmic destinations , and a left-right asymmetry at the first cell division . Using gain-of-function and loss-of-function experiments , we show that P62258 protein is likely to be an endogenous and extremely early aspect of LR patterning . These data highlight a striking conservation of signaling pathways across kingdoms , suggest common mechanisms of polarity establishment between C. elegans and vertebrate embryos , and uncover a novel entry point into the pathway of left-right asymmetry determination . Inhibition of aldehyde dehydrogenase 2 activity enhances antimycin-induced rat cardiomyocytes apoptosis through activation of MAPK signaling pathway . Aldehyde dehydrogenase 2 ( P05091 ) , a mitochondrial-specific enzyme , has been proved to be involved in oxidative stress-induced cell apoptosis , while little is known in cardiomyocytes . This study was aimed at investigating the role of P05091 in antimycin A-induced cardiomyocytes apoptosis by suppressing P05091 activity with a specific P05091 inhibitor DB02115 . Antimycin A ( 40μg/ml ) was used to induce neonatal cardiomyocytes apoptosis . DB02115 ( 60μM ) effectively inhibited P05091 activity by 50 % without own effect on cell apoptosis , and significantly enhanced antimycin A-induced cardiomyocytes apoptosis from 33.5±4.4 to 56.5±6.4 % ( Hochest method , p < 0.05 ) , and from 57.9±1.9 to 74.0±11.9 % ( FACS , p < 0.05 ) . Phosphorylation of activated MAPK signaling pathway , including extracellular signal-regulated kinase ( P27361 /2 ) , c-Jun NH2-terminal kinase ( JNK ) and p38 was also increased in antimycin A and daidzin treated cardiomyocytes compared to the cells treated with antimycin A alone . These findings indicated that modifying mitochondrial P05091 activity/expression might be a potential therapeutic option on reducing oxidative insults induced cardiomyocytes apoptosis . DB00158 transport in mouse cumulus-oocyte complexes and preimplantation embryos . Endogenous folate stores are required in preimplantation embryos of several species , but how folates are accumulated and whether they can be replenished has not been determined . Folates are generally taken up into cells by specific transporters , mainly the reduced folate carrier RFC1 ( P41440 protein ) and the high-affinity folate receptors P15328 and P14207 . Quantitative RT-PCR showed that Slc19a1 mRNA was expressed in mouse cumulus-oocyte complexes ( COCs ) and oocytes , whereas Folr1 showed expression only in preimplantation embryos , increasing from the 2-cell stage onward . The mRNAs encoding Folr2 and the intestinal folate transporter Slc46a1 were not detected . DB00563 ( MTX ) , an antifolate often used as a model substrate for folate transport , exhibited saturable transport in COCs and in preimplantation embryos starting at the 2-cell stage . However , folate transport characteristics differed between COCs and embryos . In COCs , transport of MTX and the reduced folate leucovorin was inhibited by the anion transport inhibitor SITS that blocks RFC1 but was insensitive to dynasore , a specific dynamin inhibitor that instead inhibits folate receptor-receptor mediated endocytosis , whereas the opposite was found in 2-cell embryos and blastocysts . The inhibitor profile and transport properties of MTX and leucovorin in COCs correspond to established transport characteristics of RFC1 ( P41440 ) , whereas those in 2-cell embryos and blastocysts correspond with those of P15328 , consistent with the mRNA expression patterns . Considerable folate was accumulated in COCs via RFC1 , but the presence of cumulus cells did not enhance folate accumulation in the enclosed oocyte , indicating a lack of transfer from cumulus to oocyte . A new model of cell cycle-regulated transcription : repression of the cyclin A promoter by P05231 -1 and anti-repression by E2F . Cell cycle regulation of the cyclin A gene is determined by a bipartite repressor binding site in the region of the basal promoter , termed CDE-CHR , which also controls the expression of cell cycle genes upregulated in S or G2 ( such as cdc25C ) . The CDE-CHR in the cyclin A promoter is recognized by both E2F complexes and P05231 -1 , but the contribution of each of these factors in cell cycle regulation is unknown . In the present study , we have introduced mutations into the cyclin A promoter which lead to either a loss or enhancement of E2F binding , while having only marginal effects on the interaction with P05231 -1 . Unlike mutants deficient for P05231 -1 binding , promoter variants lacking E2F binding showed an unchanged repression in G0 , thus identifying P05231 -1 as the principal repressor of the cyclin A gene . The same mutants did show , however , a delayed derepression while a mutation leading to increased E2F binding resulted in premature up-regulation . These findings clearly suggest that E2F contributes to the correct timing of cyclin A transcription , presumably by acting as an anti-repressor . In agreement with this conclusion , we find that the cyclin A promoter only poorly interacts with Q16254 , which is the major E2F family member in G0 cells , while a clear binding is seen with Q01094 and -3 , which are up-regulated in late P55008 . Use of RNA interference to elucidate the effect of P04198 on cell cycle in neuroblastoma . BACKGROUND : P04198 amplification marks poor prognosis in neuroblastoma ( NB ) tumors . In evaluating the mechanisms by which retinoic acid ( RA ) or nerve growth factor ( P01138 ) decrease cell number in P04198 amplified NB cells , we have identified a number of proteins whose expression either decreases ( E2F , P06493 , Q00534 , cyclin dependent kinase activity ) or increases ( p27 ) in association with a decrease in P04198 expression . However , it was still unclear which were P04198 dependent effects or not . PROCEDURE : This study aimed to determine which changes in cell cycle gene expression are modulated as a consequence of the decrease in P04198 . We silenced P04198 expression using siRNA targeted to the coding region of P04198 . Then , by using siRNA transient transfections , we analyzed the change of cell cycle related genes and cell cycle in P04198 amplified NB cell lines . RESULTS : We demonstrate that expression of P04198 can be suppressed by almost 60 % in P04198 amplified NB cell using siRNAs targeted to P04198 . Functionally , the decrease in P04198 leads to a decrease in cells in the S-phase of the cell cycle . Decreases in P04198 are associated with decreases in Q01094 -2 and Q02363 along with increases in p27 protein levels by post-transcriptional modification . Moreover , we find that a decrease in P04198 is accompanied by a decrease in cdk6 mRNA and protein expression . CONCLUSIONS : These results show that E2F and Q02363 expression is associated with P04198 regulation and that cdk6 is a possible new transcriptional target of P04198 . Anti- P05231 -receptor-alpha ( tocilizumab ) does not inhibit human monocyte-derived dendritic cell maturation or alloreactive T-cell responses . Significant comorbidites and lethality complicate GVHD and its treatment . Targeting the cytokine milieu may improve GVHD control ; and P05231 is an attractive candidate , given its role in dendritic cell activation and T-cell differentiation . DB06273 is a humanized mAb to P05231 -receptor-α ( P08887 -α ) , which is Food and Drug Administration-approved for treatment of rheumatoid arthritis . Mouse transplant models have demonstrated that P05231 blockade also improves GVHD scores and survival . Definitive immunologic effects of P05231 inhibition have not emerged given inconsistent alterations in regulatory T cells ( Tregs ) and suppression of T-cell proliferation . Despite on-target suppression of P08887 -α signaling in human monocyte-derived dendritic cells ( moDCs ) and T cells , our data show no effect on moDC maturation/activation , alloreactive T-cell proliferation , Treg expansion , or allogeneic Th1/Th17 responses in vitro . These findings merit attention in any clinical trials of tocilizumab for GVHD prevention or treatment and provide a rationale for evaluating more specific inhibitors of downstream O60674 / P40763 signaling as well . Activity-dependent p25 generation regulates synaptic plasticity and Aβ-induced cognitive impairment . P12004 -dependent kinase 5 regulates numerous neuronal functions with its activator , p35 . Under neurotoxic conditions , p35 undergoes proteolytic cleavage to liberate p25 , which has been implicated in various neurodegenerative diseases . Here , we show that p25 is generated following neuronal activity under physiological conditions in a Q13224 - and CaMKIIα-dependent manner . Moreover , we developed a knockin mouse model in which endogenous p35 is replaced with a calpain-resistant mutant p35 ( Δp35KI ) to prevent p25 generation . The Δp35KI mice exhibit impaired long-term depression and defective memory extinction , likely mediated through persistent P42261 phosphorylation at Ser845 . Finally , crossing the Δp35KI mice with the 5XFAD mouse model of Alzheimer 's disease ( AD ) resulted in an amelioration of β-amyloid ( Aβ ) -induced synaptic depression and cognitive impairment . Together , these results reveal a physiological role of p25 production in synaptic plasticity and memory and provide new insights into the function of p25 in Aβ-associated neurotoxicity and AD-like pathology .

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interacts_with DB01200?

multiple_choice

P11926 activity in human endometrium and endometrial cancer cells . P11926 ( ODC ) activities were significantly higher in proliferative endometrium during the estrogen-dominated follicular phase of the menstrual cycle than in secretory endometrium after the formation of the progesterone-secreting corpus luteum . The enzymatic activity was increased about fivefold by renewal of the medium during incubations of endometrial fragments or isolated endometrial glands . Endometrial adenocarcinoma cells ( O14777 -1 , O14777 -50 ) , both in monolayers and suspension , also responded to medium renewal by increasing ODC activity about 10-fold after 4 h , with subsequent reduction to control levels after 7 h . These effects were blocked by actinomycin D and cycloheximide . Endometrial stromal cells exhibited highly variable ODC activities at different passages . Difluoromethylornithine ( DB06243 ) and sodium molybdate had marked antiproliferative effects in O14777 -50 cultures , reducing cell numbers to 10 to 20 % of control values 11 d after plating and inhibiting ODC activity by approximately 80 % on Day 7 . The antiproliferative effect of DB06243 , but not that of molybdate , was reversed by 10 microM putrescine , the product of ODC activity . In contrast to DB06243 , molybdate had no effect on ODC activity of cell homogenates . Molybdate did not elicit antizyme formation in O14777 -50 cells under conditions in which putrescine did . These results indicate that ODC activity , present in both epithelial and stromal cells , as shown analytically and also by autoradiography after labeling with [3H] DB06243 , may be related to cell proliferation in vivo and that proliferation of human endometrial cancer cells in culture can be arrested by DB06243 and by molybdate . In vivo excitation of GABA interneurons in the medial prefrontal cortex through 5- Q9H205 receptors . Serotonin ( 5-hydroxytryptamine , 5-HT ) controls pyramidal cell activity in prefrontal cortex ( P27918 ) through various receptors , in particular , P08908 and 5- Q13049 receptors . Here we report that the physiological stimulation of the raphe nuclei excites local , putatively GABAergic neurons in the prelimbic and cingulate areas of the rat P27918 in vivo . These excitations had a latency of 36 +/- 4 ms and a duration of 69 +/- 9 ms and were blocked by the i.v. administration of the 5- Q9H205 receptor antagonists ondansetron and tropisetron . The latency and duration were shorter than those elicited through 5- Q13049 receptors in pyramidal neurons of the same areas . Double in situ hybridization histochemistry showed the presence of GABAergic neurons expressing 5- Q9H205 receptor mRNA in P27918 . These cells were more abundant in the cingulate , prelimbic and infralimbic areas , particularly in superficial layers . The percentages of Q99259 mRNA-positive neurons expressing 5- Q9H205 receptor mRNA in prelimbic cortex were 40 , 18 , 6 and 8 % in layers I , II-III , V and VI , respectively , a distribution complementary to that of cells expressing 5- Q13049 receptors . Overall , these results support an important role of 5-HT in the control of the excitability of apical dendrites of pyramidal neurons in the medial P27918 through the activation of 5- Q9H205 receptors in GABAergic interneurons . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . Refined localization and yeast artificial chromosome ( YAC ) contig -- mapping of genes and DNA segments in the 7q21-q32 region . The chromosome localizations for 159 gene and DNA segments have been refined to one of five intervals in the 7q21-132 region through hybridization analysis with a panel of somatic cell hybrid lines . Seventy-two of these chromosome 7 markers are also mapped on common or overlapping yeast artificial chromosome ( YAC ) clones . In addition , the breakpoints of chromosome rearrangement contained in five of the somatic cell hybrid lines have been defined by flanking probes within YAC contigs . To provide a framework for further mapping of the 7q21-q32 region , we have established the physical order of a set of reference markers : cen-( P08123 -D7S15- P08684 - P27169 )-D7S456- ( brea kpoint contained in cell hybrid 1EF2/3/K017 ) - P08236 -D7S186-ASL- ( P08183 - P21439 - P62879 -EPO- P22303 ) -D7S238- ( proximal breakpoint in GM1059-Rag5 ) -D7S240-(CUTL1- P05121 )- ( breakp oints in 1CF2/5/K016 and 2068Rag22-2 ) -( P31323 -D7S13)- P07942 - ( breakpoint in JSR-17S ) -DLD-D7S16-MET- P09544 - P13569 -D7S8-tel . Serotonin skews human macrophage polarization through P41595 and P34969 . Besides its role as a neurotransmitter , serotonin ( 5-hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT(1-7) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 production , upregulated the expression of M2 polarization-associated genes ( P05120 , P07996 , Q9NY15 , Q86Y22 ) , and reduced the expression of M1-associated genes ( P08476 , P41597 , P39900 , P05121 , P29016 , O94788 ) . Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2( P09603 ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7) , whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies . Genetic susceptibility in solvent induced neurobehavioral effects . The aim of this investigation was to study the influence of genetic polymorphisms of biotransformation enzymes and dopamine receptors on neurobehavioral effects in referents ( n = 53 ) , solvent-workers ( n = 144 ) , and chronic toxic encephalopathy ( CTE ) patients ( n = 33 ) . All participants were interviewed for exposure data and confounding factors and underwent a clinical examination . Neurobehavioral complaints ( neurotoxicity symptom checklist-60 ) and effects [ simple reaction time ( SRT ) , symbol digit substitution ( SDS ) , hand-eye coordination ( O14777 ) , and digit span backwards ( DSB ) ] were evaluated with a computer assisted test battery . The following genotypes were determined : P09488 , P30711 , P09211 , P14416 Taq1A , P14416 Taq1B , and P14416 -141Cdel . Neurotoxic effects and complaints were significantly higher in CTE patients and were related to both duration and level of exposure . An equal distribution of genotypes was found between all groups . Logistic regression analysis revealed that P30711 was negatively associated with sleep and sensorimotor complaints . P09488 had a protecting influence on the relationship between logDSB and the cumulative exposure index and between logSRT and cumulative exposure index and degree of exposure , respectively . This effect was also found when correcting for age , education level , alcohol consumption , and smoking . P14416 -141Cdel polymorphisms had a negative influence on the relationship between logSDS and the total exposure time . P30711 might be protective against sleep and sensorimotor complaints , whereas P09488 seems to decrease sustained attention and short-term memory problems in relation to solvent exposure . Individuals possessing P14416 -141Cdel variant experienced more visuomotor problems . Protein kinase A alterations in endocrine tumors . Various molecular and cellular alterations of the cyclic adenosine monophosphate ( DB02527 ) pathway have been observed in endocrine tumors . Since protein kinase A ( PKA ) is a central key component of the DB02527 pathway , studies of the alterations of PKA subunits in endocrine tumors reveal new aspects of the mechanisms of DB02527 pathway alterations in human diseases . So far , most alterations have been observed for the regulatory subunits , mainly P10644 and to a lower extent , P31323 . One of the best examples of such alteration today is the multiple neoplasia syndrome Carney complex ( CNC ) . The most common endocrine gland manifestations of CNC are pituitary GH-secreting adenomas , thyroid tumors , testicular tumors , and DB01285 -independent Cushing 's syndrome due to primary pigmented nodular adrenocortical disease ( PPNAD ) . Heterozygous germline inactivating mutations of the PKA regulatory subunit RIα gene ( P10644 ) are observed in about two-third of CNC patients , and also in patients with isolated PPNAD . P10644 is considered as a tumor suppressor gene . Interestingly , these mutations can also be observed as somatic alterations in sporadic endocrine tumors . More than 120 different P10644 mutations have been found today . Most of them lead to an unstable mutant mRNA , which will be degraded by nonsense mediated mRNA decay . In vitro and in vivo functional studies are in progress to understand the mechanisms of endocrine tumor development due to PKA regulatory subunits inactivation . P10644 mutations stimulate in most models PKA activity , mimicking in some way DB02527 pathway constitutive activation . Cross-talks with other signaling pathways summarized in this review have been described and might participate in endocrine tumorigenesis . p-Chloroamphetamine , a serotonin-releasing drug , elicited in rats a hyperglycemia mediated by the P08908 and P41595 /2C receptors . The effects of a serotonin ( 5-HT ) releasing drug , p-chloroamphetamine , on plasma glucose levels were investigated in rats . p-Chloroamphetamine elicited a significant hyperglycemia . The hyperglycemic effects of p-chloroamphetamine were completely prevented by the 5-HT synthesis inhibitor , p-chlorophenylalanine . Prior adrenodemedullation abolished the hyperglycemia elicited by p-chloroamphetamine . p-Chloroamphetamine-induced hyperglycemia was prevented by methysergide , which blocks the 5-HT1 and 5-HT2 receptor , the P08908 /1B/2C receptor antagonist , (-)-propranolol , the selective P08908 receptor antagonist , 4- ( 2'-methoxyphenyl-1- [ 2'-n-2 " pyridinyl ) -p-iodobenzamido ] -ethyl-pi perazine ( p-MPPI ) , the 5- Q13049 /2B/2C receptor antagonists , ritanserin and 4-isopropyl-7-methyl-9-(2-hydroxy-1-methyl-propoxycarbonyl)-4,6A,7 , 8,9,10,10A-octahydro-indolo[4,3-FG]quinolone maleate ( LY 53857 ) . However , the 5- Q9H205 and Q13639 receptor antagonist , tropisetron , the Q13639 receptor antagonist , 2-methoxy-4-amino-5-chloro-benzoic acid 2-(diethylamino) ethyl ester ( SDZ 205-557 ) , and the 5- Q13049 receptor antagonist , ketanserin , did not affect the p-chloroamphetamine-induced hyperglycemia . These results suggest that p-chloroamphetamine-induced hyperglycemia is elicited by an enhanced 5-HT release and facilitated adrenaline release . Moreover , our results indicate that p-chloroamphetamine-induced hyperglycemia is mediated by P08908 and P41595 /2C receptors . DB01404 saponin metabolite induces apoptosis in MCF-7 breast cancer cells through the modulation of AMP-activated protein kinase . Previous studies have shown that the ginseng saponin metabolite , Compound K ( 20-O-d-glucopyranosyl-20(S)-protopanaxadiol , IH901 ) , suppresses proliferation of various cancers and induces apoptosis . AMP-activated protein kinase ( AMPK ) is a sensor of cellular energy states and is involved in apoptosis of cancer cells . We hypothesized that Compound K may exert cytotoxicity in MCF-7 human breast cancer cells through modulation of AMPK , followed by a decrease in cyclooxygenase-2 ( P35354 ) expression . Compound K inhibited cell growth , induced apoptosis via generation of reactive oxygen species ( ROS ) , as well as decreasing P35354 expression and prostaglandin E(2) ( PGE(2) ) levels . These effects of Compound K were induced via an AMPK-dependent pathway and were abrogated by a specific AMPK inhibitor . These results suggest that Compound K induced apoptosis by modulating AMPK- P35354 signaling in MCF-7 human breast cancer cells . The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT/ P31749 phosphorylation . DB01200 ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 , official symbol Q01959 ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract . The in vitro pharmacological profile of DB05655 , a selective 5-HT(4) receptor agonist with high intrinsic activity . The in vitro pharmacological profile of DB05655 , a novel , selective 5-HT(4) receptor agonist , was compared to that of clinically efficacious gastroprokinetic 5-HT(4) receptor agonists . DB05655 produced an elevation of cyclic adenosine monophosphate in human embryonic kidney 293 cells expressing the human recombinant 5-HT(4(c)) ( h5-HT(4(c)) ) receptor ( pEC(50) = 8.3 ) and 5-HT(4) receptor-mediated relaxation of the rat esophagus ( pEC(50) = 7.9 ) and contraction of the guinea pig colon ( pEC(50) = 7.9 ) . In all in vitro assays , DB05655 was a high intrinsic activity agonist , unlike tegaserod , mosapride , and cisapride which , in the majority of test systems , had lower intrinsic activity . DB05655 had high affinity ( pK ( i ) = 7.7 ) and selectivity ( > or =25-fold ) for h5-HT(4(c)) receptors over other biogenic amine receptors . DB05655 was > 500-fold selective over other 5-HT receptors ( including h5-HT(2B) and h5-HT(3A) ) and , at 3 microM , had no effect on human ether-à-go-go-related gene K+ channels . In conclusion , DB05655 is a selective 5-HT(4) receptor agonist in vitro . The high intrinsic activity and preferential binding of DB05655 to Q13639 over other 5-HT receptors may result in an improved clinical profile for the treatment of gastrointestinal disorders of reduced motility . Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility . Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor ( P14416 ) in flexible behavioral adaptation , mostly shown in reward-based learning paradigms . Recent evidence from imaging genetics indicates that also intentional cognitive flexibility , associated with lateral frontal cortex , is affected by variations in P14416 signaling . In the present functional magnetic resonance imaging ( Q9BWK5 ) study , we tested the effects of a direct pharmacological manipulation of P14416 stimulation on intentional flexibility in a task-switching context , requiring switches between cognitive task rules and between response hands . In a double blind , counterbalanced design , participants received either a low dose of the P14416 agonist bromocriptine or a placebo in two separate sessions . DB01200 modulated the blood-oxygen-level-dependent ( BOLD ) signal during rule switching : rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo , at comparable performance levels . Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions . Hand-switching-related activity , in contrast , was reduced under bromocriptine in sensorimotor regions . Our results provide converging evidence for an involvement of P14416 signaling in fronto-striatal mechanisms underlying intentional flexibility , and indicate that the neural mechanisms underlying different types of flexibility ( cognitive vs motor ) are affected differently by increased dopaminergic stimulation . Phosphorylation of Akt/GSK-3β/ P29474 amplifies P41595 receptor blockade mediated anti-hypertrophic effect in rats . Herein , we studied the cross talk between 5-HT(2B) receptor blocker ( SB-204741 ) and GSK-3β inhibitor ( SB-216763 ) in isoproterenol-induced cardiac hypertrophy for 28 days . SB-204741 treatment significantly ameliorated ( P < 0.05 ) myocardial dysfunction , myocyte area , fibrosis and myocardial architecture in isoproterenol insulted myocardium . Moreover , this improvement in functional and morphological changes was associated with suppression of hypertrophic ( DB04899 and CK-MB ) , inflammatory ( IKK-β/NF-κB/ P01375 -α and CRP ) , and apoptotic markers ( TUNEL positivity and Bax expression ) along with phosphorylation of Akt/GSK-3β/β-catenin/ P29474 . Intriguingly , co-treatment with GSK-3β inhibitor ( P < 0.01 ) further amplified the anti-hypertrophic effect of SB-204741 ( P < 0.05 ) such that the effect was indistinguishable from that of vehicle treated rats . Thus , 5-HT(2B) receptor blockade mediated anti-hypertrophic effect is atleast in part is governed through phosphorylation of Akt/GSK-3β/β-catenin/ P29474 via attenuating inflammatory and apoptotic pathways . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . Peroxidation radical formation and regiospecificity of recombinated Anabaena sp. lipoxygenase and its effect on modifying wheat proteins . Peroxidation radical formation and the regiospecificity of recombinated lipoxygenase from Anabaena sp . PCC7120 ( ana-rLOX ) were characterized by using P03372 and HPLC-MS . It was found that ana-rLOX oxygenated at the C-13 position of the substrate linoleic acid ( LA ) ; at C-13 and C-16 of α-linolenic acid ( ALA ) ; at C-9 , C-12 , and C-15 of arachidonic acid ( AA ) ; at C-12 , C-15 , and C-18 of eicosapentaenoic acid ( EPA ) ; and at C-14 and C-16 of docosahexaenoic acid ( DB01708 ) , respectively . A total of 7 , 14 , 30 , 28 , and 18 radical adducts for LA , ALA , AA , EPA , and DB01708 were respectively identified by HPLC-MS . The functional characteristics of wheat protein , such as foaming capacity ( FC ) , foam stability ( FS ) , emulsifying activity index ( EAI ) , emulsifying stability index ( P19957 ) , increased with enzymatic reactions . However , the average particle size of wheat proteins decreased with addition of ana-rLOX/LA . The ana-rLOX was also positivele effective in improving dough properties . These results provided clear evidence that ana-rLOX from Anabaena sp. could effectively improve the quality of wheat flour , which suggested that the enzyme could be applied as flour improver . Pharmacological characterization of mitogen-activated protein kinase activation by recombinant human P28335 , 5- Q13049 , and P41595 receptors . The type 2 serotonin ( 5-HT(2) ) receptor subfamily is known to couple to phosphoinositide hydrolysis ( PI ) and the subsequent mobilization of intracellular Ca(2+) , as well as the release of arachidonic acid ( AA ) . Less is known of 5-HT(2)-mediated activation of the mitogen-activated protein kinase ( MAPK ) or extracellular signal-regulated kinase ( P27361 /2 ) signaling . The present study measured the relative efficacies and potencies of 5-HT agonists to activate P28482 in non-neuronal cells expressing recombinant human 5-HT(2A) , 5-HT(2B) , and 5-HT(2C(ISV)) receptors . 5-HT agonists stimulated P28482 activity via all three 5-HT(2) subtypes . There were no meaningful differences in the potencies or relative efficacies of these agonists to affect P28482 activity vs. PI accumulation or Ca(2+) mobilization , suggesting that these pathways may be sequentially linked . Indeed , P28482 activity was very sensitive to PKC inhibition and calcium chelation and insensitive to tyrosine kinase and P19957 -kinase inhibition . 5-HT(2) receptors efficiently couple to MAPK activation via sequential PI hydrolysis , and Ca(2+) mobilization . This profile differs from reports of " agonist-directed trafficking of receptor stimulus " between PI/Ca(2+) and AA pathways activated by 5-HT(2) receptors . Q07973 as a potential target for cancer therapy . Increasing evidence has accumulated to suggest that vitamin D may reduce the risk of cancer through its biologically active metabolite , DB00136 , which inhibits proliferation and angiogenesis , induces differentiation and apoptosis , and regulates many other cellular functions . Thus , it is plausible to assume that rapid clearance of DB00136 by highly expressed Q07973 could interrupt the normal physiology of cells and might be one cause of cancer initiation and progression . In fact , enhancement of Q07973 expression has been reported in literature for many cancers . Based on these findings , Q07973 -specific inhibitors and vitamin D analogs which are resistant to Q07973 -dependent catabolism might be useful for cancer treatment . Q07973 -specific inhibitor VID400 , which is an azole compound , markedly enhanced and prolonged the antiproliferative activity of DB00136 in the human keratinocytes . Likewise , Q07973 -resistant analogs such as 2α-(3-hydroxypropoxy)- DB00136 ( O2C3 ) and its P06681 -epimer ED-71 ( DB05295 ) , and 19nor- 2α-(3-hydroxypropyl)- DB00136 ( MART-10 ) showed potent biological effects . Our in vivo studies using rats revealed that MART-10 had a low calcemic effect , which is a suitable property as an anticancer drug . Much lower affinity of MART-10 for vitamin D binding protein ( DBP ) as compared with DB00136 may be related to its more potent cellular activities . Based on these results , we conclude that ( 1 ) high affinity for P11473 , ( 2 ) resistance to Q07973 -dependent catabolism , ( 3 ) low affinity for DBP , and ( 4 ) low calcemic effect may be required for designing potent vitamin D analogs for cancer treatment . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Neurotransmitter enzymes in telencephalon , brain stem and cerebellum during the entire life span of the mouse . Neurochemical analysis of neuronal function was undertaken by measuring the activities of cholinacetyltransferase ( CAT ) , acetylcholinesterase ( P22303 ) , and glutamic acid decarboxylase ( Q99259 ) , in the telencephalon , brain stem and cerebellum of the mouse . Cholinergic activity was first expressed in the 10-day embryonic brain stem , which showed a relatively high CAT activity at birth . Postnatal brain stem development was characterized by a rapid and parallel increase in CAT and P22303 . Although P22303 peaked at 1 month , CAT activity was no achieved until 1 year . DB03128 synthesis was initiated in the 12-day embryonic telencephalon and a steady age-related increase in CAT was maintained until birth . A lag in both CAT and P22303 activities was recorded during the first week of postnatal telencephalon development . Cerebellar CAT was low at birth , and increased irregularly to reach a maximum by 1 month . In contrast , postnatal cerebellar P22303 activity increased steadily over the same time period . The GABA-ergic neuronal system matured rapidly in each brain region , and was unaffected by aging . Although the brain stem precociously expressed cholinergic activity , it wa the region most susceptible to deterioration during aging . Telencephalon CAT activity was unaffected by aging and in the cerebellum , a significantly reduced level of CAT was only found in truly senescent animals . The decreased cholinergic function during senescence was not due to either increased proteolysis or to alteration in the molecular form of the cholinergic enzymes . Structural elucidation of DB00007 and its analogues in solution : insight into their bioactive conformation . DB00007 [ DLeu6 , NHEt10 ] DB00644 , a potent gonadotropin-releasing hormone ( DB00644 ) agonist , is used in a wide variety of hormone-related diseases like cancer and endometriosis . In this report , the conformational behaviour of DB00007 and its linear synthetic analogues , namely [ Tyr5(OMe) , DLeu6 , Aze9 , NHEt10 ] DB00644 ( 1 ) and [ Tyr5(OMe) , DLeu6 , NHEt10 ] DB00644 ( 2 ) have been studied in DB01093 and H2O solutions by means of 2D nuclear magnetic resonance ( NMR ) experiments and detailed molecular dynamics ( MD ) simulations . The aim was to identify the conformational requirements of DB00644 analogues for agonistic activity . This approach is of value as no crystallographic data are available for the P30968 ( G protein-coupled receptor , GPCR ) . The NOE data indicate the existence of a β-turn type I in the 2-5 segments of DB00007 and its linear analogues in the case of using DB01093 -d6 as solvent , whereas a β-turn type II in the 3-6 segments is indicated using D2O as solvent . The final structures fulfil the conformational requirements that are known , in the literature , to play a significant role in receptor recognition and activation . Finally , the linear analogues ( 1 ) and ( 2 ) are biologically active when tested against the human breast cancer cell line , MCF-7 . Differential regulation of the serotonin 1 A transcriptional modulators five prime repressor element under dual repression-1 and nuclear-deformed epidermal autoregulatory factor by chronic stress . Chronic stress is known to affect brain areas involved in learning and emotional responses . These changes , thought to be related to the development of cognitive deficits are evident in major depressive disorder and other stress-related pathophysiologies . The serotonin-related transcription factors ( Q6P1N0 / Q6P1N0 ; five prime repressor element under dual repression/coiled-coil P06681 domain 1a , and O75398 /Deaf-1 ; nuclear-deformed epidermal autoregulatory factor ) are two important regulators of the P08908 receptor . Using Western blotting and quantitative real-time polymerase chain reaction ( qPCR ) we examined the expression of mRNA and proteins for Q6P1N0 , O75398 , and the P08908 receptor in the prefrontal cortex ( P27918 ) of male rats exposed to chronic restraint stress ( CRS ; 6 h/day for 21 days ) . After 21 days of CRS , significant reductions in both Q6P1N0 mRNA and protein were observed in the P27918 ( 36.8 % and 32 % , respectively ; P < 0.001 ) , while the levels of both O75398 protein and mRNA did not change significantly . Consistent with reduced Q6P1N0 protein , P08908 receptor mRNA levels were equally upregulated in the P27918 , while protein levels actually declined , suggesting post-transcriptional receptor downregulation . The data suggest that CRS produces distinct alterations in the serotonin system specifically altering Q6P1N0 and the P08908 receptor in the P27918 of the male rat while having no effect on O75398 . These results point to the importance of understanding the mechanism for the differential regulation of Q6P1N0 and O75398 in the P27918 as a basis for understanding the related effects of chronic stress on the serotonin system ( serotonin-related transcription factors ) and stress-related disorders like depression . Effects of statins on nitric oxide/cGMP signaling in human umbilical vein endothelial cells . Human umbilical vein endothelial cells ( HUVECs ) were established as in vitro models for the modulation of endothelial function and cell viability by statins . Emphasis was placed on the biphasic effects of the drugs on nitric oxide ( NO ) bioavailability and cytotoxicity , as well as drug interference with the interaction of endothelial NO synthase ( P29474 ) with caveolin-1 ( Cav-1 ) . Incubation of HUVECs with fluvastatin , lovastatin or cerivastatin for 24 h caused an approximately 3-fold upregulation of P29474 expression that was associated with increased P29474 activity and accumulation of cGMP . DB00439 exhibited the highest potency with an EC50 of 13.8 +/- 2 nM after 24 h , while having no effect after only 30 min . The effects of statins on P29474 expression were similar in control and Cav-1 knockdown cells , but the increase in P29474 activity was less pronounced in Cav-1-deficient cells . Statin-triggered cytotoxicity occurred at approximately 10-fold higher drug concentrations ( maximal toxicity at 1-10 microM ) , was sensitive to mevalonate , and was significantly enhanced in the presence of NG-nitro-L-arginine . The overexpression of P29474 induced by clinically relevant concentrations of statins may contribute to the beneficial vascular effects of the drugs in patients . Stimulation of NO synthesis and cytotoxicity appear to share a common initial mechanism but involve distinct downstream signaling cascades that exhibit differential sensitivity to P04035 inhibition .

MH_train_1236

MH_train_1236

MH_train_1236

interacts_with DB06287?

multiple_choice

Epileptogenesis and reduced inward rectifier potassium current in tuberous sclerosis complex-1-deficient astrocytes . PURPOSE : Individuals with tuberous sclerosis complex ( TSC ) frequently have intractable epilepsy . To gain insights into mechanisms of epileptogenesis in TSC , we previously developed a mouse model of TSC with conditional inactivation of the Tsc1 gene in glia ( Tsc1( P14136 )CKO mice ) . These mice develop progressive seizures , suggesting that glial dysfunction may be involved in epileptogenesis in TSC . Here , we investigated the hypothesis that impairment of potassium uptake through astrocyte inward rectifier potassium ( Kir ) channels may contribute to epileptogenesis in Tsc1( P14136 )CKO mice . METHODS : Kir channel function and expression were examined in cultured Tsc1-deficient astrocytes . Kir mRNA expression was analyzed in astrocytes microdissected from neocortical sections of Tsc1( P14136 )CKO mice . Physiological assays of astrocyte Kir currents and susceptibility to epileptiform activity induced by increased extracellular potassium were further studied in situ in hippocampal slices . RESULTS : Cultured Tsc1-deficient astrocytes exhibited reduced Kir currents and decreased expression of specific Kir channel protein subunits , Kir2.1 and Kir6.1 . mRNA expression of the same Kir subunits also was reduced in astrocytes from neocortex of Tsc1( P14136 )CKO mice . By using pharmacologic modulators of signalling pathways implicated in TSC , we showed that the impairment in Kir channel function was not affected by rapamycin inhibition of the P42345 /S6K pathway , but was reversed by decreasing P24941 activity with roscovitine or retinoic acid . Last , hippocampal slices from Tsc1( P14136 )CKO mice exhibited decreased astrocytic Kir currents , as well as increased susceptibility to potassium-induced epileptiform activity . CONCLUSIONS : Impaired extracellular potassium uptake by astrocytes through Kir channels may contribute to neuronal hyperexcitability and epileptogenesis in a mouse model of TSC . Retinoid-binding proteins in plasma and in cells . Much has been learned during the past decade about the specific retinoid-binding proteins that exist in plasma , and in the intracellular compartment in a number of tissues . DB00162 is mobilized from liver stores and transported in plasma in the form of the lipid alcohol retinol , bound to a specific transport protein , retinol-binding protein ( P02753 ) . A great deal is now known about the chemical structure , metabolism , and biological roles of P02753 . DB00162 mobilization from the liver is highly regulated by factors that control the rates of P02753 production and secretion . DB00162 deficiency specifically blocks the secretion of P02753 , which can then be rapidly stimulated by intravenous retinol repletion . The cellular and molecular mechanisms that mediate these phenomena are under investigation . Delivery of retinol to peripheral tissues appears to involve specific cell surface receptors for P02753 . The retinol so delivered enters the target cell , where it may become associated with the intracellular binding protein for retinol ( P09455 ) . A number of tissues of rats , humans , and other species contain soluble proteins with binding specificity for retinol ( P09455 ) or for retinoic acid ( CRABP ) . These proteins have been purified from several tissues and partly characterized . They differ in a number of ways from plasma P02753 , and differ from each other in regard to binding specificity and immunoreactivity . It has been suggested that these intracellular proteins may play a direct role in the biological expression of vitamin A activity in the cell . Studies are in progress to explore this and other possibilities . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . PDE4B5 , a novel , super-short , brain-specific DB02527 phosphodiesterase-4 variant whose isoform-specifying N-terminal region is identical to that of DB02527 phosphodiesterase-4D6 ( PDE4D6 ) . The DB02527 -specific phosphodiesterase-4 ( DB05876 ) gene family is the target of several potential selective therapeutic inhibitors . The four DB05876 genes generate several distinct protein-coding isoforms through the use of alternative promoters and 5'-coding exons . Using mouse transcripts , we identified a novel , super-short isoform of human Q07343 encoding a novel 5' terminus , which we label PDE4B5 . The protein-coding region of the novel 5' exon is conserved across vertebrates , chicken , zebrafish , and fugu . Reverse-transcription-polymerase chain reaction ( PCR ) and quantitative ( PCR ) measurements show that this isoform is brain-specific . The novel protein is 58 +/- 2 kDa ; it has DB02527 hydrolyzing enzymatic activity and is inhibited by DB05876 -selective inhibitors rolipram and cilomilast ( DB03849 ) . Confocal and subcellular fractionation analyses show that it is distributed predominantly and unevenly within the cytosol . The 16 novel N-terminal residues of PDE4B5 are identical to the 16 N-terminal residues of the super-short isoform of Q08499 ( PDE4D6 ) , which is also brain-specific . PDE4B5 is able to bind the scaffold protein Q9NRI5 , whose gene has been linked to schizophrenia . Microarray expression profiling of the DB05876 gene family shows that specific DB05876 genes are enriched in muscle and blood fractions ; however , only by monitoring the individual isoforms is the brain specificity of the super-short Q08499 and Q07343 isoforms revealed . Understanding the distinct tissue specificity of DB05876 isoforms will be important for understanding phosphodiesterase biology and opportunities for therapeutic intervention . Predicting the possibility of two newly isolated phenetheren ring containing compounds from Aristolochia manshuriensis as P24941 inhibitors . Aristolochia manshuriensis has been used for centuries in Chinese medicinal system for their versatile medicinal uses . Recent studies have revealed two new aristolactames ( compound A and B ) with γ-lactame ring fused with the phenentherene ring as potent inhibitors of human Cycline Dependent Kinase2 ( P24941 ) . Studies on aristolactames and related compounds claim for their P24941 inhibition without delineating the involved mechanism and structural basis of interaction . Molecular structural model was used to we propose a structural basis of P24941 inhibition . We showed that these compounds ( A and B ) can successfully dock into the inhibitor binding pockets of human P24941 . Predicted binding affinities are comparable to known inhibitors of P24941 . Results were in agreement with the earlier biochemical studies . Hence , suggest that studied compounds A and B can be a promising scaffold for rational design of novel and potential drugs against cancer . ABBREVIATIONS : P24941 - cyclin-dependent kinase 2 , OLO - DB02116 , NW1 - Cyclohexylmethyloxy-5-Nitroso-Pyrimidine- 2 , 4-Diamine , CMG - DB02407 . Immoderate inhibition of estrogen by anastrozole enhances the severity of experimental polyarthritis . P11511 inhibitors have become the standard of care for the adjuvant treatment of postmenopausal , hormone-sensitive breast cancer . Meanwhile , more and more breast cancer patients who are treated with aromatase inhibitors as adjuvant therapies often experience arthralgias and musculoskeletal aching , in some cases , have necessitated discontinuation of treatment . We therefore use a rat model of human RA to test the hypothesis that anastrozole , an aromatase inhibitor , could enhance arthritis . The parameters used for analyzing the disease severity included paw volume , radiology , histopathological examination , markers for cytokine profile , immunophenotypic assays , and immune response to type II collagen . Administration of anastrozole significantly increased the severity of arthritis . DB01217 induced the increased levels of proinflammatory cytokines , P01579 , IL-12 , and the decreased levels of P05112 , P22301 secretion . We further found that anastrozole suppressed the differentiation of naive T cells to Treg cells , and it blocked the balance of IgG2a/IgG1 in peripheral blood . Meanwhile , estradiol concentration was the lowest in the anastrozole group . In a well-established model of postmenopausal RA , anastrozole potently promote the progression of arthritis and the associated development of osteoporosis . This potential problem should alert the oncologists and other health professionals . The clinical potential of temsirolimus in second or later lines of treatment for metastatic renal cell carcinoma . An impressive variety of targeted therapies has been approved for the treatment of metastatic renal cell carcinoma ( mRCC ) . Despite promising progress , there are still unmet clinical needs . The optimal sequence of these agents in the therapeutic setting has not yet been determined . Most available data address first- and second-line therapy of clear cell RCC . The P42345 inhibitor temsirolimus has been approved for first-line treatment of mRCC , and there are new data addressing the use of temsirolimus in later therapeutic lines . DB06287 has discerning features compared with other currently registered drugs , such as its intravenous administration route-providing predictable bioavailability and adherence to treatment-and potential benefit in nonclear cell histologies . Here , we review the available literature on temsirolimus , with reference to data also available for everolimus , to determine its clinical potential in mRCC . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Decreased expression of nucleophosmin/B23 increases drug sensitivity of adriamycin-resistant Molt-4 leukemia cells through mdr-1 regulation and Akt/ P42345 signaling . P06748 /B23 ( P06748 ) is a nuclear protein with prosurvival and ribosomal RNA processing functions . However , the potential role of P06748 involved in drug-resistance in leukemia has not been investigated clearly . In this study , we generated an adriamycin ( P35318 ) -resistant lymphoblastic cell line Molt-4/ADR ( MAR ) by stepwise induction . Cell proliferation , sensitivity to chemotherapy agents and expressions of drug resistance related molecules were assessed . The IC50 of Molt-4 cells were 0.58±0.11μmol/L and MAR cells were 22.56±1.94μmol/L , meaning MAR cells were 38.63 fold resistant to Molt-4 cells . Furthermore , MAR cells gained an expression of mdr-1 ( P-gp ) and a higher expression of P06748 compared to Molt-4 cells . Knockdown of P06748 by RNA interference ( RNAi ) suppressed the viability of both Molt-4 and MAR cells . After P06748 RNAi , the IC50 of MAR and Molt-4 cells were 3.83±0.38μmol/L and 0.19±0.02μmol/L respectively . Both of them revealed an increase of drug sensitivity with down-regulation of mdr-1 and Akt/ P42345 signaling . Knockdown of mdr-1 could also reverse the drug resistance , with no change in P06748 expression . It could be concluded that knockdown of P06748 reversed the drug resistance by down-regulating P-gp and Akt/ P42345 signal pathway , indicating that P06748 may serve as a potential modulator in drug resistance . Exogenous cell-permeable P13671 ceramide sensitizes multiple cancer cell lines to Doxorubicin-induced apoptosis by promoting AMPK activation and mTORC1 inhibition . New chemotherapy-enhancing strategies are needed for better cancer therapy . Previous studies suggest that exogenous cell-permeable P13671 ceramide may be a useful adjunct to the anti-tumor effects of chemotherapeutic agents ( such as DB01229 ) against multiple cancers . Here we demonstrate that exogenous cell-permeable P13671 ceramide largely sensitizes multiple progressive cancer cell lines to Doxorubicin-induced cell death and apoptosis . We found for the first time that Doxorubicin induces AMP-activated protein kinase ( AMPK ) activation in a reactive oxygen species-dependent manner . Activation of AMPK contributes to Doxorubicin-induced cancer cell death and apoptosis . Inhibition of AMPK by small interfering RNA knockdown or a pharmacological inhibitor reduces Doxorubicin-induced cancer cell apoptosis , whereas AMPK activator AICAR enhances it . Importantly , we found that P13671 ceramide largely enhances Doxorubicin-induced activation of AMPK , which leads to P42345 complex 1 inhibition and chemo-sensitization . Our data suggest that the combination of P13671 ceramide with traditional chemotherapy drugs such as Doxorubicin may have the potential to be used as a new therapeutic intervention against multiple cancers . Gene set enrichment analysis unveils the mechanism for the phosphodiesterase 4B control of glucocorticoid response in B-cell lymphoma . PURPOSE : Resistance to glucocorticoid ( GC ) is a significant problem in the clinical management of lymphoid malignancies . Addressing this issue via a mechanistic understanding of relevant signaling pathways is more likely to yield positive outcomes . EXPERIMENTAL DESIGN : We used gene set enrichment analysis ( GSEA ) , multiple genetic models of gain and loss of function in B-cell lymphoma cell lines , in vitro and in vivo , and primary patient samples to characterize a novel relationship between the cyclic AMP/phosphodiesterase 4B ( DB02527 / Q07343 ) , AKT/ P42345 activities , and GC responses . RESULTS : Starting from the GSEA , we found that overexpression of the Q07343 in diffuse large B-cell lymphoma ( DLBCL ) impinge on the same genes/pathways that are abnormally active in GC-resistant tumors . We used genetically modified cell lines to show that Q07343 modulates DB02527 inhibitory activities toward the AKT/ P42345 pathway and defines GC resistance in DLBCL . In agreement with these data , pharmacologic inhibition of DB05876 in a xenograft model of human lymphoma unleashed DB02527 effects , inhibited AKT , and restored GC sensitivity . Finally , we used primary DLBCL samples to confirm the clinical relevance and biomarker potential of AKT/ P42345 regulation by Q07343 . CONCLUSIONS : Together , these data mechanistically elucidated how DB02527 modulates GC responses in lymphocytes , defined AKT as the principal transducer of the growth inhibitory effects of DB02527 in B cells , and allowed the formulation of genomics-guided clinical trials that test the ability of DB05876 inhibitors to restore GC sensitivity and improve the outcome of patients with B-cell malignancies . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . Evaluation of the antitumor effects and mechanisms of PF00299804 , a pan-HER inhibitor , alone or in combination with chemotherapy or targeted agents in gastric cancer . Recently , P04626 -directed treatment , such as trastuzumab , has shown clinical benefit in P04626 -amplified gastric cancer . On the basis of recent studies about epidermal growth factor receptor ( P00533 ) or P04626 -targeting agents ( including gefitinib , lapatinib , and trastuzumab ) in gastric cancer , the potent effects of pan-HER inhibitors targeting the HER family are anticipated . In this study , we evaluated the activity and mechanisms of PF00299804 , an irreversible pan-HER inhibitor , in gastric cancer in vitro and in vivo models . PF00299804 showed significant growth-inhibitory effects in P04626 -amplified gastric cancer cells ( SNU216 , N87 ) , and it had lower 50 % inhibitory concentration values compared with other P00533 tyrosine kinase inhibitors , including gefitinib , lapatinib , BIBW-2992 , and DB05424 . PF00299804 induced apoptosis and G(1) arrest and inhibited phosphorylation of receptors in the HER family and downstream signaling pathways including P40763 , AKT , and extracellular signal-regulated kinases ( P29323 ) in P04626 -amplified gastric cancer cells . PF00299804 also blocked P00533 / P04626 , P04626 / P21860 , and P21860 / Q15303 heterodimer formation as well as the association of P21860 with p85α in SNU216 cells . The combination of PF00299804 with clinically relevant chemotherapeutic agents or molecular-targeted agents including trastuzumab ( an anti- P04626 monoclonal antibody ) , CP751871 ( an P08069 inhibitor ) , PD0325901 ( an P27361 /2 inhibitor ) , and PF04691502 ( a PI3K/ P42345 inhibitor ) produced synergistic effects . These findings indicate that PF00299804 can be used as a targeted therapy for the treatment of P04626 -amplified gastric cancer through inhibition of HER family heterodimer formation and may augment antitumor efficacy of chemotherapeutic and/or molecular-targeted agents . DB00162 binding protein 4 affects the adipogenesis of porcine preadipocytes through insulin signaling pathways . DB00162 binding protein 4 ( P02753 ) , a novel cytokine , is mainly secreted by hepatocytes and adipocytes . P02753 reportedly induces insulin resistance and P02753 secretion is increased in the adipocytes of animals or humans with type 2 diabetes , obesity , and metabolic syndrome , but its role in preadipocyte differentiation remains unclear . In this study , we investigated the effect of P02753 on the differentiation of porcine preadipocytes into adipocytes . The results suggest that P02753 significantly suppresses the differentiation of porcine preadipocytes into adipocytes , including those treated with the hormone cocktail methylisobutylxanthine-dexamethasone-insulin . P02753 also weakened the activity of normal threonine 308 , the phosphorylation of serine/threonine kinase AKT , and downstream insulin signaling , including the mammalian target of rapamycin ( P42345 ) and β-catenin . Moreover , the activation of insulin signaling mediated by knockdown P02753 in porcine preadipocytes was recovered in the suppression of LY294002 . P02753 also had a suppressive effect on the differentiation of porcine preadipocytes by decreasing the activation of insulin signaling pathways . O14974 and catalytic subunit δ , new members in the phosphatidylinositide 3 kinase insulin-signaling pathway . Skeletal muscle insulin resistance is an early abnormality in individuals with metabolic syndrome and type 2 diabetes ( T2D ) . P06213 substrate-1 ( P35568 ) plays a key role in insulin signaling , the function of which is regulated by both phosphorylation and dephosphorylation of tyrosine and serine/threonine residues . Numerous studies have focused on kinases in P35568 phosphorylation and insulin resistance ; however , the mechanism for serine/threonine phosphatase action in insulin signaling is largely unknown . Recently , we identified protein phosphatase 1 ( P50391 ) regulatory subunit 12A ( O14974 ) as a novel endogenous insulin-stimulated interaction partner of P35568 in Q9BTT4 myotubes . The current study was undertaken to better understand O14974 's role in insulin signaling . P01308 stimulation promoted an interaction between the P35568 /p85 complex and O14974 ; however , p85 and O14974 did not interact independent of P35568 . Moreover , kinase inhibition experiments indicated that insulin-induced interaction between P35568 and O14974 was reduced by treatment with inhibitors of phosphatidylinositide 3 kinase , PDK1 , Akt , and P42345 /raptor but not MAPK . Furthermore , a novel insulin-stimulated P35568 interaction partner , P50391 catalytic subunit ( PP1cδ ) , was identified , and its interaction with P35568 was also disrupted by inhibitors of Akt and P42345 /raptor . These results indicate that O14974 and PP1cδ are new members of the insulin-stimulated P35568 signaling complex , and the interaction of O14974 and PP1cδ with P35568 is dependent on Akt and P42345 /raptor activation . These findings provide evidence for the involvement of a particular P50391 complex , O14974 /PP1cδ , in insulin signaling and may lead to a better understanding of dysregulated P35568 phosphorylation in insulin resistance and T2D . Electrostatic steering at acetylcholine binding sites . The electrostatic environments near the acetylcholine binding sites on the nicotinic acetylcholine receptor ( nAChR ) and acetylcholinesterase were measured by diffusion-enhanced fluorescence energy transfer ( DEFET ) to determine the influence of long-range electrostatic interactions on ligand binding kinetics and net binding energy . Changes in DEFET from variously charged Tb3+ -chelates revealed net potentials of -20 mV at the nAChR agonist sites and -14 mV at the entrance to the P22303 active site , in physiological ionic strength conditions . The potential at the alphadelta-binding site of the nAChR was determined independently in the presence of DB01199 to be -14 mV ; the calculated potential at the alphagamma-site was approximately threefold stronger than at the alphadelta-site . By determining the local potential in increasing ionic strength , Debye-Hückel theory predicted that the potentials near the nAChR agonist binding sites are constituted by one to three charges in close proximity to the binding site . Examination of the binding kinetics of the fluorescent acetylcholine analog dansyl- P13671 -choline at ionic strengths from 12.5 to 400 mM revealed a twofold decrease in association rate . Debye-Hückel analysis of the kinetics revealed a similar charge distribution as seen by changes in the potentials . To determine whether the experimentally determined potentials are reflected by continuum electrostatics calculations , solutions to the nonlinear Poisson-Boltzmann equation were used to compute the potentials expected from DEFET measurements from high-resolution models of the nAChR and P22303 . These calculations are in good agreement with the DEFET measurements for P22303 and for the alphagamma-site of the nAChR . We conclude that long-range electrostatic interactions contribute -0.3 and -1 kcal/mol to the binding energy at the nAChR alphadelta- and alphagamma-sites due to an increase in association rates . Type 1 insulin-like growth factor regulates P50281 synthesis and tumor invasion via PI 3-kinase/Akt signaling . The membrane type 1 matrix metalloproteinase ( P50281 ) has been identified as a major activator of P08253 - a process involving the formation of a trimolecular complex with P16035 . We previously identified the P08069 as a positive regulator of P08253 synthesis . Here , we investigated the role of IGF-IR in the regulation of P50281 . Highly invasive Lewis lung carcinoma subline H-59 cells express P50281 and utilize it to activate their major extracellular matrix degrading proteinase- P08253 . These cells were transiently transfected with a plasmid vector expressing a luciferase reporter gene downstream of the mouse P50281 promoter . P05019 treatment increased luciferase activity in the transfected cells by up to 10-fold and augmented endogenous P50281 mRNA and protein synthesis by up to 2-3-fold , relative to controls . P50281 induction and invasion were blocked by the PI 3-kinase inhibitors LY294002 and wortmannin and by rapamycin , but not by the MEK inhibitor PD98059 . Overexpression of a dominant negative Akt mutant or of the tumor suppressor phosphatase and tensin homologue , P60484 , in these cells also caused a significant reduction in P50281 expression and invasion . The results demonstrate that IGF-IR controls tumor cell invasion by coordinately regulating P08253 expression and its P50281 -mediated activation and identify PI 3-kinase/Akt/ P42345 signaling as critical to this regulation . Differential actions of vasopeptidase inhibition versus angiotensin-converting enzyme inhibition on diuretic therapy in experimental congestive heart failure . BACKGROUND : DB00886 ( OMA ) , a vasopeptidase inhibitor , simultaneously inhibits angiotensin-converting enzyme ( P12821 ) and neutral endopeptidase , which degrades vasodilatory factors ( eg , P35318 ) and natriuretic peptides . Based on the beneficial cardiorenal and humoral properties of the natriuretic peptides , we hypothesized that an acute vasopeptidase inhibitor with or without diuretic would result in more favorable cardiorenal and hormonal actions than P12821 inhibition plus diuretic ( ACEI+D ) in congestive heart failure . METHODS AND RESULTS : We compared the actions of OMA alone and with diuretic ( OMA+D ) to ACEI+D in a model of pacing-induced congestive heart failure . OMA+D decreased pulmonary arterial and pulmonary capillary wedge pressures to a greater level than OMA alone or ACEI+D . Glomerular filtration rate was lower with ACEI+D than with either OMA group . Plasma renin activity and aldosterone immediately increased with ACEI+D , whereas OMA+D resulted in higher plasma renin activity and a delayed increase in aldosterone . OMA alone did not increase plasma renin activity and aldosterone , but resulted in a sustained increase in plasma adrenomedullin , with higher urinary atrial natriuretic peptide , adrenomedullin , and cGMP excretions than with ACEI+D . CONCLUSIONS : Acute administration of OMA with or without diuretic results in more favorable cardiorenal and humoral responses in experimental congestive heart failure than does ACEI+D . There is no acute activation of renin and aldosterone with OMA alone such as occurs with ACEI+D and OMA+D . Thus , OMA with or without a diuretic possesses beneficial cardiorenal and humoral actions comparable to those observed with ACEI+D that can be explained by potentiation of natriuretic peptides . Interferon-gamma-induced dephosphorylation of P40763 and apoptosis are dependent on the P42345 pathway . Interferon-gamma ( P01579 ) exhibits diverse biological activities , including control of cell growth and tumor suppression . Here , we report that the treatment of M12 cells , a human metastatic prostate cancer cell line , with P01579 , resulted in marked inhibition of cell proliferation and induced apoptosis . These effects were not seen with either IFN-alpha or IFN-beta . M12 cells , like many other human cancer cells , contain constitutively activated signal transducer and activator of transcription 3 ( P40763 ) . The basal levels of both Akt and P27361 /2 phosphorylation are also markedly elevated in M12 cells . Strikingly , P01579 -induced apoptosis and growth inhibition of M12 cells were associated with persistent suppression of the constitutive tyrosine-phosphorylated P40763 ( pY- P40763 ) . The P01579 -induced dephosphorylation of pY- P40763 , however , was inhibited when the P42345 pathway was specifically blocked by rapamycin . Inhibition of PI-3K with low-dose LY294002 , or MAPK with PD98059 also suppressed the P42345 / P08133 S6k pathway , and correlated with the blockage of P01579 -induced dephosphorylation of pY- P40763 . Simultaneously , treatment with LY294002 , PD98059 , or rapamycin abolished P01579 -induced apoptosis in M12 cells . The inhibition of the P42345 pathway , however , did not affect P01579 -induced activation of P42224 pathway , and suppression of P42224 expression by siRNA had no effect on P01579 -induced dephosphorylation of pY- P40763 . Taken together , these results demonstrate that an intact P42345 pathway is critical for P01579 -induced suppression of pY- P40763 and apoptosis . Our study thus provides novel insights into the contributions of signaling pathways other than the classical JAK/ P42224 pathway in the anti-proliferative , proapoptotic actions of P01579 . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling .

MH_train_1237

MH_train_1237

MH_train_1237

interacts_with DB00316?

multiple_choice

Inhibition of P51587 and DB01643 Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 mediates homologous recombination repair , and P51587 polymorphisms increase cancer risk . However , tumors with P51587 mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 synergistically potentiated drugs with mechanisms of action related to P51587 function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality. " TS knockdown induced complementary lethality to TS-targeting drugs ( 5-FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 -targeting antisense oligdeoxynucleotide ( ASO ) " BR-1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi:10.1038/mtna.2013.7 published online 12 March 2013 . Direct intracerebral delivery of cintredekin besudotox ( P35225 -PE38QQR ) in recurrent malignant glioma : a report by the DB05305 Intraparenchymal Study Group . PURPOSE : Glioblastoma multiforme ( GBM ) is a devastating brain tumor with a median survival of 6 months after recurrence . Cintredekin besudotox ( CB ) is a recombinant protein consisting of interleukin-13 ( P35225 ) and a truncated form of Pseudomonas exotoxin ( PE38QQR ) . Convection-enhanced delivery ( DB01333 ) is a locoregional-administration method leading to high-tissue concentrations with large volume of distributions . We assessed the use of intracerebral DB01333 to deliver CB in patients with recurrent malignant glioma ( MG ) . PATIENTS AND METHODS : Three phase I clinical studies evaluated intracerebral DB01333 of CB along with tumor resection . The main objectives were to assess the tolerability of various concentrations and infusion durations ; tissue distribution ; and methods for optimizing delivery . All patients underwent tumor resection followed by a single intraparenchymal infusion ( in addition to the intraparenchymal one following resection ) , with a portion of patients who had a preresection intratumoral infusion . RESULTS : A total of 51 patients with MG were treated including 46 patients with GBM . The maximum tolerated intraparenchymal concentration was 0.5 microg/mL and tumor necrosis was observed at this concentration . Infusion durations of up to 6 days were well tolerated . Postoperative catheter placement appears to be important for optimal drug distribution . CB- and procedure-related adverse events were primarily limited to the CNS . Overall median survival for GBM patients is 42.7 weeks and 55.6 weeks for patients with optimally positioned catheters with patient follow-up extending beyond 5 years . CONCLUSION : CB appears to have a favorable risk-benefit profile . DB01333 is a complex delivery method requiring catheter placement via a second procedure to achieve accurate catheter positioning , better drug distribution , and better outcome . Targeting Aurora kinase-A downregulates cell proliferation and angiogenesis in neuroblastoma . PURPOSE : O14965 ( O14965 ) overexpression is associated with poor prognosis in neuroblastoma and has been described to upregulate P15692 in gastric cancer cells . However , the exact role of O14965 in the regulation of neuroblastoma tumorigenesis remains unknown . We hypothesize that O14965 -mediated stabilization of N-Myc may affect P15692 expression and angiogenesis in neuroblastoma . Therefore , we sought to determine whether inhibition of O14965 modulates neuroblastoma angiogenesis . METHODS : Cell viability and anchorage-independent growth were determined after silencing O14965 or after treatment with DB05220 , O14965 inhibitor . Immunofluorescence was used to determine N-Myc localization . Human umbilical vein endothelial cells ( HUVECs ) were used to assess angiogenesis in vitro . Real time-PCR and ELISA were performed to determine P15692 transcription and secretion , respectively . RESULTS : Knockdown of O14965 significantly reduced cell proliferation and inhibited anchorage-independent growth . It also decreased N-Myc protein levels and nuclear localization . O14965 inhibition also decreased HUVECs tubule formation along with P15692 transcription and secretion . Similarly , DB05220 treatment decreased neuroblastoma tumorigenicity in vitro . CONCLUSIONS : Our findings demonstrate that O14965 plays a critical role in neuroblastoma angiogenesis . O14965 regulates nuclear translocation of N-Myc in neuroblastoma cells , thus potentially affecting cell proliferation , anchorage-independent cell growth , and angiogenesis . Targeting O14965 might provide a novel therapeutic strategy in treating aggressive neuroblastomas . Molecular analysis of the effect of topical imiquimod treatment of HPV 2/27/57-induced common warts . Imiquimod is effective in the treatment of genital warts and clinical studies suggest activity against common warts as well . We have analyzed the effect of topical imiquimod on gene expression and virus load in human papilloma virus ( HPV ) 2/27/57-induced common warts . mRNA was extracted from keratinocyte culture , from normal skin , from three untreated common warts and from three common warts treated topically with 5 % imiquimod cream twice daily . Differential gene expression was demonstrated by RT-PCR and by cDNA microarray hybridization . We further analyzed viral DNA content in scales from three superficially pared imiquimod-treated warts by real-time PCR . Comparison of normal skin with wart tissue revealed that HPV 2/27/57 infection led to an induction of P05231 , P22301 and interferon-gamma inducible protein ( IP10 ) and to an up-regulation of TGF-beta . We could further detect expression of PCTAIRE-3 , Q93097 , frizzled-3 , notch-2 , notch-4 and P51587 in normal skin and common warts . Analysis of imiquimod-treated warts demonstrated that imiquimod enhanced P05231 expression and induced P10145 , GM- P04141 , P05109 and P06702 . It could also be shown that imiquimod led to an infiltration of wart tissue with macrophages and to a strong decrease of viral copy number in warts within 3 months of treatment . Our data thus provide molecular proof of principle for imiquimod treatment of cutaneous common warts . Q05901 c.-57A > G functional promoter change affects Parkinson 's disease and smoking . Cigarette smoking is protective in Parkinson 's disease ( PD ) , possibly because of nicotine action on brain nicotinic-acetylcholine receptors . The β3 nicotinic-acetylcholine receptor subunit ( encoded by Q05901 ) is depleted in the striatum of PD patients and associated with nicotine dependence . Herein , the Q05901 gene was sequenced , and the c.-57G allele frequency was 0.31 and 0.26 among patients ( n = 596 ) and controls ( n = 369 ) , respectively ( p = 0.02 , odds ratio = 1.33 , 95 % confidence interval = 1.03-1.73 ) . The c.-57G allele was strongly associated with smoking in patients , as 48.4 % of c.-57G carriers compared with 32.6 % of noncarriers reported smoking history ( p < 0.0001 ) . The transcription factor P14859 binding was almost eliminated in lymphoblasts with the c.-57G/G genotype , to only 6.5 % percent , and the Q05901 promoter activity was reduced in cells with the c.-57G/G genotype by 96 % -70 % . These findings suggest that the Q05901 c.-57A > G alteration affects the promoter activity and is associated with PD and smoking in PD patients . It is therefore possible that nicotine may be valuable for patients who carry this alteration and beneficial in PD only for patients with specific genotypes . A Patient with HIV Treated with DB06186 and Stereotactic Radiosurgery for Melanoma Metastases to the Brain . Cancers , such as melanoma , that are associated with immune deficiencies are a major cause of morbidity and mortality in HIV-infected patients . Once patients develop melanoma metastases to the brain , treatment is often limited to palliative surgery and/or radiation . DB06186 , a P16410 antagonist , has been shown to improve the median survival of patients with metastatic melanoma . However , available data regarding the safety and efficacy of ipilimumab in HIV-infected patients who develop intracranial melanoma metastases is limited . Here we report our experience administering ipilimumab to a patient with HIV-AIDS who developed multiple intracranial melanoma metastases . Following treatment , our patient showed improvement in systemic tumor control without any apparent interference with antiretroviral treatment . Mechanisms involved in the nociception produced by peripheral protein kinase c activation in mice . Protein kinase C ( PKC ) is able to phosphorylate several cellular components that serve as key regulatory components in signal transduction pathways of nociceptor excitation and sensitisation . Therefore , the present study attempted to assess some of the mechanisms involved in the overt nociception elicited by peripheral administration of the PKC activator , phorbol 12-myristate 13-acetate ( PMA ) , in mice . The intraplantar ( i.pl. ) injection of PMA ( 16-1600 pmol/paw ) , but not its inactive analogue alpha-PMA , produced a long-lasting overt nociception ( up to 45 min ) , as well as the activation of PKCalpha and PKCepsilon isoforms in treated paws . Indeed , the local administration of the PKC inhibitor GF109203X completely blocked PMA-induced nociception . The blockade of NK1 , P80511 , DB01221 , beta1-adrenergic , B2 or Q8NER1 receptors with selective antagonists partially decreased PMA-induced nociception . Similarly , P23219 , P35354 , MEK or p38 Q96HU1 kinase inhibitors reduced the nociceptive effect produced by PMA . Notably , the nociceptive effect promoted by PMA was diminished in animals treated with an antagonist of IL-1beta receptor or with antibodies against TNFalpha , P01138 or P23560 , but not against P39905 . Finally , mast cells as well as capsaicin-sensitive and sympathetic fibres , but not neutrophil influx , mediated the nociceptive effect produced by PMA . Collectively , the results of the present study have shown that PMA injection into the mouse paw results in PKC activation as well as a relatively delayed , but long-lasting , overt nociceptive behaviour in mice . Moreover , these results demonstrate that PKC activation exerts a critical role in modulating the excitability of sensory neurons . F-actin involvement in guinea pig sperm motility . Sperm motility is a must for natural fertilization to occur . During their travel through the epididymis , mammalian spermatozoa gradually acquire the ability to move . This is accomplished through a sliding movement of the outer doublet microtubules of the axoneme which is energized by the dynein ATPase . Within its complex structure , the mammalian sperm flagellum contains F-actin and thus , we decided to test in the guinea pig sperm flagellum the role of F-actin in motility . During maturation , capacitation , and the acrosome reaction , a gradual decrease of the relative concentration of F-actin was observed . Motility increased as spermatozoa became able to fertilize . P06396 , phalloidin , and KI inhibited sperm motility . P06396 canceled sperm motility within 20 min of treatment while 0.6 M KI had immediate effects . Phalloidin diminished hyperactive sperm motility slightly . All three compounds significantly increased the relative concentration of F-actin . Latrunculins are conventional drugs that destabilize the F-actin cytoskeleton . DB02621 ( O43561 A ) did not affect sperm motility ; but significantly increased F-actin relative concentration . The results suggested that in guinea pig spermatozoa , randomly severing F-actin filaments inhibits flagellar motility ; while end filament alteration does not . Thus , specific filament regions seem to be important for sperm motility . Piracy of prostaglandin E2/EP receptor-mediated signaling by Kaposi 's sarcoma-associated herpes virus ( HHV-8 ) for latency gene expression : strategy of a successful pathogen . Kaposi 's sarcoma-associated herpes virus ( KSHV ) is implicated in the pathogenesis of KS , a chronic inflammation-associated malignancy . P35354 ( P35354 ) and its metabolite prostaglandin E2 ( DB00917 ) , two pivotal proinflammatory/oncogeneic molecules , are proposed to play roles in the expression of major KSHV latency-associated nuclear antigen-1 ( LANA-1 ) . Microsomal DB00917 synthase , DB00917 , and its receptors ( EP1 , EP2 , EP3 , and EP4 ) were detected in KS lesions with the distinct staining of EP2/EP4 in KS lesions . In latently infected endothelial TIVE-LTC cells , EP receptor antagonists downregulated LANA-1 expression as well as Ca(2+) , p-Src , p-PI3K , p-PKCzeta/lambda , and p-NF-kappaB , which are also some of the signal molecules proposed to be important in KS pathogenesis . Exogenous DB00917 and EP receptor agonists induced the LANA-1 promoter in 293 cells , and P25490 , Sp1 , P14859 , Q03052 , C/EBP , and c-Jun transcription factors seem to be involved in this induction . DB00917 /EP receptor-induced LANA-1 promoter activity was downregulated significantly by the inhibition of Ca(2+) , p-Src , p-PI3K , p-PKCzeta/lambda , and p-NF-kappaB . These findings implicate the inflammatory DB00917 /EP receptors and the associated signal molecules in herpes virus latency and uncover a novel paradigm that shows the evolution of KSHV genome plasticity to use inflammatory response for its survival advantage of maintaining latent gene expression . These data also suggest that potential use of anti- P35354 and anti-EP receptor therapy may not only ameliorate the chronic inflammation associated with KS but could also lead to elimination of the KSHV latent infection and the associated KS lesions . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Comparison of two polymer-based immunohistochemical detection systems : ENVISION+ and ImmPRESS . The non-specific background reaction produced in avidin-biotin-based immunohistochemistry , particularly after harsh antigen retrieval procedures , has promoted the use of non-avidin-biotin systems , yet there are few reports comparing the performance of non-avidin-biotin , polymer-based methods . In this study we compare two of these methods , ENVISION+trade mark and ImmPRESS , in animal tissues . We examined the immunoreactivity of 18 antigens in formalin-fixed , paraffin-embedded tissues . Antigens were located in the cytoplasmic membrane ( CD11d , P05107 and CD79a ) , cytoplasm ( calretinin , P23219 , P35354 , Glut-1 , HepPar 1 , P10721 , Melan A , tryptase and uroplakin III ) or nucleus ( Q2TAK8 , P09936 and thyroid transcription factor 1 ) . We also evaluated three infectious agents ( Aspergillus , calicivirus and West Nile virus ) . The staining with ENVISION+ or ImmPRESS was performed simultaneously for each antigen . The intensity of the reaction and background staining were scored . ImmPRESS yielded similar or higher reaction intensity than ENVISION+trade mark in 16/18 antigens . ImmPRESS produced abundant background with the other two antigens ( calretinin and P35354 ) , which hindered interpretation of the specific reaction . The cost of ImmPRESS was 25 % lower than for ENVISION+trade mark . Based on these results , ImmPRESS is a good polymer-based detection system for routine immunohistochemistry . The glial cell modulator and phosphodiesterase inhibitor , DB05066 ( ibudilast ) , attenuates prime- and stress-induced methamphetamine relapse . Stress and renewed contact with drug ( a " slip " ) have been linked to persisting relapse of methamphetamine abuse . Human brain microglial activation has been linked with methamphetamine abuse , and inhibitors of glial cell activation , certain phosphodiesterase ( PDE ) inhibitors , and glial cell derived neurotrophic factor ( P39905 ) have been reported to modulate drug abuse effects . Our objective was to determine whether the glial cell attenuator , 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine ( DB05066 , ibudilast ) , a non-selective PDE inhibitor and promoter of P39905 , could reduce stress- and methamphetamine prime-induced reinstatement of methamphetamine-seeking behavior . Male Long-Evans hooded rats were trained to lever press reinforced with 0.1 mg/kg i.v. methamphetamine infusion according to fixed-ratio 1 ( FR1 ) reinforcement schedules during daily , 2-hour experimental sessions . After performance had stabilized , lever pressing was extinguished for 12 consecutive sessions and doses of 0 ( vehicle ) , 2.5 and 7.5 mg/kg DB05066 were then administered intraperitoneally b.i.d. on the last 2 days of extinction and then once on the testday to separate groups of 12 rats . During testing , the rats were given 15 min of intermittent footshock or a 1 mg/kg i.p. methamphetamine prime followed by a 2-hour reinstatement test session . DB05066 significantly reduced response levels of footshock-induced ( 2.5 and 7.5 mg/kg ) and prime-induced ( 7.5 mg/kg ) reinstatement of extinguished methamphetamine-maintained responding . DB05066 has properties consistent with the ability to attenuate relapse precipitated by stress and methamphetamine " slips " during abstinence . These results thus reinforce interest in atypical neurobiological mechanisms which could be exploited for developing novel medications for treating drug abuse disorders . The receptor tyrosine kinase inhibitor amuvatinib ( DB05216 ) sensitizes tumor cells to radio- and chemo-therapies in part by inhibiting homologous recombination . BACKGROUND AND PURPOSE : Q06609 is a key protein involved in homologous recombination ( HR ) and a potential target for radiation- and chemotherapies . Amuvatinib ( formerly known as DB05216 ) is a novel receptor tyrosine kinase inhibitor that targets c- P10721 and PDGFRα and can sensitize tumor cells to ionizing radiation ( IR ) . Here , we studied amuvatinib mechanism on Q06609 and functional HR . MATERIALS AND METHODS : Protein and RNA analyses , direct repeat green fluorescent protein ( DR-GFP ) assay and polysomal fractioning were used to measure HR efficiency and global translation in amuvatinib-treated H1299 lung carcinoma cells . Synergy of amuvatinib with IR or mitomycin c ( DB00305 ) was assessed by clonogenic survival assay . RESULTS : Amuvaninib inhibited Q06609 protein expression and HR . This was associated with reduced ribosomal protein S6 phosphorylation and inhibition of global translation . Amuvatinib sensitized cells to IR and DB00305 , agents that are selectively toxic to HR-deficient cells . CONCLUSIONS : Amuvatinib is a promising agent that may be used to decrease tumor cell resistance . Our work suggests that this is associated with decreased Q06609 expression and function and supports the further study of amuvatinib in combination with chemotherapy and radiotherapy . Effects of the total saponins from Rosa laevigata Michx fruit against acetaminophen-induced liver damage in mice via induction of autophagy and suppression of inflammation and apoptosis . The effect of the total saponins from Rosa laevigata Michx fruit ( RLTS ) against acetaminophen ( DB00316 ) -induced liver damage in mice was evaluated in the present paper . The results showed that RLTS markedly improved the levels of liver SOD , CAT , DB00143 , DB00143 -Px , MDA , NO and P35228 , and the activities of serum ALT and Q9NRA2 caused by DB00316 . Further research confirmed that RLTS prevented fragmentation of DNA and mitochondrial ultrastructural alterations based on TdT-mediated dUTP nick end labeling ( TUNEL ) and transmission electron microscopy ( TEM ) assays . In addition , RLTS decreased the gene or protein expressions of cytochrome P450 ( P05181 ) , pro-inflammatory mediators ( IL-1β , P05112 , P05231 , P01375 -α , P35228 , Bax , HMGB-1 and P35354 ) , pro-inflammatory transcription factors ( NF-κB and AP-1 ) , pro-apoptotic proteins ( cytochrome C , p53 , caspase-3 , caspase-9 , p-JNK , p-p38 and p- P29323 ) , and increased the protein expressions of Bcl-2 and Bcl-xL . Moreover , the gene expression of P22301 , and the proteins including LC3 , Q14457 and Atg5 induced by DB00316 were even more augmented by the extract . These results demonstrate that RLTS has hepatoprotective effects through antioxidative action , induction of autophagy , and suppression of inflammation and apoptosis , and could be developed as a potential candidate to treat DB00316 -induced liver damage in the future . Rehmannia Glutinosa Pharmacopuncture Solution Regulates Functional Activation , FcεRI Expression , and Signaling Events in Mast Cells . OBJECTIVES : Rehmannia glutinosa pharmacopuncture solution ( RGPS ) was investigated to determine both its anti-allergic inflammatory effects on mast cells and its detailed mechanism of actions . METHODS : We investigated whether RGPS suppress cytokines , enzymes , FcεRI expression and FcεRImediated signaling in RBL-2H3 cells stimulated with anti-DNP IgE/DNP-HSA . The suppressive effects of RGPS on the levels of cytokines such as IL-1β , P05231 and GM- P04141 were measured using emzyme-linked immunospecific assay ( ELISA ) . The mRNA expression levels of cytokines , enzymes ( HDC2 , P23219 , P35354 and 5LO ) and FcεRI αβγsubunits were measured using reverse transcription polymerase chain reaction ( RTPCR ) method . The activation of FcεRI-mediated signaling was examined using Western blot analyses . RESULTS : RGPS suppressed production of proinflamm-atory cytokines ( IL-1β , P05231 , and GM- P04141 ) in stimulated RBL-2H3 cells significantly ( p < 0.05 ) . RGPS also suppressed mRNA expression of inflammatory enzymes ( HDC2 , P23219 , P35354 , 5LO ) . In addition , mRNA expression levels of FcεRIα , FcεRIβand FcεRIγ were lowered by treatment with RGPS . Finally , RGPS prevented phosphrylation of Lyn , Syk , O43561 , Gab2 , P98160 γ1/2 , PI3K , Akt , P47712 and IκBα . CONCLUSIONS : RGPS effectively suppresses mast cell activations such as degranulation and inflammatory response via down-regulation of the FcεRI-mediated signaling pathways in IgE/Ag-stimulated mast cells . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Human lung fibroblasts increase P01730 (+)CD25(+)Foxp3(+) T cells in co-cultured P01730 (+) lymphocytes . Aim of this study was to evaluate functional modifications induced by human lung fibroblasts in co-cultured P01730 (+) T lymphocytes . P01730 (+) T cells , resting or stimulated with ionomycin/PMA for 6h , were co-cultured with fibroblasts isolated from pulmonary biopsies , in contact or separated by a semi-permeable membrane . The expression of CD25 , P16410 , TGF-β , IFNγ , P60568 , P05112 , P22301 and Foxp3 was evaluated by flow cytometric analysis . Fibroblasts induced a significant increment in CD25(+) cells in co-cultured activated P01730 (+) T lymphocytes separated by a membrane . Moreover , fibroblasts treatment with a P35354 inhibitor abrogated the increment in CD25(+) cells whereas exogenous DB00917 restored it . The CD25(+) subpopulation was characterized by increased presence of Fox- P09131 , P16410 , P22301 and TGF-β positive cells while IFN-γ and P60568 positive cells were diminished . Proliferative response of P01730 (+) to the anti CD3/ P10747 -Abs was abrogated in P01730 (+) co-cultured with fibroblasts thus demonstrating a suppressive feature of the expanded CD25(+) subpopulation . Genetics of alcoholism . DB00898 use and alcohol use disorders are substantially heritable . Variants in genes coding for alcohol metabolic enzymes have long been known to influence consumption . More recent studies in family-based samples have implicated P47869 , nicotinic receptor genes such as Q05901 , and a number of other specific single genes as associated with alcohol use disorders . The growing use of genetic analyses , in particular studies using polygenic risk scores ; neurobiologic pathways ; and methods for quantifying gene × gene and gene × environment interactions have also contributed to an evolving understanding of the genetic architecture of alcohol use disorders . Additionally , the study of behavioral traits associated with alcohol dependence such as impulsivity and sensation seeking , and the influences of demographic factors ( i.e. , sex and ethnicity ) have significantly enhanced the genetics of alcoholism literature . This article provides a brief overview of the current topically relevant findings in the field to date and includes areas of research still requiring attention . All-trans retinoic acid up-regulates Prostaglandin-E Synthase expression in human macrophages . All-trans retinoic acid ( DB00755 ) is a potent retinoid , which has been used successfully in different clinical settings as a potential drug to treat P48444 and emphysema . In the present study , we analyzed genes modulated by DB00755 by performing mRNA expression array analysis on alveolar macrophages after treatment with DB00755 . Here we observed a 375-fold up-regulation of Prostaglandin-E Synthase ( microsomal O14684 -1 , NM_004878 O14684 ) which mediates the conversion of prostaglandin H(2) ( PGH(2) ) to Prostaglandin E(2) ( PGE(2) ) . We furthermore studied the expression of O14684 after treatment with DB00755 in human monocyte-derived macrophages ( MDMs ) and bronchoalveolar lavage ( BAL ) cells . DB00755 up-regulated O14684 mRNA expression in MDMs generated with P09603 by 2500-fold whereas in P09603 + P35225 macrophages the up-regulation was only 20-fold . Similarly , DB00755 up-regulated O14684 mRNA expression by factor 1524 in BAL cells . The up-regulation of O14684 mRNA expression by DB00755 is both time and dose dependent . P35225 suppressed the DB00755 induced O14684 expression at both mRNA and protein level in MDM and BAL cells . We also observed that LPS acts synergistically with DB00755 in MDMs and strongly induces O14684 expression . DB00755 had little impact on cyclooxygenase-1 and -2 ( P23219 and -2 ) expression as compared to O14684 expression under the same experimental conditions . Furthermore , we observed an induction of PGE(2) levels by DB00755 in BAL cells . These data indicate that DB00755 is a potent inducer of O14684 expression in human macrophages but not in alternatively activated macrophages and suggest that the eicosanoid pathway is important for DB00755 action in macrophages . Purification and characterization of mouse O15528 overproduced by an Escherichia coli system coexpressing molecular chaperonins GroEL/ES . The expression of mouse O15528 in Escherichia coli has been dramatically enhanced by coexpression of GroEL/ES . To reveal the enzymatic properties of O15528 , we measured its hydroxylation activity toward vitamin D3 and DB01436 ( 1alpha(OH)D3 ) in addition to the physiological substrate DB00146 . Surprisingly , O15528 converted vitamin D3 to 1alpha, DB00146 . Both 1alpha-hydroxylation activity toward vitamin D3 , and 25-hydroxylation activity toward 1alpha(OH)D3 were observed . The Km and Vmax values for 25-hydroxylation activity toward 1alpha(OH)D3 were estimated to be 1.7 microM and 0.51 mol/min/mol P450 , respectively , while those for 1alpha-hydroxylation activity toward DB00146 were 0.050 microM and 2.73 mol/min/mol P450 , respectively . Note that the substrate must be fixed in the opposite direction in the substrate-binding pocket of O15528 between 1alpha-hydroxylation and 25-hydroxylation . Based on these results and the fact that human Q02318 and Streptomyces CYP105A1 also convert vitamin D3 to 1alpha, DB00146 , 1alpha-hydroxylation , and 25-hydroxylation of vitamin D3 appear to be closely linked together . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis .

MH_train_1238

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interacts_with DB01037?

multiple_choice

Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Common variants in psychiatric risk genes predict brain structure at birth . Studies in adolescents and adults have demonstrated that polymorphisms in putative psychiatric risk genes are associated with differences in brain structure , but can not address when in development these relationships arise . To determine if common genetic variants in disrupted-in-schizophrenia-1 ( Q9NRI5 ; rs821616 and rs6675281 ) , catechol-O-methyltransferase ( P21964 ; rs4680 ) , neuregulin 1 ( Q02297 ; rs35753505 and rs6994992 ) , apolipoprotein E ( P02649 ; ε3ε4 vs. ε3ε3 ) , estrogen receptor alpha ( P03372 ; rs9340799 and rs2234693 ) , brain-derived neurotrophic factor ( P23560 ; rs6265 ) , and glutamate decarboxylase 1 ( Q99259 ; rs2270335 ) are associated with individual differences in brain tissue volumes in neonates , we applied both automated region-of-interest volumetry and tensor-based morphometry to a sample of 272 neonates who had received high-resolution magnetic resonance imaging scans . P03372 ( rs9340799 ) predicted intracranial volume . Local variation in gray matter ( GM ) volume was significantly associated with polymorphisms in Q9NRI5 ( rs821616 ) , P21964 , Q02297 , P02649 , P03372 ( rs9340799 ) , and P23560 . No associations were identified for Q9NRI5 ( rs6675281 ) , P03372 ( rs2234693 ) , or Q99259 . Of note , neonates homozygous for the Q9NRI5 ( rs821616 ) serine allele exhibited numerous large clusters of reduced GM in the frontal lobes , and neonates homozygous for the P21964 valine allele exhibited reduced GM in the temporal cortex and hippocampus , mirroring findings in adults . The results highlight the importance of prenatal brain development in mediating psychiatric risk . Increases in stimulated secretion of proinflammatory cytokines by blood monocytes following arousal of negative affect : the role of insulin resistance as moderator . We examined the effect of negative affect on changes in stimulated secretion of cytokines by blood monocytes and determined whether insulin resistance ( IR ) , as indexed by the Homeostasis Model Assessment ( HOMA ) , moderated these associations in 58 healthy men ( aged 18-65 years ) . Blood samples and ratings of negative affect were collected at rest and 15min following subjects ' participation in the Anger Recall Interview ( Q9Y4X5 ) . Assessment of lipopolysaccharide ( LPS ) -stimulated secretion of IL-1beta , P05231 , and P01375 was accomplished by ELISA of supernatant . Regression models controlling for age , body mass index , and race/ethnicity revealed that higher HOMA-IR values were associated with larger stress-induced increases in IL-1beta and P01375 ( p < .05 ) . Furthermore , arousal of negative affect during the Q9Y4X5 was differentially associated with stress-induced changes in stimulated secretion of P01375 and P05231 as a function of HOMA-IR ( p < .05 ) . Increases in stimulated cytokine secretion were associated with arousal of negative affect , but only among men with higher HOMA-IR values . Among men with lower HOMA-IR values , arousal of negative affect was associated with diminished cytokine secretion . Collectively , these data suggest that the HOMA-IR moderates the impact that arousal of negative affect has on the ability of blood monocytes to secrete inflammatory cytokines in response to LPS . Stress-induced increases in cytokine secretion among high HOMA-IR men are consistent with the role of inflammation in cardiovascular disease , hypertension , type 2 diabetes as well as the metabolic syndrome and underscore the relevance of negative affect in the etiology of these inflammatory conditions . The relationship between gastric motility and nausea : gastric prokinetic agents as treatments . Nausea is one of a cluster of symptoms described subjectively by patients with delayed gastric emptying . The mechanisms and treatments are unclear ( anti-emetic drugs are not fully effective against nausea ) . Can nausea be relieved by stimulating gastric emptying ? Physostigmine ( together with atropine ) has been shown experimentally to stimulate gastric motility , relieve nausea and restore normal gastric motility . Is this mimicked by gastric prokinetic drugs ? The answer is complicated by mixed pharmacology . DB01233 increases gastric motility by activating myenteric Q13639 receptors but also directly inhibits vomiting via D2 and 5- Q9H205 receptor antagonism ; relationships between increased gastric motility and relief from nausea are therefore unclear . Similarly , the D2 receptor antagonist domperidone has direct anti-emetic activity . Nevertheless , more selective Q13639 and motilin receptor agonists ( erythromycin , directly stimulating gastric motility ) inhibit vomiting in animals ; low doses of erythromycin can also relieve symptoms in patients with gastroparesis . Ghrelin stimulates gastric motility and appetite mostly via vagus-dependent pathways , and inhibits vomiting in animals . To date , ghrelin receptor activation has failed to consistently improve gastric emptying or symptoms in patients with gastroparesis . We conclude that nausea can be relieved by gastric prokinetic drugs , but more clinical studies are needed using drugs with selective activity . Other mechanisms ( e.g. ghrelin , vagal and central pathways , influencing a mechanistic continuum between appetite and nausea ) also require exploration . These and other issues will be further explored in a forthcoming special issue of the European Journal of Pharmacology , which focusses on mechanisms of nausea and vomiting . Potent and selective inhibition of human nitric oxide synthases . Inhibition by non-amino acid isothioureas . DB02539 was a potent competitive inhibitor of human nitric oxide synthase ( NOS ) , with Ki values of 17 , 36 , and 29 nM for the inducible ( i ) , endothelial ( e ) , and neuronal ( n ) isozymes , respectively . Unlike some potent inhibitors of NOS , no time dependence was observed . DB02539 was not a detectable substrate for P29474 . DB02539 was also a potent inhibitor of mouse P35228 ( Ki value of 5.2 nM ) , and its binding perturbed the spectrum of P35228 consistent with its altering the environment of the bound heme . The optimum binding of S-ethyl- and S-isopropylisothiourea relative to 70 other analogs suggested that these alkyl substitutions fit into a small hydrophobic pocket . Most isothioureas were 2-6-fold selective for the human P35228 ( Ki for P35228 versus Ki for P29474 ) , with one being 19-fold selective . The cyclized mimics of S-ethylisothiourea , 2-NH2-thiazoline , and 2-NH2-thiazole , were also competitive inhibitors of human NOS . A third structural class of inhibitors , bisisothioureas , were , in general , the most selective in their inhibition of human P35228 . S,S'-(1,3-Phenylenebis(1,2-ethanediyl))bisisothiourea was 190-fold selective ( Ki value of 0.047 microM against P35228 versus 9.0 microM against P29474 ) . These results demonstrate that potent and selective inhibition of human NOS isozymes is achievable . Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5-methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 -dependent enzyme methionine synthase . Q99707 specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells . Multiple pathways of apolipoprotein E signaling in primary neurons . P02649 is a genetic risk factor for Alzheimer 's disease , and the apoE protein is associated with beta-amyloid deposits in Alzheimer 's disease brain . We examined signaling pathways stimulated by apoE in primary neurons in culture . ApoE and an apoE-derived peptide activated several intracellular kinases , including prominently extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . P27361 /2 activation by apoE was blocked by an inhibitor of the low-density lipoprotein receptor family , the specific DB01221 glutamate receptor antagonist MK 801 and other calcium channel blockers . Activation of apoE receptors also induced tyrosine phosphorylation of Dab1 , an adaptor protein of apoE receptors , but experiments in Dab1 knockout neurons demonstrated that Dab1 was not necessary for P29323 activation . In contrast , apoE treatment of primary neurons decreased activation of c-Jun N-terminal kinase , a kinase that interacts with another apoE receptor adaptor protein , c-Jun N-terminal kinase-interacting protein . This change also depended on interactions with the low-density lipoprotein receptor family but was independent of calcium channels . c-Jun N-terminal kinase deactivation by apoE was blocked by gamma-secretase inhibitors and pertussis toxin . These results demonstrate that apoE affects several signaling cascades in neurons : increased disabled phosphorylation , activation of the P27361 /2 pathway ( dependent on calcium influx via the DB01221 receptor ) and inhibition of the P45983 /2 pathway ( dependent on gamma-secretase and G proteins ) . Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 [ ( R ) -(-)-2-(4-bromo-2-fluorobenzyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-4-spiro-3'-pyrrolidine-1,2',3,5'-tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K(i) = 10(-)(10) M ) 2000-fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R- and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP(+) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP(+) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP(+) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and/or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4-bromo-2-fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . [ The effect of selective cyclooxygenase-2 inhibitor nimesulide on breast cancer induced by dimethylbenzoic acid in rats ] . OBJECTIVE : To study the effect of nimesulide ( NIM ) on the tumorigenesis of mammary tumors induced by dimethylbenzoic acid ( DMBA ) , and to investigate possible mechanisms of NIM against tumors . METHODS : The studied rats were randomly divided into four groups : experimental control group , NIM group , diet and drug of NIM control group . The incidence of mammary tumor was observed . RT-PCR , Western blot were used to detect 8 cancerous tissues in every group , randomly . The expressions of cylooxygenase-2 ( P35354 ) were assessed by immunohistochemistry . The levels of prostaglandin E(2) ( PGE(2) ) in blood plasma and tumor tissues were determined by means of radio-immunity assay . The apoptosis index and the proliferation index were evaluated by TUNEL assay , immunohistochemical staining for proliferating cell nuclear antigen ( P12004 ) , respectively . RESULTS : The incidence of mammary tumor was 69.2 % in experimental control group , 40.3 % in NIM group . There was obviously decreased incidence in NIM group ; The expressions of P35354 mRNA and protein were significantly down-regulated in NIM group compared with experimental control group . The increased levels of PGE(2) synthesis in blood plasma and tumor tissues were significantly decreased by administering NIM ( P < 0.05 ) . The apoptosis index was obviously higher , the proliferation index was markedly less in NIM group than experimental control group . CONCLUSIONS : DB04743 could inhibit the tumorigenesis and development of DMBA-induced mammary tumors by inhibition of proliferation and induction of apoptosis . P35354 and P35354 -mediated PGE(2) synthesis may play an important role in rat DMBA-induced breast cancer . Cluster analysis of risk factor genetic polymorphisms in Alzheimer 's disease . Multiple genetic variants may contribute to the risk of developing Alzheimer 's disease . We have analyzed polymorphisms in 9 genes to determine whether particular combinations would contribute to this risk . The genes were P02649 , LDLr , P01034 , P07339 , P01375 , P56817 , P10636 , Q8IWL8 , P29474 , and Q12800 . Three risk groups for the disease were identified . Risk group I was younger , was heterozygous for the P01034 ( GA ) , CTSD2936 ( AG ) , P01375 -308 ( AG ) genetic variants . Risk group II was older , was homozygous for the -427 P02649 promoter polymorphism ( TT ) , and heterozygous for the P10636 deletion and for the Q8IWL8 variant ( QR ) . Group III had both the youngest and oldest subjects , were heterozygous for the -863 ( AC ) and -1031 ( CT ) P01375 promoter polymorphisms . All three groups carried the P02649 4 allele and were heterozygous for both P56817 polymorphisms . The control groups were carriers of the P02649 3 allele and were homozygous for the P56817 genetic variants . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Interaction of early environment , gender and genes of monoamine neurotransmission in the aetiology of depression in a large population-based Finnish birth cohort . Objectives Depression is a worldwide leading cause of morbidity and disability . Genetic studies have recently begun to elucidate its molecular aetiology . The authors investigated candidate genes of monoamine neurotransmission and early environmental risk factors for depressiveness in the genetically isolated population-based Northern Finland Birth Cohort 1966 ( 12 058 live births ) . Design The authors ascertained and subdivided the study sample ( n=5225 ) based on measures of early development and of social environment , and examined candidate genes of monoamine neurotransmission , many of which have shown prior evidence of a gene-environment interaction for affective disorders , namely P31645 , Q8IWU9 , P21964 , P21397 and the dopamine receptor genes P21728 - P21918 . Results and conclusion The authors observed no major genetic effects of the analysed variants on depressiveness . However , when measures of early development and of social environment were considered , some evidence of interaction was observed . Allelic variants of P21964 interacted with high early developmental risk ( p=0.005 for rs2239393 and p=0.02 for rs4680 ) so that the association with depression was detected only in individuals at high developmental risk group ( p=0.0046 and β=0.056 for rs5993883-rs2239393-rs4680 risk haplotype CGG including Val158 ) , particularly in males ( p=0.0053 and β=0.083 for the haplotype CGG ) . Rs4274224 from P14416 interacted with gender ( p=0.017 ) showing a significant association with depressiveness in males ( p=0.0006 and β=0.0023 ; p=0.00005 and β=0.069 for rs4648318-rs4274224 haplotype GG ) . The results support the role of genes of monoamine neurotransmission in the aetiology of depression conditional on environmental risk and sex , but not direct major effects of monoaminergic genes in this unselected population . DB00142 decarboxylase ( Q05329 ) autoantibodies in prediction of beta-cell function and remission in recent-onset IDDM after cyclosporin treatment . The Canadian-European Randomized Control Trial Group . We have investigated whether glutamic acid decarboxylase ( Q99259 ) autoantibodies ( Q05329 Ab ) were affected by cyclosporin therapy and were related to subsequent non-insulin-requiring remission and loss of glucagon-stimulated C-peptide response in 132 recent-onset insulin-dependent diabetes mellitus ( IDDM ) patients treated with cyclosporin or placebo for 12 months . Q05329 Ab were detected in a quantitative radioligand assay using as tracer recombinant , in vitro translated , human islet [35S]methionine-labeled Q05329 . Q05329 Ab were found at onset in 66 % ( 87 of 132 ) of IDDM patients and in 1 % ( 1 of 100 ) of healthy control subjects . The prevalence of Q05329 Ab and median Q05329 Ab levels did not change in serum samples taken 3 , 6 , 9 , and 12 months after study entry in either the cyclosporin- or the placebo-treated groups . The presence or absence of Q05329 Ab at study entry did not predict non-insulin-requiring remission in either cyclosporin- or placebo-treated patients . However , the relative ( compared with 0 months ) glucagon-stimulated C-peptide response was more than 30 % lower in Q05329 Ab+ patients receiving placebo at 9 and 12 months compared with the Q05329 Ab- placebo patients ( P < 0.035 ) . Islet cell cytoplasmic antibody ( ICA ) and Q05329 Ab+ placebo-treated patients showed no significant differences in stimulated C-peptide levels compared with those who were ICA- and Q05329 Ab+ , suggesting that ICA was not independently associated with loss of beta-cell function. ( ABSTRACT TRUNCATED AT 250 WORDS ) Antineoplastic mechanisms of niclosamide in acute myelogenous leukemia stem cells : inactivation of the NF-kappaB pathway and generation of reactive oxygen species . NF-kappaB may be a potential therapeutic target for acute myelogenous leukemia ( AML ) because NF-kappaB activation is found in primitive human AML blast cells . In this report , we initially discovered that the potent antineoplastic effect of niclosamide , a Food and Drug Administration-approved antihelminthic agent , was through inhibition of the NF-kappaB pathway in AML cells . DB06803 inhibited the transcription and DNA binding of NF-kappaB . It blocked tumor necrosis factor-induced P25963 phosphorylation , translocation of p65 , and expression of NF-kappaB-regulated genes . DB06803 inhibited the steps TAK1 --> O15111 ( IKK ) and IKK --> P25963 . DB06803 also increased the levels of reactive oxygen species ( ROS ) in AML cells . Quenching ROS by the glutathione precursor DB06151 attenuated niclosamide-induced apoptosis . Our results together suggest that niclosamide inhibited the NF-kappaB pathway and increased ROS levels to induce apoptosis in AML cells . On translational study of the efficacy of niclosamide against AML , niclosamide killed progenitor/stem cells from AML patients but spared those from normal bone marrow . DB06803 was synergistic with the frontline chemotherapeutic agents cytarabine , etoposide , and daunorubicin . It potently inhibited the growth of AML cells in vitro and in nude mice . Our results support further investigation of niclosamide in clinical trials of AML patients . The V1a vasopressin receptor is necessary and sufficient for normal social recognition : a gene replacement study . DB00067 modulates many social and nonsocial behaviors , including emotionality . We have previously reported that male mice with a null mutation in the V1a receptor ( P37288 ) exhibit a profound impairment in social recognition and changes in anxiety-like behavior . Using site-specific injections of a P37288 -specific antagonist , we demonstrate that the lateral septum , but not the medial amygdala , is critical for social recognition . Reexpressing P37288 in the lateral septum of P37288 knockout mice ( V1aRKO ) using a viral vector resulted in a complete rescue of social recognition . Furthermore , overexpression of the P37288 in the lateral septum of wild-type ( wt ) mice resulted in a potentiation of social recognition behavior and a mild increase in anxiety-related behavior . These results demonstrate that the P37288 in the lateral septum plays a critical role in the neural processing of social stimuli required for complex social behavior . Heroin addiction in African Americans : a hypothesis-driven association study . Heroin addiction is a chronic complex disease with a substantial genetic contribution . This study was designed to identify gene variants associated with heroin addiction in African Americans . The emphasis was on genes involved in reward modulation , behavioral control , cognitive function , signal transduction and stress response . We have performed a case-control association analysis by screening with 1350 variants of 130 genes . The sample consisted of 202 former severe heroin addicts in methadone treatment and 167 healthy controls with no history of drug abuse . Single nucleotide polymorphism ( SNP ) , haplotype and multi-SNP genotype pattern analyses were performed . Seventeen SNPs showed point-wise significant association with heroin addiction ( nominal P < 0.01 ) . These SNPs are from genes encoding several receptors : adrenergic ( ADRA1A ) , arginine vasopressin ( P37288 ) , cholinergic ( P08172 ) , dopamine ( P21728 ) , GABA-A ( P28472 ) , glutamate ( Q12879 ) and serotonin ( P46098 ) as well as alcohol dehydrogenase ( P40394 ) , glutamic acid decarboxylase ( Q99259 and Q05329 ) , the nucleoside transporter ( Q99808 ) and diazepam-binding inhibitor ( DBI ) . The most significant result of the analyses was obtained for the Q12879 haplotype G-A-T ( rs4587976-rs1071502-rs1366076 ) with protective effect ( P(uncorrected) = 9.6E- 05 , P(corrected) = 0.058 ) . This study corroborates several reported associations with alcohol and drug addiction as well as other related disorders and extends the list of variants that may affect the development of heroin addiction . Further studies will be necessary to replicate these associations and to elucidate the roles of these variants in drug addiction vulnerability . Cbl-b is a negative regulator of inflammatory cytokines produced by IgE-activated mast cells . c-Cbl and Cbl-b E3 ubiquitin ligases are abundantly expressed in hemopoietic cells where they negatively regulate the activity and levels of many cell surface receptors and associated signaling molecules . By comparing bone marrow-derived mast cells from c-Cbl and Cbl-b-deficient mice it has recently been shown that Cbl-b is the dominant family member for negatively regulating signaling responses from high-affinity IgE receptors . In this study , we suggest that a possible reason for the greater enhancement of IgE receptor signaling in Cbl-b-deficient mice is the relatively higher levels of Cbl-b protein over c-Cbl in mast cells compared with other hemopoietic cells . We also directly compare mast cells from c-Cbl and Cbl-b-deficient mice and find that loss of Cbl-b , but not c-Cbl , increases cell growth , retards receptor internalization , and causes the sustained tyrosine phosphorylation of Syk and its substrates . However , loss of Cbl-b does not enhance the activation of P29323 or Akt , nor does it promote a greater calcium response . Furthermore , loss of Cbl-b or c-Cbl does not increase levels of the Syk or Lyn protein tyrosine kinases . Most notable , however , is the extremely large increase in the production of proinflammatory cytokines P01375 , P05231 , and P13500 by Cbl-b(-/-) mast cells compared with levels produced by c-Cbl(-/-) or wild-type cells . This marked induction , which appears to be restricted to these three cytokines , is dependent on IgE receptor activation and correlates with enhanced O15111 phosphorylation . Thus , Cbl-b functions as a potent negative regulator of cytokines that promote allergic and inflammatory reactions . MEK inhibition in non-small cell lung cancer . P01116 mutations are the most common mutations in non-small cell lung cancer ( NSCLC ) with adenocarcinoma histology . P01116 mutations result in the activation of the RAF-MEK- P29323 pathway , and agents that target RAF-MEK- P29323 pathways have been investigated in P01116 mutant NSCLC . The two agents furthest in development are selumetinib and trametinib . DB08911 has greater binding for the Q02750 /2 allosteric site , and generally has superior pharmacokinetics . A randomized phase II trial of docetaxel with and without selumetinib revealed that the combination resulted numerically superior overall survival , and a statistically significant improvement in progression-free survival and objective response rate . However , a concerning rate of hospital admission , grade 3 or 4 neutropenia , and febrile neutropenia was observed with the combination . Trials have investigated MEK inhibitors as single agents and in combination with erlotinib , and the data do not support the further development . The activity of MEK inhibitors appears to be similar in patients with P01116 mutant and wild-type NSCLC suggesting P01116 mutation status is not a reliable biomarker for efficacy . It is possible that mutations of genes in addition to P01116 mutations impact the activity of MEK inhibitors , or specific subsets of P01116 mutations may be resistant or susceptible to MEK inhibition . Other potential explanations are gene amplifications , alternative RNA splicing of genes resulting in activation of their protein products , and deregulation of noncoding RNAs and consequent altered protein expression . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness .

MH_train_1239

MH_train_1239

MH_train_1239

interacts_with DB06616?

multiple_choice

Oral L-citrulline administration improves memory deficits following transient brain ischemia through cerebrovascular protection . L-citrulline ( L- DB00155 ) is known to increase nitric oxide ( NO ) production via the increase of L-arginine ( DB00125 ) concentration in the blood and improve endothelial dysfunction in cardiovascular diseases . However , little is known about the effects of L- DB00155 on cerebrovascular dysfunction . Here we showed that oral L- DB00155 administration prevents cerebrovascular injury following cerebral ischemia using a 20-min bilateral common carotid artery occlusion ( BCCAO ) mouse model . After BCCAO ischemia , mice were treated with L- DB00155 ( 50 , 75 , or 100 mg/kg p.o. ) for 10 days once a day . L- DB00155 administration not only prevented neuronal cell death but also prevented capillary loss in the hippocampal region following brain ischemia . The cerebrovascular protective effect of L- DB00155 was associated with the restoration of endothelial nitric oxide synthase ( P29474 ) expression in the hippocampus . In addition , we devised a novel protocol to analyze NOx(-) ( NO(2-) and NO(3-) ) productions following DB00125 infusion using in vivo microdialysis and revealed that decreased DB00125 -induced NOx(-) levels were improved in the hippocampus of BCCAO mice following repeated L- DB00155 administration . Finally , memory deficits following brain ischemia were improved by oral administration of L- DB00155 . In summary , L- DB00155 is a potential therapeutic agent that protects cerebrovascular injury and in turn prevents neuronal cell death . Thereby , oral L- DB00155 administration improves cognitive deficits following brain ischemia . Substance P-induced mitogenesis in human astrocytoma cells correlates with activation of the mitogen-activated protein kinase signaling pathway . The neuropeptide DB05875 ( SP ) regulates many biological processes through binding to and activating the SP receptor ( NK-1 subtype ) . Activation of the SP receptor induces mitogenesis in several cell types . In this study , we characterized the mitogenic response induced by SP peptide in the U-373MG astrocytoma cell line and showed that activation of the SP receptor induces [3H]thymidine incorporation into DNA . We also found that SP potently induces c-myc mRNA and protein in the U-373MG cells . Tyrphostin A25 , which blocks activity of tyrosine kinases , significantly inhibited SP-induced mitogenesis , suggesting that the mitogenic response induced by SP peptide involves phosphorylation by tyrosine kinases . Furthermore , stimulation of the SP receptor activates tyrosine phosphorylation and enzymatic activity of extracellular signal-regulated kinases ( Erk1 and Erk2 ) , also called the mitogen-activated protein kinases ( MAPKs ) . This result suggests that MAPKs participate in the SP peptide-induced signaling pathway . The addition of CP 96,345 ( [ ( 2S,3S ) -cis-2-(diphenylmethyl)-N-[(2-methoxyphenyl)-methyl]-1 -azabicyclo[2.2.2]octan-3-amine ] ; an P25103 antagonist ) or PD 098059 ( Q02750 inhibitor ) inhibited both DNA synthesis and activation of the MAPK pathway , substantiating that SP stimulates mitogenesis by activating the MAPK pathway through receptors of the NK-1 subtype . Our results demonstrate that SP peptide is a strong mitogen in the U-373MG astrocytoma cell line and establish a clear correlation between SP-induced mitogenesis and activation of MAPK signaling pathway . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Prostanoid production in Saccharomyces cerevisiae provides a novel assay for nonsteroidal anti-inflammatory drugs . Prostanoids are a large family of lipid mediators originating from prostaglandin H synthase ( PGHS ) activity on the 20-carbon polyunsaturated fatty acids dihomo-gamma-linolenic acid ( DB00154 ) , arachidonic acid ( AA ) and eicosapentaenoic acid . The two mouse PGHS isoforms , P23219 and P35354 , were expressed in Saccharomyces cerevisiae ( yeast ) , as was a signal-peptide-deleted version of P23219 ( PGHS-1MA ) . P23219 showed high activity with both AA and DB00154 as substrate , whereas P35354 activity was high with DB00154 but low with AA . Signal peptide removal reduced the activity of PGHS-1MA by > 50 % relative to P23219 , but the residual activity indicated that correct targeting to the lumen of the endoplasmic reticulum may not be necessary for enzyme function . Coexpression of P23219 with cDNAs encoding mouse prostaglandin I synthase and thromboxane A synthase , and with Trypanosoma brucei genomic DNA encoding prostaglandin F synthase in AA-supplemented yeast cultures resulted in production of the corresponding prostanoids , prostaglandin I(2) , thromboxane A(2) and prostaglandin F(2alpha) . The inhibitory effects of nonsteroidal anti-inflammatory drugs ( NSAIDs ) on prostanoid production were tested on yeast cells expressing P23219 in AA-supplemented culture . Dose-dependent inhibition of prostaglandin H(2) production by aspirin , ibuprofen and indomethacin demonstrated the potential utility of this simple expression system in screening for novel NSAIDs . Acetylbritannilactone suppresses lipopolysaccharide-induced vascular smooth muscle cell inflammatory response . To investigate the mechanism of action by which a new anti-inflammatory active compound , 1-O-acetylbritannilactone ( P00519 ) isolated from Inula britannica-F. , inhibits inflammatory responses in vascular smooth muscle cells ( VSMCs ) . Enzyme immunoassay was used to measure the levels of prostandin E(2) ( PGE(2) ) production . Immunocytochemistry staining and Western blot analysis were performed to detect the nuclear translocation of nuclear factor-kappaB ( NF-kappaB ) p65 and the expression of IkappaB-alpha , pIkappaB-alpha and cyclooxygenase-2 ( P35354 ) . Electrophoretic mobility shift assays ( EMSA ) were used to detect DNA-binding activity of NF-kappaB in VSMCs . P00519 ( 5 , 10 , 20 micrommol/l ) had several concentration-dependent effects , including inhibition of lipopolysaccharide ( LPS ) -induced PGE(2) production and P35354 expression , and blockade of NF-kappaB activation and translocation . These effects were owing to reductions in IkappaB-alpha phosphorylation and degradation induced by LPS . In addition , P00519 directly inhibited the binding of active NF-kappaB to specific DNA cis-element . These results indicate that P00519 is a potent inhibitor of LPS-stimulated VSMC inflammatory responses through blockade of NF-kappaB activity and inhibition of inflammatory gene P35354 expression . Further evidence that central tachykinin NK-1 receptors mediate the inhibitory effect of tachykinins on angiotensin-induced drinking in rats . The order of potency of tachykinin ( TK ) receptor agonists suggests that TK NK-1 receptors mediate their inhibitory effect on water intake induced by intracerebroventricular ( i.c.v. ) injection of angiotensin II ( AngII ) in rats . The present study was aimed at further evaluating which TK receptor subtype mediates the effect , using selective antagonists for the TK receptor subtypes . Pulse i.c.v. injection of the TK agonist neuropeptide gamma ( NP gamma ) , 31-250 ng/rat , markedly inhibited AngII-induced water intake . The i.c.v. injection of the P25103 antagonist SR14033 , 0.5 microgram/rat , significantly reduced , while 1 microgram/rat completely abolished the inhibitory effect of NP gamma , 125 ng/rat . The selective P21452 antagonist SR48968 and the selective P29371 antagonist R820 were devoid of any effect up to the i.c.v. dose of 2 micrograms/rat . On the other hand , i.c.v. injection of SR140333 , 1 microgram/rat , did not increase drinking induced by i.c.v. injection of AngII , 0.1-10 ng/rat , and did not increase drinking in water sated or water deprived rats . The results of the present study confirm that central TKergic mechanisms inhibit AngII-induced drinking in rats , and provide further evidence that TK NK-1 receptors mediate the effect . Failure of i.c.v. injected DB05790 to increase AngII-induced drinking , as well as water intake in sated or deprived rats suggests that brain P25103 mechanisms apparently do not exert a tonic control on AngII-induced drinking and , in general , on water intake in rats . From a pharmacological point of view , the inhibitory effect of TKs on the dipsogenic action of AngII can represent a functional test for activity at central NK-1 receptors in rats . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Agents with selective estrogen receptor ( ER ) modulator activity induce apoptosis in vitro and in vivo in ER-negative glioma cells . Tamoxifen , a member of the selective estrogen receptor modulator ( SERM ) family , is widely used in the treatment of estrogen receptor ( ER ) -expressing breast cancer . It has previously been shown that high-dose tamoxifen has cytotoxic activity against glioma cells , but whether this effect is drug specific or represents a general property of SERMs is unknown . In this study , we demonstrate that tamoxifen and DB05487 , a novel benzopyranone with SERM activity , induce glioma cell apoptosis in a dose- and time-dependent manner . Moreover , administration of tamoxifen and DB05487 suppresses tumor growth in vivo and extends animal survival in glioma xenograft models . None of the eight glioma cell lines examined express either P03372 or -beta , suggesting the mechanism for tamoxifen- and DB05487 -induced glioma cell apoptosis is independent of the ER signaling pathway . Complementary DNA microarray expression profiling allowed us to identify a subset of genes specifically regulated by tamoxifen and DB05487 , and not by other apoptotic stimuli , including nuclear factor ( NF ) -kappaB with its target genes IEX-3 , P04179 , P05231 , and P10145 . We demonstrate that suppression of NF-kappaB activation markedly enhances SERM-induced apoptosis , suggesting a role for NF-kappaB in protecting glioma cells from SERM-induced cytotoxicity . These findings demonstrate for the first time that a SERM other than tamoxifen can induce glioma cell apoptosis in vitro and in vivo and that the clinical efficacy of SERMs for the treatment of malignant gliomas could potentially be enhanced by simultaneous inhibition of the NF-kappaB pathway . Prevalence of germline mutations in the DB00024 receptor gene as a cause of juvenile thyrotoxicosis . AIM : To ascertain the prevalence of germline mutations in the DB00024 receptor gene as a cause of juvenile thyrotoxicosis ( JT ) in non-autoimmune patients . DB00024 receptor gene mutations are not seen in autoimmune-active patients . METHODS : In a nationwide study on JT , 123 patients were re-examined 10 y ( range 4 to 21 y ) after diagnosis . Two patients with toxic adenoma were excluded . In 25 patients , no P07202 , TG or P16473 antibodies were found . In 17 patients , DNA material was available for DB00024 receptor gene analysis . The entire DB00024 receptor gene was sequenced in five patients . DB00024 receptor " hot spots " for mutations in exon 9 and 10 were sequenced in the remaining 12 patients . RESULTS : A DB00024 receptor gene germline mutation was identified in only one patient of a total number of 121 patients with JT , of which 17 patients were presumed to have non-autoimmune JT by the lack of thyroid autoantibodies . CONCLUSION : In Denmark the prevalence of germline mutations in the DB00024 receptor gene is one in 121 patients with JT ( 0.8 % ; 95 % CI : 0.02-4.6 % ) and one in 17 patients with presumed non-autoimmune JT ( 6 % ; 95 % CI : 5.88 % ( 0.15-28.69 ) ) . Cytogenetic and molecular genetic aspects of essential thrombocythemia . Essential thrombocythemia ( ET ) is a chronic myeloid disorder that is characterized by thrombocytosis , thrombohemorrhagic and vasomotor symptoms , a long median survival , and a low risk of transformation to leukemia . ET can be difficult to distinguish from secondary ( reactive ) thrombocytosis , and the diagnosis of ET can only be made after the exclusion of other marrow disorders with similar features . Although ET has been assumed to be a clonal process , recent studies have suggested that a substantial number of cases classified as ET may actually not be clonal , and nonclonality may be associated with a lower risk of thrombosis . The lack of a characteristic cytogenetic marker for ET confounds analyses of clonality and offers no insight into disease pathogenesis . There is controversy over the proper classification of thrombocytosis associated with the pathological P11274 - P00519 gene rearrangement ; such cases are not clearly distinguishable from chronic myelogenous leukemia ( CML ) and should be provisionally classified as CML . New insights are emerging into the role of the megakaryocytopoiesis regulator thrombopoietin ( P07202 ) and its receptor , c-Mpl , in ET and related disorders , but P07202 -Mpl dynamics appear to be complex . In some familial thrombocythemic syndromes , mutations in the 5' untranslated region of P07202 have recently been described , but these have not yet been observed in sporadic ET . In the future , global analysis of gene expression patterns may help overcome diagnostic dilemmas , refine disease classification , and lead to an improved understanding of the pathogenesis of ET . Regional selective neuronal degeneration after protein phosphatase inhibition in hippocampal slice cultures : evidence for a Q96HU1 kinase-dependent mechanism . The regional selectivity and mechanisms underlying the toxicity of the serine/threonine protein phosphatase inhibitor okadaic acid ( OA ) were investigated in hippocampal slice cultures . Image analysis of propidium iodide-labeled cultures revealed that okadaic acid caused a dose- and time-dependent injury to hippocampal neurons . Pyramidal cells in the P07451 region and granule cells in the dentate gyrus were much more sensitive to okadaic acid than the pyramidal cells in the P00915 region . Electron microscopy revealed ultrastructural changes in the pyramidal cells that were not consistent with an apoptotic process . Treatment with okadaic acid led to a rapid and sustained tyrosine phosphorylation of the mitogen-activated protein kinases P27361 and P28482 ( Q8TCB0 /42(mapk) ) . The phosphorylation was markedly reduced after treatment of the cultures with the microbial alkaloid DB02152 ( a nonselective protein kinase inhibitor ) or the Q96HU1 kinase kinase ( Q02750 /2 ) inhibitor PD98059 . DB02152 and PD98059 also ameliorated the okadaic acid-induced cell death . Inhibitors of protein kinase C , Ca2+/calmodulin-dependent protein kinase II , or tyrosine kinase were ineffective . These results indicate that sustained activation of the Q96HU1 kinase pathway , as seen after e.g. , ischemia , may selectively harm specific subsets of neurons . The susceptibility to Q96HU1 kinase activation of the P07451 pyramidal cells and dentate granule cells may provide insight into the observed relationship between cerebral ischemia and dementia in Alzheimer 's disease . Loss of Jak2 impairs endothelial function by attenuating P04049 / Q02750 /Sp-1 signaling along with altered P29474 activities . A number of inhibitors have been used to dissect the functional relevance of Jak2 in endothelial homeostasis , with disparate results . Given that Jak2 deficiency leads to embryonic lethality , the exact role of Jak2 in the regulation of postnatal endothelial function is yet to be fully elucidated . We generated a model in which Jak2 deficiency can be induced by tamoxifen in adult mice . Loss of Jak2 significantly impaired endothelium-dependent response capacity for vasodilators . Matrigel plug assays indicated a notable decrease in endothelial angiogenic function in Jak2-deficient mice . Studies in a hindlimb ischemic model indicated that Jak2 activity is likely to be a prerequisite for prompt perfusion recovery , based on the concordance of temporal changes in Jak2 expression during the course of ischemic injury and perfusion recovery . A remarkable delay in perfusion recovery , along with reduced capillary and arteriole formation , was observed in Jak2-deficient mice . Antibody array studies indicated that loss of Jak2 led to repressed P29474 expression . In mechanistic studies , Jak2 deficiency attenuated P04049 / Q02750 signaling , which then reduced activity of Sp-1 , an essential transcription factor responsible for P29474 expression . These data are important not only for understanding the exact role that Jak2 plays in endothelial homeostasis , but also for assessing Jak2-based therapeutic strategies in a variety of clinical settings . Preclinical and clinical studies of the Q14511 scaffold protein in cancer and other diseases . Cancer progression requires a significant reprogramming of cellular signaling to support the essential tumor-specific processes that include hyperproliferation , invasion ( for solid tumors ) and survival of metastatic colonies . Q14511 ( also known as Q14511 and HEF1 ) encodes a multi-domain scaffolding protein that assembles signaling complexes regulating multiple cellular processes relevant to cancer . These include responsiveness to signals emanating from the T and B cell receptors , integrins , chemokine receptors , and receptor tyrosine kinases , as well as cytoplasmic oncogenes such as P11274 - P00519 and Q05397 - and P12931 -family kinases . Downstream , Q14511 regulation of partners including P46109 , WAVE , PI3K/AKT , P29323 , P12830 , Aurora-A ( O14965 ) , Q9UBN7 , and others allow Q14511 to influence functions as pleiotropic as migration , invasion , survival , ciliary resorption , and mitosis . In this review , we summarize a growing body of preclinical and clinical data that indicate that while Q14511 is itself non-oncogenic , changes in expression of Q14511 ( most commonly elevation of expression ) are common features of tumors , and directly impact tumor aggressiveness , metastasis , and response to at least some targeted agents inhibiting Q14511 -interacting proteins . These data strongly support the relevance of further development of Q14511 as a biomarker for therapeutic resistance . Finally , we briefly discuss emerging evidence supporting involvement of Q14511 in additional pathological conditions , including stroke and polycystic kidney disease . Antagonism of Q9GZP0 by human antibody DB05139 prevents renal scarring in experimental glomerulonephritis . Glomerular mesangial cell proliferation and/or matrix accumulation characterizes many progressive renal diseases . Q9GZP0 was identified recently as a novel mediator of mesangial cell proliferation in vitro and in vivo . This study investigated the long-term consequences of Q9GZP0 inhibition in vivo . Rats with progressive mesangioproliferative glomerulonephritis ( uninephrectomy plus anti-Thy-1.1 antibody ) received the Q9GZP0 -neutralizing , fully human mAb DB05139 on days 3 , 10 , and 17 after disease induction . Glomerular mesangioproliferative changes on day 10 were significantly reduced by anti- Q9GZP0 treatment as compared with control antibody . Eight weeks after disease induction , anti- Q9GZP0 therapy significantly ameliorated focal segmental glomerulosclerosis , podocyte damage ( de novo desmin expression ) , tubulointerstitial damage , and fibrosis as well as the accumulation of renal interstitial matrix including type III collagen and fibronectin . Treatment with anti- Q9GZP0 also reduced the cortical infiltration of monocytes/macrophages on day 56 , possibly related to lower renal cortical complement activation ( C5b-9 deposition ) and/or reduced epithelial-to-mesenchymal transition ( preserved cortical expression of P12830 and reduced expression of vimentin and alpha-smooth muscle actin ) . In conclusion , these data provide evidence for a causal role of Q9GZP0 in the pathogenesis of renal scarring and point to a new therapeutic approach to progressive mesangioproliferative renal disease . Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) -positive leukemia cell line with common-B cell phenotype , designated TMD5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD5 cells expressed 190 kDa P11274 / P00519 chimeric protein and 145 kDa P00519 protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM- P04141 , P08700 , P05231 , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia . DB01628 -induced fixed drug eruption with positive lesional patch tests . Fixed drug eruption ( FDE ) is most commonly associated with antibiotics , anticonvulsants , and nonnarcotic analgens , including nonsteroidal anti-inflammatory drugs ( NSAIDs ) . However , the newer cyclooxygenase 2 ( P35354 ) inhibitors have been rarely reported to cause FDE . We report the case of a 52-year-old Caucasian woman with erythematous pruritic plaques on the neck , left forearm , and second finger of the right hand , healing with hyperpigmentation and recurring in the same locations . The patient was sporadically taking oral etoricoxib 90 mg for her back pain and noticed the relation between administration of the drug and skin lesions , the time interval decreasing progressively from 1 week to 30 minutes . No other signs , symptoms , or drug intake was mentioned . The patch tests with etoricoxib 1 % and 5 % in petrolatum were positive at the location of the lesions and negative on the back ( nonlesional skin ) . Standard European and NSAID series were negative . Patch tests of 10 healthy controls with etoricoxib 1 % and 5 % in petrolatum were negative . After the avoidance of the drug , no relapse was mentioned . The patch test was reliable for the diagnosis of FDE , avoiding the need for subsequent oral provocation testing and therefore preventing the possible adverse effects . Despite being regarded as a safe drug , the occurrence of cutaneous adverse reactions to etoricoxib should be considered , especially in the setting of its increasing use in pain control .

MH_train_1240

MH_train_1240

MH_train_1240

interacts_with DB04908?

multiple_choice

Comparison of the effects of firocoxib , carprofen and vedaprofen in a sodium urate crystal induced synovitis model of arthritis in dogs . A randomized , placebo-controlled , four-period cross-over laboratory study involving eight dogs was conducted to confirm the effective analgesic dose of firocoxib , a selective P35354 inhibitor , in a synovitis model of arthritis . DB09217 was compared to vedaprofen and carprofen , and the effect , defined as a change in weight bearing measured via peak ground reaction , was evaluated at treatment dose levels . A lameness score on a five point scale was also assigned to the affected limb . Peak vertical ground reaction force was considered to be the most relevant measurement in this study . The firocoxib treatment group performed significantly better than placebo at the 3 h post-treatment time point and significantly better than placebo and carprofen at the 7 h post-treatment time point . Improvement in lameness score was also significantly better in the dogs treated with firocoxib than placebo and carprofen at both the 3 and 7 h post-treatment time points . [ Characteristics of abnormal behavior induced by delta 9-tetrahydrocannabinol in rats ] . delta 9- DB00470 ( THC ) , one of the active compounds of marihuana , is known to induce drug dependence and tolerance , and its action is weaker than those of other abused drugs in humans and animals . Acute effects of THC , " high " , " irritable " and " cognitive deficits " are more important than the drug dependence and tolerance . For this reason , we examined characteristics of abnormal behavior such as catalepsy-like immobilization , aggressive behavior including irritable aggression and muricide , and spatial cognition impairment induced by acute and chronic treatments of THC in rats . The catalepsy-like immobilization is related to a decrease in catecholaminergic and serotonergic neurons in the nucleus accumbens and amygdaloid nucleus and thus serves as a useful model for amotivational syndrome , one of cannabis psychoses . In aggressive behavior , muricide was determined by the housing condition . Muricide was induced if the rat was placed under an isolated housing condition within the period of the effect of single injection of THC . The behavioral change resembles exacerbation and flashback in humans . Spatial cognition is impaired by the interaction between cannabinoid ( P21554 ) and 5-HT2 receptor in the dorsal raphe-hippocampal serotonergic neurons . Thus the abnormal behavior induced by THC can be a useful model for investigating mental function in humans and new drugs for the treatment of mental disorders . A mechanism for the synergistic antimalarial action of atovaquone and proguanil . A combination of atovaquone and proguanil has been found to be quite effective in treating malaria , with little evidence of the emergence of resistance when atovaquone was used as a single agent . We have examined possible mechanisms for the synergy between these two drugs . While proguanil by itself had no effect on electron transport or mitochondrial membrane potential ( DeltaPsim ) , it significantly enhanced the ability of atovaquone to collapse DeltaPsim when used in combination . This enhancement was observed at pharmacologically achievable doses . DB01131 acted as a biguanide rather than as its metabolite cycloguanil ( a parasite dihydrofolate reductase [ P00374 ] inhibitor ) to enhance the atovaquone effect ; another P00374 inhibitor , pyrimethamine , also had no enhancing effect . DB01131 -mediated enhancement was specific for atovaquone , since the effects of other mitochondrial electron transport inhibitors , such as myxothiazole and antimycin , were not altered by inclusion of proguanil . Surprisingly , proguanil did not enhance the ability of atovaquone to inhibit mitochondrial electron transport in malaria parasites . These results suggest that proguanil in its prodrug form acts in synergy with atovaquone by lowering the effective concentration at which atovaquone collapses DeltaPsim in malaria parasites . This could explain the paradoxical success of the atovaquone-proguanil combination even in regions where proguanil alone is ineffective due to resistance . The results also suggest that the atovaquone-proguanil combination may act as a site-specific uncoupler of parasite mitochondria in a selective manner . Effect of the phosphodiesterase III inhibitor amrinone on cytokine and nitric oxide production in immunostimulated J774.1 macrophages . The level of intracellular cyclic nucleotides is a regulatory factor in a variety of immune processes . Increases in intracellular cyclic AMP ( DB02527 ) and/or cyclic GMP ( cGMP ) concentration by the inhibition of phosphodiesterase have been shown to modulate the inflammatory response . DB01427 is a clinically used positive inotropic agent which elevates intracellular DB02527 and cGMP levels by selective inhibition of the phosphodiesterase III isoenzyme . In the current study , we investigated the effect of various concentrations ( 1-300 microM ) of amrinone on lipopolysaccharide-induced production of pro- and anti-inflammatory cytokines and of nitric oxide ( NO ) in vitro . In cultured murine J774.1 macrophages , 1 ng/ml-10 microg/ml of lipopolysaccharide from Escherichia coli O55: P46977 induced production of tumor necrosis factor-alpha ( P01375 ) , interleukin-10 , and nitrite ( breakdown product of NO ) . Pretreatment of cells with amrinone caused a dose-dependent suppression of P01375 production in the concentration range of 1-100 microM . Furthermore , this drug suppressed NO production in the range of 30-300 microM . Similarly to the results in the J774.1 cells , amrinone also inhibited P01375 and NO production in the range of 10-100 microM in primary rat peritoneal macrophages . At 300 microM , but not at lower concentrations , amrinone inhibited interleukin-10 production in lipopolysaccharide-treated J774.1 macrophages . Pretreatment of the macrophages with 100 and 300 microM amrinone increased the lipopolysaccharide-elicited translocation of nuclear factor-kappa B . Taken together , our results indicate that the phosphodiesterase III inhibitor amrinone modulates the activation/production of many pro- and anti-inflammatory factors in endotoxin-stimulated cells . It remains to be further investigated how such immunomodulatory effects contribute to the clinical profile of the agent . Activation of brain prostanoid EP3 receptors via arachidonic acid cascade during behavioral suppression induced by Delta8-tetrahydrocannabinol . We have previously shown that behavioral changes induced by cannabinoid were due to an elevation of prostaglandin E2 ( DB00917 ) via the arachidonic acid cascade in the brain . In the present study , we investigated the participation of the prostanoid EP3 receptor , the target of DB00917 in the brain , in behavioral suppression induced by Delta8-tetrahydrocannabinol ( Delta8-THC ) , an isomer of the naturally occurring Delta9-THC , using a one-lever operant task in rats . Intraperitoneal administration of Delta8-THC inhibited the lever-pressing behavior , which was significantly antagonized by both the selective cannabinoid P21554 receptor antagonist SR141716A and the cyclooxygenase inhibitor diclofenac . Furthermore , intracerebroventricular ( i.c.v. ) administration of DB00917 significantly inhibited the lever-pressing performance similar to Delta8-THC . P43115 antisense-oligodeoxynucleotide ( AS-ODN ; twice a day for 3 days , i.c.v. ) significantly decreased prostanoid EP3 receptor mRNA levels as determined by the RT-PCR analysis in the cerebral cortex , hippocampus and midbrain . AS-ODN also antagonized the DB00917 -induced suppression of the lever pressing . In the same way , the suppression of lever-pressing behavior by Delta8-THC was significantly improved by AS-ODN . It is concluded that the suppression of lever-pressing behavior by cannabinoid is due to activation of the prostanoid EP3 receptor through an elevation of DB00917 in the brain . Perinatal glutathione levels in liver and brain of rats from large and small litters . In the rat , hepatic glutathione ( DB00143 ) levels increase prenatally and reach maximum concentrations at term . They are independent of the kind of delivery ( cesarean section , spontaneous labor ) . Brain levels are distinctly lower and do not show a perinatal peak . In both organs a slight increase of DB00143 concentrations can be observed in the first 3 h after birth . Different litter sizes representing different intrauterine development conditions do not influence DB00143 levels , which is in contrast to the recently reported effect of litter size on prenatal development of hepatic gamma-glutamyltranspeptidase ( P19440 ) activity . We suggest that perinatal P19440 activity and DB00143 levels are regulated by different mechanisms . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Ca2+ response of rat mesangial cells to DB00171 analogues . The aim of this investigation was to characterise the effects of DB00171 analogues and UTP on the single cell intracellular Ca2+ concentration ( [Ca2+]i ) in cultured rat mesangial cells . Typically , there were two phases in the Ca2+ response to the agonists , an initial fast transient peak and a subsequent slower decline , or plateau , phase . For the peak amplitude in [Ca2+]i the agonists had about equal effect . But when taking in consideration the percentage of responding cells and the integrated Ca2+ response over 1 min , the order of efficacy of nucleotide agonists ( 100 microM ) was UTP = DB00171 > ATPgammaS > ADP = 2MeS- DB00171 ( 2-methylthio- DB00171 ) . DB00640 , AMP and beta,gamma-Me- DB00171 ( 100 microM ) had no effect . DB04786 ( 100 microM ) and reactive blue ( 50 microM ) decreased the number of responding cells . Removing Ca2+ from the bath diminished neither the peak in [Ca2+]i nor the percentage of responding cells , but the average [Ca2+]i increase in 1 min was significantly reduced . The results indicate that P41231 receptors are present in rat mesangial cells but it can not be excluded that there are receptors distinct from P41231 which also mediate a rise in [Ca2+]i . 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 , 5- Q13049 and P21554 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 expression , but caused down-regulation of 5- Q13049 and up-regulation of P21554 receptors . Arsenic down-regulated these receptors . However , paracetamol-mediated down-regulation of 5- Q13049 was more pronounced . Arsenic did not modify paracetamol 's effect on P08908 expression , but reduced paracetamol-mediated down-regulation of 5- Q13049 and reversed up-regulation of P21554 receptors . Results suggest arsenic reduced paracetamol-induced analgesia possibly by interfering with pronociceptive 5- Q13049 and antinociceptive P21554 receptors . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Epigenetics override pro-inflammatory PTGS transcriptomic signature towards selective hyperactivation of DB00917 in colorectal cancer . BACKGROUND : Misregulation of the PTGS ( prostaglandin endoperoxide synthase , also known as cyclooxygenase or P36551 ) pathway may lead to the accumulation of pro-inflammatory signals , which constitutes a hallmark of cancer . To get insight into the role of this signaling pathway in colorectal cancer ( CRC ) , we have characterized the transcriptional and epigenetic landscapes of the PTGS pathway genes in normal and cancer cells . RESULTS : Data from four independent series of CRC patients ( 502 tumors including adenomas and carcinomas and 222 adjacent normal tissues ) and two series of colon mucosae from 69 healthy donors have been included in the study . Gene expression was analyzed by real-time PCR and Affymetrix U219 arrays . DNA methylation was analyzed by bisulfite sequencing , dissociation curves , and HumanMethylation450K arrays . Most CRC patients show selective transcriptional deregulation of the enzymes involved in the synthesis of prostanoids and their receptors in both tumor and its adjacent mucosa . DNA methylation alterations exclusively affect the tumor tissue ( both adenomas and carcinomas ) , redirecting the transcriptional deregulation to activation of prostaglandin E2 ( DB00917 ) function and blockade of other biologically active prostaglandins . In particular , Q16647 , P43115 , P43088 , and P15121 were hypermethylated in more than 40 % of all analyzed tumors . CONCLUSIONS : The transcriptional and epigenetic profiling of the PTGS pathway provides important clues on the biology of the tumor and its microenvironment . This analysis renders candidate markers with potential clinical applicability in risk assessment and early diagnosis and for the design of new therapeutic strategies . Effects of extracellular nucleotides in the thyroid : P41231 receptor-mediated P27361 /2 activation and c-Fos induction in PC Cl3 cells . Aim of the present paper was to investigate the signaling pathways of P41231 in rat thyroid PC Cl3 cell line and its effects on proliferation . This study demonstrates that P41231 activation provoked : ( a ) a cytosol-to-membrane translocation of P17252 , -betaI and -epsilon ; ( b ) the phosphorylation of the extra cellular signal-regulated kinases 1 and 2 ( P27361 /2 ) ; ( c ) the expression of c-Fos protein ; ( d ) no effects on the P55008 /S progression and overall cell proliferation . The P41231 -stimulated P27361 /2 phosphorylation was : ( a ) completely blocked by PD098059 , a mitogen-activated protein kinase ( MEK ) inhibitor or by W-7 , a Ca2+-calmodulin ( P62158 ) antagonist ; ( b ) reduced by GF109203X , inhibitor of PKCs , or AG1478 , inhibitor of P00533 tyrosine kinase , or LY294002/wortmannin , inhibitors of phosphoinositide 3-kinases , or cytochalasin D , inhibitor of actin microfilament bundles polymerization . The c-Fos induction was greatly diminished by Go6976 or PD098059 , and completely abolished when combined . In conclusion , data indicate that the P41231 -induced phosphorylation of P27361 /2 and the induction of c-Fos are due to the operation of P62158 , with PKC , PI3K , P00533 and receptor endocytosis mechanisms endorsing the signalling . On the other hand , no mitogenic effects of P41231 are whatsoever noticed in PC Cl3 cells . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . [ Influence of serotonergic transmission on response to olanzapine ] . INTRODUCTION : This study aimed to investigate associations between the response to olanzapine and genetic variations ( polymorphisms ) in serotonergic transmission related genes in a sample of prospectively studied schizophrenic patients treated with this drug . METHODOLOGY : A total of 51 non-related patients with a DSM-IV diagnosis of schizophrenia were treated with olanzapine ( mean dose : 12 mg/day ; range : 5-25 mg ) and followed-up for at least three months . Response to olanzapine was measured by the difference between baseline and post-treatment scores on the PANSS and GAS scales . The following polymorphisms were studied : serotonin receptor 5- Q13049 ( 102-T/C , His452Tyr ) , serotonin receptor P28335 ( Cys23Ser , -330- P19440 /-244-CT ) , and serotonin transporter ( VNTR , 5-HTTLPR ) . RESULTS : Global clinical improvement , measured with both the GAS and PANSS total scores , was observed . When patients were divided into responders and non-responders , the distribution of genotypic and allelic frequencies was similar to the one observed in previous studies with clozapine . When regression analyses were undertaken , polymorphism 330-GT/-244-CT of the P28335 serotonin receptor and 5-HTTLPR of the serotonin transporter showed a tendency towards the association to olanzapine response . CONCLUSIONS : The present study provides preliminary evidence of the important role of variations in serotonin transmission related genes in determining clinical response to olanzapine . Considering previous studies , it can also be concluded that olanzapine and clozapine may have similar affinities to serotonin receptors . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . Comparative effects of azimilide and ambasilide on the human ether-a-go-go-related gene ( Q12809 ) potassium channel . OBJECTIVE : To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related ( Q12809 ) channel . METHODS : Q12809 was stably transfected into Chinese hamster ovary ( CHO- P04264 ) cells and currents were measured using a whole cell , voltage-clamp technique . RESULTS : DB04957 had a ' dual effect ' , inhibiting current at voltage steps above -40 mV and augmenting current at -40 and -50 mV . Tail current inhibition following a step to +30 mV did not vary with temperature ( IC(50) 610 nM at 22 degrees C and 560 nM at 37 degrees C ) . The agonist effect at -50 mV was concentration-dependent and correlated with a hyperpolarizing shift in the V(1/2) of activation ( r=0.98 , P < 0.05 ) . Time constants of inactivation were faster and there was a -10 mV shift in the V(1/2) of steady state inactivation suggestive of open and inactivated state binding . By comparison , ambasilide inhibited Q12809 channels with lower potency ( IC(50) 3.6 microM ) , in a voltage- and time-dependent but frequency-independent manner ( 0.03-1 Hz ) . Ambasilide had no effect on activation or inactivation gating but prolonged both fast and slow components of deactivation consistent with unbinding from the open state . The net effect of both drugs was similar during a voltage ramp which simulated a cardiac action potential . CONCLUSIONS : Inhibition of Q12809 channels by azimilide and ambasilide exhibits a similar time and voltage-dependence . While both exhibit affinity for the open state , azimilide also binds to inactivated channels . Agonist-promoted down-regulation and functional desensitization in two naturally occurring variants of the human serotonin1A receptor . We recently reported two naturally occurring polymorphisms of the human serotonin1A ( P08908 ) receptor : glycine22 --> serine ( Ser22 ) and isoleucine28 --> valine ( Val28 ) in the putative aminoterminal domain of the receptor . To investigate the regulatory properties of these variants , the wild type ( WT ) and variant P08908 receptors were stably expressed in CHO- P04264 cells . WT , Ser22 , and Val28 displayed similar high-affinity binding to [ 3H ] -8-OH-DPAT . Competition experiments with P08908 agonists and antagonists demonstrated similar pharmacological profiles . Receptor agonist-promoted down-regulation was tested by exposure to 100 mumol/L 8-OH-DPAT . After 24-h exposure , WT and Val28 underwent 59.3 +/- 3.9 % and 59.5 +/- 1.4 % reduction in receptor density respectively , whereas the degree of down-regulation was significantly lower for Ser22 ( 21.4 +/- 4.2 % ) . Cell treatment for 24 h with 100 mumol/L 8-OH-DPAT reduced the 5-HT-induced inhibition of DB02527 accumulation by 24.9 +/- 5.1 % for WT and 16.4 +/- 0.8 % for Val28 , but only by 4.8 +/- 3 % for Ser22 . We conclude that the Ser22 variant is capable of attenuating agonist-mediated receptor down-regulation and desensitization .

MH_train_1241

MH_train_1241

MH_train_1241

interacts_with DB00712?

multiple_choice

Prevention and treatment of pancreatic cancer by curcumin in combination with omega-3 fatty acids . Pancreatic cancer BxPC-3 cells were exposed to curcumin , docosahexaenoic acid ( DB01708 ) , or combinations of both and analyzed for proliferation and apoptosis . Pancreatic tumor xenografts were established by injecting BxPC-3 cells into each flank of nude mice . After the tumors reached a size of approximately 190-200 mm(3) , animals were fed diets with or without 2,000 ppm curcumin in 18 % corn oil or 15 % fish oil + 3 % corn oil for 6 more wk before assessing the tumor volume and expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygeanse-2 ( P35354 ) , 5-lipoxinase ( 5- P28300 ) , and P38936 . A synergistic effect was observed on induction of apoptosis ( approximately sixfold ) and inhibition of cell proliferation ( approximately 70 % ) when cells were treated with curcumin ( 5 microM ) together with the DB01708 ( 25 microM ) . Mice fed fish oil and curcumin showed a significantly reduced tumor volume , 25 % ( P < 0.04 ) and 43 % ( P < 0.005 ) , respectively , and importantly , a combination of curcumin and fish oil diet showed > 72 % ( P < 0.0001 ) tumor volume reduction . Expression and activity of P35228 , P35354 , and 5- P28300 are downregulated , and P38936 is upregulated in tumor xenograft fed curcumin combined with fish oil diet when compared to individual diets . The preceding results evidence for the first time that curcumin combined with omega-3 fatty acids provide synergistic pancreatic tumor inhibitory properties . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Cellular distribution and contribution of cyclooxygenase P35354 to diabetogenesis in NOD mouse . Unlike most other mammalian cells , beta-cells of Langerhans constitutively express cyclooxygenase ( P36551 ) -2 rather than P23219 . P35354 is also constitutively expressed in type 1 diabetes ( T1D ) patients ' periphery blood monocytes and macrophage . To understand the role of P35354 in the beta-cell , we investigated P35354 expression in beta-cells and islet infiltrates of NOD and BALB/c mice using fluorescence immunohistochemistry and cytochemical confocal microscopy and Western blotting . Immunostaining showed that P35354 is expressed in islet-infiltrating macrophages , and that the expression of insulin and P35354 disappeared concomitantly from the beta-cells when NOD mice progressed toward overt diabetes . Also cultured P01308 -1E cells coexpressed insulin and P35354 but clearly in different subcellular compartments . Treatment with celecoxib increased insulin release from these cells in a dose-dependent manner in glucose concentrations ranging from 5 to 17 mM . Excessive P35354 expression by the islet-infiltrating macrophages may contribute to the beta-cell death during insulitis . The effects of celecoxib on P01308 -1E cells suggest that PGE(2) and other downstream products of P35354 may contribute to the regulation of insulin release from the beta-cells . Platelet-derived growth factor-D induces expression of cyclooxygenase-2 in rat mesangial cells through activation of PI3K/ P31749 and PKCs . Platelet-derived growth factor ( PDGF ) -D is suggested to be a key factor in the development of several renal pathologies , including mesangioproliferative glomerulonephritis . Cyclooxygenase ( P36551 ) -2 is a protein involved in the biosynthesis of inflammatory prostaglandins . In this study , we investigated the effect of Q9GZP0 on the regulation of P35354 expression in rat mesangial cells ( RMCs ) . Treatment with Q9GZP0 induced P35354 at both the protein and mRNA levels in RMCs , suggesting that the Q9GZP0 -mediated induction of P35354 is due to P35354 transcriptional upregulation . Q9GZP0 treatment also led to a rapid but transient activation of P31749 and extracellular signal regulated kinase ( P29323 ) -1/2 . Activities of JNK-1/2 and p38 MAPK , however , were not influenced by Q9GZP0 in RMCs . Markedly , pharmacological inhibition studies showed that pretreatment with LY294002 ( a PI3K/ P31749 inhibitor ) or GF109203X ( a pan-PKC inhibitor ) suppressed the Q9GZP0 -induced expression of P35354 protein and mRNA , while pretreatment with PD98059 ( an P27361 /2 inhibitor ) or P50391 ( an Src inhibitor ) had no effect on it . These findings collectively demonstrate for the first time that Q9GZP0 induces P35354 by transcriptional upregulation in RMCs and the induction is largely related to PI3K/ P31749 and PKCs activities . Cyclooxygenase 2-derived prostaglandin E2 production by corticotropin-releasing hormone contributes to the activated DB02527 response element binding protein content in rheumatoid arthritis synovial tissue . OBJECTIVE : To determine a mechanism by which corticotropin-releasing hormone ( P06850 ) promotes human inflammatory joint disease progression . METHODS : An ex vivo synovial tissue culture system was established to investigate the functional properties of P06850 at peripheral sites of inflammation . P06850 - and interleukin-1 beta ( P01584 ) -induced prostaglandin E(2) ( PGE(2) ) production from 10 fresh rheumatoid arthritis ( RA ) synovial tissue ( ST ) explants was quantified using a competitive enzyme-linked immunosorbent assay . Modulation of PGE(2) levels was further examined following selective and nonselective cyclooxygenase 2 ( P35354 ) inhibition . Nuclear extracts were analyzed by electrophoretic mobility shift assays to determine functional DB02527 response element binding protein ( CREB ) activity in response to P06850 and PGE(2) in isolated primary synovial cell populations . Western blot analysis measured levels of total and activated ( phosphospecific ) CREB/activating transcription factor ( P39905 ) family members prior to and following stimulation . RESULTS : P06850 , in a time- and dose-dependent manner , significantly ( P = 0.022 ) up-regulated PGE(2) production from 10 fresh RA ST explants . Costimulation of RA ST with P06850 and P01584 significantly augmented ( P = 0.036 ) the effects on PGE(2) production additively over 24 hours . We demonstrated that selective P35354 inhibitors prevent the induction of PGE(2) by both P06850 and P01584 . Further , we provided evidence that P06850 and PGE(2) signal through the induction of CREB and phosphorylated CREB/ P39905 family members in RA ST and in isolated primary RA cell populations . CONCLUSION : Our findings underscore the pathogenic role that P06850 may play in modulating inflammatory joint disease and establish the CREB/ P39905 family of transcription factors as principal effector molecules of proinflammatory mediator action in RA . On the specific interaction between the lysine-binding sites in plasmin and complementary sites in alpha2-antiplasmin and in fibrinogen . P00747 and plasminogen derivatives which contain lysine-binding sites were found to decrease the reaction rate between plasmin and alpha2-antiplasmin by competing with plasmin for the complementary site(s) in alpha2-antiplasmin . The dissocwation constant Kd for the interaction between intact plasminogen ( DB00142 -plasminogen ) and alpha2-antiplasmin is 4.0 microM but those for Lys-plasminogen or TLCK-plasmin are about 10-fold lower indicating a stronger interaction . The lysine-binding site(s) which is situated in triple-loops 1 -- 3 in the plasmin A-chain is mainly responsible for the interaction with alpha2-antiplasmin . The interaction between DB00142 -plasminogen and alpha2-antiplasmin furthermore enhances the activation of DB00142 -plasminogen by urokinase to a comparable extent as DB00513 , suggesting that similar conformational changes occur in the proenzyme after complex formation . DB09222 , fibrinogen digested with plasmin , purified fragment E and purified fragment D interfere with the reaction between plasmin and alpha2-antiplasmin by competing with alpha2-antiplasmin for the lysine-binding site(s) in the plasmin A-chain . The Kd obtained for these interactions varied between 0.2 microM and 1.4 microM ; fragment E being the most effective . Thus the fibrinogen molecule contains several complementary sites to the lysine-binding sites located both in its NH2-terminal and COOH-terminal regions ; these sites are to a large extent . Analgesic and Anti-Inflammatory Activities of Methanol Extract of Cissus repens in Mice . The aim of this study was to investigate possible analgesic and anti-inflammatory mechanisms of the CR(MeOH) . Analgesic effect was evaluated in two models including acetic acid-induced writhing response and formalin-induced paw licking . The anti-inflammatory effect was evaluated by λ-carrageenan-induced mouse paw edema and histopathologic analyses . The results showed that CR(MeOH) ( 500 mg/kg ) decreased writhing response in the acetic acid assay and licking time in the formalin test . CR(MeOH) ( 100 and 500 mg/kg ) significantly decreased edema paw volume at 4th to 5th hours after λ-carrageenan had been injected . Histopathologically , CR(MeOH) abated the level of tissue destruction and swelling of the edema paws . These results were indicated that anti-inflammatory mechanism of CR(MeOH) may be due to declined levels of NO and MDA in the edema paw through increasing the activities of SOD , GPx , and GRd in the liver . Additionally , CR(MeOH) also decreased IL-1β , P05231 , NFκB , P01375 -α , P35354 , and P35228 levels . The contents of two active ingredients , ursolic acid and lupeol , were quantitatively determined . This paper demonstrated possible mechanisms for the analgesic and anti-inflammatory effects of CR(MeOH) and provided evidence for the classical treatment of Cissus repens in inflammatory diseases . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Q9GZP0 inhibition by DB05139 ameliorates tubulointerstitial fibrosis following experimental glomerulonephritis . BACKGROUND : Arresting or regressing kidney scarring is of major clinical relevance . Q9GZP0 ( Q9GZP0 ) is widely expressed in fibrotic kidneys . Administration of the Q9GZP0 neutralizing fully human monoclonal antibody DB05139 in the acute phase of progressive anti-Thy 1.1 glomerulonephritis reduced glomerular and secondary tubulointerstitial damage . METHODS : Using this model , we now assessed the effects of DB05139 ( n=15 ) vs irrelevant control IgG ( n=17 ) administered on days 17 , 28 and 35 after disease induction , i.e. after acute glomerular damage had subsided . RESULTS : In vitro , DB05139 inhibited the Q9GZP0 - but not the PDGF-B-induced proliferation of rat renal fibroblasts . Following the first DB05139 injection on day 17 , exposure to therapeutic levels was maintained until day 49 . Proteinuria in the DB05139 -treated group was transiently reduced between days 49 and 77 ( -19 to -23 % in comparison with the controls ; P < 0.05 ) . On day 100 , DB05139 treatment reduced the number of rats that had doubled their serum creatinine ( DB05139 : 40 vs controls : 71 % ; P < 0.05 ) . Compared with controls , the DB05139 animals , on day 100 , significantly lowered glomerular expression of vimentin and collagens as well as tubulointerstitial damage scores , interstitial fibrosis , vimentin and cortical Q9GZP0 mRNA levels . CONCLUSIONS : Q9GZP0 antagonism , even after the phase of acute glomerular damage , exerts beneficial effects on the course of tubulointerstitial damage , i.e. the final common pathway of most renal diseases . Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 gene deletion and DB01997 were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells . Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention , the molecular mechanisms underlying such chemopreventive activity are not clear . Since peroxisome proliferator-activated receptor gamma ( P37231 ) has been implicated in regulating breast cancer cell differentiation and apoptosis , the effects of beta-carotene on the P37231 -mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study . The results demonstrated that beta-carotene significantly increased P37231 mRNA and protein levels in time-dependent manner . In addition , beta-carotene increased the cyclin-dependent kinase inhibitor P38936 ( P38936 /CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression . DB07863 ( GW9662 ) , an irreversible P37231 antagonist , partly attenuated the cell death caused by beta-carotene . Further , reactive oxygen species ( ROS ) production was induced by beta-carotene , resulting in mitochondrial dysfunction and cytochrome C release . DB00143 ( DB00143 ) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene . In total , these observations suggest that the synergistic effect of P37231 expression and ROS production may account for beta-carotene-mediated anticancer activities . The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor-1 ( P05121 ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline- and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai1-/- mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 ) and instead reflected proteolytic activation and release of P14210 with subsequent induction of P35354 . That the P14210 / P35354 / DB00917 axis mediates in vivo protection from fibrosis in Pai1-/- mice was demonstrated by experiments showing that a selective inhibitor of the P08581 c- DB00134 increased lung collagen to WT levels while reducing P35354 protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway . Agents with selective estrogen receptor ( ER ) modulator activity induce apoptosis in vitro and in vivo in ER-negative glioma cells . Tamoxifen , a member of the selective estrogen receptor modulator ( SERM ) family , is widely used in the treatment of estrogen receptor ( ER ) -expressing breast cancer . It has previously been shown that high-dose tamoxifen has cytotoxic activity against glioma cells , but whether this effect is drug specific or represents a general property of SERMs is unknown . In this study , we demonstrate that tamoxifen and DB05487 , a novel benzopyranone with SERM activity , induce glioma cell apoptosis in a dose- and time-dependent manner . Moreover , administration of tamoxifen and DB05487 suppresses tumor growth in vivo and extends animal survival in glioma xenograft models . None of the eight glioma cell lines examined express either P03372 or -beta , suggesting the mechanism for tamoxifen- and DB05487 -induced glioma cell apoptosis is independent of the ER signaling pathway . Complementary DNA microarray expression profiling allowed us to identify a subset of genes specifically regulated by tamoxifen and DB05487 , and not by other apoptotic stimuli , including nuclear factor ( NF ) -kappaB with its target genes IEX-3 , P04179 , P05231 , and P10145 . We demonstrate that suppression of NF-kappaB activation markedly enhances SERM-induced apoptosis , suggesting a role for NF-kappaB in protecting glioma cells from SERM-induced cytotoxicity . These findings demonstrate for the first time that a SERM other than tamoxifen can induce glioma cell apoptosis in vitro and in vivo and that the clinical efficacy of SERMs for the treatment of malignant gliomas could potentially be enhanced by simultaneous inhibition of the NF-kappaB pathway . Proliferation of endothelial cell on polytetrafluoroethylene vascular graft materials carried P15692 gene plasmid . OBJECTIVE : To investigate whether vascular endothelial growth factor ( P15692 ) gene plasmid carried by polytetrafluoroethylene ( PTFE ) vascular graft materials could transfect endothelial cells ( ECs ) and promote their growth . METHODS : PTFE vascular graft materials carried with pCDI-hVEGF(121) , pCDI or pEGFP were incubated in DB03754 -buffer solution and the values of optical density of 260 nm at different time were plotted , then the DNA controlled release curve was made . ECs derived from human umbilical vein were seeded on the pCDI-hVEGF(121)/pCDI/pEGFP-PTFE materials or tissue culture plates , ECs numbers were counted and P15692 protein concentrations at different time were measured by enzyme-linked immunoadsorbent assay method . Green fluorescent protein ( GFP ) expression in ECs on pEGFP-PTFE materials was examined with fluorescence microscopy . RESULTS : The controlled release curve showed that the gene released from PTFE materials was rapid within 8 h , then slowed down and that the gene released continuously even after 72 h . At 24 , 72 and 120 h , ECs number and proliferation rate of pCDI-hVEGF(121)-PTFE materials were higher than those of pCDI or pEGFP-PTFE materials ( P < 0.05 ) . P15692 protein concentration of pCDI-hVEGF(121)-PTFE materials was higher than that of pCDI or pEGFP-PTFE materials at 6 , 24 , 72 and 120 h ( P < 0.01 ) . GFP expression in ECs on the pEGFP-PTFE materials could be detected by fluorescence microscopy . CONCLUSION : PTFE graft can be used as a carrier of P15692 gene plasmid , P15692 gene carried by PTFE can transfect ECs and promote ECs growth . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . A Nile blue based infrared fluorescent probe : imaging tumors that over-express cyclooxygenase-2 . The first Golgi-localized cyclooxygenase-2 ( P35354 ) -specific near-infrared ( Q9Y3T9 ) fluorescent probe , Niblue- P13671 -IMC , able to detect cancer cells , was designed . Importantly , Niblue- P13671 -IMC preferentially labeled the tumors in a mouse tumor model with deep tissue penetration capacity . It may be a promising molecular tool for guiding tumor resection during surgery . The impact of biological agents interfering with receptor/ligand binding in the immune system . We herein discuss the impact of biological agents based on the ability of monoclonal antibodies to target specific molecules . This approach has given to clinical immunologists a spectrum of drugs able to manipulate the immune system . In the first session , we discuss drugs targeting T-cell function by : ( 1 ) targeting P10747 mediated costimulation ( DB01281 and DB06681 ) ; ( 2 ) interfering with interleukin-2 receptor ( DB00074 and DB00111 ) ; ( 3 ) blocking cell adhesion and homing ( DB00092 , DB00095 , DB00108 ) . The second session is dedicated to drugs targeting cytokines or their receptors . The best known and largely experimented case is represented by drugs targeting tumor necrosis factor ( P01375 ) ( DB00065 , Adalilumab , Certolizumab ) or its p75 receptor ( DB00005 ) . However , newer products are now available to target other inflammatory cytokines including P05231 , P10145 , IL-12 , P40933 , Q14116 , IL-23 . These agents have the potential to become powerful tools in the control of several immune-mediated diseases , especially auto-immune and inflammatory ones . They traslate into reality the prediction that antibodies will eventually become " magic bullets which seek their own target " ( P. Ehrich , 1906 ) .

MH_train_1242

MH_train_1242

MH_train_1242

interacts_with DB06271?

multiple_choice

Ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in human aqueous humor . PURPOSE : The aim of this study was to evaluate the ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in subjects undergoing routine cataract surgery with intraocular lens implantation . METHODS : An open-label , phase II confirmatory study of 54 subjects undergoing cataract surgery with intraocular lens implantation . A single drop of bromfenac sodium ophthalmic solution 0.1 % was administered at 30 , 60 , 90 , 120 , 180 , or 240 min prior to the initiation of cataract surgery . Samples of aqueous humor were aspirated through a paracentesis and analyzed by using high-performance liquid chromatography . Based upon these data , predicted concentrations of bromfenac in the aqueous humor over 24 h were generated by using computer simulation and compared with the IC(50) for bromfenac as a measure of anti-inflammatory efficacy . RESULTS : Peak aqueous-humor concentrations of bromfenac occurred between 150 and 180 min following ophthalmic dosing , with a mean concentration of 78.7 ng/mL . The level of bromfenac decreased in a log-linear fashion with an elimination-rate constant of 1.4 . DB00963 remained above the IC(50) value of cyclo-oxygenase-2 ( P35354 ) during the evaluated time points and over the 12-h dosing interval , using a computer model of extrapolated time points and assuming first-order elimination . CONCLUSIONS : Pharmacokinetic modeling , based upon samples collected over 240 min after a single dose of bromfenac sodium ophthalmic solution 0.1 % suggests that aqueous-humor concentrations remain at clinically effective levels ( above its IC(50) value for P35354 ) for over 12 h . Based upon this rationale , these results supported clinical trials that demonstrated the efficacy of twice-daily dosing of bromfenac sodium ophthalmic solution 0.1 % to manage patients with postoperative ocular pain and inflammation . Effects of an alpha 7-nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3-(2,4-dimethoxybenzylidene)-anabaseine ( DB05708 ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 , the α7-nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 genotype . CONCLUSIONS : The observed DB05708 -related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7-nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents . Gene regulation by hypoxia and the neurodevelopmental origin of schizophrenia . Neurodevelopmental changes may underlie the brain dysfunction seen in schizophrenia . While advances have been made in our understanding of the genetics of schizophrenia , little is known about how non-genetic factors interact with genes for schizophrenia . The present analysis of genes potentially associated with schizophrenia is based on the observation that hypoxia prevails in the embryonic and fetal brain , and that interactions between neuronal genes , molecular regulators of hypoxia , such as hypoxia-inducible factor 1 ( Q9BYW2 ) , and intrinsic hypoxia occur in the developing brain and may create the conditions for complex changes in neurodevelopment . Consequently , we searched the literature for currently hypothesized candidate genes for susceptibility to schizophrenia that may be subject to ischemia-hypoxia regulation and/or associated with vascular expression . Genes were considered when at least two independent reports of a significant association with schizophrenia had appeared in the literature . The analysis showed that more than 50 % of these genes , particularly P31749 , P23560 , O75052 , P32238 , P36544 , P21554 , P21964 , DNTBP1 , Q99259 , Q14832 , P22301 , MLC1 , Q99466 , Q02297 , P43354 / P43354 , O43272 , P78509 , P49798 , Q9NQC3 / Q9NQC3 and P01375 , are subject to regulation by hypoxia and/or are expressed in the vasculature . Future studies of genes proposed as candidates for susceptibility to schizophrenia should include their possible regulation by physiological or pathological hypoxia during development as well as their potential role in cerebral vascular function . Polymorphism identification in the P11310 , P01008 , P22301 , P15173 and P01222 genes of cattle . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Defects in auxiliary redox proteins lead to functional methionine synthase deficiency . Q99707 catalyzes a methyl transfer reaction from methyltetrahydrofolate to homocysteine to form methionine and DB00116 and is dependent on methylcobalamin , a derivative of vitamin B12 , for activity . Due to the lability of the intermediate , cob(I)alamin , the activity of methionine synthase is additionally dependent on a redox activation system . In bacteria , two flavoproteins , NADPH-flavodoxin reductase and flavodoxin , shuttle electrons from NADPH to methionine synthase . Their mammalian counterparts are unknown , and a putative intrinsic thiol oxidase activity of the mammalian methionine synthase has been proposed to be involved . We demonstrate that the mammalian methionine synthase can be activated in an NADPH-dependent reaction and requires a minimum of two redox proteins . This model is consistent with our results from biochemical complementation studies between cblG and cblE cell lines and mutation detection analysis in cblG cell lines . These demonstrate that the cblG cell line has defects affecting methionine synthase directly , whereas the cblE cell line has defects in the redox proteins . We have also identified a P1173L mutation in the activation domain of methionine synthase in the cblG cell line WG1505 . Effects of phenacetin and its metabolite p-phenetidine on P23219 and P35354 activities and expression in vitro . The present study was aimed to test the possible cyclooxygenase ( P36551 ) -1/ P35354 selectivity of the old analgesic drug phenacetin and its metabolite p-phenetidine , which exhibits high renal toxicity . DB00316 ( acetaminophen ) , the main metabolite of phenacetin with low renal toxicity , and indomethacin were selected as reference compounds . Collagen-stimulated platelet thromboxane B2 ( TxB2 ) production and phorbol 12-myristate-13-acetate ( PMA ) -induced neutrophil prostaglandin E2 ( DB00917 ) synthesis were used as indicators for P23219 and P35354 activity , respectively . DB03783 was even less potent than paracetamol to reduce the production of both TxB2 and DB00917 , and no clear preference for either of the P36551 -enzymes was seen . P-phenetidine was a more potent inhibitor , already at nanomolar level , of the synthesis of these prostanoids than indomethacin and showed some preference to P35354 inhibition . Somewhat higher , micromolar , concentrations of p-phenetidine also reduced P35354 expression in neutrophils . We suggest that the very potent inhibitory activity of p-phenetidine on DB00917 synthesis combined with the reduction of P35354 expression could explain the renal papillary necrosis in phenacetin kidney . [ DB09043 ( Eperzan ) : a new once-weekly agonist of glucagon-like peptide-1 receptors ] . DB09043 ( Eperzan ) is a new once-weekly agonist of Glucagon-Like Peptide-1 ( P0C6A0 ) receptors that is indicated in the treatment of type 2 diabetes . Two doses are available , 30 mg and 50 mg , to be injected subcutaneously once a week . It has been extensively evaluated in the HARMONY programme of eight large randomised controlled trials that were performed at different stages of type 2 diabetes , in comparison with placebo or an active comparator . The endocrine and metabolic effects of albiglutide are similar to those of other P43220 agonists : stimulation of insulin secretion ( incretin effect ) and inhibition of glucagon secretion , both in a glucose-dependent manner , retardation of gastric emptying and increase of satiety . These effects lead to a reduction in glycated haemoglobin ( HbA(1c) ) levels , combined with a weight reduction . The overall tolerance profile is good . DB09043 is currently reimbursed in Belgium after failure ( HbA(1c) > 7.5 % ) of and in combination with a dual therapy with metformin and a sulfonylurea as well as in combination with a basal insulin ( with or without oral antidiabetic drugs ) . To avoid hypoglycaemia , a reduction in the dose of sulfonylurea or insulin may be recommended . A once-weekly administration should increase patient 's acceptance of injectable therapy and improve compliance . A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 and P35354 ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg . [ Activation of coagulation cascade in children during an idiopathic nephrotic syndrome relapse ] . The objective of this study was to assess concentrations of selected markers of coagulation in children with relapse of idiopathic nephrotic syndrome during a 6-week therapy . Study groups : 22 subjects ( 32 relapses ) -- 14 males , 8 females ( mean age 7.15 +/- 1.5 y. ) with no thrombotic complications were included into the study . All children were clinically steroid-sensitive . METHODS : Coagulation markers ( platelet count , thrombin time , APTT , INR , fibrinogen 1 + 2 fragments ( F1 + 2 ) , thrombin-antithrombin complexes ( TAT ) , serum levels of D-dimer ( DD ) , fibrin monomers ( FM ) and antithrombin activity ( P01008 ) ) were measured three times : on admission , after 2 and 6 weeks . The control group consisted of 13 healthy children . RESULTS : Serum concentration of TAT or F1 + 2 did not differ between 3 stages ( p > 0.05 ) . However , values at 0 and 2 weeks were significantly higher than in control group ( p < 0.05 ) . We found no correlation between TAT or F1 + 2 and FBG , ALB , TCH , TG levels . [ table : see text ] CONCLUSIONS : The coagulation cascade in relapse of NS was activated during first 6 weeks of therapy whereas metabolic disturbances ( low ALB , high P02675 , TCH , TG , high platelets ) normalized . It is speculative whether it was caused by active immunological process but definitely it resulted in " prothrombotic state " in P01308 patients . Genetic polymorphisms for the study of multifactorial stroke . Single-gene disorders explain only a minority of stroke cases . Stroke represents a complex trait , which is usually assumed to be polygenic . On this topic , the role of a wide number of candidate genes has been investigated in stroke through association studies , with controversial results . Therefore , it is difficult for the clinician to establish the validity and the level of clinical applicability of the previously reported associations between genetic factors and stroke . This review is an update and an extensive analysis of the more recent association studies conducted in stroke . We evaluated a number of studies on several candidate genes ( including P12259 , F2 , P02671 / P02675 / P02679 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / Q9UKI9 / P04054 / P08514 , P17301 , P07359 , P12821 , AGT , NOS3 , P02649 , P06858 , P27169 , Q08499 , P20292 , P42898 , Q99707 , and P35520 ) , providing a final panel of genes and molecular variants . We categorized this panel in relation to the degree of association with stroke , supported by the results of meta-analyses and case-control studies . Our findings could represent a useful tool to address further molecular investigations and to realize more detailed meta-analyses . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Arundic acid ( DB05343 ) ameliorates delayed ischemic brain damage by preventing astrocytic overproduction of P04271 . After focal cerebral ischemia , the infarct volume increases rapidly within acute infarct expansion ( initial 12 to 24 h ) and continues slowly during delayed infarct expansion ( 25 to 168 h ) . While acute infarct expansion represents progressive necrosis within the ischemic core , delayed infarct expansion starts as disseminated apoptotic cell death in a narrow rim surrounding the infarct border , which gradually coalesces to form a larger infarct . Discovery of a distinct correlation between reactive astrogliosis along the infarct border and delayed infarct expansion in the rodent ischemia model led us to investigate the possible causal relationship between the two events . Specifically , the calcium binding protein P04271 exerts detrimental effects on cell survival through activation of various intracellular signaling pathways , resulting in altered protein expression . Arundic acid [ ( R ) -(-)-2-propyloctanoic acid , DB05343 ] is a novel agent that inhibits P04271 synthesis in cultured astrocytes . In the rodent ischemia model , this agent was shown to inhibit both the astrocytic overexpression of P04271 and the subsequent activation of signaling pathways in the peri-infarct area . Concurrently , delayed infarct expansion was prevented , and neurologic deficits were promptly ameliorated . The results of subsequent studies suggest that the efficacy of arundic acid is mediated by restoring the activity of astroglial glutamate transporters via enhanced genetic expression . Antithrombin III concentrate in the acute phase of thermal injury . BACKGROUND : Thermal injury disrupts homeostasis by inducing subclinical disseminated intravascular coagulation , fibrinolysis. and an acquired deficiency of Antithrombin III ( P01008 ) , a natural anticoagulant . As a result , thermally injured patients have a high incidence of hypercoagulability and thrombosis . OBJECTIVE : P01008 ( Human ) concentrate was given to a thermally injured patient to evaluate safety , and dosage requirements in this setting . DESIGN : The patient was a 40 yr old male with a 68 % total burn surface area , right femoral comminuted fracture , and P01031 - P13671 subluxation sustained in a vehicular crash . He received nine infusions of AT III ( H ) concentrate ( 100-50 u/kg ) within the first four days of injury . RESULT : The P01008 plasma level increased from 45 % on admission ( normal = 100+/-20 % ) to 120+/-25 % in the next four days . During the 64 day hospitalization , there were 11 grafting procedures with an estimated blood loss ( EBL ) /procedure : 1140 cc ; and EBL/grafted surface area ratio : 0.6 cc cm2 . The average time to healing of the meshed autograft was 6.4 days . CONCLUSION : P01008 ( H ) concentrate can be safely utilized in the acute phase of thermal injury : no excessive bleeding or prolongation of wound healing was documented . Thrombin antithrombin complex and Q14116 serum levels in stroke patients . The complex picture of inflammation and coagulation alterations comes to life in acute stroke phases . Increasing evidence points to a strong interaction and extensive crosstalk between the inflammation and coagulation systems : the interest towards this relationship has increased since recent experimental research showed that the early administration of antithrombin III ( P01008 ) decreases the volume of ischemia in mice and might be neuroprotective , playing an antiinflammatory role.We aimed to establish the extent of the relationship among markers of inflammation ( P04271 and Q14116 ) and procoagulant and fibrinolytic markers ( P01008 , thrombin-antithrombin III complex ( TAT ) , Fibrin Degradation Products ( Q9NRC9 ) , D-dimer ) in 13 comatose patients affected by focal cerebral ischemia.Plasma levels of TAT , D-dimer and Q9NRC9 , Q14116 and P04271 were increased . Q14116 and P04271 high serum levels in ischemic patients suggest an early activation of the inflammatory cascade in acute ischemic injury.The basic principles of the interaction between inflammatory and coagulation systems are revised , from the perspective that simultaneous modulation of both coagulation and inflammation , rather than specific therapies aimed at one of these systems could be more successful in stroke therapy . A P43220 agonist liraglutide inhibits endothelial cell dysfunction and vascular adhesion molecule expression in an ApoE-/- mouse model . The glucagon like peptide-1 receptor ( P43220 ) agonist liraglutide attenuates induction of plasminogen activator inhibitor type-1 ( P05121 ) and vascular adhesion molecule ( VAM ) expression in human vascular endothelial cells ( hVECs ) in vitro and may afford protection against endothelial cell dysfunction ( O95905 ) , an early abnormality in diabetic vascular disease . Our study aimed to establish the dependence of the in vitro effects of liraglutide on the P43220 and characterise its in vivo effects in a mouse model of O95905 . In vitro studies utilised the human vascular endothelial cell line P10144 - Q8IWL8 and enzyme-linked immunosorbent assays ( ELISA ) for determination of P05121 and VAM expression . In vivo studies of vascular reactivity and immunohistochemical analysis were performed in the ApoE(-/-) mouse model . In vitro studies demonstrated P43220 -dependent liraglutide-mediated inhibition of stimulated P05121 and VAM expression . In vivo studies demonstrated significant improvement in endothelial function in liraglutide treated mice , a P43220 dependent effect . DB06655 treatment also increased endothelial nitric oxide synthase ( P29474 ) and reduced intercellular adhesion molecule-1 ( P05362 ) expression in aortic endothelium , an effect again dependent on the P43220 . Together these studies identify in vivo protection , by the P43220 agonist liraglutide , against O95905 and provide a potential molecular mechanism responsible for these effects . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . [ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity .

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interacts_with DB01267?

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Oxidative stress in susceptibility to breast cancer : study in Spanish population . BACKGROUND : Alterations in the redox balance are involved in the origin , promotion and progression of cancer . Inter-individual differences in the oxidative stress regulation can explain a part of the variability in cancer susceptibility.The aim of this study was to evaluate if polymorphisms in genes codifying for the different systems involved in oxidative stress levels can have a role in susceptibility to breast cancer . METHODS : We have analyzed 76 single base polymorphisms located in 27 genes involved in oxidative stress regulation by SNPlex technology . First , we have tested all the selected SNPs in 493 breast cancer patients and 683 controls and we have replicated the significant results in a second independent set of samples ( 430 patients and 803 controls ) . Gene-gene interactions were performed by the multifactor dimensionality reduction approach . RESULTS : Six polymorphisms rs1052133 ( O15527 ) , rs406113 and rs974334 ( GPX6 ) , rs2284659 ( P08294 ) , rs4135225 ( P10599 ) and rs207454 ( P47989 ) were significant in the global analysis . The gene-gene interactions demonstrated a significant four-variant interaction among rs406113 ( GPX6 ) , rs974334 ( GPX6 ) , rs105213 ( O15527 ) and rs2284659 ( P08294 ) ( p-value = 0.0008 ) with high-risk genotype combination showing increased risk for breast cancer ( OR = 1.75 [ 95 % CI ; 1.26-2.44 ] ) . CONCLUSIONS : The results of this study indicate that different genotypes in genes of the oxidant/antioxidant pathway could affect the susceptibility to breast cancer . Furthermore , our study highlighted the importance of the analysis of the epistatic interactions to define with more accuracy the influence of genetic variants in susceptibility to breast cancer . Low pH stimulates vasopressin V2 receptor promoter activity and enhances downregulation induced by V1a receptor stimulation . DB00067 ( AVP ) plays a key role in the urine concentration mechanism via the vasopressin V2 receptor ( P30518 ) and aquaporin 2 ( P41181 ) in the kidney . It is well known that P30518 is localized on the basolateral side and the V1a receptor ( P37288 ) is distributed on the luminal side of the collecting ducts . Previously , we reported an increase of P37288 mRNA and a decrease of P30518 mRNA in the collecting ducts under chronic metabolic acidosis . However , the regulatory mechanism of P30518 in acidic conditions has not yet been determined . In the present study , we investigated the effect of changes in pH on P30518 promoter activity , using the LLC-PK(1) cell line stably expressing rat P37288 ( LLC-PK(1)/rV1aR ) . The rV2R promoter activity was significantly increased at 12 h after the incubation in low-pH conditions , which was sustained for 24 h . mRNA and protein expressions of P30518 were also increased in low-pH conditions . P37288 stimulation suppressed rV2R promoter activity in a pH-dependent manner . PKA and JNK inhibitors suppressed rV2R promoter activity in both neutral and low-pH conditions without FBS . However , a JNK inhibitor prevented the increase of P30518 promoter activity only in low-pH conditions in the presence of FBS . In summary , P30518 expression is increased at transcriptional , mRNA , and protein levels in LLC-PK(1)/rV1aR cells under low-pH conditions . Acidic condition-induced P30518 enhancement was suppressed by P37288 stimulation , suggesting the crucial role of P37288 in water and electrolyte homeostasis in acidosis . DB01233 stimulates catecholamine- and granin-derived peptide secretion from pheochromocytoma cells through activation of serotonin type 4 ( Q13639 ) receptors . The gastroprokinetic agent metoclopramide is known to stimulate catecholamine secretion from pheochromocytomas . The aim of the study was to investigate the mechanism of action of metoclopramide and expression of serotonin type 4 ( 5-HT(4) ) receptors in pheochromocytoma tissues . Tissue explants , obtained from 18 pheochromocytomas including the tumor removed from a 46-year-old female patient who experienced life-threatening hypertension crisis after metoclopramide administration and 17 additional pheochromocytomas ( 9 benign and 8 malignant ) were studied . Cultured pheochromocytoma cells derived from the patient who previously received metoclopramide were incubated with metoclopramide and various 5-HT(4) receptor ligands . In addition , total mRNAs were extracted from all the 18 tumors . Catecholamine- and granin-derived peptide concentrations were measured in pheochromocytoma cell incubation medium by HPLC and radioimmunological assays . In addition , expression of 5-HT(4) receptor mRNAs in the 18 pheochromocytomas was investigated by the use of reverse transcriptase-PCR . RESULTS : DB01233 and the 5-HT(4) receptor agonist cisapride were found to activate catecholamine- and granin-derived peptide secretions by cultured tumor cells . DB01233 - and cisapride-evoked catecholamine- and granin-derived peptide productions were inhibited by the 5-HT(4) receptor antagonist GR 113808 . 5-HT(4) receptor mRNAs were detected in the patient 's tumor and the series of 17 additional pheochromocytomas . This study shows that pheochromocytomas express functional 5-HT(4) receptors that are responsible for the stimulatory action of metoclopramide on catecholamine- and granin-derived peptide secretion . All 5-HT(4) receptor agonists must therefore be contraindicated in patients with proven or suspected pheochromocytoma . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Signatures of positive selection in genes associated with human skin pigmentation as revealed from analyses of single nucleotide polymorphisms . Phenotypic variation between human populations in skin pigmentation correlates with latitude at the continental level . A large number of hypotheses involving genetic adaptation have been proposed to explain human variation in skin colour , but only limited genetic evidence for positive selection has been presented . To shed light on the evolutionary genetic history of human variation in skin colour we inspected 118 genes associated with skin pigmentation in the Perlegen dataset , studying single nucleotide polymorphisms ( SNPs ) , and analyzed 55 genes in detail . We identified eight genes that are associated with the melanin pathway ( Q9UMX9 , Q04671 , P17643 , P40126 , P21583 , P00533 , P14416 and Q03181 ) and presented significant differences in genetic variation between Europeans , Africans and Asians . In six of these genes we detected , by means of the EHH test , variability patterns that are compatible with the hypothesis of local positive selection in Europeans ( Q04671 , P17643 and P21583 ) and in Asians ( Q04671 , P40126 , P21583 , P00533 and P14416 ) , whereas signals were scarce in Africans ( P40126 , P00533 and P14416 ) . Furthermore , a statistically significant correlation between genotypic variation in four pigmentation candidate genes and phenotypic variation of skin colour in 51 worldwide human populations was revealed . Overall , our data also suggest that light skin colour is the derived state and is of independent origin in Europeans and Asians , whereas dark skin color seems of unique origin , reflecting the ancestral state in humans . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Nanocell targeting using engineered bispecific antibodies . There are many design formats for bispecific antibodies ( BsAbs ) , and the best design choice is highly dependent on the final application . Our aim was to engineer BsAbs to target a novel nanocell ( EnGeneIC Delivery Vehicle or EDV(TM)nanocell ) to the epidermal growth factor receptor ( P00533 ) . EDV(TM)nanocells are coated with lipopolysaccharide ( LPS ) , and BsAb designs incorporated single chain Fv ( scFv ) fragments derived from an anti-LPS antibody ( 1H10 ) and an anti- P00533 antibody , DB01269 . We engineered various BsAb formats with monovalent or bivalent binding arms and linked scFv fragments via either glycine-serine ( P15848 ) or Fc-linkers . Binding analyses utilizing ELISA , surface plasmon resonance , bio-layer interferometry , flow cytometry and fluorescence microscopy showed that binding to LPS and to either soluble recombinant P00533 or MDA-MB-468 cells expressing P00533 , was conserved for all construct designs . However , the Fc-linked BsAbs led to nanocell clumping upon binding to EDV(TM)nanocells . Clumping was eliminated when additional disulfide bonds were incorporated into the scFv components of the BsAbs , but this resulted in lower BsAb expression . The P15848 -linked tandem scFv BsAb format was the optimal design with respect to EDV binding and expression yield . Doxorubicin-loaded EDV(TM)nanocells actively targeted with tandem scFv BsAb in vivo to MDA-MB-468-derived tumors in mouse xenograft models enhanced tumor regression by 40 % compared to passively targeted EDV(TM)nanocells . BsAbs therefore provide a functional means to deliver EDV(TM)nanocells to target cells . DB05229 sodium , a stable prostacyclin analogue , elicits dilation of isolated porcine retinal arterioles : roles of P29474 and potassium channels . PURPOSE : DB01240 ( DB01240 ) is usually described as an endoEDRFsthelium-derived relaxing factor , but the vasoreactivity to DB01240 in the retinal arterioles and the underlying mechanisms are not fully understood . We examined the effects of DB01240 on the retinal microcirculation using beraprost sodium ( BPS ) , a stable DB01240 analogue , and the signaling mechanisms involved in this vasomotor activity . METHODS : Porcine retinal arterioles were isolated , cannulated , and pressurized without flow in vitro . Video microscopic techniques recorded the diametric responses to BPS . RESULTS : DB05229 sodium elicited dose-dependent ( 0.1 pM-0.1 μM ) vasodilation of the retinal arterioles that was abolished by the P43119 ( IP ) antagonist CAY10441 . DB05229 sodium-induced vasodilation decreased by 50 % after the endothelium was removed and was inhibited by the nitric oxide ( NO ) synthase inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) comparable with denudation . Inhibition of soluble guanylyl cyclase by 1H-1,2,4-oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) and blockage of protein kinase A ( PKA ) by Rp-8-Br-cAMPS were comparable to L-NAME . DB05229 sodium-induced vasodilation was also inhibited by the nonselective potassium channel inhibitor , tetraethylammonium , and the adenosine triphosphate-sensitive potassium ( KATP ) channel blocker , glibenclamide . Residual vasodilation in the presence of glibenclamide decreased further with subsequent application of ODQ . CONCLUSIONS : DB05229 sodium , a stable DB01240 analogue , causes vasodilation of the retinal arterioles mediated via the IP receptor . The current findings suggest that BPS elicits endothelium-dependent and -independent dilation of the retinal arterioles mediated by NO induced by activation of PKA in the endothelium and the KATP channel activation in the vascular smooth muscle , respectively . Circulating levels of immunoreactive cytokines in women with preeclampsia . PROBLEM : Circulating inflammatory cytokines have been implicated in the pathogenesis of preeclampsia . To test this hypothesis , we measured plasma levels of immunoreactive tumor necrosis factor ( P01375 ) -alpha and -beta , interleukin ( IL ) -1 alpha and -beta , and P05231 and -10 in women with preeclampsia , in women with transient gestational hypertension , and throughout normal pregnancy . METHOD OF STUDY : Enzyme-linked immunosorbent assays were used and subjected to extensive validation studies . RESULTS : The median concentration of plasma P01375 was increased by twofold in women with preeclampsia compared with that in normal third-trimester pregnancy ( P < 0.001 ) and in women with gestational hypertension ( P < 0.04 ) . The median concentration of plasma P05231 was increased by threefold in women with preeclampsia compared with that in normal third-trimester pregnancy ( P < 0.001 ) and increased twofold compared with that in women with gestational hypertension ( P < 0.1 ) . There were no significant differences observed in the levels of plasma P01584 and P22301 between the preeclamptic and other subject groups . The level of P01584 , but not the levels of P22301 , P01375 , or P05231 , was significantly changed during normal pregnancy compared with the nonpregnant condition manifesting an overall decline ( P < 0.04 ) . P01374 and P01583 were not detected in any samples , possibly because of the low sensitivity of these particular immunoassays . CONCLUSION : Elevated levels of P01375 and P05231 may contribute to the putative endothelial dysfunction of preeclampsia . Inhibition of p38alpha MAPK disrupts the pathological loop of proinflammatory factor production in the myelodysplastic syndrome bone marrow microenvironment . Myelodysplastic syndromes ( P43034 ) are common causes of ineffective hematopoiesis and cytopenias in the elderly . Various myelosuppressive and proinflammatory cytokines have been implicated in the high rates of apoptosis and hematopoietic suppression seen in P43034 . We have previously shown that p38 MAPK is overactivated in P43034 hematopoietic progenitors , which led to current clinical studies of the selective p38alpha inhibitor , DB05412 , in this disease . We now demonstrate that the myelosuppressive cytokines TNFalpha and IL-1beta are secreted by bone marrow ( BM ) cells in a p38 MAPK-dependent manner . Their secretion is stimulated by paracrine interactions between BM stromal and mononuclear cells and cytokine induction correlates with P28906 + stem cell apoptosis in an inflammation-simulated in vitro bone marrow microenvironment . Treatment with DB05412 inhibits P01375 secretion in primary P43034 bone marrow cells and protects cytogenetically normal progenitors from apoptosis ex vivo . Furthermore , p38 inhibition diminishes the expression of TNFalpha or IL-1beta-induced proinflammatory chemokines in BM stromal cells . These data indicate that p38 inhibition has anti-inflammatory effects on the bone marrow microenvironment that complements its cytoprotective effect on progenitor survival . These findings support clinical investigation of p38alpha as a potential therapeutic target in P43034 and other related diseases characterised by inflammatory bone marrow failure . Comprehensive proteomic study identifies serpin and cystatin antiproteases as novel correlates of HIV-1 resistance in the cervicovaginal mucosa of female sex workers . Not all individuals exposed to HIV-1 become infected , and evidence from HIV-1 highly exposed seronegative women ( HIV-1-resistant ) suggests that mucosal factors in the female genital tract , the first site of contact for the virus , are playing a role . To better understand factors mediating protection from HIV-1 , we performed a large clinical study using the tools of systems biology to fully characterize the cervicovaginal mucosa proteome in HIV-1-resistant women . Cervicovaginal lavage fluid was collected from 293 HIV-1-resistant , uninfected , and infected sex workers and analyzed by 2D-LC LTQ-FT-MS . Of the more than 360 unique proteins identified , 41 were differentially abundant ( > 3-fold cutoff ) in HIV-1-resistant women . The majority of over-abundant proteins were antiproteases ( > 40 % ) , some with described anti-inflammatory and anti-HIV-1 activity . Quantification of specific anti-HIV-1 antiproteases P01009 , P01011 , and Cystatin B and an epithelial antiprotease A8K2U0 found them to be significantly over-abundant in HIV-1-resistant women ( p = 0.004 ; p = 0.046 ; p = 0.0003 ; and p = 0.04 , respectively ) . Expression levels were not correlated to sexual practices or other epidemiological factors . Mucosal antiprotease levels correlated with pro-inflammatory cytokine concentration ( p = < 0.0001 ) , but independently of pro-inflammatory cytokine levels in HIV-1-resistant women including P01375 , P01583 , P01584 , P05231 , and P10145 . This comprehensive systems biology approach identifies mucosal serpins and cystatins as novel correlates of HIV-1-resistance . This represents the first study characterizing these factors in the female genital tract . P35354 -derived prostacyclin confers atheroprotection on female mice . Female gender affords relative protection from cardiovascular disease until the menopause . We report that estrogen acts on estrogen receptor subtype alpha to up-regulate the production of atheroprotective prostacyclin , DB01240 , by activation of cyclooxygenase 2 ( P35354 ) . This mechanism restrained both oxidant stress and platelet activation that contribute to atherogenesis in female mice . Deletion of the P43119 removed the atheroprotective effect of estrogen in ovariectomized female mice . This suggests that chronic treatment of patients with selective inhibitors of P35354 could undermine protection from cardiovascular disease in premenopausal females . Crystal structures of human glutaryl- DB01992 dehydrogenase with and without an alternate substrate : structural bases of dehydrogenation and decarboxylation reactions . Acyl- DB01992 dehydrogenases ( ACDs ) are a family of flavoenzymes that metabolize fatty acids and some amino acids . Of nine known ACDs , glutaryl- DB01992 dehydrogenase ( Q92947 ) is unique : in addition to the alpha,beta-dehydrogenation reaction , common to all ACDs , Q92947 catalyzes decarboxylation of glutaryl- DB01992 to produce CO(2) and crotonyl- DB01992 . Crystal structures of Q92947 and its complex with 4-nitrobutyryl- DB01992 have been determined to 2.1 and 2.6 A , respectively . The overall polypeptide folds are the same and similar to the structures of other family members . The active site of the unliganded structure is filled with water molecules that are displaced when enzyme binds the substrate . The structure strongly suggests that the mechanism of dehydrogenation is the same as in other ACDs . The substrate binds at the re side of the DB03147 ring . Glu370 abstracts the P06681 pro-R proton , which is acidified by the polarization of the thiolester carbonyl oxygen through hydrogen bonding to the 2'-OH of DB03147 and the amide nitrogen of Glu370 . The P01024 pro-R proton is transferred to the N(5) atom of DB03147 . The structures indicate a plausible mechanism for the decarboxylation reaction . The carbonyl polarization initiates decarboxylation , and Arg94 stabilizes the transient crotonyl- DB01992 anion . Protonation of the crotonyl- DB01992 anion occurs by a 1,3-prototropic shift catalyzed by the conjugated acid of the general base , Glu370 . A tight hydrogen-bonding network involving gamma-carboxylate of the enzyme-bound glutaconyl- DB01992 , with Tyr369 , Glu87 , Arg94 , Ser95 , and Thr170 , optimizes orientation of the gamma-carboxylate for decarboxylation . Some pathogenic mutations are explained by the structure . The mutations affect protein folding , stability , and/or substrate binding , resulting in inefficient/inactive enzyme . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . [ Effectiveness of the screening programme for galactosemia. New strategy in Poland ] . Galactosemia is an autosomal recessive disease related to deficiency of one of three different enzymes involved in the metabolism of galactose : galactokinase ( P51570 ) , galactoso-J-phosphate uridyltransferase ( P07902 ) or DB03501 -4-epimerase ( Q14376 ) . Classic galactosemia is due to P07902 deficiency and is the most common . Longitudinal studies have shown that in spite of early diagnosis and early treatment of children with galactosemia detected in the mass screening programme , the results are poor and mental retardation as well as other complications are of similar severity as in children diagnosed clinically without screening . In many investigations it was also proved that some impairments developed already in the prenatal period . Therefore , many countries among them also Poland , stopped mass screening for galactosemia . At present , in Poland the procedure strategy in galactosemic children and their families include : diagnosis of new cases on the basis of clinical symptoms , selective screening in high-risk families , prophylactic lactose-free diet for mothers during pregnancy . Such management can help to prevent clinical manifestations in newborns and prevent death in the early period of life . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . Identification of DNA polymorphisms associated with the V type alpha1-antitrypsin gene . alpha1-Antitrypsin ( alpha1-AT ) is a highly polymorphic protein . The V allele of alpha1-AT has been shown to be associated with focal glomerulosclerosis ( FGS ) in Negroid and mixed race South African patients . To identify mutations and polymorphisms in the gene for the V allele of alpha1-AT in five South African patients with FGS nephrotic syndrome DNA sequence analysis and restriction fragment length polymorphisms of the coding exons were carried out . Four of the patients were heterozygous for the BstEII RFLP in exon III [ M1(Val213)(Ala213) ] and one patient was a M1(Ala213) homozygote . The mutation for the V allele was identified in exon II as DB00145 -148 ( GGG ) --> DB00125 ( AGG ) and in all patients was associated with a silent mutation at position 158 ( AAC --> P01009 ) . The patient who was homozygous for ( Ala213 ) also had a silent mutation at position 256 in exon III ( Q6IB77 --> GAC ) which was not present in any of the other four patients . Although the V allele of alpha1-AT is not associated with severe plasma deficiency , it may be in linkage disequilibrium with other genes on chromosome 14 that predispose to FGS . Furthermore , the associated silent mutation at position 158 and the Ala213 polymorphism are of interest , as these could represent an evolutionary intermediate between the M1(Ala213) and M1(Val213) subtypes . Effects of inhaled prostacyclin analogue on chronic hypoxic pulmonary hypertension . Inhaled DB01240 has been reported to elicit pulmonary vasodilation , but whether it is also effective in treating chronic hypoxic pulmonary hypertension is still uncertain . We designed this study to address the in vivo effectiveness of inhaled DB05229 , a stable DB01240 analogue , on pulmonary vascular tone during hypoxic exposure in normoxic ( N ) and chronically hypoxic ( CH ) rats . Pulmonary vasodilation was observed by low-dose inhaled DB05229 in N rats , but not in CH rats . It was not until higher doses of DB05229 were given that pulmonary vasodilation was obtained in CH rats . When the agent was continuously administered by an intravascular route at the inhaled dose , it elicited no vasodilation in N rats . On the contrary , it elicited profound vasodilation in CH rats , although a concomitant systemic hypotension was observed . The P43119 mRNA expression was unchanged in the lungs of CH rats compared with that of N rats . We conclude that low doses of aerosolized DB05229 may reduce pulmonary vascular tone in rats without preexisting lung diseases . In contrast , when hypoxic pulmonary hypertension is present , the threshold of DB05229 inhalation was elevated to provoke pulmonary vasodilation . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . DB01411 , a cysteinyl leukotriene type 1 receptor antagonist , attenuates the progression but not the onset of silica-induced pulmonary fibrosis in mice . BACKGROUND : Although cysteinyl leukotrienes ( CysLTs ) have been implicated in the etiology of acute inflammatory diseases , recent studies have suggested that they also directly stimulate fibroblasts . However , their precise role in the pathogenesis of pulmonary fibrosis is unclear . METHODS : In this study , we evaluated the effect of both short- and long-term treatment with pranlukast , a CysLT type 1 ( CysLT(1) ) receptor antagonist , on silica-induced pulmonary fibrosis in mice , which is characterized by persistent progression of fibrosis in the chronic phase . DB01411 ( 30 mg/kg/day ) was administered orally to mice for 2 or 10 weeks after intratracheal silica instillation . RESULTS : DB01411 treatment for 10 weeks significantly attenuated the progression of pulmonary fibrosis , and decreased the content of CysLTs and Q06643 (4) , which were markedly increased in the bronchoalveolar lavage fluid ( BALF ) and lung tissues of silica-instilled mice in the chronic phase . However , pranlukast treatment for 2 weeks neither affected the acute inflammatory response induced by silica instillation nor inhibited the onset of fibrosis . The expression of TGF-β1 and P01375 -α was not affected by pranlukast treatment for either 2 or 10 weeks . CONCLUSIONS : DB01411 attenuates the progression of pulmonary fibrosis in the chronic phase but has no effect on the acute inflammatory response or on the onset of pulmonary fibrosis . The antifibrotic effect of pranlukast may be exhibited by antagonizing the direct profibrotic effect of CysLTs , without affecting the expression of other profibrotic cytokines such as TGF-β1 and P01375 -α , and also by decreasing the production of CysLTs and Q06643 (4) . Structural analysis and molybdenum-dependent expression of the pAO1-encoded nicotine dehydrogenase genes of Arthrobacter nicotinovorans . The genes of nicotine dehydrogenase ( NDH ) were identified , cloned and sequenced from the catabolic plasmid pAO1 of Arthrobacter nicotinovorans . In immediate proximity to this gene cluster is the beginning of the 6-hydroxy-L-niotine oxidase ( 6-HLNO ) gene . NDH is composed of three subunits ( A , B and C ) of M(r) 30,011 , 14,924 and 87,677 . It belongs to a family of bacterial hydroxylases with a similar subunit structure ; they have molybdopterin dinucleotide , DB03147 and Fe-S clusters as cofactors . Here the first complete primary structure of a bacterial hydroxylase is provided . Sequence alignments of each of the NDH subunits show similarities to the sequences of eukaryotic xanthine dehydrogenase ( P47989 ) but not to other known molybdenum-containing bacterial enzymes . Based on alignment with P47989 it is inferred that the smallest subunit ( NDHB ) carries an iron-sulphur cluster , that the middle-sized subunit ( NDHA ) binds DB03147 , and that the largest NDH subunit ( NDHC ) corresponds to the molybdopterin-binding domain of P47989 . Expression of both the ndh and the 6-hino genes required the presence of nicotine and molybdenum in the culture medium . Tungsten inhibited enzyme activity but not the synthesis of the enzyme protein . The enzyme was found in A. nicotinovorans cells in a soluble form and in a membrane-associated form . In the presence of tungsten the fraction of membrane-associated NDH increased . DB01269 : the evidence for its use in the treatment of metastatic colorectal cancer . DB01269 is the first fully human monoclonal antibody to Epidermal Growth Factor Receptor ( P00533 ) to enter clinical trials for the treatment of solid tumors . The anti-tumor activity of panitumumab has been tested in vitro and in vivo , and inhibition of tumor growth has been observed in numerous cancer models , particularly lung , kidney and colorectal ( CRC ) . Preclinical and clinical studies have established a role for panitumumab in metastatic colorectal cancer ( mCRC ) refractory to multiple chemotherapeutic regimens . Based on these encouraging findings , panitumumab was approved by the US Food and Drug Administration for the treatment of patients with epidermal growth factor receptor-expressing mCRC refractory to fluoropyrimidine- , oxaliplatin- , and/or irinotecan-containing chemotherapeutic regimens . The improvement in progression free survival ( PFS ) and response rate ( RR ) produced by panitumumab monotherapy was significantly greater in patients with non mutated ( wild-type ) K- DB01367 than in those with mutant K- DB01367 . Therefore implementing routine K- DB01367 screening and limiting the use of P00533 inhibitors to patients with wild-type K- DB01367 appears the better strategy for select only the patients who could benefit from the therapy with panitumumab and also may have the potential for cost savings . The purpose of this review was to evaluate the patient-related , disease-related and economic-related evidence for the use of panitumumab in the treatment of metastatic colorectal cancer in clinical practice . Positive correlation between galactose-1-phosphate uridyltransferase ( P07902 ) and DB03501 -4'-epimerase ( Q14376 ) activities . OBJECTIVES : The aim of our study was to determine whether the activities of galactose-1-phosphate uridyltransferase and DB03501 -4'-epimerase are correlated , and in what way they may influence one another . DESIGN AND METHODS : Enzyme activities were measured in red blood cells from 214 individuals . RESULTS : A statistically significant ( p < 0.001 ) positive correlation was observed between P07902 and Q14376 activities . CONCLUSIONS : Our results suggest that P07902 and Q14376 activities are correlated and that Q14376 activity has a greater impact on P07902 activity than vice versa . DB00067 -independent targeting of aquaporin-2 by selective E-prostanoid receptor agonists alleviates nephrogenic diabetes insipidus . In the kidney , the actions of vasopressin on its type-2 receptor ( P30518 ) induce increased water reabsorption alongside polyphosphorylation and membrane targeting of the water channel aquaporin-2 ( P41181 ) . Loss-of-function mutations in the P30518 cause X-linked nephrogenic diabetes insipidus . Treatment of this condition would require bypassing the P30518 to increase P41181 membrane targeting , but currently no specific pharmacological therapy is available . The present study examined specific E-prostanoid receptors for this purpose . In vitro , prostaglandin E2 ( DB00917 ) and selective agonists for the E-prostanoid receptors EP2 ( butaprost ) or EP4 ( CAY10580 ) all increased trafficking and ser-264 phosphorylation of P41181 in Madin-Darby canine kidney cells . Only DB00917 and butaprost increased DB02527 and ser-269 phosphorylation of P41181 . Ex vivo , DB00917 , butaprost , or CAY10580 increased P41181 phosphorylation in isolated cortical tubules , whereas DB00917 and butaprost selectively increased P41181 membrane accumulation in kidney slices . In vivo , a P30518 antagonist caused a severe urinary concentrating defect in rats , which was greatly alleviated by treatment with butaprost . In conclusion , EP2 and EP4 agonists increase P41181 phosphorylation and trafficking , likely through different signaling pathways . Furthermore , EP2 selective agonists can partially compensate for a nonfunctional P30518 , providing a rationale for new treatment strategies for hereditary nephrogenic diabetes insipidus . Follicle epithelial M cells are a source of interleukin-1 in Peyer 's patches . The production of interleukin-1 ( IL-1 ) by rabbit Peyer 's patch M cells populating the follicle-associated epithelium ( FAE ) was studied . Sorted 5D9+ phagocytic epithelial M cells synthesized IL-1 after stimulation with lipopolysaccharide ( LPS ) in vitro , as evidenced by the ability of culture supernatants to induce the proliferation of the T-cell line D10.G4.1 . Fixed LPS-stimulated M cells were less effective at mediating T-cell proliferation than supernatants from LPS-activated M cells . The magnitude of T-cell proliferation was M-cell concentration dependent , and proportional to the dose of LPS . The M-cell-mediated D10.G4.1 cell proliferation was inhibited > 75 % with anti- P01583 , but < 50 % with similar concentrations of anti- P01584 . The results show that M cells secrete IL-1 , and suggest the participation of M cells in the delivery of a localized co-stimulatory signal for T-cell and B-cell proliferation in the microenvironment of gut-associated lymphoid tissue ( P07902 ) . P14416 desensitization by dopamine or corticotropin releasing factor in ventral tegmental area neurons is associated with increased glutamate release . Neurons of the ventral tegmental area ( VTA ) are the source of dopaminergic ( DAergic ) input to important brain regions related to addiction . Prolonged exposure of these VTA neurons to moderate concentrations of dopamine ( DA ) causes a time-dependent decrease in DA-induced inhibition , a complex desensitization called DA inhibition reversal ( P30518 ) . P30518 is mediated by conventional protein kinase C ( cPKC ) through concurrent stimulation of D2 and D1-like DA receptors , or by D2 stimulation concurrent with activation of some Gq-linked receptors . DB01285 releasing factor ( CRF ) acts via Gq , and can modulate glutamater neurotransmission in the VTA . In the present study , we used brain slice electrophysiology to characterize the interaction of DA , glutamate antagonists , and CRF agonists in the induction and maintenance of P30518 in the VTA . Glutamate receptor antagonists blocked induction but not maintenance of P30518 . Putative blockers of neurotransmitter release and store-operated calcium channels blocked and reversed P30518 . CRF and the CRF agonist urocortin reversed inhibition produced by the D2 agonist quinpirole , consistent with our earlier work indicating that Gq activation reverses quinpirole-mediated inhibition . In whole cell recordings , the combination of urocortin and quinpirole , but not either agent alone , increased spontaneous excitatory postsynaptic currents ( sEPSCs ) in VTA neurons . Likewise , the combination of a D1-like receptor agonist and quinpirole , but not either agent alone , increased sEPSCs in VTA neurons . In summary , desensitization of D2 receptors induced by dopamine or CRF on DAergic VTA neurons is associated with increased glutamatergic signaling in the VTA . DB01411 inhibits NF-kappaB activation and Q02817 gene expression in cultured human epithelial cells . DB01411 is a selective cysteinyl leukotriene ( 1 ) (cysLT(1)) receptor antagonist , and is now widely used in the treatment of asthma . The anti-asthmatic effect of pranlukast may be rendered not only by antileukotriene activity , but also by other pharmacological activity . This study was designed to investigate whether pranlukast had inhibitory effects on nuclear factor-kappaB ( NF-kappaB ) activation and mucin gene expression in cultured human epithelial cells . Luciferase assay was mainly used for analysis . Cultured epithelial cells were transfected with NF-kappaB luciferase vector , Q02817 or P98088 luciferase vectors . Lipopolysaccharide ( LPS ) significantly increased NF-kappaB activation in NCI-H292 cells , which was inhibited by the pretreatment by pranlukast in a dose-dependent manner . Either LTD(4) or pranlukast alone did not increase NF-kappaB activation in NCI-H292 cells . DB01411 also inhibited NF-kappaB activation induced by phorbol 12-myristate 13-acetate ( PMA ) . DB01411 also significantly inhibited LPS-induced Q02817 mRNA expression by reverse transcription-polymerase chain reaction ( RT-PCR ) analysis in NCI-H292 cells . DB01411 also inhibited LPS-induced Q02817 gene expression in HM3- Q02817 cells . However , pranlukast did not inhibit P98088 gene transcription activity induced by lipoteichoic acid ( P01374 ) in NCI-H292 cells . These results suggest that pranlukast may inhibit NF-kappaB activation and Q02817 gene transcription through pathways distinct from cysLT(1) receptor antagonism in cultured human epithelial cells . Association study between dopamine D3 receptor gene variant and personality traits . Dopamine receptor gene variation has been hypothesized to influence personality traits characterized by novelty seeking and related traits . We analyzed a dopamine D(3) receptor gene ( P35462 ) variant in a Swedish population ( n = 373 ) investigated with one or more of several personality questionnaires . No significant relationships were found between P35462 genotypes and any of the 15 Karolinska Scales of Personality ( KSP ) and five Health-relevant Personality 5 factor inventory ( HP5i ) scales . The P35462 variant was associated with some scales related to novelty seeking : the Swedish universities Scales of Personality ( SSP ) Adventure Seeking and the revised NEO personality inventory ( NEO-PI-R ) Fantasy ( O1 ) and Order ( P06681 ) scales . There were also associations with the Temperament and Character Inventory ( TCI ) Cooperativeness and Compassion ( C4 ) scales . After correction for multiple testing , however , no significant difference remained . We conclude that the investigated P35462 polymorphism does not have a major impact on personality in the investigated population . P01583 and beta , P01375 and HTTLPR gene variants study on alcohol toxicity and detoxification outcome . Genetic factors may influence the liability to treatment outcome and medical complications in alcoholism . In the present study we investigated the IL-1A rs1800587 , IL-1B rs3087258 , P01375 rs1799724 and the HTTLPR variants in a sample of 64 alcohol dependents and 47 relatives versus a set of clinical parameters and outcome measures . DB00898 dependents had a less favorable clinical profile compared to relatives ( higher cholesterol , triglycerides , glucose , glutamic oxaloacetic transaminase , glutamic pyruvic transaminase , gamma-glutamyltransferase ) . After detoxification , all clinical indexes improved and hepatic enzyme levels were similar in alcohol dependents and relatives , except for the P19440 that remained significantly higher in alcohol dependents . Alcoholic depressive and anxiety scores were significantly reduced after detoxification . IL-1A , IL-1B , P01375 and HTTLPR variants were not associated with any baseline clinical index or change after detoxification . In our sample IL-1A , IL-1B , P01375 and HTTLPR do not appear as liability factors for alcohol toxicity or detoxification outcome , however the small sample size may influence the observed results . p38 Q96HU1 kinase regulates stem cell apoptosis in human hematopoietic failure . Myelodysplastic syndromes ( P43034 ) are clonal stem cell disorders that lead to ineffective hematopoiesis and are common causes of low blood counts in the elderly . The exact molecular mechanisms regulating increased stem apoptosis in these disorders are not well defined . p38 MAPK activation is important in regulating the growth inhibitory signals of P01375 , TGF-beta and Interferons on human hematopoiesis . Our findings show that p38 MAPK is overactivated in myelodysplasia bone marrows and regulates hematopoietic stem cell apoptosis . Inhibition of p38 MAPK by genetic or pharmacologic means decreases apoptosis and stimulates in vitro hematopoiesis from primary P43034 hematopoietic progenitors . These studies point to the potential efficacy of selective p38alpha inhibitor , DB05412 , in human bone marrow failure . Joint effects of P29474 gene T-786C and P00325 Arg47His polymorphisms on the risk of premature coronary artery disease . INTRODUCTION : Both T-786C mutation in endothelial nitric oxide synthase ( P29474 ) gene and alcohol dehydrogenase ( DB00067 ) gene polymorphism such as P00326 gamma1/gamma2 have been reportedly associated with coronary artery disease ( CAD ) . Since P00325 Arg47His polymorphism is common in Asian population , the aim of this present study was to assess the interaction between P29474 gene T-786C and P00325 Arg47His polymorphisms on premature CAD risk . MATERIALS AND METHODS : Hospital-based case-control study was conducted with 167 premature CAD and 235 late-onset CAD patients . Polymerase chain reaction restriction fragment length polymorphism was used to detect the polymorphisms . Multivariate logistic regression model was performed to adjust the potential confounders and estimate odds ratios ( ORs ) with 95 % confidence intervals ( CIs ) . Synergy index ( S ) was the measure to assess the interaction as departure from additivity . RESULTS : After the adjustment for the potential confounders , and compared with the carriers of TT and DB00125 / DB00125 as the reference , the ORs with 95 % CIs in parentheses of premature CAD were that 1.13 ( 0.19-6.59 ) for CT or CC and DB00125 / DB00125 carriers ; 2.24 ( 0.77-6.49 ) for TT and DB00125 / DB00117 or DB00117 / DB00117 carriers ; 4.18 ( 1.32-13.22 ) for CT or CC and DB00125 / DB00117 or DB00117 / DB00117 carriers , respectively . Based on those ORs , S was 2.32 ( 95 % CI : 0.37-14.72 ) . CONCLUSIONS : The mutant genotypes of P29474 gene T-786C mutation and the fast form of P00325 Arg47His polymorphism had an additive interaction on the risk of premature CAD in Chinese population . Further investigations with big sample size are necessary for confirming this additive interaction . Detection of K-ras mutation in sputum by mutant-allele-specific amplification ( Q9UHY7 ) . From 10 to 30 % of lung carcinomas examined to date contain mutant K-ras genes . We report here that the mutant-allele-specific amplification ( Q9UHY7 ) method may be useful for detection of the K-ras mutations in cells obtained from the sputum of patients with lung cancer . The PCR product from one of five patients revealed an alteration when mixed oligonucleotides representing variants of the second letter at codon 12 of this gene were used as 5' primers , and further experiments showed a mutation of P19440 ( DB00145 ) to Q6IB77 ( DB00128 ) at codon 12 . The Q9UHY7 system could also be applied to an examination of metastatic lung carcinomas , particularly from adenocarcinomas in colon and pancreas in which frequent K-ras mutations are detected , and to mass-screening for colorectal tumors using DNA isolated from feces as template .

MH_train_1244

MH_train_1244

MH_train_1244

interacts_with DB00086?

multiple_choice

Dynamics of P08514 /IIIa-mediated platelet-platelet interactions in platelet adhesion/thrombus formation on collagen in vitro as revealed by videomicroscopy . The conventional description of platelet interactions with collagen-coated surfaces in vitro , based on serial static measurements , is that platelets first adhere and spread to form a monolayer and then recruit additional layers of platelets . To obtain dynamic information , we studied gravity-driven platelet deposition in vitro on purified type 1 collagen by video phase-contrast microscopy at 22 degrees C . With untreated human and wild-type mouse platelets , soon after the initial adhesion of a small number of " vanguard " platelets , " follower " platelets attached to the spread-out vanguard platelets . Follower platelets then adhered to and spread onto nearby collagen or over the vanguard platelets . Thus , thrombi formed as a concerted process rather than as sequential processes . Treatment of human platelets with monoclonal antibody ( mAb ) DB00054 ( anti- P08514 /IIIa ( alphaIIbbeta3 ) + alphaVbeta3 ) or tirofiban ( anti- P08514 /IIIa ) did not prevent platelet adhesion but nearly eliminated the deposition of follower platelets onto vanguard platelets and platelet thrombi . Similar results were obtained with Glanzmann thrombasthenia platelets . Wild-type mouse platelets in the presence of mAb 1B5 ( anti- P08514 /IIIa ) and platelets from beta3-null mice behaved like human platelets in the presence of 7E3 or tirofiban . Deposition patterns of untreated human and wild-type mouse platelets were consistent with random distributions under a Poisson model , but those obtained with 7E3- and tirofiban-treated human platelets , 1B5-treated mouse platelets , or beta3-null platelets demonstrated a more uniform deposition than predicted . Thus , in this model system , absence or blockade of P08514 /IIIa receptors interferes with thrombus formation and alters the pattern of platelet deposition . Single-stranded DNA-binding proteins and neuron-restrictive silencer factor participate in cell-specific transcriptional control of the Q05586 gene . Our previous studies revealed that a proximal region of the N-methyl-D-aspartate receptor 1 ( Q05586 ) promoter is important for cell-type-specific expression . We have now explored the contributions of several regulatory elements to this specificity . Deletion of the neuron-restrictive silencer element partially relieved the suppression of promoter activity in P13671 glioma and HeLa cells . An overlapping G(C/G)G/tandem Sp1-containing region crucial for both basal and nerve growth factor ( P01138 ) -regulated promoter activity specifically bound nuclear proteins on its purine-rich sense strand . A faster migrating complex , single-stranded binding protein complex 1 ( SBPC1 ) , was highly enriched in HeLa cells , whereas a slower migrating complex , SBPC2 , was enriched in PC12 cells . A high ratio of 2/1 complex correlated with a high level of promoter activity . P01138 treatment of PC12 cells reduced SBPC1 but increased SBPC2 . Competition experiments showed that the SBPC1 binding required a dG4 sequence and the SBPC2 needed a core of TG3A plus a 5'-flanking sequence . Single-stranded DNA encompassing TG3A and/or dG4 specifically suppressed cotransfected Q05586 promoter activity . UV cross-linking studies indicated that a 31.5-kDa protein mainly formed SBPC1 , whereas SBPC2 contained several larger proteins . Our results suggest that neuron-restrictive silencer factor and single-stranded DNA-binding proteins may both play a role in cell-type specificity of the Q05586 gene , and the latter may also be involved in basal and P01138 -regulated activity . Dexamethasone potentiates serotonin-2 receptor-mediated intracellular Ca2+ mobilization in P13671 glioma cells . Serotonin ( 5-hydroxytryptamine ; 5-HT ) caused a transient increase in intracellular Ca2+ in C6BU-1 glioma cells in a concentration-dependent manner ; half maximally at 73 nM . The 5-HT2 agonist 1-(4-iodo-2,5-dimethoxyphenyl)-2- aminopropane also increased the levels of intracellular Ca2+ , whereas the P28335 agonist 1-(3-chlorophenyl)piperazine and P08908 agonist 8-hydroxy-2- (di-n-propylamino)tetralin were completely ineffective . Ketanserin and spiperone blocked the response to 5-HT at a nanomolar concentration , but the 5- Q9H205 antagonist MDL 72222 had no effect on it . Thus 5-HT2 receptors are responsible for activating Ca2+ mobilization in P13671 glioma cells . Treatment of P13671 glioma cells with dexamethasone potentiated the ability of 5-HT to cause intracellular Ca2+ mobilization in both a dose- and time-dependent manner . The dose-response curve for 5-HT was shifted 9-fold to the left compared to controls , and the Vmax value was also significantly enhanced . This enhanced Ca2+ mobilization was completely inhibited by ketanserin dose-dependently . In addition , the treatment with dexamethasone enhanced fluoride-activated Ca2+ mobilization , suggesting that the enhanced GTP binding protein function is one of the mechanisms responsible for the enhancement of the 5-HT response induced by dexamethasone treatment . This enhancement of agonist activity was mediated by the type II glucocorticoid receptor ( GR ) since RU 38486 , an inhibitor of the type II GR , antagonized the dexamethasone-induced enhancement . P43490 / P43490 /visfatin and cancer . P43490 / P43490 /visfatin is the rate-limiting enzyme that catalyzes the first step in NAD biosynthesis from nicotinamide and regulates growth , apoptosis and angiogenesis of mammalian cells . This enzyme was originally cloned as a putative cytokine shown to enhance the B cell precursor maturation in the presence of P13232 and stem cell factor . A number of cancers have increased expression of P43490 / P43490 /visfatin , which regulates a variety of different signaling pathways such as PI3K/Akt , P27361 /2 and P40763 . FK866/APO866 and CHS828/ DB05217 are two known inhibitors of P43490 / P43490 /visfatin and have been evaluated as anticancer agents in the clinic . This review will focus on its role in carcinogenesis and cancer progression and its inhibitors as therapeutic target for cancer treatment . P18509 effects in pituitary cells : modulation by gonadotropin-releasing hormone in alpha DB00279 -1 cells . P18509 ( PACAP ) acts via type I receptors in the pituitary to stimulate DB02527 production . Gonadotropes are likely target cells for PACAP action , and we have recently shown alpha DB00279 -1 cells , a clonal gonadotrope-derived cell line , to be PACAP responsive . Here we have explored the influence of DB00644 on PACAP action in alpha DB00279 -1 cells and show that PACAP38-stimulated DB02527 production is inhibited by DB00644 in both the presence and the absence of a phosphodiesterase inhibitor . This effect appears not to be Ca++ mediated but is mimicked by protein kinase C activation with phorbol 12-myristate 13-acetate . However , DB00644 and phorbol 12-myristate 13-acetate do not inhibit binding of [125I]PACAP27 to intact alpha DB00279 -1 cells , nor do they inhibit forskolin- or cholera toxin-stimulated DB02527 accumulation , implying that the inhibitory effects are exerted at early stages in the PACAP receptor signaling pathway but distal to receptor occupancy . When cells were preincubated with PACAP38 , extensive washing failed to prevent the stimulatory effect of the polypeptide presumably because of the slow rate of receptor-ligand dissociation . However , when the time course of PACAP38-stimulated effects on intracellular DB02527 was assessed , the stimulatory effect of PACAP38 could be rapidly reversed by DB00644 addition , and the inhibitory effect of DB00644 was rapidly be reversed by a P30968 antagonist . The data provide the first demonstration of cross-talk between phospholipase C and adenylate cyclase-activating peptides in gonadotrope-derived cells and establish the potential for hormonal modulation of PACAP action . We suggest that this inhibitory effect of DB00644 might enable the releasing hormone to control the kinetics of DB02527 signaling in gonadotropes in vivo . [ Effects of plasminogen and streptokinase on the vital functions of nervous tissue cells in culture ] . In the protein-deficient media plasminogen stimulated the vital functions of cells and in concentrations 10(-7)-10(-10) M it protected cells of sympathetic ganglia , neocortex and continues cell lines under damaging actions of H2O2 ( 0.0001 M ) , NH4CI ( 0.01 M ) and cooling . DB00086 essentially influenced the mode of damaging effect of DB00171 ( 0.001 M ) . Even a short-term exposition ( 20 min ) of PC12 cells with both proteins ( each in the concentration 10(-9) M ) led to sharp alterations in intracellular DB00171 - or Ca(2+)-activated proteolysis . In some cases plasminogen and streptokinase provided acceleration of cultured tissue maturation , improvement of cell adhesion , high survival rate , the increase in quantity and length of processes and their arborisation . Electronic microscopy established the character of structural rearrangements of nervous tissue cells ( neurons , astrocytes , oligodendrocytes ) , reflecting the protective action of plasminogen and streptokinase . In the presence of plasminogen and especially streptokinase , the total number of cultured glioma P13671 and neuroblastoma IMR-32 cells , the intracellular contents of protein , RNA and DNA increased several-fold . Addition of plasminogen promoted formation of processes by neuroblastoma cells , this suggests initiation of differentiation of cellular elements . In cultures of sensitive and sympathetic ganglia streptokinase increased proliferation of Schwann cells . These proteins did not cause transformation of PC12 enterochromaffine cells to neurons , though plasminogen facilitated it . P00747 addition to cell cultures did not increase fibrinolytic activity of the culture medium in the culture medium , and streptokinase did not lose its plasminogen-activating capacity . Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells . Dendritic cells ( DC ) play a key role in antigen presentation and activation of specific immunity . Much current research focuses on harnessing the potency of DC for vaccines , gene therapy , and cancer immunotherapy applications . However , DC are not readily transfected in vitro by traditional nonviral techniques . A novel DNA vaccine formulation was used to determine if DC are transfected in vitro . The formulation consists of plasmid DNA adsorbed on to cationic microparticles composed of the biodegradable polymer polylactide-co-glycolide ( P00747 ) and the cationic surfactant , cetyltrimethylammonium bromide ( CTAB ) . Using preparations of fluorescent-labeled plasmid DNA formulated on P00747 -CTAB microparticles to study internalization by macrophages and dendritic cells in vitro and in vivo , we found that most , but not all , of the fluorescence was concentrated in endosomal compartments . Furthermore , uptake of plasmid DNA encoding HIV p55 gag adsorbed to P00747 -CTAB microparticles by murine bone marrow-derived dendritic cells resulted in target gene expression , as detected by RT-PCR . The antigen was subsequently processed and presented , resulting in stimulation of an H-2kd-restricted , gag-specific T cell hybridoma . Activation of the hybridoma , detected by P60568 production , was dose-dependent in the range of 0.1-20 microg DNA ( 10-2000 microg P00747 ) and was sustained up to 5 days after transfection . Thus , adsorption of plasmid DNA on P00747 -CTAB microparticles provides a potentially useful nonviral approach for in vitro transfection of dendritic cells . Gene Therapy ( 2000 ) 7 , 2105-2112 . Plant methionine sulfoxide reductase A and B multigenic families . DB00134 oxidation to methionine sulfoxide ( MetSo ) , which results in modification of activity and conformation for many proteins , is reversed by an enzyme present in most organisms and termed as methionine sulfoxide reductase ( Q9UBK8 ) . On the basis of substrate stereospecificity , two types of Q9UBK8 , A and B , that do not share any sequence similarity , have been identified . In the present review , we first compare the multigenic Q9UBK8 families in the three plant species for which the genome is fully sequenced : Arabidopsis thaliana , Oryza sativa , and Populus trichocarpa . The Q9UBK8 gene content is larger in A. thaliana ( five MSRAs and nine MSRBs ) compared to P. trichocarpa ( five MSRAs and four MSRBs ) and O. sativa ( four MSRAs and three MSRBs ) . A complete classification based on gene structure , sequence identity , position of conserved reactive cysteines and predicted subcellular localization is proposed . On the basis of in silico and experimental data originating mainly from Arabidopsis , we report that some Q9UBK8 genes display organ-specific expression patterns and that those encoding plastidic MSRs are highly expressed in photosynthetic organs . We also show that the expression of numerous Q9UBK8 genes is enhanced by environmental conditions known to generate oxidative stress . Thioredoxins ( TRXs ) constitute very likely physiological electron donors to plant Q9UBK8 proteins for the catalysis of MetSO reduction , but the specificity between the numerous TRXs and methionine sulfoxide reductases ( MSRs ) present in plants remains to be investigated . The essential role of plant MSRs in protection against oxidative damage has been recently demonstrated on transgenic Arabidopsis plants modified in the content of cytosolic or plastidic Q9UJ68 . Induction of cytokine gene expression in human thyroid epithelial cells irradiated with HZE particles ( iron ions ) . Gene expression profiles were examined using cDNA microarray technology in human thyroid epithelial ( Htori-3 ) cells exposed to a low , non-toxic dose ( 10 cGy ) of radiation from HZE particles in the form of iron ions in the absence or presence of selenomethionine ( SeM ) . A total of 215 genes were differentially regulated 2 h after exposure to a 10-cGy dose of iron-ion radiation . In the microarray analysis , SeM had profound effects on the radiation-induced expression of several specific genes , which includes P00749 , P17936 , P15328 , P15291 and P02452 . Of particular interest to us was a gene cluster , " secreted proteins " , that was up-regulated after radiation exposure . Seven up-regulated genes of this gene cluster fall within the chemokine/cytokine gene cluster , namely , P09341 , P19875 , P05231 , P20809 , P10145 , Q13007 and TGFbeta2 . In microarray studies , the radiation-induced up-regulated expression of some these genes encoding cytokine/chemokine proteins was significantly decreased by SeM treatment . For P10145 , TGFbeta2 , P09341 and P19875 , these observations were validated by qPCR techniques . It is concluded that SeM can regulate ionizing radiation-induced gene expression and may serve as an effective countermeasure for some of the acute inflammatory/immune responses induced by low-dose HZE-particle radiation . P18509 modulates plasminogen activator expression in rat granulosa cell . P18509 ( PACAP ) is a bioactive peptide isolated from ovine hypothalamus . It has been demonstrated to be transiently expressed in preovulatory follicles and to positively affect several parameters correlated with the ovulatory process . The aim of the present study was to investigate whether PACAP influences the plasminogen/plasmin system in rat ovary . P00747 activators ( PAs ) are serine proteases , modulated by gonadotropins and several peptides in preovulatory follicles , that appear to be involved in ovulation . Granulosa cells obtained from immature eCG-treated rats were cultured for 24 h in the presence of increasing concentrations of PACAP and vasoactive intestinal peptide ( P01282 ) . A significant , dose-dependent increase in tissue-type PA ( tPA ) activity and decrease in urokinase-type ( uPA ) PA activity were observed in PACAP-treated cells . These effects were exerted at the mRNA level . The use of cycloheximide , a protein synthesis inhibitor , suggested that PACAP requires an intermediary protein to decrease uPA-mRNA , but not to induce tPA-mRNA . However , no significant modulation of PAs was observed in the presence of P01282 . When granulosa cells were stimulated within the intact follicle ( i.e. , maintaining the three-dimensional structure and in the presence of the theca cell layers ) , both PACAP and P01282 dose-dependently stimulated tPA . These data suggest that , in addition to the P41586 present on granulosa cells , different subtypes of PACAP receptors are present in the different ovarian compartments . DB05595 , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC(50) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 did not block membrane binding of radiolabeled folic acid , 5-methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 had a minimal effect on the cytoplasmic accumulation of 5-methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC(50) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated . Two bifunctional enzymes with ferric reduction ability play complementary roles during magnetosome synthesis in Magnetospirillum gryphiswaldense Q9UBK8 -1 . The bacterial strain Magnetospirillum gryphiswaldense Q9UBK8 -1 does not produce siderophores , but it absorbs a large amount of ferric iron and synthesizes magnetosomes . We demonstrated previously the presence of six types of ferric reductase isozymes ( termed FeR1 through FeR6 ) in Q9UBK8 -1 . Of these isozymes , FeR5 was the most abundant and FeR6 showed the highest ferric reductase activity . In the present study , we cloned the fer5 and fer6 genes from Q9UBK8 -1 and expressed them separately in Escherichia coli . FeR5 and FeR6 were shown to be bifunctional enzymes through analysis of amino acid sequence homologies , structural predictions ( using data from GenBank ) , and detection of enzyme activities . FeR5 is a thioredoxin reductase and FeR6 is a flavin reductase , in addition to being ferric reductases . To elucidate the functions of the enzymes , we constructed two single-gene-deletion mutant strains ( Δfer5 and Δfer6 mutants ) and a double-gene-deletion mutant strain ( Δfer5 Δfer6 [ Δfer5+6 ] mutant ) along with its complemented strains ( P01031 and P13671 ) . An evaluation of phenotypic and physiological properties did not reveal significant differences between the wild-type and single-gene-deletion strains , whereas the double-gene-deletion strain showed reduced iron absorption and no magnetosome synthesis . Complementation of the double-gene-deletion strain using either fer5 or fer6 resulted in the partial recovery of magnetosome synthesis . Quantitative real-time PCR analysis of fer5 and fer6 transcriptional levels in the wild-type and complemented strains demonstrated consistent transcription of the two genes and confirmed that FeR5 and FeR6 are bifunctional enzymes that play complementary roles during the process of magnetosome synthesis in Q9UBK8 -1 . Neutrophil apoptosis : selective regulation by different ligands of integrin alphaMbeta2 . Neutrophils undergo spontaneous apoptosis , but their survival can be extended during inflammatory responses . alpha(M)beta(2) is reported either to delay or accelerate neutrophil apoptosis , but the mechanisms by which this integrin can support such diametrically opposed responses are poorly understood . The abilities of closely related alpha(M)beta(2) ligands , plasminogen and angiostatin , derived from plasminogen , as well as fibrinogen and its two derivative alpha(M)beta(2) recognition peptides , P1 and P2-C , differed markedly in their effects on neutrophil apoptosis . P00747 , fibrinogen , and P2-C suppressed apoptosis via activation of Akt and P27361 /2 kinases , while angiostatin and P1 failed to activate these prosurvival pathways and did not prevent neutrophil apoptosis . Using cells transfected with alpha(M)beta(2) or its individual alpha(M) or beta(2) subunits , and purified receptors and its constituent chains , we show that engagement of both subunits with prosurvival ligands is essential for induction of the prosurvival response . Hence , engagement of a single integrin by closely related ligands can induce distinct signaling pathways , which can elicit distinct cellular responses . Effect of ifenprodil on Q05586 / Q13224 N-methyl-D-aspartate receptor gating . DB08954 is an allosteric inhibitor of Q05586 / Q13224 N-methyl-D-aspartate receptors . Despite its widespread use as a prototype for drug development and a subtype-selective tool for physiologic experiments , its precise effect on Q05586 / Q13224 gating is yet to be fully understood . Interestingly , recent crystallographic evidence identified that ifenprodil , unlike zinc , binds at the interface of the Q05586 / Q13224 amino terminal domain dimer by an induced-fit mechanism . To delineate the effect of this unique binding on Q05586 / Q13224 receptor gating , we recorded steady-state currents from cell-attached and outside-out patches . At pH 7.9 in cell-attached patches , ifenprodil increased the occupancy of the long-lived shut conformations , thereby reducing the open probability of the receptor with no change in the mean open time . In addition , ifenprodil selectively affected the area of shut time constants , but not the time constants themselves . Kinetic analyses suggested that ifenprodil prevents the transition of the receptor to an open state and increases its dwell time in an intrinsically occurring closed conformation or desensitized state . We found distinct differences in the action of ifenprodil at Q05586 / Q13224 in comparison with previous studies on the effect of zinc on Q05586 / Q12879 gating , which may arise due to their unique binding sites . Our data also uncover the potential pH-dependent action of ifenprodil on gating . At a low pH ( pH 7.4 ) , but not pH 7.9 , ifenprodil reduces the mean open time of Q05586 / Q13224 receptors , which may be responsible for its usefulness as a context-dependent inhibitor in conditions like ischemia and stroke , when the pH of the extracellular milieu becomes acidic . Isoenzyme-specific cyclooxygenase inhibitors : a whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6 . NSAIDs inhibit the conversion of arachidonic acid into DB03866 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase ( P36551 ) . Two genetically distinct isoforms have been discovered , P23219 and P35354 . While P23219 is thought to account for homeostatic amounts of eicosanoids , P35354 is induced during inflammation leading to pathologic amounts of eicosanoids . Since NSAIDs inhibit both P36551 isoforms , antiinflammatory drug research has refocused to discovering P35354 inhibitors that do not inhibit P23219 . For this purpose , we have developed a whole cell assay system using the human erythroleukemic cell line HEL as a source for P23219 and the human monocytic cell line Mono Mac 6 as a source for P35354 . Mono Mac 6 cells express high amounts of P35354 upon stimulation with lipopolysaccharide ( LPS ) in the absence of any detectable P23219 protein . On the other hand , we find HEL cells to naturally express P23219 protein , but not P35354 . Testing of a panel of NSAIDs as well as some P35354 specific inhibitors showed that this assay system is suitable for identifying compounds that selectively inhibit either P23219 or P35354 . This test system offers the advantage of assessing P23219 and P35354 inhibitors within the human species , within a similar test set-up , and circumvents the need for tedious purification of either platelets or peripheral blood monocytes . The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor-1 ( P05121 ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline- and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai1-/- mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 ) and instead reflected proteolytic activation and release of P14210 with subsequent induction of P35354 . That the P14210 / P35354 / DB00917 axis mediates in vivo protection from fibrosis in Pai1-/- mice was demonstrated by experiments showing that a selective inhibitor of the P08581 c- DB00134 increased lung collagen to WT levels while reducing P35354 protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway . P00747 interacts with human platelets through two distinct mechanisms . DB00142 -plasminogen , the native form of plasminogen , interacts in a specific and saturable manner with unstimulated human platelets , and the binding is enhanced fivefold by thrombin stimulation ( Miles and Plow , 1985. J. Biol. Chem. 260:4303 ) . This study characterizes the nature of the DB00142 -plasminogen binding sites by analyzing platelets deficient in selected proteins and functions . Platelets from patients with afibrinogenemia , Gray platelet syndrome , and the Cam Variant of thrombasthenia , a form of thrombasthenia with near normal levels of glycoprotein IIb/IIIa ( P08514 /IIIa ) , showed minimal augmentation of plasminogen binding to thrombin-stimulated platelets but normal binding to unstimulated platelets . This selective deficiency indicates that two distinct mechanisms are involved in the interaction of plasminogen with platelets . These abnormal platelets share a deficiency in fibrinogen . Surface expression of platelet fibrinogen , however , was not sufficient for enhanced plasminogen binding to stimulated platelets , and experiments with alpha-thrombin and gamma-thrombin indicated that fibrin formation on the platelet surface is necessary for the augmented plasminogen binding . Unstimulated and stimulated thrombasthenic platelets deficient in P08514 /IIIa bound markedly reduced levels of plasminogen , which suggests a role for P08514 /IIIa in plasminogen binding to unstimulated platelets . Treatment of platelets to dissociate the heterodimeric complex of P08514 /IIIa did not significantly perturb plasminogen binding to unstimulated platelets , but the complex may be necessary for thrombin-stimulated plasminogen binding via its interaction with platelet fibrin . A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma . DB00004 , a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 ( P60568 ) , efficiently targets lymphoma cells expressing the high-affinity P60568 receptor ( IL-2R ) consisting of the alpha/p55/CD25 , beta/p75/CD122 , and gamma/ P31785 /CD132 chains . In vitro studies demonstrated that the retinoid X receptor ( RXR ) retinoid , bexarotene , at biologically relevant concentrations of 10(-6) M to 10(-8) M , upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox . To determine whether this biomodulatory effect could be recapitulated in vivo , we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene ( 75 mg/day-300 mg/day ) and denileukin diftitox ( 18 mcg/kg per day x 3 days every 21 days ) in a phase 1 trial . Overall response was 67 % ( 4 complete responses , 4 partial responses ) . Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day . Four patients experienced grade 2 or 3 leukopenia , and 2 had grade 4 lymphopenia . Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses ( 150 mg/day ) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells . Reduced satiating effect of d-fenfluramine in serotonin 5-HT(2C) receptor mutant mice . RATIONALE : d- DB00574 stimulates the release of serotonin ( 5-HT ) and is a potent inhibitor of the re-uptake of 5-HT into nerve terminals . Administration of d-fenfluramine suppresses food intake in both animals and humans . OBJECTIVE : We have investigated the role of the P28335 receptor in mediating the effect of d-fenfluramine on mouse food intake and the behavioural satiety sequence . METHODS : Mutant mice lacking serotonin P28335 receptors and wild-type animals were habituated to a daily presentation of wet mash . Animals were non-deprived and received d-fenfluramine ( 3-30 mg/kg ) 30 min prior to being assessed for the presence of stereotypy and presented with wet mash . The behaviour of animals was observed for the subsequent 40 min and food intake was recorded . RESULTS : d- DB00574 dose-dependently inhibited the consumption of a palatable wet mash by the mice . d- DB00574 ( 3 mg/kg ) significantly reduced the amount of wet mash consumed by wild-type mice and induced a temporal advance in the behavioural satiety sequence consistent with an enhancement of satiety . Mutant mice were less sensitive to the satiating effects of 3 mg/kg d-fenfluramine . Hence , this dose of d-fenfluramine had a reduced effect on both food consumption and the behavioural satiety sequence in the P28335 mutant mice . In contrast , mutant mice showed an increased sensitivity to the stereotypy induced by high doses of d-fenfluramine ( 10 , 30 mg/kg ) compared to that of wild-type littermates . CONCLUSION : These data demonstrate a role for the P28335 receptor in mediating d-fenfluramine-induced satiety . 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 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries .

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interacts_with DB00338?

multiple_choice

Involvement of cyclin-dependent kinase activities in CD437-induced apoptosis . A novel synthetic retinoid , 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid ( CD437 ) , is a selective ligand of the RARgamma nuclear receptor . We examined the in vitro effects of CD437 and found that CD437 induces S phase arrest within 24 to 48 h , followed by cell death , in the p53-negative Hep3B and the p53-positive HepG2 human hepatoma cell lines . Based on observations of cellular and nuclear fragmentation , chromatin condensation , and DNA fragmentation , the CD437-mediated cell-killing effect appears to be due to apoptosis . On morphological examination , a number of CD437-treated cells were found to have increased 5- to 10-fold in size and persisted as single giant cells without cell division , while the remainder underwent nuclear division ( multiple nuclei ) but were unable to complete cytokinesis , and finally all died by apoptosis . In HepG2 cells that possessed wild-type p53 , CD437-induced S phase arrest and apoptosis were accompanied by the up-regulation of cyclin A , cyclin B , p53 , P38936 (CIP1/Waf1) , Bad , and Bcl-Xs proteins and by a decrease in Bcl-2 protein levels . In Hep3B cells , CD437-mediated S phase arrest and apoptosis were also associated with a concomitant up-regulation of cyclin A , cyclin B , Bad , and Bcl-Xs . However , Hep3B cells did not express p53 or Bcl-2 messages . DB02116 and roscovitine , the potent p34(cdc2) and P24941 inhibitors , effectively blocked CD437-mediated cyclin A- and B-dependent kinase activation and prevented CD437-induced cell death . Furthermore , antisense oligonucleotide complementary to cyclin A and B mRNA significantly rescued CD437-induced apoptosis . These findings suggest that activation of cyclin A- and B-dependent kinases is a critical determinant of apoptotic death mediated by CD437 . Tanshinone IIA inhibits constitutive P40763 activation , suppresses proliferation , and induces apoptosis in rat P13671 glioma cells . P40763 ( P40763 ) is usually constitutively activated in a variety of malignancies . Thus , P40763 may be a promising target for treatment of tumor cells . Recently , Tanshinone IIA ( Tan IIA ) , a major active constituent from the root of Salvia miltiorrhiza Bunge , was reported to have apoptosis inducing effects on a large variety of cancer cells . In this study , we evaluate the anti-proliferation and apoptosis inducing effects of Tan IIA on P13671 glioma cells . Cell growth and proliferation were measured by MTT assay , cell apoptosis was observed by flow cytometry and DNA-fragmentation analysis . Further more , we investigated inhibitory effects of Tan IIA on P40763 activity and its downstream targets : Bcl-XL , cyclin D1 . Alteration of P40763 activity was examined by measuring their DNA binding activity and tyrosine phosphorylation . Changes in the expression levels of Bcl-XL and cyclin D1 were examined by Western blot analysis . We found that the cellular growth were inhibited and cell apoptosis were observed after the treatment with Tan IIA . The P40763 activity was significantly reduced by Tan IIA parallel with a significant attenuation of expression of Bcl-XL and cyclin D1 . These results suggest that Tan IIA may serve as an effective adjunctive reagent in the treatment of glioma for its targeting of constitutive P40763 signaling . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . The thioredoxin reductase inhibitor auranofin triggers apoptosis through a Bax/Bak-dependent process that involves peroxiredoxin 3 oxidation . P30044 ( TrxR ) is a key selenoprotein antioxidant enzyme and a potential target for anti-cancer drugs . One potent inhibitor of TrxR is the gold ( I ) compound auranofin , which can trigger mitochondrial-dependent apoptosis pathways . The exact mechanism of apoptosis induction by auranofin is not yet clear , but there are indications that mitochondrial oxidative stress is a central event . We assessed the redox state of the peroxiredoxins ( Prxs ) in Jurkat T-lymphoma cells treated with auranofin , and found that mitochondrial Prx3 was considerably more sensitive to oxidation than the cytosolic Prx1 and 2 , indicating selective mitochondrial stress . Prx3 oxidation was detected at apoptotic doses of auranofin in several cell types , and occurred before other mitochondrial events including cytochrome c release and mitochondrial depolarisation . DB00995 was also able to sensitise U937 cells to P01375 -mediated apoptosis . DB00995 -induced apoptosis was effectively blocked by the overexpression of Bcl-2 , and Bax/Bak deficient mouse embryonic fibroblasts were also resistant to apoptosis , indicating a central role for the pro-apoptotic proteins of this family in auranofin-triggered apoptosis . DB00995 exposure inhibited the proliferation of apoptosis-resistant cells , and at higher doses of auranofin could cause cell death through necrosis . We conclude that auranofin induces apoptosis in cells through a Bax/Bak-dependent mechanism associated with selective disruption of mitochondrial redox homeostasis in conjunction with oxidation of Prx3 . Treatment with arimoclomol , a coinducer of heat shock proteins , delays disease progression in P35858 mice . Amyotrophic lateral sclerosis ( P35858 ) is a fatal neurodegenerative condition in which motoneurons of the spinal cord and motor cortex die , resulting in progressive paralysis . This condition has no cure and results in eventual death , usually within 1-5 years of diagnosis . Although the specific etiology of P35858 is unknown , 20 % of familial cases of the disease carry mutations in the gene encoding Cu/Zn superoxide dismutase-1 ( P00441 ) . Transgenic mice overexpressing human mutant P00441 have a phenotype and pathology that are very similar to that seen in human P35858 patients . Here we show that treatment with arimoclomol , a coinducer of heat shock proteins ( HSPs ) , significantly delays disease progression in mice expressing a P00441 mutant in which glycine is substituted with alanine at position 93 ( P00441 (G93A) ) . DB05025 -treated P00441 (G93A) mice show marked improvement in hind limb muscle function and motoneuron survival in the later stages of the disease , resulting in a 22 % increase in lifespan . Pharmacological activation of the heat shock response may therefore be a successful therapeutic approach to treating P35858 , and possibly other neurodegenerative diseases . Matrix proteinase inhibition by DB05387 , a multifunctional antiangiogenic compound . BACKGROUND : Matrix metalloproteinases ( MMPs ) play an important role in tissue remodeling under normal physiological and pathological conditions and are thus attractive targets for both diagnostic and therapeutic purposes . Here , we examined the effect of DB05387 , an orally bioavailable standardized extract made of cartilage that shows significant antiangiogenic and antimetastatic properties in vivo , on the activity of various members of the MMP family . MATERIALS AND METHODS : The effect of DB05387 on the activity of MMPs was assessed by fluorimetric assays and by substrate gel zymography . RESULTS : DB05387 markedly inhibits the gelatinolytic activity of P08253 and to a lesser extent those of P03956 , P09237 , P14780 and P45452 . DB05387 also inhibited the elastinolytic activities of P08253 and P14780 as well as P39900 ( metalloelastase ) , porcine pancreatic elastase ( PPE ) , and human leukocyte elastase ( P08246 ) . Western blot analysis revealed the presence within DB05387 of immunoreactive P01033 -like proteins , suggesting that these proteins may be at least partly responsible for the observed MMP inhibition . CONCLUSIONS : Taken together , these results demonstrate that DB05387 contains P01033 -like proteins that could be responsible for the specific inhibition of MMPs . Given the recent studies suggesting the presence within this compound of specific inhibitor(s) of endothelial cell proliferation , DB05387 appears as a pleotropic agent able to interfere with several biochemical steps leading to angiogenesis and to other physiopathological conditions . Since DB05387 is currently under Phase III clinical investigations , these findings are also of considerable importance for our understanding of its anticancer properties . Crystallization and preliminary X-ray diffraction analysis of the catalytic domain of recombinant human phosphodiesterase 3B . The catalytic domain of human phosphodiesterase 3B has been cloned , expressed in Escherichia coli and purified in the presence of the PDE3 inhibitors DB07954 ( 3-isobutylmethylxanthine ) or MERCK1 by affinity chromatography . Initial screening of crystallization conditions for these complexes in the hanging-drop vapor-diffusion mode resulted in three different crystal forms , all characterized by quite large unit-cell parameters , elevated solvent content and poor diffraction quality . Subsequent optimization of these conditions led to crystals that diffract to 2.4 A and belong to space group P06681 , with unit-cell parameters a = 146.7 , b = 121.5 , c = 126.3 A , beta = 100.6 degrees . Rotation-function analysis indicates that the asymmetric unit contains four copies of the monomeric enzyme , corresponding to a solvent content of 64 % . To solve the structure of the Q13370 catalytic domain , molecular replacement as well as multiple isomorphous replacement methods are currently being utilized . Inhibition of rat renal fibroblast proliferation by halofuginone . BACKGROUND/AIM : Interstitial fibrosis is the final common pathway of renal damage and represents an important therapeutic target . DB04866 is a nontoxic alkaloid , used as a coccidiostat , and is a potent inhibitor of collagen alpha(1)(I) and matrix metalloproteinase-2 ( P08253 ) expression . We thus studied the effects of halofuginone on proliferation , collagen I synthesis , and P08253 activity of rat renal papillary fibroblasts in culture . METHODS : Fibroblasts were isolated from rat renal papillae and studied during passages 3-4 . The cell proliferation was studied in the presence of varying concentrations of halofuginone . The collagen synthesis was studied by [3H]proline uptake , before and after collagenase digestion , at varying concentrations of halofuginone . The P08253 activity was determined by zymography . The gelatinolytic activity was determined on gelatin-impregnated polyacrylamide gels containing samples of cell medium after incubation for 24 h with different halofuginone doses . RESULTS : We studied a phenotype of papillary fibroblasts which stained positive for alpha smooth muscle actin . These cells are phenotypically myofibroblasts . Halufuginone inhibited the proliferation of these cells in a dose-related and reversible manner . Platelet-derived growth factor is known to stimulate fibroblast proliferation . DB04866 at a concentration of 250 ng/ml almost completely abolished the effect of platelet-derived growth factor . It also almost completely inhibited the P08253 activity at doses of 250-350 ng/ml , as shown by zymography . CONCLUSIONS : DB04866 exhibits antifibrotic effects in rat renal papillary fibroblasts in culture , in terms of inhibition of proliferation and inhibition of P08253 . These findings could have therapeutic potential . P62937 facilitates translocation of the Clostridium botulinum P06681 toxin across membranes of acidified endosomes into the cytosol of mammalian cells . The binary Clostridium botulinum P06681 toxin consists of the binding/translocation component C2IIa and the separate enzyme component C2I , which mono-ADP-ribosylates actin in eukaryotic cells . Pore formation of C2IIa in early endosomal membranes facilitates translocation of unfolded C2I into the cytosol . We discovered earlier that translocation of C2I depends on the activity of the host cell chaperone heat shock protein Hsp90 . Here , we demonstrate that cyclosporin A , which inhibits the peptidyl-prolyl cis/trans isomerase activity of cyclophilins , inhibited intoxication of cells with P06681 toxin and prevented uptake of C2I into the cytosol . DB00091 blocked the pH-dependent translocation of C2I activity across membranes of intact cells and of partially purified early endosomes . In vitro , the addition of cytosol to P06681 toxin-loaded endosomes induced translocation of C2I activity into the cytosol , which was prevented by pretreatment of the cytosol with an antibody against cyclophilin A . Pull-down experiments with lysates from P06681 toxin-treated cells revealed specific binding of cyclophilin A to the N-terminal domain of C2I . In conclusion , our results suggest an essential role of cyclophilin A for translocation of C2I across endosomal membranes during the uptake of P06681 toxin into mammalian cells . DB09130 metabolism and oxidative stress in chronic inflammatory and cholestatic liver diseases in dogs . Inherited defects of copper metabolism resulting in hepatic copper accumulation and oxidative-stress might cause breed-associated forms of hepatitis . Biliary excretion is the major elimination route of copper , therefore increased hepatic copper concentrations could also be caused by cholestasis . The aim of this study was to find criteria to determine whether copper-accumulation is primary or occurs secondary to hepatitis . Liver samples of Bedlington Terriers with copper toxicosis ( CT ) , breeds with non-copper-associated chronic extrahepatic cholestasis ( EC ) or chronic hepatitis ( CH ) , and healthy dogs were used . DB09130 metabolism was analyzed by means of histochemical staining ( copper concentration ) and quantitative reverse transcriptase polymerase chain reaction ( Q-PCR ) on copper excretion/storage ( O00244 , Q14061 , Q04656 , P35670 , CP , P04731 , Q8N668 , P98170 ) . Oxidative stress was measured by determining DB00143 /GSSG ratios and gene-expression ( P00441 , CAT , GSHS , P07203 , O14618 , p27KIP , Bcl-2 ) . Results revealed 5+ copper in CT , but no or 1-2+ copper in EC and CH . Most gene products for copper metabolism remained at concentrations similar to healthy dogs . Three clear exceptions were observed in CT : 3-fold mRNA increase of Q04656 and P98170 and complete absence of MURRI . The only quantitative differences between the diseased and the control groups were in oxidative stress , evidenced by reductions in all DB00143 /GSSG ratios . We conclude that 3+ or higher histochemical detection of copper indicates a primary copper storage disease . The expression profile of copper-associated genes can be used as a reference for future studies on copper-associated diseases . All 3 diseases have reduced protection against oxidative stress , opening a rationale to use antioxidants as possible therapy . Oxidative stress-induced p53 activity is enhanced by a redox-sensitive Q96A56 SUMOylation . Tumor Protein p53-Induced Nuclear Protein 1 ( Q96A56 ) is a tumor suppressor that modulates the p53 response to stress . Q96A56 is one of the key mediators of p53 antioxidant function by promoting the p53 transcriptional activity on its target genes . Q96A56 expression is deregulated in many types of cancers including pancreatic ductal adenocarcinoma in which its decrease occurs early during the preneoplastic development . In this work , we report that redox-dependent induction of p53 transcriptional activity is enhanced by the oxidative stress-induced SUMOylation of Q96A56 at lysine 113 . This SUMOylation is mediated by Q9Y6X2 and O00257 , two SUMO ligases especially related to the p53 activation upon DNA damage . Importantly , this modification is reversed by three P63165 -specific proteases Q9P0U3 , 2 and 6 . Moreover , Q96A56 SUMOylation induces its binding to p53 in the nucleus under oxidative stress conditions . Q96A56 mutation at lysine 113 prevents the pro-apoptotic , antiproliferative and antioxidant effects of Q96A56 by impairing the p53 response on its target genes P38936 , Bax and PUMA . We conclude that Q96A56 SUMOylation is essential for the regulation of p53 activity induced by oxidative stress . Slow overmethylation of housekeeping genes in the body mucosa is associated with the risk for gastric cancer . Helicobacter pylori infection increases age-related diverse overmethylation in gene-control regions , which increases the risk of gastric cancer . The H. pylori-associated overmethylation changes subsequently disappear when gastric atrophy and cancer develop . To identify cancer-risk epigenotypes , we traced dynamic methylation changes in the background mucosa of the stomach depending on the extent of gastric atrophy . Paired biopsy specimens were obtained from the noncancerous antrum and body mucosa of 102 patients with cancer and 114 H. pylori-positive and 112 H. pylori-negative controls . The grade of gastric atrophy was evaluated using the endoscopic atrophic border score . The methylation-variable sites at the CpG-island margins and near the transcriptional start sites lacking CpG islands were semiquantitatively analyzed by radioisotope-labeling methylation-specific PCR . We selected eight housekeeping genes adjacent to Alu ( CDH1 , Q8NCT1 , P37231 , and Q9UL33 ) or LTR retroelements ( P08253 , CDKN2A , Q13950 , and Q13761 ) and eight stomach-specific genes ( Q03403 , P20142 , P51164 , P04155 , Q07654 , Q9UBU3 , DB00158 , and P20648 ) . Analysis of age-related methylation in the H. pylori-positive controls revealed slow overmethylation in the body and in the LTR-adjacent genes . A high-frequency overmethylation defined based on the slowly overmethylated genes was frequently observed in the body of patients with gastric cancer with open-type atrophy ( OR , 12.7 ; 95 % confidence interval , 3.2-49.8 ) . The rapidly changing methylation of Alu-adjacent genes was barely increased in the antrum of patients with gastric cancer . Among diverse methylation changes associated with H. pylori infection , an increase in slowly changing methylation could serve as a cancer-risk marker . Electrostatic steering at acetylcholine binding sites . The electrostatic environments near the acetylcholine binding sites on the nicotinic acetylcholine receptor ( nAChR ) and acetylcholinesterase were measured by diffusion-enhanced fluorescence energy transfer ( DEFET ) to determine the influence of long-range electrostatic interactions on ligand binding kinetics and net binding energy . Changes in DEFET from variously charged Tb3+ -chelates revealed net potentials of -20 mV at the nAChR agonist sites and -14 mV at the entrance to the P22303 active site , in physiological ionic strength conditions . The potential at the alphadelta-binding site of the nAChR was determined independently in the presence of DB01199 to be -14 mV ; the calculated potential at the alphagamma-site was approximately threefold stronger than at the alphadelta-site . By determining the local potential in increasing ionic strength , Debye-Hückel theory predicted that the potentials near the nAChR agonist binding sites are constituted by one to three charges in close proximity to the binding site . Examination of the binding kinetics of the fluorescent acetylcholine analog dansyl- P13671 -choline at ionic strengths from 12.5 to 400 mM revealed a twofold decrease in association rate . Debye-Hückel analysis of the kinetics revealed a similar charge distribution as seen by changes in the potentials . To determine whether the experimentally determined potentials are reflected by continuum electrostatics calculations , solutions to the nonlinear Poisson-Boltzmann equation were used to compute the potentials expected from DEFET measurements from high-resolution models of the nAChR and P22303 . These calculations are in good agreement with the DEFET measurements for P22303 and for the alphagamma-site of the nAChR . We conclude that long-range electrostatic interactions contribute -0.3 and -1 kcal/mol to the binding energy at the nAChR alphadelta- and alphagamma-sites due to an increase in association rates . Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Cytokine signaling via various transcription factors regulates receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) -mediated osteoclast differentiation from monocyte/macrophage lineage cells involved in propagation and resolution of inflammatory bone destruction . Protein inhibitor of activated P40763 ( Q9Y6X2 ) was initially identified as a molecule that inhibits DNA binding of P40763 and regulates many transcription factors through distinct mechanisms . To analyze Q9Y6X2 function in osteoclasts in vivo , we have generated transgenic mice in which Q9Y6X2 is specifically expressed in the osteoclast lineage using the tartrate-resistant acid phosphatase ( TRAP ) gene promoter . Q9Y6X2 transgenic mice showed an osteopetrotic phenotype due to impairment of osteoclast differentiation . Overexpression of Q9Y6X2 in RAW264.7 cells suppressed O14788 -induced osteoclastogenesis by inhibiting the expression of c-Fos and O95644 . Interestingly , Q9Y6X2 inhibits the transcriptional activity of microphthalmia-associated transcription factor ( O75030 ) independent of sumoylation . Down-regulation of Q9Y6X2 markedly enhances O14788 -mediated osteoclastogenesis in RAW264.7 cells . Furthermore , overexpression of Q9Y6X2 in mouse primary osteoblast ( DB00925 ) , down-regulates O14788 expression induced by interleukin-6 ( P05231 ) cytokine family , and inhibits osteoclast formation from bone marrow macrophages ( BMMs ) in vitro coculture system . Down-regulation of Q9Y6X2 leads to the accelerated expression of O14788 in DB00925 stimulated with P05231 and soluble P05231 receptor ( sIL-6R ) . Taken together , our results clearly indicate that Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Uptake of particulate vaccine adjuvants by dendritic cells activates the Q96P20 inflammasome . Many currently used and candidate vaccine adjuvants are particulate in nature , but their mechanism of action is not well understood . Here , we show that particulate adjuvants , including biodegradable poly(lactide-co-glycolide) ( P00747 ) and polystyrene microparticles , dramatically enhance secretion of interleukin-1beta ( IL-1beta ) by dendritic cells ( DCs ) . The ability of particulates to promote IL-1beta secretion and caspase 1 activation required particle uptake by DCs and Q96P20 . Uptake of microparticles induced lysosomal damage , whereas particle-mediated enhancement of IL-1beta secretion required phagosomal acidification and the lysosomal cysteine protease cathepsin B , suggesting a role for lysosomal damage in inflammasome activation . Although the presence of a Toll-like receptor ( TLR ) agonist was required to induce IL-1beta production in vitro , injection of the adjuvants in the absence of TLR agonists induced IL-1beta production at the injection site , indicating that endogenous factors can synergize with particulates to promote inflammasome activation . The enhancement of antigen-specific antibody production by P00747 microparticles was independent of Q96P20 . However , the ability of P00747 microparticles to promote antigen-specific P05231 production by T cells and the recruitment and activation of a population of CD11b(+)Gr1(-) cells required Q96P20 . Our data demonstrate that uptake of microparticulate adjuvants by DCs activates the Q96P20 inflammasome , and this contributes to their enhancing effects on innate and antigen-specific cellular immunity . Prion protein stimulates tissue-type plasminogen activator-mediated plasmin generation via a lysine-binding site on kringle 2 . Recombinant human prion-protein ( PrP23-231 ) stimulates plasminogen activation by tissue-type plasminogen activator ( t-PA ) . The stimulatory activity is conserved in the N-terminal fragment ( PrP23-110 ) . It has further been shown by others that P04156 (c) binds to kringle-domains of plasminogen . We compared the stimulatory activity of recombinant PrP23-231 and PrP23-110 on plasminogen activation catalyzed by t-PA , urokinase ( u-PA ) , streptokinase and Desmodus salivary plasminogen activator ( DSPAalpha1 ) . As these plasminogen activators are distinct , with respect to their kringle domains we studied their binding to immobilized PrP23-110 . P00747 activation was measured in a chromogenic assay in vitro and binding studies were carried out using surface plasmon resonance technology . We found that recombinant full-length prion protein , PrP23-231 , and PrP23-110 specifically stimulate t-PA mediated plasminogen activation . Two hundred nanomoles per liter of PrP23-110 stimulated 1.8 nmol L(-1) t-PA 48-fold , 180 nmol L(-1) DB04925 (alpha1) 2.5-fold , 1.8 nmol L(-1) u-PA 1.1-fold , and 1.8 nmol L(-1) streptokinase 1.8-fold . Our data show no specific binding for streptokinase . In contrast all plasminogen activators carrying a kringle domain bound to PrP23-110 . We further studied the effect of lysine on binding to PrP23-110 and on plasminogen activation by DB04925 (alpha1) or t-PA . DB00123 decreased both the binding of t-PA to PrP23-110 and the stimulation of plasmin generation by t-PA . Both binding and plasminogen activation of DB04925 (alpha1) were not influenced by the presence of lysine . All plasminogen activators tested bearing kringle domains bind to PrP23-110 . Binding to PrP23-110 is not sufficient for stimulation of plasmin generation . Thus the lysine-binding site of kringle 2 that is unique to t-PA appears to mediate the specific stimulation of plasminogen activation by the cellular prion protein . Secreted cyclophilin A mediates P55008 /S phase transition of cholangiocarcinoma cells via CD147/ P27361 /2 pathway . P62937 ( CypA ) was shown to be upregulated in human cholangiocarcinoma ( CCA ) tissues . Suppression of intracellular CypA ( inCypA ) significantly reduces cell proliferation in vitro and tumor growth in nude mice . In the present study , the effect and potential mechanism of secreted CypA ( sCypA ) on cell proliferation of CCA cell lines were further investigated . CCA cells were treated with sCypA-containing conditioned media ( CM ) or with purified recombinant human CypA ( rhCypA ) . Cell proliferation , cell cycle , P27361 /2 , p38 MAPK , NF-κB, and P40763 activities were examined by MTS assay , flow cytometry , and Western blot . sCypA was detected in CM from MMNK1 ( an immortalized human cholangiocyte cell line ) and six CCA cell lines . The sCypA levels corresponded to the inCypA levels indicating the intracellular origin of sCypA . Both sCypA-containing CM and rhCypA significantly increased proliferation of CCA cells . CD147 depletion by shRNA-knockdown or neutralizing with a CD147-monoclonal antibody significantly reduced sCypA- , and rhCypA-mediated cell proliferation . Upon rhCypA treatment , P27361 /2 was rapidly phosphorylated ; whereas neutralizing CD147 inhibited P27361 /2 phosphorylation . Cell cycle analysis showed a significant increase in S phase and decrease in P55008 population in rhCypA-treated cells . The expression levels of cyclin D1 and phosphorylated-retinoblastoma protein in the rhCypA-treated cells were increased compared with those in the non-treated control cells . p38 MAPK pathway was shown to be suppressed in siCypA-treated cells . In summary , CypA is secreted from CCA cells and enhances cell proliferation in an autocrine/paracrine manner , at least via direct binding with CD147 , which may activate the P27361 /2 and p38 MAPK signaling pathways . Inhibition of JAKs in macrophages increases lipopolysaccharide-induced cytokine production by blocking P22301 -mediated feedback . Macrophages are an important source of cytokines following infection . Stimulation of macrophages with TLR agonists results in the secretion of P01375 -α , P05231 , and IL-12 , and the production of these cytokines is controlled by multiple feedback pathways . Macrophages also produce P22301 , which acts to inhibit proinflammatory cytokine production by macrophages via a JAK/ P40763 -dependent pathway . We show in this paper that , DB08877 , a recently described selective inhibitor of JAKs , increases P01375 , P05231 , and IL-12 secretion in mouse bone marrow-derived macrophages stimulated with LPS . This effect is largely due to its ability to block P22301 -mediated feedback inhibition on cytokine transcription in macrophages . Similar results were also obtained with a second structurally unrelated Jak inhibitor , DB08895 . In addition , LPS induced the production of IFN-β , which was then able to activate JAKs in macrophages , resulting in the stimulation of P42224 phosphorylation . The initial induction of P22301 was independent of JAK signaling ; however , inhibition of JAKs did reduce P22301 secretion at later time points . This reflected a requirement for the IFN-β feedback loop to sustain P22301 transcription following LPS stimulation . In addition to P22301 , IFN-β also helped sustain P05231 and IL-12 transcription . Overall , these results suggest that inhibition of JAKs may increase the inflammatory potential of macrophages stimulated with O00206 agonists . A role of phosphodiesterase-3B pathway in mediating leptin action on proopiomelanocortin and neurotensin neurons in the hypothalamus . Leptin signaling in the hypothalamus is required for normal food intake and body weight homeostasis . Recent evidence suggests that besides the signal transducer and activator of transcription-3 ( P40763 ) pathway , several non- P40763 pathways mediate leptin signaling in the hypothalamus . We have previously demonstrated that leptin stimulates phosphodiesterase-3B ( Q13370 ) activity in the hypothalamus , and PDE3 inhibitor cilostamide reverses anorectic and body weight reducing effects of leptin . To establish the physiological role of Q13370 signaling in the hypothalamus , we examined if leptin signaling through the Q13370 pathway is responsible for the activation of proopiomelanocortin ( P01189 ) and neurotensin ( NT ) neurons , which are known to play a critical role in energy homeostasis . To this end , we assessed the effect of cilostamide on leptin-induced P01189 and NT gene expression in the rat hypothalamus . Results showed that while central injection of leptin significantly increased both P01189 and NT mRNA levels in the medial basal hypothalamus , cilostamide completely reversed this effect of leptin suggesting a Q13370 -activation dependent induction of P01189 and NT gene expression by leptin . This result further suggests that the Q13370 pathway plays an important role in mediating leptin action in the hypothalamus .

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interacts_with DB00009?

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NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC . Tuberous sclerosis complex : disease modifiers and treatments . PURPOSE OF REVIEW : Tuberous sclerosis complex ( TSC ) is an autosomal dominant neurocutaneous disorder involving benign growths in multiple organ systems of affected patients . Variable phenotypes from mild to severe have been reported for related as well as unrelated patients affected by TSC . The two causative genes , Q92574 and P49815 , which code for hamartin and tuberin respectively , play central roles in regulating cell survival and proliferation signaling pathways . The severity of disease phenotypes of TSC patients is influenced by the activities of genes both up and down-stream in the associated pathways . RECENT FINDINGS : The high-expressing12CA repeat variant of the P01579 gene was suggested to contribute lower risk for kidney angiomyolipomas in patients with P49815 gene mutations . Genetic modifiers for TSC have been localized on chromosomes 3 and 5 of the rat genome . We performed association studies linking the c.68C allele of the P28335 gene to lower seizure risk in TSC-affected individuals . SUMMARY : Genetic and epigenetic factors affecting the activity of each and every interacting partner of the tuberin-hamartin complex could potentially alter the disease presentation . Identifying functional polymorphic variants of interacting partners affecting TSC gene functions will delineate the mechanisms leading to TSC disease severity , ultimately resulting in treatment strategies . Identification of flavopiridol analogues that selectively inhibit positive transcription elongation factor ( P-TEFb ) and block HIV-1 replication . The positive transcription elongation factor ( P-TEFb ; P50750 /cyclin T1 ) regulates RNA polymerase II-dependent transcription of cellular and integrated viral genes . It is an essential cofactor for HIV-1 Tat transactivation , and selective inhibition of P-TEFb blocks HIV-1 replication without affecting cellular transcription ; this indicates that P-TEFb could be a potential target for developing anti-HIV-1 therapeutics . DB03496 , a small molecule CDK inhibitor , blocks HIV-1 Tat transactivation and viral replication by inhibiting P-TEFb kinase activity , but it is highly cytotoxic . In the search for selective and less cytotoxic P-TEFb inhibitors , we prepared a series of flavopiridol analogues and evaluated their kinase inhibitory activity against P-TEFb and P24941 /cyclin A , and tested their cellular antiviral potency and cytotoxicity . We identified several analogues that selectively inhibit P-TEFb kinase activity in vitro and show antiviral potency comparable to that of flavopiridol , but with significantly reduced cytotoxicity . These compounds are valuable molecular probes for understanding P-TEFb-regulated cellular and HIV-1 gene transcription and provide potential anti-HIV-1 therapeutics . The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor-1 ( P05121 ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline- and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai1-/- mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 ) and instead reflected proteolytic activation and release of P14210 with subsequent induction of P35354 . That the P14210 / P35354 / DB00917 axis mediates in vivo protection from fibrosis in Pai1-/- mice was demonstrated by experiments showing that a selective inhibitor of the P08581 c- DB00134 increased lung collagen to WT levels while reducing P35354 protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway . Görög Thrombosis Test : a global in-vitro test of platelet function and thrombolysis . This is the first laboratory evaluation of a new instrument , designed to test both platelet function and thrombolytic activity from a native blood sample , in vitro . The inventor assumed that the reduction and arrest of blood flow was due to activation , aggregation and stabilized thrombus formation by shear-activated platelets , and that re-establishment of flow was due to thrombolysis . Morphologic and functional studies presented here confirm these mechanisms . In vitro tests provided incontestable evidence for the principal role of platelets in the obstruction of flow ( occlusion time ) and for thrombolysis as the principal mechanism underlying the restoration of blood flow ( lysis time ) . In addition to aggregation , it is the explosive generation of thrombin by shear-activated platelets that results in the formation of an occlusive haemostatic thrombus . Anticoagulation of blood completely prevented occlusion . Platelet-rich thrombus formation ( occlusion time ) was dose-dependently inhibited by monoclonal antibody against platelet glycoprotein ( GP ) Ib ( 6B4 and 12E4 ) , aurin tricarboxylic acid , monoclonal antibody against platelet P08514 /IIIa ( MA-16N7C2 and abciximab ) , a synthetic P08514 /IIIa antagonist ( P50750 -029 ) , thrombin inhibitor ( argatroban ) , and anti- P04275 , but not by anti-fibrinogen . P00747 activator streptokinase ( Varidase ) and tissue-type plasminogen activator ( Monteplase ) dose-dependently enhanced thrombolysis ( lysis time ) without affecting platelet function ( occlusion time ) . The test is specific for thrombolysis . The plasmin inhibitor tranexamic acid prevented plasminogen activator-induced thrombolysis , while inhibition of clot retraction by cytochalasin B did not affect the lysis time . This rapid and sensitive global test of platelet function and thrombolytic activity could be of great value both in research and in clinical practice . Chronic treatment with agomelatine or venlafaxine reduces depolarization-evoked glutamate release from hippocampal synaptosomes . BACKGROUND : Growing compelling evidence from clinical and preclinical studies has demonstrated the primary role of alterations of glutamatergic transmission in cortical and limbic areas in the pathophysiology of mood disorders . Chronic antidepressants have been shown to dampen endogenous glutamate release from rat hippocampal synaptic terminals and to prevent the marked increase of glutamate overflow induced by acute behavioral stress in frontal/prefrontal cortex . DB06594 , a new antidepressant endowed with MT1/ P02795 agonist and P28335 serotonergic antagonist properties , has shown efficacy at both preclinical and clinical levels . RESULTS : Chronic treatment with agomelatine , or with the reference drug venlafaxine , induced a marked decrease of depolarization-evoked endogenous glutamate release from purified hippocampal synaptic terminals in superfusion . No changes were observed in GABA release . This effect was accompanied by reduced accumulation of SNARE protein complexes , the key molecular effector of vesicle docking , priming and fusion at presynaptic membranes . CONCLUSIONS : Our data suggest that the novel antidepressant agomelatine share with other classes of antidepressants the ability to modulate glutamatergic transmission in hippocampus . Its action seems to be mediated by molecular mechanisms located on the presynaptic membrane and related with the size of the vesicle pool ready for release . Activated factor B ( Bb ) of the alternative pathway of complement activation cleaves and activates plasminogen . Activated Factor B ( Bb ) , the central serine esterase of the alternative pathway of complement activation , exhibits restricted substrate specificity in the complement system for P01024 and P01031 . The results presented here indicate that Bb can cleave and activate plasminogen in an experimental system containing purified plasminogen and Bb ; complement cellular intermediate bearing the Bb-enzyme ; or cobra venom factor-stabilized Bb-enzyme ( CVF,Bb ) . Cleavage of plasminogen by Factor Bb generated 2 disulfide-linked polypeptides with apparent m.w. of 64,000 and 25,000 to 32,000 ( SDS-PAGE ) . Complement cellular intermediates containing the C3b , Bb-enzyme cleave 40 to 80 % of 4.5 micrograms of 125I-labeled plasminogen during 30 min of incubation at 37 degrees C ; native Factor B was inactive ; and anti-Factor Blg inhibited by 100 % the plasminogen cleavage mediated by complement cellular intermediates bearing the Bb-enzyme . Fibrinolytic activity was detected in plasminogen activator ( PA ) assays when purified plasminogen and 125I-labeled fibrin tubes were incubated with Bb , CVF , Bb , or complement cellular intermediates bearing the C3b,Bb-enzyme : 10 micrograms Bb released 40 to 65 % of the 125I-fibrin released by 5 micrograms urokinase in 4 hr at 37 degrees C . P00747 activator activity of the C3b,Bb-enzyme was found to be regulated in serum . At dilutions of Q6T4R5 1:50 , the PA-activity of 1.6 micrograms Bb was 100 % inhibited , and at a 1:250 dilution , 50 % inhibition was observed . This report describes a novel activity for the Bb-enzyme , which constitutes the P01024 / P01031 -convertase of the alternative pathway of complement activation . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . Q07973 as a potential target for cancer therapy . Increasing evidence has accumulated to suggest that vitamin D may reduce the risk of cancer through its biologically active metabolite , DB00136 , which inhibits proliferation and angiogenesis , induces differentiation and apoptosis , and regulates many other cellular functions . Thus , it is plausible to assume that rapid clearance of DB00136 by highly expressed Q07973 could interrupt the normal physiology of cells and might be one cause of cancer initiation and progression . In fact , enhancement of Q07973 expression has been reported in literature for many cancers . Based on these findings , Q07973 -specific inhibitors and vitamin D analogs which are resistant to Q07973 -dependent catabolism might be useful for cancer treatment . Q07973 -specific inhibitor VID400 , which is an azole compound , markedly enhanced and prolonged the antiproliferative activity of DB00136 in the human keratinocytes . Likewise , Q07973 -resistant analogs such as 2α-(3-hydroxypropoxy)- DB00136 ( O2C3 ) and its P06681 -epimer ED-71 ( DB05295 ) , and 19nor- 2α-(3-hydroxypropyl)- DB00136 ( MART-10 ) showed potent biological effects . Our in vivo studies using rats revealed that MART-10 had a low calcemic effect , which is a suitable property as an anticancer drug . Much lower affinity of MART-10 for vitamin D binding protein ( DBP ) as compared with DB00136 may be related to its more potent cellular activities . Based on these results , we conclude that ( 1 ) high affinity for P11473 , ( 2 ) resistance to Q07973 -dependent catabolism , ( 3 ) low affinity for DBP , and ( 4 ) low calcemic effect may be required for designing potent vitamin D analogs for cancer treatment . [ Use of 2-DE and MALDI-TOF-MS to screen differentially expressed serum proteins in patients with oral lichen planus before and after hydroxychloroquine treatment ] . PURPOSE : To establish 2-dimensional gel electrophoresis images and compare the differences of serum proteins of oral lichen planus patients before and after hydroxychloroquine therapy . METHODS : The serum of oral lichen planus patients before and after hydroxychloroquine therapy were collected , and total protein were extracted . Differential proteome profiles were established and analysed by means of 2-DE and MALDI-TOF-MS . The types and functions of protein were analyzed . SAS 9.12 software package was used for statistical analysis . RESULTS : Six proteins were well characterized including plasminogen precursor,Apo A-IV precursor , P0C0L4 / P0C0L5 complement , P06681 precursor , Vitamin D binding protein and hypothetical protein . The differences were statistically significant . CONCLUSIONS : P00747 precursor , Apo A-IV precursor , P0C0L4 / P0C0L5 complement , P06681 precursor , Vitamin D binding protein and hypothetical protein are differentially expressed in oral lichen planus patients before and after hydroxychloroquine therapy , but the results need to be validated by other biochemical technologies . Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin-1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA/uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 ) or interleukin-1 beta ( P01584 ) , are present in epidermal wounds . We have therefore tested P01584 and P01375 for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 and P01375 induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing . Management of ocular inflammation and pain following cataract surgery : focus on bromfenac ophthalmic solution . Recently , several new ophthalmic NSAID products have been introduced for commercial use in the United States . The purpose of this review is to briefly overview the ophthalmic NSAIDs currently in use and to discuss the management of postoperative ocular inflammation and pain following cataract surgery with a particular focus on bromfenac ophthalmic solution 0.09 % . DB00963 ophthalmic solution 0.09 % is indicated for the reduction of ocular pain and inflammation following cataract surgery . Studies have shown that bromfenac ophthalmic solution 0.09 % has equivalent efficacy to the other topical NSAIDs in reducing postsurgical inflammation and controlling pain . The unique chemical structure of bromfenac makes it both a potent inhibitor of the P35354 enzyme and a highly lipophilic molecule that rapidly penetrates to produce early and sustained drug levels in all ocular tissues . Clinically , these pharmacokinetic features are manifested in a rapid reduction of postsurgical inflammation and pain with bid dosing . DB00963 ophthalmic solution 0.09 % is a versatile agent and is effective when used as either monotherapy or as an adjunct therapy to steroids . z-VAD-fmk-induced non-apoptotic cell death of macrophages : possibilities and limitations for atherosclerotic plaque stabilization . Macrophages play a pivotal role in atherosclerotic plaque destabilization in contrast to smooth muscle cells ( SMCs ) . As a consequence , removal of macrophages from plaques via selective induction of cell death represents a promising approach to stabilize non-obstructive , rupture-prone atherosclerotic lesions . However , the mechanisms to initiate cell death in macrophages but not in other cell types of the plaque , in particular SMCs , are unknown . Recently , we have shown that the pan-caspase inhibitor z-VAD-fmk induces autophagy and necrotic cell death in J774A.1 and RAW264.7 macrophages as well as in P01579 primed primary mouse peritoneal macrophages , but not in vascular SMCs or C2C12 myoblasts . The different sensitivity to z-VAD-fmk is largely based on differential expression of receptor-interacting protein 1 ( Q13546 ) . This finding suggests that caspase inhibition activates Q13546 which in turn initiates autophagy , although other explanations should be taken into account . z-VAD-fmk-treated J774A.1 macrophages overexpress and secrete several chemokines and cytokines , including TNFalpha . The combination of z-VAD-fmk and TNFalpha , but not TNFalpha alone , induces SMC necrosis . In this regard , z-VAD-fmk is detrimental and not beneficial for atherosclerotic plaque stability due to stimulation of inflammatory responses and indirect induction of SMC death . Future work is needed to determine the mechanism(s) that selectively trigger non-apoptotic cell death in plaque macrophages without evoking inflammation and SMC death . A multicenter , open-label , prospective , randomized , dose-ranging pharmacokinetic study of the anti- P01375 antibody afelimomab in patients with sepsis syndrome . OBJECTIVE : To investigate the pharmacokinetics and safety of afelimomab , a murine antibody fragment against human tumor necrosis factor ( P01375 ) -alpha in patients with sepsis . DESIGN : Multicenter , randomized , open-label , placebo-controlled phase I/II clinical trial . SETTING : Intensive care units of six academic medical centers in the United States . PATIENTS : Forty-eight patients with a clinical diagnosis of sepsis who received standard supportive care and antimicrobial therapy . INTERVENTIONS : Patients received 0.3 , 1.0 , or 3.0 mg/kg afelimomab or placebo intravenously over 20 min . Three patients in each dose group received single doses ; the remaining nine patients in each group received multiple ( nine ) doses at 8-h intervals over 72 h . MEASUREMENTS AND MAIN RESULTS : DB04956 appeared safe and well tolerated . Single- and multiple-dose kinetics were predictable and dose related . The elimination half-life was 44.7 h . DB04956 treatment resulted in increased serum concentrations of P01375 ( includes P01375 -antibody complexes ) and decreased serum interleukin-6 concentrations , whereas no discernible trends were observed in placebo-treated patients . There was no significant treatment effect on 28-day mortality as was expected given the small number of patients . However , overall mortality was significantly ( p = 0.001 ) associated with baseline interleukin-6 concentration . All patients experienced adverse events , but the vast majority were considered unrelated to the study drug and demonstrated no apparent relationship to afelimomab dose . Although 41 % of patients developed human anti-murine antibodies , there were no clinical sequelae . CONCLUSIONS : Multidose therapy with afelimomab was safe , well tolerated , and had predictable linear kinetics . A large randomized trial comparing afelimomab to placebo in patients with well defined sepsis has recently been completed . Pharmacological activities of the MIF-1 analogues Pro- DB00149 - DB00145 , DB00135 -Pro- DB00149 - DB00145 and pareptide . The pharmacological activities of the related free acid analogues of MIF-1 , Pro- DB00149 - DB00145 ( P00747 ) and DB00135 -Pro- DB00149 - DB00145 ( YPLG ) , were investigated because of the possibility that they may be formed during the digestion of milk and wheat proteins in vivo . The amino acid sequences - DB00135 -Pro- DB00149 - DB00145 - and -Pro- DB00149 - DB00145 - are present in proteins from these foods . Chronic administration of either P00747 ( 0.25 mg/kg , SC , P55957 ) or the control substance , pareptide ( 0.25 mg/kg , SC , P55957 ) , antagonized the development of tolerance to the cataleptic effects of haloperidol in mice . The effect of YPLG ( 0.25 mg/kg , SC , P55957 ) on the development of this tolerance was borderline and not statistically significant . Nanomolar concentrations of P00747 , YPLG , and pareptide each increased the in vitro binding of 3H-apomorphine to rat striatal receptors . In this in vitro system , bell-shaped dose response curves were observed for each peptide . The effects of these peptides on tolerance development and apomorphine binding are similar to those previously reported for MIF-1 and demonstrate that amidation at the carboxyl terminus is not required for biological activity . Can P35354 inhibitor-induced increase in cardiovascular disease risk be modified by essential fatty acids ? Selective P35354 inhibitors increase the risk of myocardial infarction and stroke . This has been attributed to their ability to inhibit endothelial P35354 derived prostacyclin ( DB01240 ) but not platelet P23219 derived thromboxane A2 ( TXA2 ) . On the other hand , aspirin blocks both P23219 and P35354 enzymes without decreasing DB01240 but blocks TXA2 synthesis that explains its beneficial action in the prevention of coronary heart disease ( Q8NE62 ) . The inhibitory action of aspirin on P23219 and P35354 enzymes enhances the tissue concentrations of dihomo-gamma-linolenic acid ( DB00154 ) , arachidonic acid , eicosapentaenoic acid ( EPA ) , and docosahexaenoic acid ( DB01708 ) . These fatty acids form precursors to PGE1 , DB01240 , PGI3 , lipoxins ( LXs ) , and resolvins that have anti-inflammatory actions . In contrast , increase in the concentrations of DB00154 , AA , EPA , and DB01708 is much less with specific P35354 inhibitors since they do not block the formation of eicosanoids through P23219 pathway . P35354 inhibitors interfere with the formation of LXs and resolvins that have neuroprotective and cardioprotective actions . EPA and DB01240 have anti-arrhythmic action . EPA , DB01708 , and AA augment eNO formation that prevents atherosclerosis . This suggests that P35354 inhibitors increase cardiovascular and stroke risk by interfering with the formation of eNO , DB01240 , LXs , and resolvins and implies that combining EFAs with P35354 inhibitors could prevent these complications . Mutational analysis of the mitochondrial P47985 of Saccharomyces cerevisiae . II . Biochemical characterization of temperature-sensitive Q13546 - mutations . Although the function of the P47985 is generally understood , little is known of how the structure of this protein supports its mechanistic role in electron transfer in the cytochrome bc1 complex . To better understand the structural basis of iron-sulfur protein function , we have undertaken a mutational analysis of the gene encoding this protein and initially isolated five temperature-sensitive iron-sulfur protein mutants ( Beckmann , J. D. , Ljungdahl , P. O. , and Trumpower , B. L. ( 1989 ) J. Biol. Chem. 264 , 3713-3722 ) . Each of the five ts-rip1- mutants exhibited pleiotropic effects . Although the mutant iron-sulfur proteins manifest several in vitro phenotypes in common , each exhibited unique characteristics . All of the ts-rip1- mutations resulted in membranes with decreased ubiquinol-cytochrome c oxidoreductase activities and decreased thermostability compared to membranes containing wild type iron-sulfur protein . All of the mutations conferred slight but significant resistance to the respiratory inhibitor myxothiazol , and one mutant was hypersensitive to inhibition by DB04799 , a structural analog of ubiquinone . In addition , one of the mutations completely blocks post-translational processing of the iron-sulfur protein , leading to accumulation of pre-iron-sulfur protein in mitochondrial membranes at nonpermissive temperatures . Finally , a mutation 12-amino acid residues away from the carboxyl terminus ( 203S ) results in an extremely unstable protein . This region of the protein may be essential in blocking degradation of pre-iron-sulfur protein by cytoplasmic proteases as the protein is imported into the mitochondria , or may be a " degradation signal , " which tags the iron-sulfur protein for turnover . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . An open label clinical trial of complement inhibition in multifocal motor neuropathy . Human and animal studies on antibody-mediated neuropathy implicate complement in pathogenesis . In animal models complement inhibition is therapeutically beneficial . The monoclonal antibody , eculizumab ( Soliris™ , Alexion Pharmaceuticals , Cheshire , CT ) , prevents cleavage of P01031 and thus inhibits terminal complement activation . In an open label study , 13 multifocal motor neuropathy patients received eculizumab for 14 weeks , 10 of whom were concomitantly receiving intravenous immunoglobulin . The primary outcome was safety of eculizumab , and the secondary outcomes included change in intravenous immunoglobulin ( DB00028 ) dosing frequency , performance , and electrophysiological parameters . Adverse events were minor during the study . Nine of 10 patients on DB00028 maintenance continued to require DB00028 . DB00028 dosing interval was not different between the run-in and the treatment period . There were improvements in patient-rated subjective scores and selected clinical and electrophysiological measurements . Overall , a small treatment effect occurred in some patients that appeared supplementary to and independent of the DB00028 treatment effect , and occurred more frequently in patients with higher baseline motor function .

MH_train_1247

MH_train_1247

MH_train_1247

interacts_with DB01109?

multiple_choice

Endothelial protective genes induced by statin are mimicked by Q13164 activation as triggered by a drug combination of FTI-277 and GGTI-298 . BACKGROUND : Statins are potent inhibitors of cholesterol biosynthesis and are clinically beneficial in preventing cardiovascular diseases , however , the therapeutic utility of these drugs is limited by myotoxicity . Here , we explored the mechanism of statin-mediated activation of Q13164 in the human endothelium with the goal of identifying compounds that confer endothelial protection but are nontoxic to muscle . METHODS : An Q13164 -one hybrid luciferase reporter transfected into COS-7 cells with pharmacological and molecular manipulations dissected the signaling pathway leading to statin activation of Q13164 . qRT-PCR of HUVEC cells documented the transcriptional activation of endothelial-protective genes . Lastly , morphological and cellular DB00171 analysis , and induction of atrogin-1 in C2C12 myotubes were used to assess statin-induced myopathy . RESULTS : Statin activation of Q13164 is dependent on the cellular reduction of GGPPs . Furthermore , we found that the combination of FTI-277 ( inhibitor of farnesyl transferase ) and GGTI-298 ( inhibitor of geranylgeranyl transferase I ) mimicked the statin-mediated activation of Q13164 . FTI-277 and GGTI-298 together recapitulated the beneficial effects of statins by transcriptionally upregulating anti-inflammatory mediators such as P29474 , P07204 , and Q9Y5W3 . Finally , C2C12 skeletal myotubes treated with both FTI-277 and GGTI-298 evoked less morphological and cellular changes recognized as biomarkers of statin-associated myopathy . CONCLUSIONS : Statin-induced endothelial protection and myopathy are mediated by distinct metabolic intermediates and co-inhibition of farnesyl transferase and geranylgeranyl transferase I confer endothelial protection without myopathy . GENERAL SIGNIFICANCE : The combinatorial FTI-277 and GGTI-298 drug regimen provides a promising alternative avenue for endothelial protection without myopathy . DB01628 -induced fixed drug eruption with positive lesional patch tests . Fixed drug eruption ( FDE ) is most commonly associated with antibiotics , anticonvulsants , and nonnarcotic analgens , including nonsteroidal anti-inflammatory drugs ( NSAIDs ) . However , the newer cyclooxygenase 2 ( P35354 ) inhibitors have been rarely reported to cause FDE . We report the case of a 52-year-old Caucasian woman with erythematous pruritic plaques on the neck , left forearm , and second finger of the right hand , healing with hyperpigmentation and recurring in the same locations . The patient was sporadically taking oral etoricoxib 90 mg for her back pain and noticed the relation between administration of the drug and skin lesions , the time interval decreasing progressively from 1 week to 30 minutes . No other signs , symptoms , or drug intake was mentioned . The patch tests with etoricoxib 1 % and 5 % in petrolatum were positive at the location of the lesions and negative on the back ( nonlesional skin ) . Standard European and NSAID series were negative . Patch tests of 10 healthy controls with etoricoxib 1 % and 5 % in petrolatum were negative . After the avoidance of the drug , no relapse was mentioned . The patch test was reliable for the diagnosis of FDE , avoiding the need for subsequent oral provocation testing and therefore preventing the possible adverse effects . Despite being regarded as a safe drug , the occurrence of cutaneous adverse reactions to etoricoxib should be considered , especially in the setting of its increasing use in pain control . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . Q9UEF7 gene deficiency causes salt-sensitive hypertension via monocyte chemotactic protein-1/CC chemokine receptor 2-mediated inflammation . Q9UEF7 ( KL ) is a newly discovered aging suppressor gene . In mice , the KL gene extends the lifespan when overexpressed and shortens the lifespan when disrupted . This study investigated if KL deficiency affects BP and salt sensitivity using KL mutant heterozygous ( +/- ) mice and wild-type ( WT ) mice ( 9 weeks of age , 16 mice per group ) . Notably , systolic BP in KL(+/-) mice began to increase at the age of 15 weeks , reached a peak level at the age of 17 weeks , and remained elevated thereafter , whereas systolic BP remained consistent in WT mice . High salt ( HS ) intake further increased BP in KL(+/-) mice but did not affect BP in WT mice . Blockade of CC chemokine receptor 2 ( P41597 ) , involved in monocyte chemotaxis , by a specific P41597 antagonist ( DB05130 ) abolished the HS-induced increase in BP in KL(+/-) mice . Furthermore , HS loading substantially increased the expression of monocyte chemotactic protein-1 and the infiltration of macrophages and T cells in kidneys in KL(+/-) mice , and treatment with DB05130 abolished these effects . Treatment of KL(+/-) mice with DB05130 also attenuated the increased renal expressions of serum glucocorticoid-regulated kinase 1 , thiazide-sensitive NaCl cotransporter , and DB00171 synthase β along with the renal structural damage and functional impairment induced by HS loading . In conclusion , KL deficiency caused salt-sensitive hypertension and renal damage by P41597 -mediated inflammation . Implicating P21583 complexes in organogenesis in Caenorhabditis elegans . Development of the Caenorhabditis elegans foregut ( pharynx ) is regulated by a network of proteins that includes the Retinoblastoma protein ( P06400 ) ortholog LIN-35 ; the ubiquitin pathway components P0CG48 -18 and Q9Y4X5 ; and PHA-1 , a cytoplasmic protein . Loss of pha-1 activity impairs pharyngeal development and body morphogenesis , leading to embryonic arrest . We have used a genetic suppressor approach to dissect this complex pathway . The lethality of pha-1 mutants is suppressed by loss-of-function mutations in sup-35/ztf-21 and sup-37/ztf-12 , which encode Zn-finger proteins , and by mutations in sup-36 . Here we show that sup-36 encodes a divergent Skp1 family member that binds to several F-box proteins and the microtubule-associated protein Q02083 -1/τ . Like P60880 -35 , P60880 -36 levels were negatively regulated by P0CG48 -18- Q9Y4X5 . We also found that P60880 -35 and P60880 -37 physically associated and that P60880 -35 could bind microtubules . Thus , P60880 -35 , P60880 -36 , and P60880 -37 may function within a pathway or complex that includes cytoskeletal components . Additionally , P60880 -36 may regulate the subcellular localization of P60880 -35 during embryogenesis . We carried out a genome-wide RNAi screen to identify additional regulators of this network and identified 39 genes , most of which are associated with transcriptional regulation . Twenty-three of these genes acted via the LIN-35 pathway . In addition , several S-phase kinase-associated protein (Skp)1-Cullin-F-Box ( P21583 ) components were identified , further implicating P21583 complexes as part of the greater network controlling pharyngeal development . Investigation of a potential protective mechanism against heparin-induced thrombocytopenia in patients on chronic intermittent hemodialysis . BACKGROUND : DB01109 -induced thrombocytopenia ( HIT ) develops as a result of platelet ( Q02083 ) activation by anti-platelet factor 4 ( P02776 ) /heparin complex antibodies . Despite repeated exposure to heparin , patients undergoing chronic intermittent hemodialysis ( HD ) rarely develop HIT . We investigated the possibility that HD decreases/removes P02776 from Q02083 surfaces and/or plasma , thereby disfavoring immune complex formation as a mechanism of protection against HIT . MATERIALS AND METHODS : We enrolled 20 patients undergoing chronic HD at the Penn Presbyterian Medical Center . Blood samples were drawn before , during and after treatment in the presence and absence of heparin . P02776 , anti- P02776 /heparin antibody , heparin , and P16109 levels were measured . RESULTS : No patients demonstrated clinical symptoms of HIT . Q02083 surface P02776 levels decreased and plasma P02776 levels increased concurrently with the increase in plasma heparin concentration . In the absence of heparin , Q02083 surface and plasma P02776 levels were unchanged . Anti- P02776 /heparin antibodies , which were non-functional by the serotonin release assay , were detectable in 8 patients . Q02083 surface P16109 levels did not change during treatment . CONCLUSIONS : Removal of Q02083 surface and/or plasma P02776 as a mechanism of protection against HIT in patients undergoing HD is not supported by the results of our study , although the transient decrease in Q02083 surface P02776 in the presence of large amounts of heparin remains a candidate mechanism . The small sample size , single type of dialyzer membrane , and early sampling time points may have led to the inability to detect changes in P02776 levels . Future studies should explore other potential protective mechanisms . [ Effect of the monophase oral contraceptive combination with 20 ug ethinyl estradiol/150 ug desogestrel on haemostasis ] . The authors examined the changes in the haemostasis during the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel at 35 women , a basic group , who used the oral contraceptive in the duration of 12 months and a control group ( n=35 ) , who do not use the pills . We found statistically significant increase of Antithrombin III ( P01008 ) ( p < 0.011 ) , Cofactor II of DB01109 ( HCII ) ( p < 0.001 ) , the activity of plasminogen ( p < 0.026 ) and beta2-antiplasmin ( 0.026 ) , significant decrease of P02810 ( PrC ) ( p < 0.0001 ) and of total Protein S ( TPrS ) ( p < 0.03 ) in the basic group in comparision with the control one . We do not observe significant changes in the rest of the haemostatic variables between the two groups . During the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel the changes in the system of the natural inhibitors are balanced by these in the system of fibrinolysis . DB00114 ( PLP ) deficiency might contribute to the onset of type I diabetes . The incidence of type I diabetes is rising worldwide , particularly in young children . Type I diabetes is considered a multifactorial disease with genetic predisposition and environmental factors participating . Currently , despite years of research , there is no consensus regarding the factors that initiate the autoimmune response . Type I diabetes is preceded by autoimmunity to islet antigens , among them the protein glutamic acid decarboxylase , Q05329 . DB00114 ( PLP ) is formed from vitamin B6 by the action of pyridoxal kinase . Interaction of Q05329 with PLP is necessary for Q05329 -mediated synthesis of the neurotransmitter γ-aminobutyric acid ( GABA ) . PLP is also a required cofactor for dopamine synthesis by L-aromatic decarboxylase ( L- P20711 ) . Both Q05329 and L- P20711 are expressed in pancreatic islets . Here it is proposed that lack of the vitamin B6 derivative pyridoxal 5'-phosphate might contribute to the appearance of pancreatic islet autoimmunity and type I diabetes onset . The polyglutamine neurodegenerative protein ataxin 3 regulates aggresome formation . The polyglutamine-containing neurodegenerative protein ataxin 3 ( P01008 ) has deubiquitylating activity and binds ubiquitin chains with a preference for chains of four or more ubiquitins . Here we characterize the deubiquitylating activity of P01008 in vitro and show it trims/edits K48-linked ubiquitin chains . P01008 also edits polyubiquitylated (125)I-lysozyme and decreases its degradation by proteasomes . Cellular studies show that endogenous P01008 colocalizes with aggresomes and preaggresome particles of the misfolded cystic fibrosis transmembrane regulator ( P13569 ) mutant CFTRDeltaF508 and associates with histone deacetylase 6 and dynein , proteins required for aggresome formation and transport of misfolded protein . Small interfering RNA knockdown of P01008 greatly reduces aggresomes formed by CFTRDeltaF508 , demonstrating a critical role of P01008 in this process . Wild-type P01008 restores aggresome formation ; however , P01008 with mutations in the active site or ubiquitin interacting motifs can not restore aggresome formation in P01008 knockdown cells . These same mutations decrease the association of P01008 and dynein . These data indicate that the deubiquitylating activity of P01008 and its ubiquitin interacting motifs as well play essential roles in CFTRDeltaF508 aggresome formation . Therapeutic effect of P23560 -overexpressing human neural stem cells ( HB1. P13726 . P23560 ) in a rodent model of middle cerebral artery occlusion . Ischemic stroke mainly caused by middle cerebral artery occlusion ( MCAo ) represents the major type of stroke ; however , there are still very limited therapeutic options for the stroke-damaged patients . In this study , we evaluated the neurogenic and therapeutic potentials of human neural stem cells ( NSCs ) overexpressing brain-derived neurotrophic factor ( HB1. P13726 . P23560 ) following transplantation into a rodent model of MCAo . P13726 . P23560 human NSCs ( P13726 . P23560 ) were transplanted into the contralateral side of striatum at 7 days after MCAo , and the transplanted animals were monitored up to 8 weeks using animal Q9BWK5 and various behavioral tests before they were sacrificed for immunohistochemical analysis . Interestingly , animal Q9BWK5 results indicate that the majority of contralaterally transplanted neural stem cells were migrated to the peri-infarct area , showing a pathotropism . Transplanted animals exhibited significant behavioral improvements in stepping , rotarod , and modified neurological severity score ( mNSS ) tests . We also found that the transplanted human cells were colocalized with nestin , O43602 , P11137 , Q9UD71 , TH , Q05329 /67-positive cells , of which results can be correlated with neural regeneration and behavioral recovery in the transplanted animals . More importantly , we were able to detect high levels of human P23560 protein expression , presumably derived from the transplanted P13726 . P23560 . Taken together , these results provide strong evidence that human neural stem cells ( P13726 . P23560 ) are effective in treating stroke animal models . Subunit complementation of thymidylate synthase in Plasmodium falciparum bifunctional dihydrofolate reductase-thymidylate synthase . P04818 of Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) functions as a dimeric enzyme with extensive contact between the two TS domains . Structural data of PfDHFR-TS shows that the formation of the two TS active sites involves contribution of the amino acid residues from both TS domains . DB00125 -470 donated from the adjoining domain is shown to hydrogen-bond to DB03800 , while DB00151 -490 is a key nucleophile for TS catalysis by attacking C-6 of DB03800 . However , mutants of the two series could complement one another , giving rise to active enzyme . By means of subunit complementation assay using DB00125 -470 and DB00151 -490 mutants , it is shown that co-transformants of both TS-inactive DB00125 -470 and DB00151 -490 mutants can complement the growth of thymidine auxotroph chi2913RecA(DE3) by formation of a functional TS heterodimer contributing from both DB00125 -470 and DB00151 -490 mutant subunits . 6-[3H]-FdUMP thymidylate synthase activity assay further elaborate the essence of restoration of TS activity . The TS k(cat) value of the R470D+C490A heterodimer is decreased by half from that of the wild-type PfDHFR-TS . However , the Km values for DB03800 and CH2H4folate of the R470D+C490A heterodimer are similar to those of wild-type enzyme , indicating that the catalytic efficiency of the functional TS from the R470D+C490A heterodimer is similar to the wild-type TS enzyme in P. falciparum P00374 -TS . Selective expression of the large neutral amino acid transporter at the blood-brain barrier . Amino acid supply in brain is regulated by the activity of the large neutral amino acid transporter ( O43561 ) at the brain capillary endothelial cell , which forms the blood-brain barrier ( BBB ) in vivo . Bovine BBB poly(A)(+) RNA was isolated from 2.0 kg of fresh bovine brain and size fractionated on a sucrose density gradient , and a size-fractionated bovine BBB cDNA library in the pSPORT vector was prepared . The full-length cDNA encoding the bovine BBB O43561 was isolated from this library , and the predicted amino acid sequence was 89-92 % identical to the Q01650 isoform . The bovine BBB Q01650 mRNA produced a 10-fold enhancement in tryptophan transport into frog oocytes coinjected with bovine BBB Q01650 mRNA and the mRNA for P08195 , which encodes the heavy chain of the heterodimer . DB00150 transport into the mRNA-injected oocytes was sodium independent and was specifically inhibited by other large neutral amino acids , and the K(m) of tryptophan transport was 31.5 +/- 5.5 microM . Northern blotting with the bovine BBB Q01650 cDNA showed that the Q01650 mRNA is 100-fold higher in isolated bovine brain capillaries compared with P13671 rat glioma cells or rat brain , and the Q01650 mRNA was not detected in rat liver , heart , lung , or kidney . These studies show that the Q01650 transcript is selectively expressed at the BBB compared with other tissues , and the abundance of the Q01650 mRNA at the BBB is manyfold higher than that of transcripts such as the P08195 antigen , actin , or the Glut1 glucose transporter . The GLU298ASP variant of nitric oxide synthase interacts with asymmetric dimethyl arginine in determining cardiovascular mortality in patients with end-stage renal disease . OBJECTIVES : Impaired nitric oxide generation and accumulation of the endogenous inhibitor of nitric oxide synthase ( NOS ) , asymmetric dimethylarginine ( DB01686 ) , have been identified as strong predictors of cardiovascular outcomes in patients with end-stage renal disease ( ESRD ) . We evaluated the role of endothelial NOS ( P29474 ) gene polymorphisms and its interaction with plasma DB01686 in the high cardiovascular complications rate of these patients . METHODS : The relationship between the Glu298Asp variant of P29474 and all-cause and cardiovascular mortality was assessed in a cohort study including 261 ESRD patients that were followed up for an average of 42 months . RESULTS : During the follow-up period , 138 patients died , 81 of them ( i.e. 59 % of total deaths ) of cardiovascular causes . On univariate Cox 's regression analysis , P29474 genotype tended to be related to all-cause death but failed to reach formal statistical significance ( P for trend=0.11 ) . However , P29474 genotype showed a significant association with cardiovascular mortality in statistical models , including traditional risk factors and factors peculiar to ESRD , and became even stronger when plasma DB01686 was forced into the Cox model ( P=0.006 ) . Furthermore , the risk of cardiovascular death was maximum in heterozygotes and homozygotes patients carrying the risk allele and in those having high DB01686 levels ( hazard ratio=2.71 , 95 % confidence interval 1.38-5.35 , P=0.004 ) compared to those having just one of these two risk factors . CONCLUSIONS : The T allele of the Glu298Asp polymorphism predicts cardiovascular mortality and interacts with plasma DB01686 in determining this outcome in dialysis patients . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . Repair of benzo(a)pyrene diol epoxide- and UV-induced DNA damage in dihydrofolate reductase and adenine phosphoribosyltransferase genes of CHO cells . Using Uvr proteins we have quantified benzo(a)pyrene diol epoxide ( BPDE ) -DNA adduct formation and repair at the dihydrofolate reductase ( P00374 ) and adenine phosphoribosyltransferase ( P07741 ) genes in two Chinese hamster ovary cell lines : B-11 cells , which are 50-fold amplified for P00374 , and P01008 -2 cells , which are diploid for P00374 . We have found that : 1 ) BPDE-DNA adduct formation in different regions of the P00374 gene is proportional to the concentration of BPDE . 2 ) There is no significant difference in the repair of BPDE-DNA adducts between the coding and noncoding regions in either amplified or nonamplified P00374 gene domains . 3 ) Repair in the nonamplified P00374 gene is more efficient ( 30-40 % ) than in the amplified P00374 genes . 4 ) There are no significant differences of repair in the transcribed or nontranscribed strands of the P00374 gene . 5 ) BPDE-DNA adduct formation and repair in the P07741 gene in B-11 and P01008 -2 cells are the same . These results contrast those for the repair of cyclobutane pyrimidine dimers , which occurs preferentially in the transcribed strand of the P00374 gene and in which gene amplification appears to play no role . DB00159 inhibits prostaglandin D2 generation by inhibiting cyclo-oxygenase-2 in cultured human mast cells . BACKGROUND : DB00159 ( EPA ) is catalysed by cyclo-oxygenase ( P36551 ) , as is arachidonic acid , and is a competitive inhibitor of arachidonate metabolism . OBJECTIVES : We examined the effect of EPA on prostaglandin ( PG ) D2 generation in the cultured human mast cells with IgE-anti-IgE challenge incubation . METHODS : Cultured human mast cells were incubated with EPA ( 1 micromol/L ) for 20 h , then challenged with anti-IgE incubation after treatment with IgE . At the same time , P36551 inhibitors were tested to identify P23219 and P35354 activity . PGD2 synthetic activity was also assayed in a cell-free homogenate of cultured mast cells with P36551 inhibitors and EPA . DB11320 in the culture medium and in cells was assayed with the HPLC-fluorescent method . PGD2 and PGD3 were assayed with gas chromatography-mass spectrometry and the stable isotope dilution method . RESULTS : Although EPA incubation did not affect histamine release by cultured human mast cells in response to IgE-anti-IgE challenge incubation , it did decrease PGD2 generation by inhibiting the P35354 pathway . In contrast , in the cell-free homogenate of cultured human mast cells , EPA inhibited both P23219 and P35354 activities . CONCLUSION : Pre-incubation with EPA primarily affects the P35354 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE-anti-IgE challenge incubation . These findings suggest that P23219 and P35354 have different substrate flow systems in mast cells . They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti-inflammatory substrate in human mast cells . Stimulation of human endothelium with P08700 induces selective basophil accumulation in vitro . Basophils have been shown to accumulate in allergic airways and other extravascular sites . Mechanisms responsible for the selective recruitment of basophils from the blood into tissue sites remain poorly characterized . In this study , we characterized human basophil rolling and adhesion on HUVECs under physiological shear flow conditions . Interestingly , treatment of endothelial cells with the basophil-specific cytokine P08700 ( 0.01-10 ng/ml ) promoted basophil and eosinophil , but not neutrophil , rolling and exclusively promoted basophil adhesion . Preincubation of HUVECs with an IL-3R-blocking Ab ( CD123 ) before the addition of P08700 inhibited basophil rolling and adhesion , implicating IL-3R activation on endothelial cells . Incubation of basophils with neuraminidase completely abolished both rolling and adhesion , indicating the involvement of sialylated structures in the process . Abs to the beta(1) integrins , CD49d and CD49e , as well as to P16109 and Q14242 , inhibited basophil rolling and adhesion . Furthermore , blocking chemokine receptors expressed by basophils , such as P41597 , P51677 , and P32248 , demonstrated that P32248 was involved in the observed recruitment of basophils . These data provide novel insights into how P08700 , acting directly on endothelium , can cause basophils to preferentially interact with blood vessels under physiological flow conditions and be selectively recruited to sites of inflammation . DB01109 's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins . DB01109 has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 and P14151 . DB01109 and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N-acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P- or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P- and L-selectins . We conclude that ( a ) heparin 's anti-inflammatory effects are mainly mediated by blocking P- and P14151 -initiated cell adhesion ; ( b ) the sulfate groups at P13671 on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Enhancement of L-cystine transport activity and its relation to Q9UPY5 gene induction at the blood-brain barrier by diethyl maleate treatment . The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier ( BBB ) . The uptake of [(14)C]L-cystine and [(3)H]L-glutamic acid ( L- DB00142 ) was determined using a mouse brain endothelial cell line ( MBEC4 ) as an in vitro BBB model . The mRNA levels of L-cystine/L- DB00142 exchanger , system x(c)(-) , which consists of Q9UPY5 and P08195 , were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis . The [(14)C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na(+)-independent saturable process . The corresponding Michaelis-Menten constant ( K(m) ) was 63.7 microM . In the presence of L- DB00142 , there was competitive inhibition with an inhibition constant ( K(i) ) of 83.5 microM . [(3)H]L- DB00142 uptake in the absence of Na(+) was saturable with a K(m) of 48.1 microM , and it exhibited competitive inhibition with a K(i) of 24.9 microM in the presence of L-cystine . The mutual inhibition between L-cystine and L- DB00142 and the type of inhibition suggest that system x(c)(-) operates in MBEC4 cells . The Q9UPY5 and P08195 mRNAs were expressed in MBEC4 cells and , following diethyl maleate ( DEM ) treatment , the Q9UPY5 mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration ( up to 500 microM ) . Concomitantly , the glutathione concentration in MBEC4 cells was increased . In conclusion , system x(c)(-)-mediated L-cystine uptake takes place in MBEC4 cells . DB00138 transport via system x(c)(-) at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the Q9UPY5 gene . Decreasing Poly(ADP- DB01936 ) Polymerase Activity Restores ΔF508 P13569 Trafficking . Most cystic fibrosis is caused by mutations in P13569 that prevent its trafficking from the ER to the plasma membrane and is associated with exaggerated inflammation , altered metabolism , and diminished responses to oxidative stress . P09874 is activated by oxidative stress and causes energy depletion and cell dysfunction . Inhibition of this enzyme protects against excessive inflammation and recent studies have also implicated it in intracellular protein trafficking . We hypothesized that P09874 activity is altered in CF and affects trafficking and function of the most common CF mutant ΔF508 P13569 . Indeed , P09874 activity was 2.9-fold higher in CF ( ΔF508/ΔF508 ) human bronchial epithelial primary cells than in non-CF cells , and similar results were obtained by comparing CF vs. non-CF bronchial epithelial cell lines ( 2.5-fold higher in CFBE41o(-) vs. 16HBE14o(-) , P < 0.002 ) . A P09874 inhibitor ( ABT-888 , DB07232 ) partially restored P13569 channel activity in CFBE41o(-) cells overexpressing ΔF508 P13569 . Similarly , reducing P09874 activity by 85 % in ileum from transgenic CF mice ( Cftr(tm1)Eur ) partially rescued ΔF508 P13569 activity to 7 % of wild type mouse levels , and similar correction ( 7.8 % ) was observed in vivo by measuring salivary secretion . Inhibiting P09874 with ABT-888 or siRNA partially restored ΔF508 P13569 trafficking in cell lines , and most ΔF508 P13569 was complex glycosylated when heterologously expressed in P09874 (-/-) mouse embryonic fibroblasts . Finally , levels of the mature glycoform of P13569 were reduced by peroxynitrite , a strong activator of P09874 . These results demonstrate that P09874 activity is increased in CF , and identify a novel pathway that could be targeted by proteostatic correctors of P13569 trafficking . Inhibition of a thrombin anion-binding exosite-2 mutant by the glycosaminoglycan-dependent serpins protein C inhibitor and heparin cofactor II . Antithrombin ( P01008 ) , heparin cofactor II ( HCII ) and protein C inhibitor ( P05154 ; also named plasminogen activator inhibitor-3 ) are serine protease inhibitors ( serpins ) whose thrombin inhibition activity is accelerated in the presence of glycosaminoglycans . We compared the inhibition properties of P05154 and HCII to P01008 using R93A/R97A/R101A thrombin , an anion-binding exosite-2 ( exosite-2 ) mutant that has greatly reduced heparin-binding properties . DB01109 -enhanced P05154 inhibition of R93A/R97A/R101A thrombin was only approximately 2-fold compared to 40-fold enhancement with wild-type recombinant thrombin . P07204 ( TM ) ( with or without the chondroitin sulfate moiety ) accelerated P05154 inhibition of both wild-type and R93A/R97A/R101A thrombins . HCII achieved the same maximum activity in the presence of heparin with both wild-type and R93A/R97A/R101A thrombins ; however , the optimum heparin concentration was 20 times greater than the reaction with wild-type thrombin , indicative of a decrease in heparin affinity . Dermatan sulfate ( DSO4 ) -catalyzed HCII thrombin inhibition was unchanged in R93A/R97A/R101A thrombin compared to wild-type recombinant thrombin . These results suggest that P05154 is similar to P01008 and depends upon ternary complex formation with heparin and these specific thrombin exosite-2 residues to accelerate thrombin inhibition . In contrast , HCII does not require DB00125 (93) , DB00125 (97) and DB00125 (101) of thrombin exosite-2 and further supports the hypothesis that HCII uses an allosteric process following glycosaminoglycan binding to inhibit thrombin . Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 [ ( R ) -(-)-2-(4-bromo-2-fluorobenzyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-4-spiro-3'-pyrrolidine-1,2',3,5'-tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K(i) = 10(-)(10) M ) 2000-fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R- and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP(+) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP(+) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP(+) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and/or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4-bromo-2-fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) .

MH_train_1248

MH_train_1248

MH_train_1248

interacts_with DB00514?

multiple_choice

Putative role of HIF transcriptional activity in melanocytes and melanoma biology . Hypoxia-inducible factor-1α ( HIF-1α ) is a highly oxygen sensitive bHLH protein that is part of the heterodimeric Q9BYW2 transcription factor . Under hypoxic stress , Q9BYW2 activity is induced to control expression of multiple downstream target genes , including vascular endothelial growth factor ( P15692 ) . The normal epidermis exists in a constant mild hypoxic microenvironment and constitutively expresses HIF-1α and HIF-2α . Expression of HIF-1α and/or HIF-2α has been suggested to correlate with the increased malignant potential of melanocytes , therefore , failures of melanoma therapies may be partially linked to high HIF activity . Notably , melanomas that have the V600E P15056 mutation exhibit increased HIF-1α expression . We have utilized a bioinformatics approach to identify putative hypoxia response elements ( HREs ) in a set of genes known to participate in the process of melanogenesis ( includingTRPM1 , Q9UMX9 , P01112 , C- P10721 , P40967 and P06850 ) . While some of the mechanistic links between these genes and the HIF pathway have been previously explored , others await further investigation . Although agents targeting HIF activity have been proposed as novel treatment modalities for melanoma , there are currently no clinical trials in progress to test their efficacy in melanoma . Conditional P40337 gene deletion activates a local NO- P15692 axis in a balanced manner reinforcing resistance to endothelium-targeted glomerulonephropathy . BACKGROUND/AIMS : We have reported that tubular epithelial cell injury caused by renal ischemia-reperfusion is attenuated in conditional P40337 knockout ( P40337 -KO ) mice and also that induction of hypoxia-inducible factor ( HIF ) suppresses angiotensin II-accelerated Habu snake venom ( HV ) glomerulonephropathy in rats . However , it remains unknown whether P40337 knockdown protects glomerular endothelial cells from endothelium-targeted glomerulonephritis . METHODS AND RESULTS : P40337 -KO mice with HV glomerulonephropathy ( HV GN ) had fewer injured glomeruli , a lower mesangiolysis score and reduced blood urea nitrogen levels . Immunoreactivity of vascular endothelial growth factor ( P15692 ) in the glomerular capillaries was enhanced by P40337 knockdown and was conserved even in P40337 -KO mice with HV GN , despite HV-attenuating endothelial P15692 expression in vitro . P40337 -KO mice showed enhanced nitric oxide ( NO ) production in glomerular endothelial cells and tubular cells , associated with activated P15692 expression in the kidney ( i.e. an activated NO- P15692 axis ) . The levels of NO in glomeruli and tubules were conserved even in mice with HV GN . In contrast , suppressing NO production in glomerular endothelial cells by an NO synthase inhibitor , N(ϖ)-nitro-L-arginase , completely blunted the protection of P40337 -KO from HV GN . The activated NO- P15692 axis in the kidney of P40337 -KO mice was also associated with an elevation in Flk-1 phosphorylation and increased levels of P22301 and P02778 . CONCLUSION : Conditional P40337 knockdown may enhance the NO- P15692 axis and protect glomerular endothelial cells from HV GN , thereby providing resistance to injury of tubular epithelial cells and glomerular endothelial cells . DB01404 Berry Extract Prevents Atherogenesis via Anti-Inflammatory Action by Upregulating Phase II Gene Expression . DB01404 berry possesses higher ginsenoside content than its root , which has been traditionally used in herbal medicine for many human diseases , including atherosclerosis . We here examined the antiatherogenic effects of the Korean ginseng berry extract ( KGBE ) and investigated its underlying mechanism of action in vitro and in vivo . Administration of KGBE decreased atherosclerotic lesions , which was inversely correlated with the expression levels of phase II genes to include heme oxygenase-1 ( P09601 ) and glutamine-cysteine ligase ( GCL ) . Furthermore , KGBE administration suppressed NF-κB-mediated expression of atherogenic inflammatory genes ( P01375 -α , IL-1β , P35228 , P35354 , P05362 , and P19320 ) , without altering serum cholesterol levels , in ApoE(-/-) mice fed a high fat-diet . Treatment with KGBE increased phase II gene expression and suppressed lipopolysaccharide-induced reactive oxygen species production , NF-κB activation , and inflammatory gene expression in primary macrophages . Importantly , these cellular events were blocked by selective inhibitors of P09601 and GCL . In addition , these inhibitors reversed the suppressive effect of KGBE on P01375 -α-mediated induction of P05362 and P19320 , resulting in decreased interaction between endothelial cells and monocytes . These results suggest that KGBE ameliorates atherosclerosis by inhibiting NF-κB-mediated expression of atherogenic genes via upregulation of phase II enzymes and thus has therapeutic or preventive potential for atherosclerosis . Functional autoradiography of neuropeptide Y Q03519 and P49146 subtypes in rat brain using agonist stimulated [35S]GTPgammaS binding . Neuropeptide Y , one of the most abundant brain peptides , has been found to modulate several important biological functions via a family of G-protein coupled receptors . To investigate the localization of functional P01303 receptor subtypes in the rat brain , we performed agonist-induced [35S]GTPgammaS autoradiography . The Q03519 /Y4/Y5 agonist DB00149 (31) , Pro(34)- P01303 increased [35S]GTPgammaS binding in several brain areas with a regional distribution consistent with that produced when labeling adjacent sections with [ 125I ] - DB00149 (31) , Pro(34)- P10082 . The Q03519 selective antagonist BIBP3226 antagonized the DB00149 (31) , Pro(34)- P01303 stimulated increase in [35S]GTPgammaS binding in all areas examined . The P28062 agonist P06681 - P01303 stimulated [35S]GTPgamma binding in numerous brain areas with a regional distribution similar to the binding observed with [ 125I ] - DB05004 . No increase in [35S]GTPgammaS binding above basal was observed in any brain area evaluated using Y4 and Y5 selective agonists . This study demonstrates abundant Q03519 and P49146 activation in the rat brain , while evidence for functional Y4 and Y5 receptors was not observed . Increased interleukin-6 receptor expression in the paraventricular nucleus of rats with heart failure . Activation of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis and augmented plasma and tissue levels of P05231 are hallmarks of heart failure ( HF ) . Within the forebrain , cardiovascular homeostasis is mediated in part by the paraventricular nucleus ( PVN ) of the hypothalamus . P05231 , via binding to the P05231 receptor ( IL-6R ) /glycoprotein 130 ( P40189 ) complex influences cellular and physiological responses . Thus , in the current study , we hypothesized that PVN IL-6R protein and gene expression are upregulated in HF vs. sham-operated rats , whereas P40189 levels in the same tissues remain stable . Six weeks after coronary ligation surgery , hemodynamic measurements were obtained , and HF rats were divided into moderate noncongestive and severe chronic congestive groups based on cardiac indices . Plasma P05231 levels were determined and changes in gene and protein expression of IL-6R and P40189 between sham-operated and HF rats were determined via real-time PCR and Western blot analyses , respectively . Plasma levels of P05231 were elevated in rats with severe , but not moderate , HF compared with sham-operated controls . In both moderate and severe HF rats , protein but not gene expression of IL-6R was significantly increased in PVN tissue but not in non-PVN tissue , compared with sham-operated controls . Gene and protein levels of the P40189 subunit were not altered by HF in either tissue analyzed . Collectively , these data suggest that within the brain of HF rats , IL-6R expression is not a global change . Rather the increased P05231 levels characteristic of HF may alter PVN-mediated physiological responses via enhanced expression of the IL-6R . Differential regulation of human antigen-specific Th1 and Th2 lymphocyte responses by isozyme selective cyclic nucleotide phosphodiesterase inhibitors . Our study explores the relative efficacy of phosphodiesterase ( PDE ) inhibitors on antigen-specific Th1 and Th2 clonal responses . Proliferative responses for both phenotypes were down-regulated by the DB05876 inhibitor , rolipram , but not the PDE3 inhibitor , siguazodan . The Th2 clones were more sensitive than the Th1 clones to DB05876 inhibition ( P < .05 at 10 and 100 microM rolipram ) . The addition of 1 microM of the adenylyl cyclase activator , isoproterenol , significantly decreased both the EC50 and IC50 of rolipram in both phenotypes ( P < .05 ) . Gene expression for interleukin-4 , interleukin-5 , or interferon-gamma , assessed by reverse transcription-polymerase chain reaction , was down-regulated by the DB05876 inhibitor , but not the PDE3 inhibitor , in each respective clone . Cytokine protein secretion paralleled the results of reverse transcription-polymerase chain reaction for P05112 and interferon-gamma ( P < .01 for each ) . No differential efficacy on cytokine generation parameters between T helper phenotypes was apparent . DB01954 treatment significantly elevated intracellular cyclic AMP ( adenosine 3',5'-cyclic monophosphate ) in clonal T cells ( P < .01 for Th1 or Th2 clones ) ; these elevations were consistently greater in the Th2 clones ( P < .05 ) . Finally , Th1 cells showed reduced gene expression for the Q08493 isoform and a lack of gene expression for the Q08499 isoform by reverse transcription-polymerase chain reaction , compared to the Th2 cells . These data demonstrate the potent immunomodulatory efficacy of DB05876 inhibition on antigen-specific T cell clones . The enhanced sensitivity of Th2 cells to DB05876 inhibition may be due , in part , to the differential expression of DB05876 isoforms between Th1 and Th2 cells . Alterations of respiratory chain complexes in sporadic pheochromocytoma . DB00139 dehydrogenase ( SDH ) has been associated with carcinogenesis in hereditary pheochromocytoma ( PC ) and paraganglioma . We investigated if a similar association applies to sporadic pheochromocytoma . No genetic alteration was found in the P21912 , Q99643 or O14521 genes of sporadic PC . However , in eight of nine sporadic PCs the SDH activity was , on average , reduced by 40 % ; moreover , the activities of the other oxidative phosphorylation ( OXPHOS ) complexes and citrate synthase were significantly lower compared to normal kidney tissue . Furthermore , immunohistochemical staining revealed a significant down-regulation of respiratory chain complexes . Since no pathogenic mutations were detected in the von Hippel-Lindau ( P40337 ) gene , we can rule out that P40337 deficiency is causing the general reduction of OXPHOS enzymes observed in the PCs investigated . In contrast to the single enzyme defects found in a subset hereditary PCs , a more generalized reduction of mitochondrial respiration seems to be present in most sporadic PCs . Strikingly , one of the nine PCs showed specific loss of complex I and a compensatory up-regulation of complexes II-V , which is a phenotype usually characteristic of oncocytic tumors . Pharmacological characterization of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel selective P09917 -activating protein inhibitor that reduces acute and chronic inflammation . Leukotrienes ( LTs ) are proinflammatory lipid mediators synthesized by the conversion of arachidonic acid ( AA ) to P01374 (4) by the enzyme P09917 ( P09917 ) in the presence of P09917 -activating protein ( P20292 ) . 3-[3-tert-Butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) is a novel selective P20292 inhibitor in development for the treatment of respiratory conditions such as asthma . In a rat ex vivo whole-blood calcium ionophore-induced Q06643 (4) assay , DB05225 ( administered orally at 1 mg/kg ) displayed > 50 % inhibition for up to 6 h with a calculated EC(50) of approximately 60 nM . When rat lung was challenged in vivo with calcium ionophore , DB05225 inhibited Q06643 (4) and cysteinyl leukotriene ( CysLT ) production with ED(50) values of 0.8 and 1 mg/kg , respectively . In this model , the EC(50) derived from plasma DB05225 was approximately 330 nM for inhibition of both Q06643 (4) and CysLT . In an acute inflammation setting , DB05225 displayed dose-dependent inhibition of Q06643 (4) , CysLT , and plasma protein extravasation induced by peritoneal zymosan injection . In a model of chronic lung inflammation using ovalbumin-primed and challenged BALB/c mice , DB05225 reduced the concentrations of eosinophil peroxidase , CysLTs , and interleukin-5 in the bronchoalveolar lavage fluid . Finally , DB05225 increased survival time in mice exposed to a lethal intravenous injection of platelet-activating factor . In summary , DB05225 is a novel , potent and selective P20292 inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute and chronic inflammation and in a model of lethal shock . Drug evaluation : DB06684 -- a combined SSRI and P08908 partial agonist for the treatment of depression . DB06684 is a combined selective serotonin reuptake inhibitor ( SSRI ) and a 5-HT(1A) receptor partial agonist that is being developed by Clinical Data Inc for the treatment of depression . In preclinical studies , vilazodone compared favorably to other antidepressants such as paroxetine and fluoxetine . Orally administered vilazodone inhibited ultrasonic vocalization in the rat after electrical foot shock ( a model of anxiolytic activity ) . Yet , in the forced swimming test model of depression in rats , vilazodone administered intraperitoneally was active at 1 mg/kg but not at 3 or 10 mg/kg . During clinical trials , vilazodone completely abolished O75628 sleep for 8 h and demonstrated antidepressant efficacy that was equal to that of current antidepressant therapeutics . The author concludes that the success of vilazodone as an effective antidepressant agent will depend on whether the drug can produce a more rapid antidepressant effect than other SSRI agents , or if specific genetic markers of patients can be associated with clinical efficacy . Lectins and antibodies to blood group antigens as markers for the basal cells of the human respiratory epithelium . We used a pattern of 30 lectins and antibodies against antigens of the P16442 -blood group system to find specific and sensitive markers for the basal cells of the human respiratory surface epithelium . Three lectins always stained the basal cells : Aaptos papillata agglutinin I ( APA I ) , peanut agglutinin ( PNA ) , and wheat germ agglutinin ( WGA ) : Other lectins and the antibodies gave positive results only in tissue of secretors ( blood group antigens in secretions ) and these were dependent on the P16442 -blood group . Griffonia simplicifolia agglutinin ( GSA I B4 ) bound to basal cells of humans with blood group B and AB , Helix pomatia agglutinin ( Q9Y251 ) , Soy bean agglutinin ( SBA ) , and Dolichos biflorus agglutinin ( DBA ) bound to blood group A and AB , Lens tetragonolobus agglutinin ( P01374 ) and Ulex europaeus agglutinin I ( UEA ) bound to secretors in every case , and strongly to blood group O . The antibodies bound to basal cells only in the tissue of secretors , dependent on the P16442 -blood group . The results show that lectins and antibodies may be used as markers for the detection of basal cells in the human respiratory epithelium . Furthermore they suggest that the glycosylation of some glycocomponents of the basal cells is under the control of the genes of the secretor- and P16442 -blood group system . Immunomodulatory activity of red ginseng against influenza A virus infection . DB01404 herbal medicine has been known to have beneficial effects on improving human health . We investigated whether red ginseng extract ( RGE ) has preventive effects on influenza A virus infection in vivo and in vitro . RGE was found to improve survival of human lung epithelial cells upon influenza virus infection . Also , RGE treatment reduced the expression of pro-inflammatory genes ( P05231 , P10145 ) probably in part through interference with the formation of reactive oxygen species by influenza A virus infection . Long-term oral administration of mice with RGE showed multiple immunomodulatory effects such as stimulating antiviral cytokine IFN-γ production after influenza A virus infection . In addition , RGE administration in mice inhibited the infiltration of inflammatory cells into the bronchial lumens . Therefore , RGE might have the potential beneficial effects on preventing influenza A virus infections via its multiple immunomodulatory functions . Multiple antigenic polypeptide composed of heparanase B‑cell epitopes shrinks human hepatocellular carcinoma in mice . The purpose of this study was to evaluate the anti‑growth effect of the self‑designed multiple antigenic polypeptide ( Q96HU1 ) vaccine comprising B‑cell epitopes of heparanase ( Q9Y251 ) on HCC97‑H hepatocellular carcinoma ( HCC ) in mice . The polyclonal antibodies against the B‑cell epitopes of Q9Y251 were prepared by immunizing rabbits with freshly synthesized Q96HU1 vaccine . HCC‑bearing models were constructed on BALB/c nude mice . Anti‑ Q96HU1 antibodies were administrered to the models to assess the effects on Q9Y251 activity , HCC growth , the expression of P15692 / P09038 and the value of micro‑vessel density ( P53602 ) . The anti‑ Q96HU1 antibodies were harvested , purified and identified . These antibodies were able to specifically bind with the dominant epitopes of the precursor protein and large subunit monomer of Q9Y251 , decrease Q9Y251 activity , suppress the expressions of P15692 and P09038 , reduce the P53602 , and markedly shrink the HCC volume . Based on these findings , Q96HU1 vaccine based on the B‑cell epitopes of Q9Y251 seemed to provide theoretical evidence for further study of the synthesized Q9Y251 Q96HU1 vaccine in the treatment of HCC . P15121 inhibitor fidarestat attenuates leukocyte-endothelial interactions in experimental diabetic rat retina in vivo . PURPOSE : Dysregulation of the polyol pathway has been implicated as a major cause of diabetic retinopathy . The aldose reductase inhibitor fidarestat was recently reported to prevent retinal oxidative stress and overexpression of vascular endothelial growth factor ( P15692 ) protein in diabetic rats . In this study , we investigated the effect of fidarestat on leukocyte-endothelial cell interactions in an in vivo experimental model for diabetic retina . MATERIALS AND METHODS : Diabetes was induced in six-week-old male Long-Evans rats by intraperitoneal injection of streptozotocin ( Q11206 ) ( 75 mg/kg ) . The rats were divided into four experimental groups : non-diabetic control rats , untreated diabetic rats , and diabetic rats treated with a low ( 4 mg/kg/day ) or high ( 16 mg/kg/day ) oral dose of fidarestat . After four weeks of treatment , accumulated leukocytes in the retina were counted in vivo by acridine orange digital fluorography . Intercellular adhesion molecule-1 ( P05362 ) and P15692 -164 mRNA levels in the retina were analyzed using the quantitative reverse transcription-polymerase chain reaction . P05362 protein expression in the retina was investigated by immunohistochemistry . RESULTS : DB02021 treatment significantly decreased concentrations of sorbitol and fructose in the retinas of Q11206 -induced diabetic rats . Leukocyte accumulation in the retinas of fidarestat-treated rats was significantly less than in the untreated diabetic group ( P < 0.01 ) . DB02021 treatment significantly reduced the expression P05362 mRNA , but not P15692 -164 mRNA , in the retina of diabetic rats . Immunohistochemical study also revealed the suppressive effect of fidarestat on expression of P05362 . CONCLUSIONS : Oral administration of fidarestat attenuated leukocyte accumulation in the retina of Q11206 induced-diabetic rats , suggesting that fidarestat may have a therapeutic role in preventing the progression of diabetic retinopathy . 5-Lipoxygenase-activating protein ( P20292 ) inhibitors . Part 4 : development of 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid ( AM803 ) , a potent , oral , once daily P20292 inhibitor . The potent P09917 -activating protein ( P20292 ) inhibitor 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid 11cc is described ( AM803 , now GSK2190915 ) . Building upon DB05225 ( 1 ) ( Hutchinson et al. J. Med Chem.2009 , 52 , 5803-5815 ; Stock et al. Bioorg. Med. Chem. Lett. 2010 , 20 , 213-217 ; Stock et al. Bioorg. Med. Chem. Lett.2010 , 20 , 4598-4601 ) , SAR studies centering around the pyridine moiety led to the discovery of compounds that exhibit significantly increased potency in a human whole blood assay measuring Q06643 (4) inhibition with longer drug preincubation times ( 15 min vs 5 h ) . Further studies identified 11cc with a potency of 2.9 nM in P20292 binding , an IC(50) of 76 nM for inhibition of Q06643 (4) in human blood ( 5 h incubation ) and excellent preclinical toxicology and pharmacokinetics in rat and dog . 11cc also demonstrated an extended pharmacodynamic effect in a rodent bronchoalveolar lavage ( BAL ) model . This compound has successfully completed phase 1 clinical studies in healthy volunteers and is currently undergoing phase 2 trials in asthmatic patients . Key residues at the riboflavin kinase catalytic site of the bifunctional riboflavin kinase/ Q8NFF5 from Corynebacterium ammoniagenes . Many known prokaryotic organisms depend on a single bifunctional enzyme , encoded by the RibC of RibF gene and named DB03147 synthetase ( FADS ) , to convert DB00140 ( RF ) , first into Q68DA7 and then into DB03147 . The reaction occurs through the sequential action of two activities present on a single polypeptide chain where the N-terminus is responsible for the DB00171 : Q8NFF5 ( FMNAT ) activity and the C-terminus for the DB00171 : riboflavin kinase ( Q969G6 ) activity . Sequence and structural analysis suggest that T208 , N210 and E268 at the C-terminus Q969G6 module of Corynebacterium ammoniagenes FADS ( CaFADS ) might be key during RF phosphorylation . The effect of site-directed mutagenesis on the Q969G6 activity , as well as on substrates and products binding , indicates that T208 and N210 provide the Q969G6 active-site geometry for binding and catalysis , while E268 might be involved in the catalytic step as catalytic base . These data additionally suggest concerted conformational changes at the Q969G6 module of CaFADS during its activity . Mutations at the Q969G6 site also modulate the binding parameters at the FMNAT active site of CaFADS , altering the catalytic efficiency in the transformation of Q68DA7 into DB03147 . This observation supports the hypothesis that the hexameric assembly previously revealed by the crystal structure of CaFADS might play a functional role during catalysis . [ Serotonin transporter gene and stress reactivity in unipolar depression. Role of the Q9Y251 system as endophenotype of the P31645 gene ] . BACKGROUND : A length polymorphism in the promoter region of the serotonin transporter gene ( 5-HTTLPR ) is associated with both depression and hypothalamic-pituitary-adrenal ( Q9Y251 ) system activity . A dysregulation of the Q9Y251 system is considered to be a candidate endophenotype of depression . The objective of the present study was an investigation of a possible gene-endophenotype-interaction between 5-HTTLPR and Q9Y251 system activity in a sample of inpatients with major depression . MATERIALS AND METHODS : A total of 237 inpatients with major depression were genotyped for 5-HTTLPR and participated in a combined dexamethasone-corticotropin-releasing hormone test ( DB00514 - P06850 test ) as well as using the Hamilton score ( Hamilton rating scale for depression ) to determine the severity of the psychopathology . RESULTS : Patients with the ss-genotype showed a significantly higher Q9Y251 -system activity in comparison to patients with the lI-genotype , but no association between 5-HTTLPR and the severity of psychopathology could be detected . CONCLUSIONS : The results of the current study demonstrate an influence of 5-HTTLPR on dysregulation of the Q9Y251 system in patients with major depression and support the hypothesis that 5-HTTLPR- and Q9Y251 -system-interaction constitutes an important component in the pathogenesis of depression . Effects of the inhibition of cyclo-oxygenase 1 or 2 or P09917 on the activation of the hypothalamic-pituitary-adrenal axis induced by interleukin-1beta in the male Rat . The limited entry of interleukin-1beta ( IL-1beta ) into the central nervous system has led to the hypothesis that IL-1beta acts , through IL-1beta receptors located notably on endothelial cells , on the release of prostaglandins which in turn stimulate the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . We used cyclo-oxygenase-1 ( P23219 ) and cyclo-oxygenase-2 ( P35354 ) and P09917 ( 5- P28300 ) inhibitors , before the injection of IL-1beta , to explore the role of arachidonic acid metabolic pathways on Q9Y251 axis activation . Adult male rats were i.m injected 20 min before i.p injection of IL-1beta , with ( i ) : a P23219 / P35354 inhibitor ( ketoprofen ) ; ( ii ) a P35354 selective inhibitor ( NS 398 ) ; or ( iii ) a 5- P28300 inhibitor ( BW A4C ) . Following this , rats were killed 90 min after i.p . IL-1beta injection and analysis for plasma adrenocorticotropic hormone ( DB01285 ) and corticosterone concentrations and determination of anterior pituitary pro-opio melanocortin ( P01189 ) gene transcription was conducted . Administration of the P23219 / P35354 inhibitor led to a complete blockage of DB01285 and corticosterone secretion and P01189 gene transcription . The P35354 inhibitor led to a complete blockade of DB01285 secretion and P01189 gene transcription but had no effect on corticosterone secretion . The 5- P28300 inhibitor had no significant effect on any parameter . These results demonstrate the crucial role of eicosanoid pathways in mediating the stimulation of the Q9Y251 axis induced by IL-1beta . Moreover , we found a clear dissociation of the effect of the blockage of COXs upon DB01285 and corticosterone secretion , suggesting that IL-1beta may act at the brain as well as at the adrenal cortex to stimulate the secretion of corticosterone . Biological and enzymatic features of human melanoma clones with different invasive potential . Cell clones derived from a human melanoma metastasis selected for different integrin profiles were examined in vitro for invasive potential and biological and biochemical features potentially related to this process . Clones which expressed high levels of integrins showed high invasive potential , extracellular matrix degradation , and adhesion to gelatin-coated substrates . A correlation was also found between invasiveness and intracellular and extracellular plasminogen activator activity . Q9Y251 and collagenase type IV activities were apparently unrelated to invasiveness . gamma-Glutamyl transferase ( P19440 ) activity was high in highly invasive clones , whereas melanin content was high in slightly invasive clones . Heterogeneity was also observed in cellular parameters such as cell dimensions , growth features and DNA index . The intrinsic biological and biochemical heterogeneity of a cell population derived from a single metastasis may be responsible for the different behaviour of clones , regardless of their invasive potential . Since slightly invasive cells are more differentiated than highly invasive cells , malignancy and differentiation are inversely correlated in such human melanoma clones . SDH mutations establish a hypermethylator phenotype in paraganglioma . Paragangliomas are neuroendocrine tumors frequently associated with mutations in P07949 , P21359 , P40337 , and succinate dehydrogenase ( SDHx ) genes . Methylome analysis of a large paraganglioma cohort identified three stable clusters , associated with distinct clinical features and mutational status . SDHx-related tumors displayed a hypermethylator phenotype , associated with downregulation of key genes involved in neuroendocrine differentiation . DB00139 accumulation in SDH-deficient mouse chromaffin cells led to DNA hypermethylation by inhibition of 2-OG-dependent histone and DNA demethylases and established a migratory phenotype reversed by decitabine treatment . Epigenetic silencing was particularly severe in P21912 -mutated tumors , potentially explaining their malignancy . Finally , inactivating FH mutations were identified in the only hypermethylated tumor without SDHx mutations . These findings emphasize the interplay between the Krebs cycle , epigenomic changes , and cancer . A Novel Medical Treatment of Cushing 's Due to Ectopic DB01285 in a Patient With Neurofibromatosis Type 1 . A 64-year-old male presented with neurofibromatosis 1 and Cushing 's syndrome . Clinically he was over weight , depressed with extensive skin bruising and hypertension . DB00117 24 hours urinary metanephrines , urinary 5HIAA , gut peptides and chromgranin levels were normal . DB00117 renal function and renal Q9BWK5 scan was also normal . DB00117 cortisol failed to suppress on overnight dexamethsone suppression test . DB00117 low dose dexamethasone suppression with P06850 stimulation showed failure of suppression of cortisol to < 50 nmol/L and DB01285 was measurable at 10 ng/L on day 3 . There was no response of DB01285 or cortisol to P06850 stimulation . DB00117 DB01285 precursors were high at 126 pmol/L consistent with defective pro-opiomelanocortin ( P01189 ) processing suggesting an ectopic source of DB01285 production . The Q9BWK5 scan of his pituitary and CT scan of the adrenal glands was normal . DB00117 octreotide scan was negative . The source of his ectopic DB01285 was most likely a large retroperitoneal plexiform neurofibroma seen on CT abdomen that had undergone malignant peripheral nerve sheath tumour transformation on histology . He was a poor surgical risk for tumour debulking procedure . In view of the available literature and role of c-kit signalling in neurofibromatosis , he was treated with Imitinib . Four months after the treatment his Cushings had resolved on biochemical testing . After a year his plexiform neurofibroma has not increased in size . To our knowledge , this is the first case of P21359 associated with clinical and biochemical features of Cushing 's secondary to ectopic DB01285 due to MPNST in a plexiform neurofibroma and its resolution on treatment with imatinib . [ Proteolytic processing by dipeptidyl aminopeptidase IV generates receptor selectivity for peptide YY ( P10082 ) ] . Two receptor subtypes , Q03519 and P28062 , are known to mediate P10082 biological activity . P10082 1-36 binds to Q03519 and P28062 receptors with equal affinity , whereas the second endogenous form of P10082 , DB05004 , selectively binds to P28062 receptors . Dipeptidyl cleavage thus transforms an unselective Y agonist into a highly selective P28062 agonist , DB05004 . The enzyme responsible for this processing is unknown . Since P10082 has a proline in the penultimate position it is protected from the attack of most unspecific exopeptidases . Only a few exopeptidases are theoretically capable of generating DB05004 from P10082 1-36 . Of the enzymes tested only the dipeptidyl aminopeptidase IV ( P27487 , E.C. 3.4.14.5 ) cleaved DB00135 -Pro from P10082 1-36 with high activity . Since P27487 is found on the endothelial surface and brush border membranes it can be considered a candidate enzyme for generating DB05004 in vivo , thereby regulating the ratio of Q03519 / P49146 stimulation by P10082 . Leptin signaling in the hypothalamus : emphasis on energy homeostasis and leptin resistance . Leptin , the long-sought satiety factor of adipocytes origin , has emerged as one of the major signals that relay the status of fat stores to the hypothalamus and plays a significant role in energy homeostasis . Understanding the mechanisms of leptin signaling in the hypothalamus during normal and pathological conditions , such as obesity , has been the subject of intensive research during the last decade . It is now established that leptin action in the hypothalamus in regulation of food intake and body weight is mediated by a neural circuitry comprising of orexigenic and anorectic signals , including P01303 , P20382 , galanin , orexin , Q9UBC7 , alpha-MSH , NT , and P06850 . In addition to the conventional O60674 - P40763 pathway , it has become evident that PI3K- Q13370 - DB02527 pathway plays a critical role in leptin signaling in the hypothalamus . It is now established that central leptin resistance contributes to the development of diet-induced obesity and ageing associated obesity . Central leptin resistance also occurs due to hyperleptinimia produced by exogenous leptin infusion . A defective nutritional regulation of leptin receptor gene expression and reduced P40763 signaling may be involved in the development of leptin resistance in DIO . However , leptin resistance in the hypothalamic neurons may occur despite an intact O60674 - P40763 pathway of leptin signaling . Thus , in addition to defective O60674 - P40763 pathway , defects in other leptin signaling pathways may be involved in leptin resistance . We hypothesize that defective regulation of PI3K- Q13370 - DB02527 pathway may be one of the mechanisms behind the development of central leptin resistance seen in obesity . The immunological effects of electrolyzed reduced water on the Echinostoma hortense infection in C57BL/6 mice . Electrolyzed reduced water ( ERW ) is widely used for drinking by people in Asia . The purpose of this study was to examine the immunological effect of ERW on the immunity of animals by supplying ERW to C57BL/6 mice infected with Echinostoma hortense metacercariae . In the non-infected groups , interleukin ( IL ) -4 ( p < 0.001 ) , P05113 , P22301 , IL-1beta , tumor necrosis factor ( P01375 ) -alpha and immunoglobulin ( Ig ) A expression of the group fed ERW ( ERW group ) increased in small intestine compared with the normal control group . In the case of infected groups , the group fed ERW ( ERW+E. hortense group ) showed the result that P05112 , P05113 , P22301 and Ig A expression increased , but IL-1beta and P01375 ( p < 0.001 ) decreased , and the number of goblet cells ( p < 0.001 ) and helix pomatia agglutinin ( Q9Y251 ) positive cells increased compared with the group without feeding ERW . However , adult worm recovery rate was markedly increased ( p < 0.05 ) . On the other hand , the expression of all the cytokines except P22301 in spleen was mildly increased but not significant statistically , and there was no significant difference in the numerical changes of white blood cell ( WBC ) . These results indicate that feeding ERW may have influence on the local immune response ( Th-1 type cytokines such as IL-1beta , P01375 ) in the small intestine but not on the systemic immune response . Pharmacodynamics and pharmacokinetics of DB05225 , a novel inhibitor of P09917 -activating protein ( P20292 ) . The P09917 -activating protein ( P20292 ) gene and an increase in leukotriene ( LT ) production are linked to the risk of asthma , myocardial infarction , and stroke . We evaluated the pharmacodynamics , pharmacokinetics , and tolerability of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel P20292 inhibitor , in healthy subjects . Single and multiple doses of DB05225 demonstrated dose-dependent inhibition of blood Q06643 (4) production and dose-related inhibition of urinary LTE(4) . After a single oral dose ( 50-1,000 mg ) of DB05225 , the maximum concentration ( C(max) ) and area under the curve ( AUC ) in plasma increased in a dose-dependent manner . After multiple-dose administration ( 50-1,000 mg once daily for 11 days ) , there were no significant differences in the pharmacokinetic parameters between the first and last days of treatment . DB05225 was well tolerated at all doses in both the single- and multiple-dose cohorts . Further clinical trials with DB05225 in inflammatory diseases are warranted . P08908 receptor responsivity in unipolar depression . Evaluation of ipsapirone-induced DB01285 and cortisol secretion in patients and controls . The selective P08908 receptor ligand ipsapirone ( IPS ) induces corticotropin ( DB01285 ) and cortisol secretion in humans . To explore P08908 receptor-mediated hypothalamic-pituitary-adrenal ( Q9Y251 ) system activation in depression , 24 subjects ( 12 patients with unipolar depression and 12 individually matched controls ) were given 0.3 mg/kg IPS or placebo in random order . Compared with controls , the depressed patients exhibited significantly decreased DB01285 and cortisol responses to IPS in association with increased basal cortisol secretion . The impaired Q9Y251 response following P08908 receptor challenge in unipolar depression could have resulted from glucocorticoid-dependent subsensitivity of the ( post-synaptic ) P08908 receptor itself and/or from a defective postreceptor signaling pathway [ inhibitory guanine nucleotide-binding protein ( Gi ) -adenylate cyclase complex function ] , thus supporting the hypothesis that a disintegrated 5-HT and Q9Y251 system interaction may be present in depression . Future studies of the Q9Y251 response to direct-acting P08908 ligands , such as IPS , should facilitate the assessment of 5-HT/ Q9Y251 system integrity in various affective disorders and its involvement in psychotropic drug effects . Recurrent epimutation of Q99643 in gastrointestinal stromal tumors . DB00139 dehydrogenase ( SDH ) is a conserved effector of cellular metabolism and energy production , and loss of SDH function is a driver mechanism in several cancers . SDH-deficient gastrointestinal stromal tumors ( dSDH GISTs ) collectively manifest similar phenotypes , including hypermethylated epigenomic signatures , tendency to occur in pediatric patients , and lack of P10721 / P16234 mutations . dSDH GISTs often harbor deleterious mutations in SDH subunit genes ( P31040 , P21912 , Q99643 , and O14521 , termed SDHx ) , but some are SDHx wild type ( WT ) . To further elucidate mechanisms of SDH deactivation in SDHx-WT GIST , we performed targeted exome sequencing on 59 dSDH GISTs to identify 43 SDHx-mutant and 16 SDHx-WT cases . Genome-wide DNA methylation and expression profiling exposed Q99643 promoter-specific CpG island hypermethylation and gene silencing in SDHx-WT dSDH GISTs [ 15 of 16 cases ( 94 % ) ] . Six of 15 Q99643 -epimutant GISTs occurred in the setting of the multitumor syndrome Carney triad . We observed neither P21912 promoter hypermethylation nor large deletions on chromosome 1q in any SDHx-WT cases . Deep genome sequencing of a 130-kbp ( kilo-base pair ) window around Q99643 revealed no recognizable sequence anomalies in Q99643 -epimutant tumors . More than 2000 benign and tumor reference tissues , including stem cells and malignancies with a hypermethylator epigenotype , exhibit solely a non-epimutant Q99643 promoter . Mosaic constitutional Q99643 promoter hypermethylation in blood and saliva from patients with Q99643 -epimutant GIST implicates a postzygotic mechanism in the establishment and maintenance of Q99643 epimutation . The discovery of Q99643 epimutation provides a unifying explanation for the pathogenesis of dSDH GIST , whereby loss of SDH function stems from either SDHx mutation or Q99643 epimutation . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . P15121 inhibition counteracts oxidative-nitrosative stress and poly(ADP-ribose) polymerase activation in tissue sites for diabetes complications . This study evaluated the effects of aldose reductase inhibition on diabetes-induced oxidative-nitrosative stress and poly(ADP-ribose) polymerase ( PARP ) activation . In animal experiments , control and streptozotocin-induced diabetic rats were treated with or without the aldose reductase inhibitor ( Q9Y4X5 ) fidarestat ( 16 mg . kg(-1) . day(-1) ) for 6 weeks starting from induction of diabetes . DB09391 pathway intermediate , but not glucose , accumulation in sciatic nerve and retina was completely prevented in diabetic rats treated with fidarestat . Sciatic motor nerve conduction velocity , hindlimb digital sensory nerve conduction velocity , and sciatic nerve concentrations of two major nonenzymatic antioxidants , glutathione and ascorbate , were reduced in diabetic versus control rats , and these changes were prevented in diabetic rats treated with fidarestat . DB02021 prevented the diabetes-induced increase in nitrotyrosine ( a marker of peroxynitrite-induced injury ) and poly(ADP-ribose) immunoreactivities in sciatic nerve and retina . DB02021 counteracted increased superoxide formation in aorta and epineurial vessels and in in vitro studies using hyperglycemia-exposed endothelial cells , and the DCF test/flow cytometry confirmed the endothelial origin of this phenomenon . DB02021 did not cause direct inhibition of PARP activity in a cell-free system containing PARP and NAD(+) but did counteract high-glucose-induced PARP activation in Schwann cells . In conclusion , aldose reductase inhibition counteracts diabetes-induced nitrosative stress and PARP activation in sciatic nerve and retina . These findings reveal the new beneficial properties of fidarestat , thus further justifying the ongoing clinical trials of this specific , potent , and low-toxic Q9Y4X5 . [ Effect of Jiawei Sini Decoction on Q9Y251 function of rats with restraint psychological stress ] . OBJECTIVE : To observe the effect of Jiawei Sini Decoction ( JWSND ) on Q9Y251 function of rats with restraint psychological stress and the pathway of thymus immunosuppressive function . METHODS : The rats were randomly divided into 4 groups , control group ( C ) , model group ( M ) , group treated by JWSND ( C1 ) and ginsenosides ( P06681 ) . Hypothalamus P06850 , blood plasma DB01285 and O00230 , the numberofthy moeyte GCR sites and the GCR nuclear translocation rate were detected by radioimmunoassay . RESULTS : Compared with C , in M , Hypothalamus P06850 , blood plasma DB01285 and O00230 increased significantly ( P < 0.01 ) , but all these of C1 and P06681 lowered significantly ( P < 0.05 , P < 0.01 ) . Compared with C , the GCR nuclear translocation rate was increased significantly ( P < 0. 01 ) . Compared with M , the GCR nuclear translocation rate lowered significantly ( P < 0.01 ) . Moreover , no significant difference was found in all indexes between C1 and C . CONCLUSION : JWSND participates effectively in the adjustment of Q9Y251 of the rats with restraint psychological stress , and relieves significantly the inhibitory effect of glucocorticoid on the thymus , the pathway closely relates to the translocation of GCR . Death receptors 4 and 5 activate Nox1 NADPH oxidase through riboflavin kinase to induce reactive oxygen species-mediated apoptotic cell death . Stimulation of the proapoptotic tumor necrosis factor ( P01375 ) -related apoptosis-inducing ligand ( P50591 ) receptors , death receptors 4 ( DR4 ) and 5 ( DR5 ) , conventionally induces caspase-dependent apoptosis in tumor cells . Here we report that stimulation of DR4 and/or DR5 by the agonistic protein KD548-Fc , an Fc-fused DR4/DR5 dual-specific Kringle domain variant , activates plasma membrane-associated Nox1 NADPH oxidase to generate superoxide anion and subsequently accumulates intracellular reactive oxygen species ( ROS ) , leading to sustained c-Jun N-terminal kinase activation and eventual apoptotic cell death in human HeLa and Jurkat tumor cells . KD548-Fc treatment induces the formation of a DR4/DR5 signaling complex containing riboflavin kinase ( Q969G6 ) , Nox1 , the Nox1 subunits ( Rac1 , Noxo1 , and Noxa1 ) , P01375 receptor-associated death domain ( Q15628 ) , and Q12933 ( TRAF2 ) . Depletion of Q969G6 , but not the Nox1 subunits , Q15628 and TRAF2 , failed to recruit Nox1 and Rac1 to DR4 and DR5 , demonstrating that Q969G6 plays an essential role in linking DR4/DR5 with Nox1 . Knockdown studies also reveal that Q969G6 , Q15628 , and TRAF2 play critical , intermediate , and negligible roles , respectively , in the KD548-Fc-mediated ROS accumulation and downstream signaling . Binding assays using recombinantly expressed proteins suggest that DR4/DR5 directly interact with cytosolic Q969G6 through Q969G6 -binding regions within the intracellular death domains , and Q15628 stabilizes the DR4/DR5- Q969G6 complex . Our results suggest that DR4 and DR5 have a capability to activate Nox1 by recruiting Q969G6 , resulting in ROS-mediated apoptotic cell death in tumor cells . Phosphodiesterase 4B gene transcription is activated by lipopolysaccharide and inhibited by interleukin-10 in human monocytes . There are four different genes encoding the DB02527 -specific phosphodiesterase ( DB05876 ) isozymes ( A , B , C , and D ) . DB02527 has been the only agent known to induce DB05876 gene expression . In the present study , we demonstrate , for the first time , that lipopolysaccharide ( LPS ) significantly and selectively stimulated Q07343 mRNA production in human monocytes . The LPS stimulation occurred very rapidly ( in 30-45 min ) and in a dose-dependent manner ( 0.01-100 ng/ml ) . We also demonstrate that LPS induction of Q07343 mRNA expression was inhibited strongly by interleukin ( IL ) -10 . The inhibition with P22301 was dose-dependent ( 0.1-10 ng/ml ) . P05112 also inhibited the LPS induction , but to a lesser extent than P22301 . Q07343 mRNA expression was also stimulated by dibutyryl- DB02527 . Interestingly , unlike LPS induction , the dibutyryl- DB02527 induction of Q07343 mRNA expression was not inhibited by P22301 . By performing nuclear run-on and mRNA stability assays , we demonstrate further that P22301 inhibited LPS-stimulated Q07343 mRNA synthesis by abolishing the gene transcription rather than by enhancing mRNA degradation . The present study suggests that Q07343 , as the only LPS-inducible DB05876 subtype , may be an appropriate target for discovering anti-inflammatory drugs . DB00160 aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis . Plant peroxisomal glyoxylate aminotransferases play central roles within the photorespiratory pathway . Genes encoding glyoxylate aminotransferases have been isolated from several animals and microbes , but only recently have plant homologs been identified . Three Arabidopsis homologs of alanine (Ala):glyoxylate aminotransferase 2 ( Q9BYV1 ) contain a putative type 1 peroxisomal targeting signal ( PTS1 ) , but the metabolic significance of these Q9BYV1 homologs is unknown . P19440 and P36268 are Ala aminotransferase ( AlaAT ) homologs from Arabidopsis that represent another type of glyoxylate aminotransferase . These proteins are class I aminotransferases , each containing a putative PTS1 . P19440 and P36268 are members of a small family of AlaATs in Arabidopsis . When expressed as recombinant proteins in Escherichia coli , P19440 and P36268 displayed biochemical characteristics very similar to one another , and to the Arabidopsis protein purified from leaves . Four aminotransferase activities were specifically associated with P19440 and P36268 , using the substrate pairs glutamate ( DB00142 ):glyoxylate , Ala:glyoxylate , DB00142 :pyruvate , and Ala:2-oxoglutarate . P19440 and P36268 may have partially redundant functions ; transcripts of both genes were detected in many of the same tissues . Although DB00142 :glyoxylate aminotransferase ( P19440 ) activity has been observed in several locations in different plants and algae , including the cytoplasm and mitochondria , our subcellular fractionation data indicate that P19440 activity was exclusively peroxisomal in Arabidopsis . Thus , glyoxylate aminotransferase reactions in plant peroxisomes appear to be catalyzed by at least two distinct types of aminotransferases : an AGT1 homolog with serine:glyoxylate aminotransferase activity ( A.H. Liepman , L.J. Olsen [ 2001 ] Plant J 25 : 487-498 ) , and a pair of closely related , potentially redundant AlaAT homologs with P19440 activity . The role of the polyol pathway in acute kidney injury caused by hindlimb ischaemia in mice . The polyol pathway , a collateral glycolytic process , previously considered to be active in high glucose milieu , has recently been proposed to play a crucial role in ischaemia/reperfusion tissue injury . In this study , we explored the role of the polyol pathway in acute kidney injury ( AKI ) , a life-threatening condition , caused by hindlimb ischaemia , and determined if inhibition of the polyol pathway by aldose reductase ( AR ) inhibitor is beneficial for this serious disorder . Mice 8 weeks of age rendered hindlimb ischaemic for 3 h by the clipping of major supporting arteries revealed marked muscle necrosis with accumulation of sorbitol and fructose in ischaemic muscles . Serum concentrations of blood urea nitrogen ( BUN ) , creatinine phosphokinase ( CPK ) , creatinine , tumour necrosis factor ( P01375 ) -alpha as well as interleukin ( IL ) -6 were all elevated in these mice . Treatment with AR inhibitor ( Q9Y4X5 ) effectively suppressed muscle necrosis and accompanying inflammatory reactions and prevented renal failure . Similar to Q9Y4X5 -treated mice , AR-deficient mice were protected from severe ischaemic limb injury and renal failure , showing only modest muscle necrosis and significant suppression of serum markers of renal failure and inflammation . Thus , these findings suggest that the polyol pathway is implicated in AKI caused by ischaemic limb injury and that AR may be a potential therapeutic target for this condition . Neuroendocrine pathways in benzodiazepine dependence : new targets for research and therapy . Benzodiazepines are known to modulate the activity of the hypothalamo-pituitary-adrenocortical ( Q9Y251 ) axis by antagonizing the effects of DB05394 ( P06850 ) . Besides regulating the Q9Y251 axis P06850 evolves properties of a neurotransmitter in the limbic system that is closely involved in the delivery of the emotional consequences of the stress response . At a superordinated level Neuropeptide Y ( P01303 ) and Cholecystokinin ( CCK ) affect the release of P06850 and modulate thereby the intensity of the physiological stress response . Benzodiazepine treatment interferes not only with the release of P06850 but also with the release of P01303 and CCK . Alterations in the intracortical ratio of P01303 , CCK and P06850 are correlated with behavioural changes like increased respectively decreased anxiety and subsequent alterations in the activity of the Q9Y251 axis . Recent research offers the possibility that the alterations of plasma levels of these neuropeptides are not only a secondary phenomenon due to drug intake , but that low levels of those neuropeptides that modulate anxiety and fear can possibly explain addiction to substances that counterbalance these deficits . Depending on the available results possible implications of P01303 and CCK on benzodiazepine addiction and withdrawal symptoms are reviewed , thereby providing topics for further research . Inhibition of Q13370 augments DB05876 inhibitor-induced apoptosis in a subset of patients with chronic lymphocytic leukemia . PURPOSE : DB02527 phosphodiesterase ( PDE ) 4 is a family of enzymes the inhibition of which induces chronic lymphocytic leukemia ( CLL ) apoptosis . However , leukemic cells from a subset of CLL patients are relatively resistant to treatment with the DB05876 inhibitor rolipram , particularly when this drug is used in the absence of an adenylate cyclase stimulus such as forskolin . Elevated DB02527 levels induce compensatory up-regulation of several cyclic nucleotide PDE families in other model systems . We here examine the hypothesis that CLL cells that survive treatment with rolipram do so as a result of residual PDE activity that is not inhibited by this drug . EXPERIMENTAL DESIGN : We examined by Western analysis the effect of rolipram treatment on CLL expression of Q13370 , P27815 , Q07343 , Q08499 , and Q13946 . We also examined the ability of rolipram ( DB05876 inhibitor ) or cilostamide ( PDE3 inhibitor ) , alone or together , to induce apoptosis or elevate cyclic AMP in leukemic cells from patients with CLL . RESULTS : DB01954 increased levels of Q07343 and , to a variable extent , Q08499 . When combined with forskolin , rolipram also increased levels of a second family of PDEs , Q13370 . Addition of the specific PDE3 inhibitor , cilostamide , modestly augmented rolipram-induced apoptosis in five of seven " rolipram-resistant " CLL samples . CONCLUSIONS : Although this work confirms that DB05876 appears to be the most important PDE target for induction of apoptosis in CLL , combination therapy with PDE3 and DB05876 inhibitors or use of dual-selective drugs may be of benefit in a subset of relatively DB05876 -inhibitor resistant CLL patients . Consequences of missense mutations for dimerization and turnover of alanine:glyoxylate aminotransferase : study of a spectrum of mutations . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme , deficiency of which results in primary hyperoxaluria type 1 ( P78364 ) . More than 65 P78364 -related mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have generated a spectrum of 15 missense changes including the most common P78364 mutation , G170R , and expressed them on the appropriate background of the major or minor allele , in an Escherichia coli overexpression system and in a rabbit reticulocyte transcription/translation system . We have investigated their effects on enzyme activity , dimerization , aggregation , and turnover . The effect of pyridoxal phosphate ( PLP ) on dimerization and stability was also investigated . Although all 15 mutant AGTs were expressed as intact proteins in E. coli , only three : G41R and G41V on the major allele , and the common mutation G170R , resulted in significant amounts of enzymatic activity . Dimerization failure was a frequent observation ( 13/15 ) except for G41V and D183N . Dimerization was poor with S187F but was substantially improved with PLP . Proteasome-mediated protein degradation was observed for all the mutations except G41R on the major allele , G41V , D183N , G170R , and S218L . Increases in the stability of the mutant enzymes in the presence of PLP were small ; however , G41R on the minor allele showed a direct relationship between its half life and the concentration of PLP . The minor allele AGT product and many of the mutants were subject to a limited non-proteasomal proteolytic cleavage when DB00171 was depleted . DB06684 : a P08908 receptor agonist/serotonin transporter inhibitor for the treatment of affective disorders . DB06684 ( P50402 68843 ; 5-{4-[4-(5-cyano-3-indolyl)-butyl]-1-piperazinyl}-benzofuran-2-carboxamide hydrochloride ) is a combined serotonin specific reuptake inhibitor ( SSRI ) and P08908 receptor partial agonist currently under clinical evaluation for the treatment of major depression . This molecule was designed based on the premise that negative feedback circuitry , mediated via 5-HT1 receptors , limits the acute SSRI-induced enhancements in serotonergic neurotransmission . If the hypothesis is correct , combination of SSRI with P08908 partial agonism should temporally enhance the neuroplastic adaptation and subsequently hasten therapeutic efficacy compared to current treatments . Preclinical in vitro evaluation has confirmed vilazodone 's primary pharmacological profile both in clonal and native systems , that is , serotonin reuptake blockade and P08908 partial agonism . However , in vivo and in contrast to combination of 8-OH-DPAT and paroxetine , vilazodone selectively enhanced serotonergic output in the prefrontal cortex of rats . Behavioral evaluations , in the ultrasonic vocalization model of anxiety in rats , demonstrated anxiolytic efficacy . In the forced swim test ( a putative model of depression ) , vilazodone also showed efficacy but at a single dose only . In man , vilazodone abolished O75628 sleep and demonstrated clinical antidepressant efficacy equivalent to an SSRI . Ongoing clinical evaluations will hopefully reveal whether the founding hypothesis was valid and if vilazodone will produce a more rapid onset of antidepressant efficacy . Involvement of neuropeptide Y Q03519 receptors in the regulation of neuroendocrine corticotropin-releasing hormone neuronal activity . The neuroendocrine parvocellular P06850 neurons in the paraventricular nucleus ( PVN ) of the hypothalamus are the main integrators of neural inputs that initiate hypothalamic-pituitary-adrenal ( Q9Y251 ) axis activation . Neuropeptide Y ( P01303 ) expression is prominent within the PVN , and previous reports indicated that P01303 stimulates P06850 mRNA levels . The purpose of these studies was to examine the participation of P01303 receptors in Q9Y251 axis activation and determine whether neuroendocrine P06850 neurons express P01303 receptor immunoreactivity . Infusion of 0.5 nmol P01303 into the third ventricle increased plasma corticosterone levels in conscious rats , with the peak of hormone levels occurring 30 min after injection . This increase was prevented by pretreatment with the Q03519 receptor antagonist BIBP3226 . Immunohistochemistry showed that P06850 -immunoreactive neurons coexpressed Q03519 receptor immunoreactivity ( Y1r-ir ) in the PVN , and a majority of these neurons ( 88.8 % ) were neuroendocrine as determined by ip injections of FluoroGold . Bilateral infusion of the Q03519 /Y5 agonist , [leu(31)pro(34)] P01303 ( 110 pmol ) , into the PVN increased c-Fos and phosphorylated DB02527 response element-binding protein expression and elevated plasma corticosterone levels . Increased expression of c-Fos and phosphorylated DB02527 response element-binding protein was observed in populations of P06850 /Y1r-ir cells . The current findings present a comprehensive study of P01303 Q03519 receptor distribution and activation with respect to P06850 neurons in the PVN . The expression of P01303 Y1r-ir by neuroendocrine P06850 cells suggests that alterations in P01303 release and subsequent activation of P01303 Q03519 receptors plays an important role in the regulation of the Q9Y251 . Role of key residues at the flavin mononucleotide ( Q68DA7 ):adenylyltransferase catalytic site of the bifunctional riboflavin kinase/flavin adenine dinucleotide ( DB03147 ) Synthetase from Corynebacterium ammoniagenes . In mammals and in yeast the conversion of DB00140 ( RF ) into flavin mononucleotide ( Q68DA7 ) and flavin adenine dinucleotide ( DB03147 ) is catalysed by the sequential action of two enzymes : an DB00171 :riboflavin kinase ( Q969G6 ) and an DB00171 : Q8NFF5 ( FMNAT ) . However , most prokaryotes depend on a single bifunctional enzyme , DB03147 synthetase ( FADS ) , which folds into two modules : the C-terminal associated with Q969G6 activity and the N-terminal associated with FMNAT activity . Sequence and structural analysis suggest that the 28-HxGH-31 , 123-Gx(D/N)-125 and 161-xxSSTxxR-168 motifs from FADS must be involved in DB00171 stabilisation for the adenylylation of Q68DA7 , as well as in DB03147 stabilisation for DB03147 phyrophosphorolysis . Mutants were produced at these motifs in the Corynebacterium ammoniagenes FADS ( CaFADS ) . Their effects on the kinetic parameters of CaFADS activities ( Q969G6 , FMNAT and DB03147 pyrophosphorilase ) , and on substrates and product binding properties indicate that H28 , H31 , N125 and S164 contribute to the geometry of the catalytically competent complexes at the FMNAT-module of CaFADS . Inhibition of canine exocrine pancreatic secretion by peptide YY is mediated by P10082 -preferring P28062 receptors . It is still unclear , which receptor subtype , Q03519 and/or P28062 , mediates the inhibitory action of P10082 on exocrine pancreatic secretion . The present study was undertaken to characterize functionally the Y receptor subtype that mediates the inhibition of exocrine pancreatic secretion by peptide YY ( P10082 ) . In eight conscious dogs with chronic gastric and pancreatic fistulas , we compared the action of intravenous infusion of 200 and 400 pmol/kg/h of the Y receptor agonists P10082 1-36 , DB05004 , P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 on the pancreatic secretory response to secretin ( 20.5 pmol/kg/h ) and cerulein ( 29.6 pmol/kg/h ) . P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 were also studied by giving a fivefold dose ( 1,000 and 2,000 pmol/kg/h ) . P10082 1-36 and the P49146 agonist DB05004 significantly inhibited pancreatic secretory responses to secretin and cerulein , whereas inhibition by P01303 1-36 and the P49146 agonist P10082 13-36 was attainable only at doses of 1,000 and 2,000 pmol/kg/h . The Q03519 receptor agonist Pro34PYY 1-36 was without effect on pancreatic secretion . We conclude that in dogs the inhibition of exocrine pancreatic secretion by P10082 is mediated via P28062 receptors of a P10082 -preferring subtype . A review of vilazodone , serotonin , and major depressive disorder . OBJECTIVE : To review the mechanism of selective serotonin reuptake inhibitor ( SSRI ) -mediated serotonergic neurotransmission , focusing on serotonin 1A ( P08908 ) autoreceptors , which are proposed to be involved in delaying therapeutic efficacy . DB06684 was specifically designed to function both as an SSRI and a partial agonist at P08908 receptors . This combined mechanism is proposed to decrease time to efficacy , minimize sexual side effects , and provide concomitant anxiolytic properties . DATA SOURCES : A PubMed search of all English-language articles from January 1990 to January 2013 was conducted using the search terms depression and 5-HT 1A , depression and buspirone , depression and pindolol , and vilazodone . STUDY SELECTION : We found 47 articles and abstracts that were selected for inclusion on the basis of information about the pharmacology of P08908 receptors and the clinical data on pindolol , buspirone , and vilazodone in depression . DATA EXTRACTION : This review summarizes current literature involving antidepressant activity , the role of P08908 autoreceptors , and clinical trials involving serotonin reuptake inhibition in conjunction with P08908 agonists and partial agonists , with a focus on vilazodone . RESULTS : DB06684 has demonstrated efficacy in 2 large , randomized , double-blind , placebo-controlled trials in major depressive disorder . RESULTS suggest that vilazodone has a low incidence of sexual side effects and is effective in patients with high levels of anxiety . A pooled analysis shows evidence of significant symptom reduction after only 1 week of therapy . CONCLUSIONS : If future studies corroborate the clinical benefits attributed to its mechanism of action , vilazodone may show potential advantages in terms of onset of action , sexual side effects , and anxiolytic activity in patients with major depressive disorder . Synthesis of pyrrolo[2,3-d]pyridazinones as potent , subtype selective DB05876 inhibitors . A series of pyrrolo [2,3-d]pyridazinones was synthesized and tested for their inhibitory activity on DB05876 subtypes A , B and D and selectivity toward DB01954 high affinity binding site ( HARBS ) . New agents with interesting profile were reported ; in particular compound 9e showed a good DB05876 subtype selectivity , being 8 times more potent ( IC50 = 0.32 microM ) for Q07343 ( anti-inflammatory ) than for Q08499 ( IC50 = 2.5 microM ) , generally considered the subtype responsible for emesis . Moreover the ratio HARBS/ Q07343 was particularly favourable for 9e ( 147 ) , suggesting that the best arranged groups around the pyrrolopyridazinone core are an isopropyl at position-1 , an ethoxycarbonyl at position-2 , together with an ethyl group at position-6 . For compounds 8 and 15a the ability to inhibit TNFalpha production in PBMC was evaluated and the results are consistent with their DB05876 inhibitory activity . DB00140 ( vitamin B2 ) deficiency impairs P04839 ( Nox2 ) priming and defense against Listeria monocytogenes . DB00140 , also known as vitamin B2 , is converted by riboflavin kinase ( Q969G6 ) into flavin mononucleotide ( Q68DA7 ) and flavin adenine dinucleotide ( DB03147 ) , which are essential cofactors of dehydrogenases , reductases , and oxidases including the phagocytic P04839 ( Nox2 ) . DB00140 deficiency is common in young adults and elderly individuals , who are at the coincidental risk for listeriosis . To address the impact of acute riboflavin deficiency on host defense against Listeria monocytogenes ( L.m. ) , we generated conditional Q969G6 knockout ( KO ) strains of mice . Phagocyte-specific Q969G6 KO impaired the capability of phagocytes to control intracellular L.m. , which corresponded to a greater susceptibility of mice to in vivo challenge with L.m . The oxidative burst of Q969G6 -deficient phagocytes in response to L.m. infection was significantly reduced . Mechanistically , P01375 -induced priming of Nox2 , which is needed for oxidative burst , was defective in Q969G6 -deficient phagocytes . Lack of riboflavin in wild-type macrophages for only 6 h shut down P01375 -induced , Q969G6 -mediated de novo Q68DA7 / DB03147 generation , which was accompanied by diminished ROS production and impaired anti-listerial activity . Vice versa , ROS production by riboflavin-deprived macrophages was rapidly restored by riboflavin supplementation . Our results suggest that acute riboflavin deficiency immediately impairs priming of Nox2 , which is of crucial relevance for an effective phagocytic immune response in vivo . The inhibitory effect of ginseng saponins on the stress-induced plasma interleukin-6 level in mice . The effect of ginseng saponins on plasma interleukin-6 ( P05231 ) in non-stressed and immobilization-stressed mice were investigated . DB01404 total saponins , ginsenosides Rb2 , Rg1 and Rd administered intraperitoneally attenuated the immobilization stress-induced increase in plasma P05231 level . But , intracerebroventricular injection of each ginsenoside did not affect plasma P05231 level induced by immobilization stress . Ginsenosides Rb2 , Rd and Rg1 significantly decreased norepinephrine and/or epinephrine-induced increase of P05231 level in macrophage cell line ( RAW 264.7 ) . Thus , it can be suggested that the inhibitory action of ginseng saponins against the immobilization stress-induced increase of plasma P05231 level would be in periphery ; at least in part , mediated by blocking norepinephrine- and/or epinephrine-induced increase of P05231 level in macrophage rather than in the brain . DB01404 saponins might be proposed as a possible candidate in the research or therapeutic modulation of stress-related disorders . Partial trypsin digestion as an indicator of mis-folding of mutant alanine:glyoxylate aminotransferase and chaperone effects of specific ligands . Study of a spectrum of missense mutants . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme whose deficiency results in primary hyperoxaluria type 1 ( P78364 ) . More than 75 P78364 mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have previously demonstrated that many such mutants expressed by transcription/translation are subject to generalized degradation by the proteasome and a specific limited trimming by an endogenous DB00171 -independent protease activity . Here , we report the results of partial digestion using trypsin as a mimic for the endogenous non-proteasomal protease and the use of N-terminal protein sequencing to determine the sensitive site . Partial trypsin digestion also provided an indicator of proper folding of the mutant enzyme . For selected mutations the sensitivity to trypsin could be ameliorated by addition of pyridoxal phosphate or aminooxy acetic acid as specific pharmacological chaperones . [ Protective effects of penehyclidine hydrochloride against acute renal injury induced by hemorrhagic shock and lipopolysaccharides in rats ] . OBJECTIVE : To investigate the effect of penehyclidine hydrochloride ( Q00325 ) in a rat model of renal injury induced by hemorrhagic shock and lipopolysaccharides ( LPS ) . METHODS : Forty-five healthy Wistar rats were randomized into sham operated group , model group , and 3 penehyclidine hydrochloride ( Q00325 ) dose ( 1 , 2 and 3 mg/kg ) groups ( P78364 , Q8IXK0 , and Q8NDX5 groups , respectively ) . The arterial blood samples were collected to determine the concentrations of serum tumor necrosis factor-α ( P01375 -α ) , interleukin-8 ( P10145 ) , interleukin-1 ( IL-1 ) , urine creatinine ( Cr ) and blood urine nitrogen ( BUN ) , and the renal tissues were collected to measure the expressions of P05362 and nuclear factor-κB ( NF-κB ) and observe the pathological changes . RESULTS : P01375 -α , P10145 , IL-1 , Cr , BUN , P05362 and NF-κB in the 3 Q00325 groups were significantly lower than those in the model group ( P < 0.05 ) . P01375 -α , P10145 , IL-1 , Cr and BUN were significantly lower in P78364 ( P < 0.05 ) than in the Q8IXK0 and Q8NDX5 groups , and P05362 and NF-κB were similar between 3 Q00325 groups ( P > 0.05 ) . Compared with the model group , the 3 Q00325 groups showed lessened pathological changes in the renal tubules . CONCLUSION : Q00325 has protective effects against renal injury induced by hemorrhagic-endotoxin shock in rats , and treatment with 1 mg/kg Q00325 produces the most significant protective effect . P15121 inhibition suppresses azoxymethane-induced colonic premalignant lesions in C57BL/KsJ-db/db mice . Type-2 diabetes and obesity-related metabolic abnormalities are major risk factors for the development of colon cancer . In the present study , we examined the effects of polyol pathway enzyme aldose reductase ( AR ) inhibitor , fidarestat , on the development of azoxymethane ( AOM ) -induced colonic premalignant lesions in C57BL/KsJ-db/db obese mice . Our results indicate that fidarestat given in the drinking water caused a significant reduction in the total number of colonic premalignant lesions in the AOM treated obese mice . Further , the expression levels of PKC-β2 , AKT , P35354 and P35228 in the colonic mucosa of AOM-treated mice were significantly decreased by fidarestat . The serum levels of IL-1α , P02778 , Q07325 , P01375 -α and P15692 are significantly suppressed in AOM + fidarestat treated obese mice . DB02021 also decreased the expression of P35354 , P35228 , P98170 , survivin , β-catenin and NF-κB in high glucose-treated HT29 colon cancer cells . In conclusion , our results indicate that fidarestat inhibits the development of colonic premalignant lesions in an obesity-related colon cancer and is chemopreventive to colorectal carcinogenesis in obese individuals .

MH_train_1249

MH_train_1249

MH_train_1249

interacts_with DB00734?

multiple_choice

Investigation of mechanisms mediating 8-OH-DPAT-induced impairment of spatial memory : involvement of P08908 receptors in the dorsal hippocampus in rats . The purpose of this study was to identify mechanisms that mediate the impairment of spatial memory induced by 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 / P34969 receptor agonist , in the eight-arm radial maze in rats . WAY-100635 and NAN-190 , P08908 receptor antagonists , reversed the impairment of spatial memory induced by systemic injection of 8-OH-DPAT ( 1 mg/kg , i.p. ) . On the other hand , the alpha1-adrenoceptor antagonist prazosin and a selective P34969 receptor antagonist SB269970 had no effect on 8-OH-DPAT-induced impairment of spatial memory . Bilateral microinjection of 8-OH-DPAT ( 4 microg/side ) impaired spatial memory when injected into the dorsal hippocampus ( DH ) . Contrastingly , spatial memory was unaffected by microinjections of 8-OH-DPAT into the other six areas examined : ventral hippocampus ( VH ) , central amygdaloid nucleus ( P12821 ) , lateral hypothalamus ( LH ) , nucleus accumbens ( NAc ) , and dorsal ( DR ) and median ( MR ) raphe nucleus . Furthermore , NAN-190 significantly reversed the impairment of spatial memory induced by intra-DH injection of 8-OH-DPAT . These findings suggest that P08908 receptors in the DH play an important role in the mechanisms underlying the 8-OH-DPAT-induced impairment of spatial memory in rats . DB09341 transporter-2 ( P11168 ) promoter mediated transgenic insulin production reduces hyperglycemia in diabetic mice . P01308 production afforded by hepatic gene therapy ( HGT ) retains promise as a potential treatment for type 1 diabetes , but successful approaches have been limited . We employed a novel and previously untested promoter for this purpose , glucose transporter-2 ( P11168 ) to drive insulin production via delivery by recombinant adeno-associated virus ( rAAV ) . In vitro , the P11168 promoter was capable of robust glucose-responsive expression in transduced HepG2 human hepatoma cells . Therefore , rAAV constructs were designed to express the furin-cleavable human preproinsulin B10 gene , under the control of the murine P11168 promoter and packaged for delivery with rAAV expressing the type 5 capsid . DB00428 -induced diabetic mice were subjected to hepatic portal vein injection immediately followed by implantation of a sustained-release insulin pellet to allow time for transgenic expression . All mice injected with the rAAV5- P11168 -fHPIB10 virus remained euglycemic for up to 35 days post-injection , with 50 % euglycemic after 77 days post-injection . In contrast , mock-injected mice became hyperglycemic within 15 days post-injection following dissolution of the insulin pellet . Serum levels of both human insulin and C-peptide further confirmed successful transgenic delivery by the rAAV5- P11168 -fHPIB10 virus . These findings indicate that the P11168 promoter may be a potential candidate for regulating transgenic insulin production for hepatic insulin gene therapy in the treatment of type I diabetes . Effects of short- and long-term risperidone treatment on prolactin levels in children with autism . BACKGROUND : The effects of short- and long-term risperidone treatment on serum prolactin were assessed in children and adolescents with autism . METHODS : Patients with autism ( N = 101 , 5-17 years of age ) were randomized to an 8-week trial of risperidone or placebo and 63 then took part in a 4-month open-label follow-up phase . Serum samples were obtained at Baseline and Week-8 ( N = 78 ) , and at 6-month ( N = 43 ) and 22-month ( N = 30 ) follow-up . Serum prolactin was determined by immunoradiometric assay ; dopamine type-2 receptor ( P14416 ) polymorphisms were genotyped . RESULTS : Baseline prolactin levels were similar in the risperidone ( N = 42 ) and placebo ( N = 36 ) groups ( 9.3 +/- 7.5 and 9.3 +/- 7.6 ng/ml , respectively ) . After 8 weeks of risperidone , prolactin increased to 39.0 +/- 19.2 ng/ml , compared with 10.1 +/- 8.8 ng/ml for placebo ( p < .0001 ) . P01236 levels were also significantly increased at 6 months ( 32.4 +/- 17.8 ng/ml ; N = 43 , p < .0001 ) and at 22 months ( N = 30 , 25.3 +/- 15.6 ng/ml , p < .0001 ) . P01236 levels were not associated with adverse effects and P14416 alleles ( Taq1A , -141C Ins/Del , C957T ) did not significantly influence baseline levels or risperidone-induced increases in prolactin . CONCLUSIONS : DB00734 treatment was associated with two- to four-fold mean increases in serum prolactin in children with autism . Although risperidone-induced increases tended to diminish with time , further research on the consequences of long-term prolactin elevations in children and adolescents is needed . The missing link in coenzyme A biosynthesis : PanM ( formerly YhhK ) , a yeast Q92830 acetyltransferase homologue triggers aspartate decarboxylase ( PanD ) maturation in Salmonella enterica . DB01992 ( DB01992 ) is an essential cofactor for all forms of life . The biochemistry underpinning the assembly of DB01992 in Escherichia coli and other enterobacteria is well understood , except for the events leading to maturation of the L-aspartate-α-decarboxylase ( PanD ) enzyme that converts pantothenate to β-alanine . PanD is synthesized as pro-PanD , which undergoes an auto-proteolytic cleavage at residue Ser25 to yield the catalytic pyruvoyl moiety of the enzyme . Since 1990 , it has been known that E. coli yhhK strains are pantothenate auxotrophs , but the role of YhhK in pantothenate biosynthesis remained an enigma . Here we show that Salmonella enterica yhhK strains are also pantothenate auxotrophs . In vivo and in vitro evidence shows that YhhK interacts directly with PanD , and that such interactions accelerate pro-PanD maturation . We also show that S. enterica yhhK strains accumulate pro-PanD , and that not all pro-PanD proteins require YhhK for maturation . For example , the Corynebacterium glutamicum panD(+) gene corrected the pantothenate auxotrophy of a S. enterica yhhK strain , supporting in vitro evidence obtained by others that some pro-PanD proteins autocleave at faster rates . We propose the name PanM for YhhK to reflect its role as a trigger of pro-PanD maturation by stabilizing pro-PanD in an autocleavage-prone conformation . DB00659 : recent findings and future research directions . This article explores the mechanisms of action and the potential responder profile of acamprosate , a compound efficacious in relapse prevention of alcoholism . New evidence at the molecular and cellular level suggests that acamprosate attenuates hyper-glutamatergic states that occur during early abstinence and involves iono ( DB01221 ) - and metabotrotropic ( P41594 ) glutamate receptors along with augmented intracellular calcium release and electrophysiological changes . Thus mutant mice with enhanced glutamate levels exhibit higher alcohol consumption than wild type mice and respond better to acamprosate , demonstrating that acamprosate acts mainly on a hyper-glutamatergic system . This mode of action further suggests that acamprosate exhibits neuroprotective properties . In rats , cue-induced reinstatement behavior is significantly reduced by acamprosate treatment whereas cue-induced craving responses in alcohol-dependent patients seem not to be affected by this treatment . An ongoing study ( " Project Predict " ) defines specific responder profiles for an individualized use of acamprosate and naltrexone . Neurophysiological as well as psychometric data are used to define 2 groups of patients : " reward cravers " and " relief cravers " . While naltrexone should work better in the first group , acamprosate is hypothesized to be efficacious in the latter where withdrawal associated and/or cue induced hyper-glutamatergic states are thought to trigger relapse . Further research should target the definition of subgroups applying endophenotypic approaches , e.g. by detecting a hyperglutamatergic syndrome using MR spectroscopy . Neuronal ablation of p-Akt at Ser473 leads to altered P08908 /2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5-HT ) signaling . To explore how impairment in Akt function regulates 5-HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser473 . Cortical P08908 and 5- Q13049 receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 receptor agonist 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 receptor density was associated with higher P08908 receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5- Q13049 /C agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , with evidence of impaired 5- Q13049 /C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 receptor expression and attenuated DOI-induced 5- Q13049 receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5-HT receptor regulation . These data reveal that defective central Akt function alters 5-HT signaling as well as 5-HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5-HT receptor function . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . P12004 D1-Cdk4 controls glucose metabolism independently of cell cycle progression . P01308 constitutes a principal evolutionarily conserved hormonal axis for maintaining glucose homeostasis ; dysregulation of this axis causes diabetes . P20142 -1α ( peroxisome-proliferator-activated receptor-γ coactivator-1α ) links insulin signalling to the expression of glucose and lipid metabolic genes . The histone acetyltransferase Q92830 ( general control non-repressed protein 5 ) acetylates P20142 -1α and suppresses its transcriptional activity , whereas sirtuin 1 deacetylates and activates P20142 -1α . Although insulin is a mitogenic signal in proliferative cells , whether components of the cell cycle machinery contribute to its metabolic action is poorly understood . Here we report that in mice insulin activates cyclin D1-cyclin-dependent kinase 4 ( Cdk4 ) , which , in turn , increases Q92830 acetyltransferase activity and suppresses hepatic glucose production independently of cell cycle progression . Through a cell-based high-throughput chemical screen , we identify a Cdk4 inhibitor that potently decreases P20142 -1α acetylation . P01308 /GSK-3β ( glycogen synthase kinase 3-beta ) signalling induces cyclin D1 protein stability by sequestering cyclin D1 in the nucleus . In parallel , dietary amino acids increase hepatic cyclin D1 messenger RNA transcripts . Activated cyclin D1-Cdk4 kinase phosphorylates and activates Q92830 , which then acetylates and inhibits P20142 -1α activity on gluconeogenic genes . Loss of hepatic cyclin D1 results in increased gluconeogenesis and hyperglycaemia . In diabetic models , cyclin D1-Cdk4 is chronically elevated and refractory to fasting/feeding transitions ; nevertheless further activation of this kinase normalizes glycaemia . Our findings show that insulin uses components of the cell cycle machinery in post-mitotic cells to control glucose homeostasis independently of cell division . Inhibition of central angiotensin converting enzyme ameliorates scopolamine induced memory impairment in mice : role of cholinergic neurotransmission , cerebral blood flow and brain energy metabolism . Evidences indicate that inhibition of central P00797 angiotensin system ( DB01367 ) ameliorates memory impairment in animals and humans . Earlier we have reported involvement of central angiotensin converting enzyme ( P12821 ) in streptozotocin induced neurodegeneration and memory impairment . The present study investigated the role of central P12821 in cholinergic neurotransmission , brain energy metabolism and cerebral blood flow ( Q03701 ) in model of memory impairment induced by injection of scopolamine in mice . DB00790 ( 0.05 and 0.1 mg/kg , PO ) was given orally for one week before administration of scopolamine ( 3mg/kg , IP ) . Then , memory function was evaluated by Morris water maze and passive avoidance tests . Q03701 was measured by laser Doppler flowmetry . Biochemical and molecular parameters were estimated after the completion of behavioral studies . DB00747 caused impairment in memory which was associated with reduced Q03701 , acetylcholine ( ACh ) level and elevated acetylcholinesterase ( P22303 ) activity and malondialdehyde ( MDA ) level . DB00790 ameliorated scopolamine induced amnesia in both the behavioral paradigms . Further , perindopril prevented elevation of P22303 and MDA level in mice brain . There was a significant increase in Q03701 and ACh level in perindopril treated mice . However , scopolamine had no significant effect on DB00171 level and mRNA expression of angiotensin receptors and P12821 in cortex and hippocampus . But , perindopril significantly decreased P12821 activity in brain without affecting its mRNA expression . The study clearly showed the interaction between P12821 and cholinergic neurotransmission and beneficial effect of perindopril can be attributed to improvement in central cholinergic neurotransmission and Q03701 . Zebrafish express the common parathyroid hormone/parathyroid hormone-related peptide receptor ( Q03431 ) and a novel receptor ( PTH3R ) that is preferentially activated by mammalian and fugufish parathyroid hormone-related peptide . To further explore the evolution of receptors for parathyroid hormone ( PTH ) and PTH-related peptide ( P12272 ) , we searched for zebrafish ( z ) homologs of the PTH/ P12272 receptor ( Q03431 ) . In mammalian genes encoding this receptor , exons M6/7 and M7 are highly conserved and separated by 81-84 intronic nucleotides . Genomic polymerase chain reaction using degenerate primers based on these exons led to two distinct DNA fragments comprising portions of genes encoding the zPTH1R and the novel zPTH3R . Sequence comparison of both full-length teleost receptors revealed 69 % similarity ( 61 % identity ) , but less homology with zPTH2R . When compared with hPTH1R , zPTH1R showed 76 % and zPTH3R 67 % amino acid sequence similarity ; similarity with hPTH2R was only 59 % for both teleost receptors . When expressed in COS-7 cells , zPTH1R bound [ DB00135 (34)] DB05829 -(1-34)-amide ( DB05829 ) , [ DB00135 (36)]hPTHrP-(1-36)-amide ( hPTHrP ) , and [ Ala(29), DB00142 (30) , Ala(34), DB00142 (35) , DB00135 (36) ] fugufish P12272 -(1-36)-amide ( fuguPTHrP ) with a high apparent affinity ( IC(50) : 1.2-3.5 nM ) , and was efficiently activated by all three peptides ( EC(50) : 1.1-1.7 nM ) . In contrast , zPTH3R showed higher affinity for fuguPTHrP and hPTHrP ( IC(50) : 2.1-11.1 nM ) than for DB05829 ( IC(50) : 118.2-127.0 nM ) ; DB02527 accumulation was more efficiently stimulated by fugufish and human P12272 ( EC(50) : 0.47 +/- 0.27 and 0.45 +/- 0.16 , respectively ) than by DB05829 ( EC(50) : 9.95 +/- 1.5 nM ) . Agonist-stimulated total inositol phosphate accumulation was observed with zPTH1R , but not zPTH3R . Innovative approaches for the development of antidepressant drugs : current and future strategies . Depression is a highly debilitating disorder that has been estimated to affect up to 21 % of the world population . Despite the advances in the treatment of depression with selective serotonin reuptake inhibitors ( SSRIs ) and serotonin and norepinephrine reuptake inhibitors ( SNRIs ) , there continue to be many unmet clinical needs with respect to both efficacy and side effects . These needs range from efficacy in treatment resistant patients , to improved onset , to reductions in side effects such as emesis or sexual dysfunction . To address these needs , there are numerous combination therapies and novel targets that have been identified that may demonstrate improvements in one or more areas . There is tremendous diversity in the types of targets and approaches being taken . At one end of a spectrum is combination therapies that maintain the benefits associated with SSRIs but attempt to either improve efficacy or reduce side effects by adding additional mechanisms ( P08908 , P28222 , P28221 , P28335 , alpha-2A ) . At the other end of the spectrum are more novel targets , such as neurotrophins ( P23560 , IGF ) , based on recent findings that antidepressants induce neurogenesis . In between , there are many approaches that range from directly targeting serotonin receptors ( P28335 , P50406 ) to targeting the multiplicity of potential mechanisms associated with excitatory ( glutamate , DB01221 , Q14416 , P41594 ) or inhibitory amino acid systems ( GABA ) or peptidergic systems ( neurokinin 1 , DB05394 1 , melanin-concentrating hormone 1 , V1b ) . The present review addresses the most exciting approaches and reviews the localization , neurochemical and behavioral data that provide the supporting rationale for each of these targets or target combinations . Regulation of milk lipid secretion : effects of oxytocin , prolactin and ionomycin on triacylglycerol release from rat mammary gland slices . A system for the study of the regulation of the release of triacylglycerols by mammary gland slices was developed . By prelabelling the triacylglycerol pool with [3H]oleate measurements of release of both mass of triacylglycerol and of newly synthesized triacylglycerol have been made . DB00107 and ovine prolactin stimulated release of triacylglycerol and protein , but the former was 40-fold more effective . Recombinant bovine prolactin was even less active than ovine prolactin , suggesting that contamination of the latter with oxytocin and/or vasopressin was partly responsible for its stimulatory effect on release . The findings support the view that the major effect of oxytocin is to stimulate contraction of myoepithelial cells and thus release secreted lipid stored in the lumen of the mammary gland alveoli . Ionomycin , a Ca2+ ionophore , also stimulated lipid release , but probably not by the usual apocrine route . P12272 , a peptide produced by the mammary gland , did not stimulate release or antagonize the effects of oxytocin . Release of lipid was also measured in mammary gland slices from late-pregnant , early- and mid-lactating rats and lactating rats made prolactin-deficient . Hormonal stimulation in vitro showed the maturation of response seen in vivo on transition from late pregnancy to peak lactation . P01236 deficiency resulted in decreased release of newly synthesized lipid in response to oxytocin . Interaction between fatty acid synthase- and ErbB-systems in ovarian cancer cells . P49327 ( P49327 ) represents a metabolic oncogene . It produces phospholipids for membrane microdomains that accommodate receptor tyrosine kinases including Epidermal Growth Factor-Receptor ( P00533 , ErbB1 ) and ErbB2 ( P04626 /neu ) . P49327 and ErbBs are overexpressed in ovarian cancer . We examined the effect of P49327 and ErbB inhibition on A2780 and SKOV3 ovarian cancer cells . Growth assays reveal that P49327 inhibitor C75 sensitizes tumor cells against anti-ErbB drugs ( pelitinib [ Q9Y259 -569 ] , canertinib [ DB05424 ] , erlotinib , cetuximab , matuzumab , trastuzumab ) suggesting P49327 /ErbB cooperation . qRT-PCR and Western blotting revealed that C75 represses P49327 , P00533 , ErbB2 , and AKT suggesting that P49327 -induced membrane microdomains accommodate/stabilize ErbBs and facilitate AKT recruitment/activation . Our data indicate that AKT is crucial for ErbB/ P49327 interaction , AKT cross-inhibits P29323 and feeds loops that boost P49327 and P00533 transcription , and P00533 and ErbB2 must be co-silenced for maximal P49327 downregulation . Taken together , interference with P49327 and ErbB abrogates their oncogenicity and should be exploited for ovarian cancer treatment . Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . DB00640 , dopamine and serotonin receptors imbalance in lymphocytes of Lesch-Nyhan patients . Lesch-Nyhan disease ( LND ) is caused by complete deficiency of the hypoxanthine-guanine phosphoribosyltransferase enzyme . It is characterized by overproduction of uric acid , jointly with severe motor disability and self-injurious behaviour which physiopathology is unknown . These neurological manifestations suggest a dysfunction in the basal ganglia , and three neurotransmitters have been implicated in the pathogenesis of the disease : dopamine , adenosine and serotonin . All of them are implicated in motor function and behaviour , and act by binding to specific G-protein coupled receptors in the synaptic membrane where they seem to be integrated through receptor-receptor interactions . In this work we have confirmed at protein level the previously reported increased expression of P21918 and the variably aberrant expression of P29274 , in LND PBL respect to control PBL . We have also described , for the first time , a decreased expression and protein level of 5- P08908 in LND PBL respect to control PBL . If these results were confirmed in the Lesch-Nyhan patients basal ganglia cells , this would support the hypothesis that pathogenesis of neurological manifestations of Lesch-Nyhan patients may be related to an imbalance of neurotransmitters , rather than to the isolated disturbance of one of the neurotransmitters , and this fact should be taken into account in the design of pharmacologic treatment for their motor and behavioural disturbances . Role of antispasmodics in the treatment of irritable bowel syndrome . Irritable bowel syndrome ( IBS ) is a long-lasting , relapsing disorder characterized by abdominal pain/discomfort and altered bowel habits . Intestinal motility impairment and visceral hypersensitivity are the key factors among its multifactorial pathogenesis , both of which require effective treatment . Voltage-gated calcium channels mediate smooth muscle contraction and endocrine secretion and play important roles in neuronal transmission . Antispasmodics are a group of drugs that have been used in the treatment of IBS for decades . DB01616 citrate , a spasmolytic , decreases the sensitivity of smooth muscle contractile proteins to calcium , and it is a selective P08908 receptor antagonist . DB01616 , in combination with simethicone , has been demonstrated to effectively reduce abdominal pain and discomfort in a large placebo-controlled trial . Mebeverine is a musculotropic agent that potently blocks intestinal peristalsis . Non-placebo-controlled trials have shown positive effects of mebeverine in IBS regarding symptom control ; nevertheless , in recent placebo-controlled studies , mebeverine did not exhibit superiority over placebo . Otilonium bromide is poorly absorbed from the GI tract , where it acts locally as an L-type calcium channel blocker , an antimuscarinic and a tachykinin NK2 receptor antagonist . Otilonium has effectively reduced pain and improved defecation alterations in placebo-controlled trials in IBS patients . DB09090 bromide is also an L-type calcium channel blocker that acts locally in the GI tract . DB09090 improves motility disorders and consequently reduces stool problems in IBS patients . Phloroglucinol and trimethylphloroglucinol are non-specific antispasmodics that reduced pain in IBS patients in a placebo-controlled trial . Antispasmodics have excellent safety profiles . T-type calcium channel blockers can abolish visceral hypersensitivity in animal models , which makes them potential candidates for the development of novel therapeutic agents in the treatment of IBS . Association of adenosine receptor gene polymorphisms and in vivo adenosine A1 receptor binding in the human brain . DB00640 A1 receptors ( A1ARs ) and the interacting adenosine A2A receptors are implicated in neurological and psychiatric disorders . Variants within the corresponding genes P30542 and P29274 were shown associated with pathophysiologic alterations , particularly increased anxiety . It is unknown so far , if these variants might modulate the A1AR distribution and availability in different brain regions . In this pilot study , the influence of P30542 and P29274 variants on in vivo A1AR binding was assessed with the A1AR-selective positron emission tomography ( PET ) radioligand [(18)F]CPFPX in brains of healthy humans . Twenty-eight normal control subjects underwent PET procedures to calculate the binding potential BPND of [(18)F]CPFPX in cerebral regions and to assess P30542 and P29274 single nucleotide polymorphism ( SNP ) effects on regional BPND data . Our results revealed SNPs of both genes associated with [(18)F]CPFPX binding to the A1AR . The strongest effects that withstood even Bonferroni correction of multiple SNP testing were found in non-smoking subjects ( N=22 ) for P29274 SNPs rs2236624 and rs5751876 ( corr. Pall < 0.05 ) . SNP alleles previously identified at risk for increased anxiety like the rs5751876 T-allele corresponded to consistently higher A1AR availability in all brain regions . Our data indicate for the first time that variation of A1AR availability was associated with ADORA SNPs . The finding of increased A1AR availability in regions of the fear network , particularly in P29274 risk allele carriers , strongly warrants evaluation and replication in further studies including individuals with increased anxiety .

MH_train_1250

MH_train_1250

MH_train_1250

interacts_with DB00222?

multiple_choice

Relationship of five type 2 diabetes candidate gene polymorphisms to the age at diagnosis of diabetes in the Slovakian population . OBJECTIVES : To examine the relationship between polymorphisms of five candidate genes for type 2 diabetes mellitus ( T2DM ) and the age at diagnosis of T2DM . METHODS : 538 Slovakian patients with T2DM were included and their age at diagnosis of T2DM retrieved from their medical records . Polymorphisms of genes encoding peroxisome proliferator activated receptor gamma ( P37231 ) , P37231 -coactivator-1 ( PGC1 ) , insulin-receptor substrate 1 ( P35568 ) , the subunit Kir 6.2 of the DB00171 -dependent potassium-channel ( Q14654 ) and transcription factor 7-like 2 ( Q9NQB0 ) were detected by PCR-RFLP methods . RESULTS : No significant relationship between the risk alleles of the examined gene polymorphisms to the lower mean age at diagnosis of T2DM was observed . The carriers of the TT-genotype of Q9NQB0 rs7903146 polymorphism had significantly increased odds ratio for diagnosis of diabetes before the age of 40 years [ OR 3.02 ( 1.34 , 6.81 ) , p = 0.008 ] , in comparison with the CC/CT genotype carriers . CONCLUSION : No significant association of P37231 , PGC1 , P35568 , Q14654 and Q9NQB0 gene polymorphisms and the age at diagnosis of T2DM was observed in the present study . Homozygotes for the risk allele of Q9NQB0 had more frequently early onset of T2DM , before age of 40 years ( Tab. 4 , Ref. 15 ) . 2(3H)-benzoxazolone and bioisosters as " privileged scaffold " in the design of pharmacological probes . The 2(3H)-benzoxazolone heterocycle and its bioisosteric surrogates ( such as 2(3H)-benzothiazolinone , benzoxazinone , etc. ) have received considerable attention from the medicinal chemists owing to their capacity to mimic a phenol or a catechol moiety in a metabolically stable template . These heterocycles and pyrocatechol have indeed similar pKa 's , electronic charge distribution , and chemical reactivity . Therapeutic applications of this template are very broad , and range from analgesic anti-inflammatory compounds ( including P37231 antagonists ) to antipsychotic and neuroprotective anticonvulsant compounds . High affinity ligands have been obtained also for dopaminergic ( D2 and D4 ) , serotoninergic ( P08908 and P28223 ) , sigma-1 and sigma-2 receptors . Owing to the high number of positive hits encountered with this heterocycle and its congeners , 2(3H)-benzoxazolone template certainly deserves the title of " privileged scaffold " in medicinal chemistry . Genetic determinants of low high-density lipoprotein cholesterol . PURPOSE OF REVIEW : High-density lipoprotein cholesterol ( HDL-C ) has been well established as an inverse predictor of coronary heart disease ( Q8NE62 ) , and in recent years , investigations have focused on the genetic regulation of high-density lipoprotein . Although numerous candidate genes contribute to the low HDL-C phenotype , their impact on Q8NE62 is heterogeneous , reflecting diverse gene-gene interactions and gene-environmental relationships . This review summarizes recent data involving HDL regulatory genes and their role in atherothrombosis . RECENT FINDINGS : The primary genetic determinants associated with relative HDL-C deficiency states are the DB00171 binding cassette protein , O95477 ; apolipoprotein ( APO ) A1 ; and lecithin cholesteryl acyl transferase . Other potentially important candidates invoked in low HDL-C syndromes in humans include P02656 , lipoprotein lipase , sphingomyelin phosphodiesterase 1 , and glucocerebrosidase . Molecular variation in ABCAI and APOAI and , in selected cases , lecithin cholesteryl acyl transferase deficiency have been associated with increased Q8NE62 , whereas two notable variants , APOAIMilano and APOAIParis , are associated with reduced risk . SUMMARY : Low HDL-C syndromes have generally been correlated with an increased risk of Q8NE62 . However , single-gene abnormalities responsible for HDL-C deficiency states may have variable effects on atherothrombotic risk . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Dissecting the first transcriptional divergence during human embryonic development . The trophoblast cell lineage is specified early at the blastocyst stage , leading to the emergence of the trophectoderm and the pluripotent cells of the inner cell mass . Using a double mRNA amplification technique and a comparison with transcriptome data on pluripotent stem cells , placenta , germinal and adult tissues , we report here some essential molecular features of the human mural trophectoderm . In addition to genes known for their role in placenta ( P01215 , P49763 , P10696 and Q9UNQ0 ) , human trophectoderm also strongly expressed Laminins , such as P25391 , and the GAGE Cancer/Testis genes . The very high level of Q9UNQ0 expression in trophectoderm , 7.9-fold higher than in placenta , suggests a major role of this gene in shielding the very early embryo from xenobiotics . Several genes , including P32239 and Q9UJW3 , were specifically up-regulated only in trophectoderm , indicating that the trophoblast cell lineage shares with the germinal lineage a transient burst of Q9UJW3 expression . A trophectoderm core transcriptional regulatory circuitry formed by 13 tightly interconnected transcription factors ( P49715 , P23769 , P23771 , Q9NP62 , Q13887 , O60675 , P35548 , Q05195 , Q03181 , P37231 , Q8WUF5 , Q92754 and Q9H3D4 ) , was found to be induced in trophectoderm and maintained in placenta . The induction of this network could be recapitulated in an in vitro trophoblast differentiation model . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . P62158 -mediated effects of loperamide on chloride transport by brush border membrane vesicles from human ileum . We investigated whether the synthetic opiate loperamide-HCl is able to regulate specific transport systems for sodium and chloride in brush border membrane vesicles ( BBMVs ) from human ileum and whether such activities are mediated by calcium/calmodulin . In BBMVs we studied Na+/H+ antiport , Cl+/OH- antiport , Na+/Cl- cotransport , and the Cl- conductive pathway . Brush border membrane vesicles were incubated with 10 microM loperamide over 4 h at 5 degrees C before the uptake experiments . In ileal BBMVs , loperamide stimulated intravesicular accumulation of Na+ in the presence of Cl- and vice versa . After 1 min of incubation , the stimulatory effect was 35 % +/- 5 % ( p less than 0.005 ) of the control without loperamide . DB00836 also stimulated Cl-/OH- antiport by 30 % +/- 5 % ( p less than 0.005 ) in BBMVs of ileum . In addition , we studied the role of Ca2+/calmodulin in the action of loperamide on chloride transport by human BBMVs . In loperamide-pretreated BBMVs , calmodulin activity was significantly decreased ( 12 +/- 2 vs. 38 +/- 4 pmol/mg protein ) . When loperamide-pretreated vesicles were incubated with 2 microM calcium ( free concentration ) plus 5 microM calmodulin for 1 h at 5 degrees C , complete inhibition of the stimulatory effect of loperamide on Cl-/OH- antiport and Na+/Cl- cotransport was observed . Increasing the Ca2+/calmodulin activity of loperamide-pretreated BBMVs with 2 microM calcium plus 5 microM calmodulin led to a significant inhibition of Cl-/OH- antiport and Na+/Cl- cotransport by 40 % +/- 5 % ( p less than 0.005 ) . P01308 augments gonadotropin-releasing hormone induction of translation in LbetaT2 cells . The integrated signaling of insulin and gonadotropin-releasing hormone in the pituitary gonadotropes may have a profound bearing on reproductive function , although the cross-receptor signaling mechanisms are unclear . We demonstrate that the insulin receptor is constitutively localized to non-caveolar lipid raft microdomains in the pituitary gonadotrope cell line LbetaT2 . The localization to rafts is consistent with similar localization of the P30968 . P06213 phosphorylation occurs in raft domains and activates the downstream signaling targets P01308 Receptor Substrate1 and Akt/Protein Kinase B . Although insulin alone does not strongly activate the extracellular signal-regulated kinase second messenger cascade , co-stimulation potentiates the phosphorylation of the extracellular signal-regulated kinase by gonadotropin-releasing hormone . The co-stimulatory effect of insulin and gonadotropin-releasing hormone is also evident in increased activation of cap-dependent translation . In contrast , co-stimulation attenuates Akt/Protein Kinase B activation . Our results show that both gonadotropin-releasing hormone and insulin are capable of mutually altering their respective regulatory signaling cascades . We suggest that this provides a mechanism to integrate neuropeptide and energy homeostatic signals to modulate reproductive function . Domain organization of the DB00171 -sensitive potassium channel complex examined by fluorescence resonance energy transfer . K( DB00171 ) channels link cell metabolism to excitability in many cells . They are formed as tetramers of Kir6.2 subunits , each associated with a Q09428 subunit . We used mutant GFP-based FRET to assess domain organization in channel complexes . Q8N1N2 -length Kir6.2 subunits were linked to YFP or cyan fluorescent protein ( P27918 ) at N or C termini , and all such constructs , including double-tagged YFP-Kir6.2- P27918 ( Y6.2C ) , formed functional K( DB00171 ) channels . In intact COSm6 cells , background emission of YFP excited by 430-nm light was ∼6 % , but the Y6.2C construct expressed alone exhibited an apparent FRET efficiency of ∼25 % , confirmed by trypsin digestion , with or without Q09428 co-expression . Similar FRET efficiency was detected in mixtures of P27918 - and YFP-tagged full-length Kir6.2 subunits and transmembrane domain only constructs , when tagged at the C termini but not at the N termini . The FRET-reported Kir6.2 tetramer domain organization was qualitatively consistent with Kir channel crystal structures : C termini and M2 domains are centrally located relative to N termini and M1 domains , respectively . Additional FRET analyses were performed on cells in which tagged full-length Kir6.2 and tagged Q09428 constructs were co-expressed . These analyses further revealed that 1 ) NBD1 of Q09428 is closer to the C terminus of Kir6.2 than to the N terminus ; 2 ) the Kir6.2 cytoplasmic domain is not essential for complexation with Q09428 ; and 3 ) the N-terminal half of Q09428 can complex with itself in the absence of either the C-terminal half or Kir6.2 . [ Low doses of sulphonyluria as a successful replacement for insulin therapy in a patient with neonatal diabetes due to a mutation of Q14654 gene encoding Kir6.2 ] . Neonatal diabetes mellitus is a rare metabolic disorder with an estimated incidence of 1:300.000 to 400.000 newborns , and less than 50 % of the neonates have permanent neonatal diabetes mellitus ( PNDM ) . Recently , activating mutation in the Q14654 gene encoding Kir6.2 subunit of the adenosin triphosphate-sensitive potassium ( K( DB00171 ) ) channel has been described as the most frequent cause of PNDM . Under physiological circumstances K( DB00171 ) channel closure plays a central role in glucose-stimulated insulin secretion from pancreatic beta cells . Sulphonylurea drugs stimulate insulin secretion by binding to and closing K( DB00171 ) channels and thus bypassing beta cell metabolism stimulate the same chain of reactions as glucose . We describe a boy diagnosed with PNDM at the age of 3 months when insulin therapy was started , and at the age of 4.5 years Q14654 gene was sequenced and found that the boy carried a de novo activating R201H mutation . P01308 therapy was successfully switched to low doses of oral glibenclamide . Accordingly , it is important to emphasize that every person diagnosed with diabetes before six months of life , however old they actually are , should be tested for K( DB00171 ) mutations which is offered via the website www.diabetesgenes.org . Development of DB00644 antagonists for prostate cancer : new approaches to treatment . Prostate cancer has become the most common cancer among American men and is second only to lung cancer as a cause of male cancer-related death . Several treatment options exist for different stages of prostate cancer including observation , prostatectomy , radiation therapy , chemotherapy , and hormone therapy . Hormone therapy has evolved from the use of estrogens to gonadotropin-releasing hormone ( DB00644 ) agonists and recently , investigational DB00644 antagonists . P30968 agonists such as leuprolide , bruserelin and goserelin have been used for the treatment of prostate cancer . These agonists eventually cause the inhibition of lutenizing hormone production , which in turn causes a suppression of testosterone and dihydrotestosterone , on which continued growth of prostate cancer cells depend . Several comparative studies of leuprorelin administered as daily injections or monthly depot injections have been reported . Disease progression was prevented in more than 72 % of men administered daily leuprorelin , and in 82 % to 89 % of those receiving monthly depots . Another synthetic DB00644 analog , goserelin , has been studied in a similar population of men with daily injections producing partial responses in 60 % to 80 % of men with previously untreated prostate cancer . DB00106 , a peptide antagonist of P30968 , is also being studied for the treatment of prostate cancer . The discovery and development of DB00644 antagonists may provide an important advance for patients with prostate cancer . Clearly the studies described herein , as well as many others , outline an exciting era of research to define the optimal use of hormonal therapy in prostate cancer . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . P62158 kinase II inhibition enhances ischemic preconditioning by augmenting DB00171 -sensitive K+ current . Mice with genetic inhibition ( O60266 -I ) of the multifunctional Ca(2+)/calmodulin dependent protein kinase II ( CaMKII ) have improved cardiomyocyte survival after ischemia . Some K(+) currents are up-regulated in O60266 -I hearts , but it is unknown if CaMKII inhibition increases the DB00171 sensitive K(+) current ( I(KATP) ) that underlies ischemic preconditioning ( IP ) and confers resistance to ischemia . We hypothesized increased I(KATP) was part of the mechanism for improved ventricular myocyte survival during ischemia in O60266 -I mice . O60266 -I hearts were protected against global ischemia due to enhanced IP compared to wild type ( WT ) and transgenic control ( O60266 -C ) hearts . Q14654 was significantly increased , while the negative regulatory dose-dependence of DB00171 was unchanged in O60266 -I compared to WT and O60266 -C ventricular myocytes , suggesting that CaMKII inhibition increased the number of functional I(KATP) channels available for IP . We measured increased sarcolemmal Kir6.2 , a pore-forming I(KATP) subunit , but not a change in total Kir6.2 in cell lysates or single channel I(KATP) opening probability from O60266 -I compared to WT and O60266 -C ventricles , showing CaMKII inhibition increased sarcolemmal I(KATP) channel expression . There were no differences in mRNA for genes encoding I(KATP) channel subunits in O60266 -I , WT and O60266 -C ventricles . The I(KATP) opener pinacidil ( 100 microM ) reduced MI area in WT to match O60266 -I hearts , while the I(KATP) antagonist HMR1098 ( 30 microM ) increased MI area to an equivalent level in all groups , indicating that increased I(KATP) and augmented IP are important for reduced ischemic cell death in O60266 -I hearts . Our study results show CaMKII inhibition enhances beneficial effects of IP by increasing I(KATP) . Cholecystokinin ( CCK ) stimulates aldosterone secretion from human adrenocortical cells via CCK2 receptors coupled to the adenylate cyclase/protein kinase A signaling cascade . Cholecystokinin ( CCK ) IS a regulatory peptide that acts via two receptor subtypes , P32238 and P32239 . RT-PCR demonstrated the expression of both P32238 and P32239 in the zona glomerulosa ( ZG ) , but not zona fasciculata-reticularis cells of the human adrenal cortex . CCK and the P32239 agonist pentagastrin enhanced basal aldosterone secretion from ZG cells without affecting cortisol production from zona fasciculata-reticularis cells . The aldosterone response to CCK and pentagastrin was suppressed by a P32239 antagonist , but not by a P32238 antagonist . DB00183 evoked a sizeable DB02527 , but not inositol triphosphate , response from ZG cells , whereas CCK plus P32239 antagonist was ineffective . The DB02527 response to pentagastrin was abrogated by P32239 antagonist or the adenylate cyclase inhibitor SQ-22536 , and the aldosterone response was abolished by both SQ-22536 and the protein kinase A inhibitor H-89 . Both CCK and pentagastrin increased steroidogenic acute regulatory protein mRNA expression in ZG cells ; the effect was abrogated by P32239 antagonist . We conclude that CCK exerts secretagogue action on human ZG cells , acting through CCK2-Rs coupled to the adenylate cyclase/protein kinase A signaling cascade , which , in turn , stimulates the expression of steroidogenic acute regulatory protein , the rate-limiting step of steroidogenesis . Effects of LSD on Ca++ currents in central 5-HT-containing neurons : P08908 receptors may play a role in hallucinogenesis . Drugs that influence the activity of central serotonergic neurons by activating a 5-hydroxytryptamine subtype of receptor ( P08908 ) alter mood and perception . Previously , we demonstrated with whole-cell recordings from acutely isolated 5-HT-containing dorsal raphe ( DR ) neurons from the adult rat that 5-HT inhibited Ca++ current and activated K+ current in DR neurons . We now show that DB04829 ( LSD ) mimics the actions of 5-HT ; it dramatically suppresses Ca++ current in a dose-dependent manner and activates an inwardly rectifying K+ conductance . Spiperone ( 0.2 microM ) , a P08908 /5-HT2 antagonist , blocks the effect of both LSD and 5-HT . The nonhallucinogenic structural analog 2-bromo-LSD ( 2-Bol ) at 10 microM has no effect on either Ca++ or K+ current by itself , but it competitively antagonizes both effects of LSD . Inhibition of 5-HT release resulting from P08908 receptor activation may play an integral role in the hallucinogenic actions of LSD by reducing competition between 5-HT and LSD for the postsynaptic 5-HT receptors . Molecular mechanisms governing different pharmacokinetics of ginsenosides and potential for ginsenoside-perpetrated herb-drug interactions on Q9NPD5 . BACKGROUND AND PURPOSE : Ginsenosides are bioactive saponins derived from Panax notoginseng roots ( Sanqi ) and ginseng . Here , the molecular mechanisms governing differential pharmacokinetics of 20(S)-protopanaxatriol-type ginsenoside Rg1 , ginsenoside Re and notoginsenoside Q96GN5 and 20(S)-protopanaxadiol-type ginsenosides Rb1 , Rc and Rd were elucidated . EXPERIMENTAL APPROACH : Interactions of ginsenosides with human and rat hepatobiliary transporters were characterized at the cellular and vesicular levels . A rifampin-based inhibition study in rats evaluated the in vivo role of organic anion-transporting polypeptide (Oatp)1b2 . Plasma protein binding was assessed by equilibrium dialysis . Drug-drug interaction indices were calculated to estimate potential for clinically relevant ginsenoside-mediated interactions due to inhibition of human OATP1Bs . KEY RESULTS : All the ginsenosides were bound to human Q9NPD5 and rat Oatp1b2 but only the 20(S)-protopanaxatriol-type ginsenosides were transported . Human multidrug resistance-associated protein (MRP)2/breast cancer resistance protein ( Q9UNQ0 ) /bile salt export pump ( O95342 ) /multidrug resistance protein-1 and rat Mrp2/Bcrp/Bsep also mediated the transport of the 20(S)-protopanaxatriol-type ginsenosides . Glomerular-filtration-based renal excretion of the 20(S)-protopanaxatriol-type ginsenosides was greater than that of the 20(S)-protopanaxadiol-type counterparts due to differences in plasma protein binding . DB01045 -impaired hepatobiliary excretion of the 20(S)-protopanaxatriol-type ginsenosides was effectively compensated by the renal excretion in rats . The 20(S)-protopanaxadiol-type ginsenosides were potent inhibitors of Q9NPD5 . CONCLUSION AND IMPLICATIONS : Differences in hepatobiliary and in renal excretory clearances caused markedly different systemic exposure and different elimination kinetics between the two types of ginsenosides . Caution should be exercised with the long-circulating 20(S)-protopanaxadiol-type ginsenosides as they could induce hepatobiliary herb-drug interactions , particularly when patients receive long-term therapies with high-dose i.v. Sanqi or ginseng extracts . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling . Effects of DB01454 on Extracellular Dopamine and Serotonin Levels in Mice Lacking Dopamine and/or Serotonin Transporters . 3,4-Methylendioxymethamphetamine ( DB01454 ) has both stimulatory and hallucinogenic properties which make its psychoactive effects unique and different from those of typical psychostimulant and hallucinogenic agents . The present study investigated the effects of DB01454 on extracellular dopamine ( DA(ex) ) and serotonin ( 5-HT(ex) ) levels in the striatum and prefrontal cortex ( P27918 ) using in vivo microdialysis techniques in mice lacking DA transporters ( Q01959 ) and/or 5-HT transporters ( P31645 ). subcutaneous injection of DB01454 ( 3 , 10 mg/kg ) significantly increased striatal DA(ex) in wild-type mice , P31645 knockout mice , and Q01959 knockout mice , but not in Q01959 / P31645 double-knockout mice . The DB01454 -induced increase in striatal DA(ex) in P31645 knockout mice was significantly less than in wildtype mice . In the P27918 , DB01454 dose-dependently increased DA(ex) levels in wildtype , Q01959 knockout , P31645 knockout and Q01959 / P31645 double-knockout mice to a similar extent . In contrast , DB01454 markedly increased 5-HT(ex) in wildtype and Q01959 knockout mice and slightly increased 5-HT(ex) in P31645 -KO and Q01959 / P31645 double-knockout mice . The results confirm that DB01454 acts at both Q01959 and P31645 and increases DA(ex) and 5-HT(ex) . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . The PEPvIII-KLH ( DB05374 ) vaccine in glioblastoma multiforme patients . Conventional therapies for glioblastoma multiforme ( GBM ) fail to target tumor cells exclusively , resulting in non-specific toxicity . Immune targeting of tumor-specific mutations may allow for more precise eradication of neoplastic cells . P00533 variant III ( EGFRvIII ) is a tumor-specific mutation that is widely expressed in GBM and other neoplasms and its expression enhances tumorigenicity . This in-frame deletion mutation splits a codon , resulting in a novel glycine at the fusion junction producing a tumor-specific epitope target for cellular or humoral immunotherapy . We have previously shown that vaccination with a peptide that spans the EGFRvIII fusion junction ( PEPvIII-KLH/ DB05374 ) is an efficacious immunotherapy in syngeneic murine models . In this review , we summarize our results in GBM patients targeting this mutation in multiple , multi-institutional Phase II immunotherapy trials . These trials demonstrated that a selected population of GBM patients who received vaccines targeting EGFRvIII had an unexpectedly long survival time . Further therapeutic strategies and potential pitfalls of using this approach are discussed . Diabetes susceptibility in Mayas : Evidence for the involvement of polymorphisms in Q03014 , HNF4α , Q14654 , PPARγ , CDKN2A/2B , Q8IWU4 , O75794 / Q8IU85 , Q9NQB0 , O95477 and Q8NCK7 genes . Association of type 2 diabetes ( T2D ) with common variants in Q03014 , HNF4α , Q14654 , PPARγ , CDKN2A/2B , Q8IWU4 , O75794 / Q8IU85 , Q9NQB0 , O95477 and Q8NCK7 genes have been reported , mainly in populations of European and Asian ancestry and to a lesser extent in Latin Americans . Thus , we aimed to investigate the contribution of rs1111875 ( Q03014 ) , rs1800961 ( HNF4α ) , rs5219 ( Q14654 ) , rs1801282 ( PPARγ ) , rs10811661 ( CDKN2A/2B ) , rs13266634 ( Q8IWU4 ) , rs12779790 ( O75794 / Q8IU85 ) , rs7903146 ( Q9NQB0 ) , rs9282541 ( O95477 ) and rs13342692 ( Q8NCK7 ) polymorphisms in the genetic background of Maya population to associate their susceptibility to develop T2D . This is one of the first studies designed specifically to investigate the inherited component of T2D in the indigenous population of Mexico . SNPs were genotyped by allelic discrimination method in 575 unrelated Maya individuals . Two SNPs rs10811661 and rs928254 were significantly associated with T2D after adjusting for BMI ; rs10811661 in a recessive and rs9282541 in a dominant model . Additionally , we found phenotypical alterations associated with genetic variants : HDL to rs9282541 and insulin to rs13342692 . In conclusion , these findings support an association of genetic polymorphisms to develop T2D in Maya population .

MH_train_1251

MH_train_1251

MH_train_1251

interacts_with DB08910?

multiple_choice

Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 ( Q16739 ) is an approved drug target for the treatment of Gaucher disease and is considered as a valid target for combating other human pathologies , including type 2 diabetes . The clinical drug N-butyldeoxynojirimycin ( DB00419 ) is thought to inhibit through mimicry of its substrate , ceramide . In this work we demonstrate that , in contrast to what is proposed in this model , the P06681 -hydroxyl of the deoxynojirimycin core is important for Q16739 inhibition . Here we show that P13671 -OH appears of less important , which may set guidelines for the development of Q16739 inhibitors that have less affinity ( in comparison with DB00419 ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide . DB06016 ( RGH-188 ) , a potent D3/D2 dopamine receptor partial agonist , binds to dopamine D3 receptors in vivo and shows antipsychotic-like and procognitive effects in rodents . We investigated the in vivo effects of orally administered cariprazine ( RGH-188 ; trans-N-{4-[2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-ethyl]-cyclohexyl}-N',N'-dimethyl-urea ) , a D(3)/ P14416 partial agonist with ∼10-fold preference for the D(3) receptor . Oral bioavailability of cariprazine at a dose of 1mg/kg in rats was 52 % with peak plasma concentrations of 91ng/mL . DB06016 10mg/kg had good blood-brain barrier penetration , with a brain/plasma AUC ratio of 7.6:1 . In rats , cariprazine showed dose-dependent in vivo displacement of [ (3)H ] (+)-PHNO , a dopamine D(3) receptor-preferring radiotracer , in the D(3) receptor-rich region of cerebellar lobules 9 and 10 . Its potent inhibition of apomorphine-induced climbing in mice ( ED(50)=0.27mg/kg ) was sustained for 8h . DB06016 blocked amphetamine-induced hyperactivity ( ED(50)=0.12mg/kg ) and conditioned avoidance response ( CAR ) ( ED(50)=0.84mg/kg ) in rats , and inhibited the locomotor-stimulating effects of the noncompetitive DB01221 antagonists MK-801 ( ED(50)=0.049mg/kg ) and phencyclidine ( ED(50)=0.09mg/kg ) in mice and rats , respectively . It reduced novelty-induced motor activity of mice ( ED(50)=0.11mg/kg ) and rats ( ED(50)=0.18mg/kg ) with a maximal effect of 70 % in both species . DB06016 produced no catalepsy in rats at up to 100-fold dose of its CAR inhibitory ED(50) value . DB06016 0.02-0.08mg/kg significantly improved the learning performance of scopolamine-treated rats in a water-labyrinth learning paradigm . Though risperidone , olanzapine , and aripiprazole showed antipsychotic-like activity in many of these assays , they were less active against phencyclidine and more cataleptogenic than cariprazine , and had no significant effect in the learning task . The distinct in vivo profile of cariprazine may be due to its higher affinity and in vivo binding to D(3) receptors versus currently marketed typical and atypical antipsychotics . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Integrative gene network analysis provides novel regulatory relationships , genetic contributions and susceptible targets in autism spectrum disorders . Autism spectrum disorders ( ASDs ) are a group of diseases exhibiting impairment in social drive , communication/language skills and stereotyped behaviors . Though an increased number of candidate genes and molecular interactions have been identified by various approaches , the pathogenesis remains elusive . Based on clinical observations , data from accessible GWAS and expression datasets we identified ASDs gene candidates . Integrative gene network and a novel CNV-centric Node Network ( CNN ) analysis method highlighted ASDs-associated key elements and biological processes . Functional analysis identified neurological functions including synaptic cholinergic receptor ( CHRNA ) families , dopamine receptor ( P14416 ) , and correlations between social behavior and oxytocin related pathways . CNN analysis of genome-wide genetic and expression data identified inheritance-related clusters related to P60484 / Q92574 / Q06787 and mTor/PI3K regulation . Integrative analysis identified potential regulators of networks , specifically P01375 and beta-estradiol , suggesting a potential central role in ASDs . Our data provide information on potential disease mechanisms , and key regulators that may generate novel postulations , and diagnostic molecular biomarkers . Enhanced expression of heat shock protein 70 ( hsp70 ) and heat shock factor 1 ( Q00613 ) activation in rheumatoid arthritis synovial tissue . Differential regulation of hsp70 expression and hsf1 activation in synovial fibroblasts by proinflammatory cytokines , shear stress , and antiinflammatory drugs . Heat shock proteins ( hsp ) have been repeatedly implicated to participate in the pathogenesis of rheumatoid arthritis ( RA ) . Herein , we investigated the regulation of synovial hsp70 expression by analyzing the DNA-binding activity of heat shock transcription factor 1 ( Q00613 ) as well as inducible hsp70 expression . Experiments were performed both on synovial tissue and on synovial fibroblast-like cells ( SFC ) . Gel mobility shift analysis revealed increased Q00613 activation , and Western blotting and immunohistochemistry revealed increased hsp70 expression in RA synovial tissue , but not in synovial tissue derived from patients with osteoarthritis . Proinflammatory cytokines ( P01375 , IL-1alpha , P05231 ) , but not P01579 or TGF-beta , induced activation of Q00613 -DNA binding and hsp70 expression in cultivated SFC . Activation of Q00613 in SFC was accompanied by hyperphosphorylation and nuclear translocation of Q00613 . Furthermore , shear stress also induced a complete heat shock response in cultivated synovial cells . In contrast , nonsteroidal antiinflammatory drugs triggered only an incomplete heat shock response , with Q00613 activation but not hsp70 induction , whereas steroids and immunosuppressive drugs did not affect the heat shock response at all . In summary , these data suggest that induction of hsp70 expression in rheumatoid synovial tissue is based on transcriptional activation of Q00613 due to the presence of proinflammatory cytokines ( and possibly also shear stress ) . P01375 -like weak inducer of apoptosis ( O43508 ) activates proinflammatory signaling pathways and gene expression through the activation of O43318 . O43508 , P01375 -like weak inducer of apoptosis , is a relatively recently identified proinflammatory cytokine that functions through binding to Fn14 receptor in target cells . Although O43508 has been shown to modulate several biological responses , the O43508 -induced signaling pathways remain poorly understood . In this study , we tested the hypothesis that TAK1 ( O43318 ) is involved in O43508 -induced activation of NF-kappaB and MAPK and expression of proinflammatory protein . O43508 increased the phosphorylation and kinase activity of TAK1 in cultured myoblast and fibroblast cells . The activation of NF-kappaB was significantly inhibited in TAK1-deficient ( TAK1(-/-) ) mouse embryonic fibroblasts ( MEF ) compared with wild-type MEF . Deficiency of TAK1 also inhibited the O43508 -induced activation of O15111 and the phosphorylation and degradation of P25963 protein . However , there was no difference in the levels of Q9ULW0 protein in O43508 -treated wild-type and TAK1(-/-) MEF . Furthermore , O43508 -induced transcriptional activation of NF-kappaB was significantly reduced in TAK1(-/-) MEF and in C2C12 myoblasts transfected with a dominant-negative TAK1 or TAK1 short interfering RNA . TAK1 was also required for the activation of AP-1 in response to O43508 . Activation of P45983 and p38 MAPK , but not P27361 /2 or Akt kinase , was significantly inhibited in TAK1(-/-) MEF compared with wild-type MEF upon treatment with O43508 . O43508 -induced expression of proinflammatory genes such as P14780 , DB00833 -2 , and P19320 was also reduced in TAK1(-/-) MEF compared with wild-type MEF . Furthermore , the activation of NF-kappaB and the expression of P14780 in response to O43508 involved the upstream activation of Akt kinase . Collectively , our study demonstrates that TAK1 and Akt are the important components of O43508 -induced proinflammatory signaling and gene expression . Anti-tumor effect of rutin on human neuroblastoma cell lines through inducing G2/M cell cycle arrest and promoting apoptosis . AIMS : To further investigate the antineuroblastoma effect of rutin which is a type of flavonoid . METHODS : The antiproliferation of rutin in human neuroblastoma cells LAN-5 were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ( MTT ) assay . Chemotaxis of LAN-5 cells was assessed using transwell migration chambers and scratch wound migration assay . The cell cycle arrest and apoptosis in a dose-dependent manner was measured by flow cytometric and fluorescent microscopy analyses . The apoptosis-related proteins Q07812 and P10415 as well as P04198 mRNA express were determined by RT-PCR analysis . Secreted P01375 - α level were determined using specific enzyme-linked immunosorbent assay kits . RESULTS : DB01698 significantly inhibited the growth of LAN-5 cells and chemotactic ability . Flow cytometric analysis revealed that rutin induced G2/M arrest in the cell cycle progression and induced cell apoptosis . The RT-PCR showed that rutin could decrease P10415 expression and P10415 / Q07812 ratio . In the meantime , the P04198 mRNA level and the secretion of P01375 - α were inhibited . CONCLUSION : These results suggest that rutin produces obvious antineuroblastoma effects via induced G2/M arrest in the cell cycle progression and induced cell apoptosis as well as regulating the expression of gene related to apoptosis and so on . It supports the viability of developing rutin as a novel therapeutic prodrug for neuroblastoma treatment , as well as providing a new path on anticancer effect of Chinese traditional drug . Antiemetic and motor-depressive actions of CP55,940 : cannabinoid P21554 receptor characterization , distribution , and G-protein activation . Dibenzopyran ( Delta(9)-tetrahydrocannabinol ) and aminoalkylindole [ R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrolol[1,2,3-de]-1,4-benzoxazin-yl]-(1-naphthalenyl) methanone mesylate ; ( WIN55,212-2 ) ] cannabinoids suppress vomiting produced by cisplatin via cannabinoid CB(1) receptors . This study investigates the antiemetic potential of the " nonclassical " cannabinoid CP55,940 [ 1alpha,2beta-(R)-5alpha ] -(-)-5-(1,1-dimethyl)-2- [ 5-hydroxy-2-(3-hydroxypropyl) cyclohexyl-phenol ] against cisplatin-induced vomiting and assesses the presence and functionality of cannabinoid CB(1) receptors in the least shrew ( Cryptotis parva ) brain . CP55,940 ( 0.025-0.3 mg/kg ) reduced both the frequency of cisplatin-induced emesis ( ID(50)=0.025 mg/kg ) and the percentage of shrews vomiting ( ID(50)=0.09 mg/kg ) . CP55,940 also suppressed shrew motor behaviors ( ID(50)=0.06- 0.21 mg/kg ) at such doses . The antiemetic and motor-suppressant actions of CP55,940 were countered by SR141716A [ N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)- DB01213 -3-carboxamide ] , indicating both effects are cannabinoid CB(1) receptor-mediated . Autoradiographic studies with [ 3H ] -SR141716A and [ 35S ] -GTPgammaS binding revealed that the distribution of the cannabinoid CB(1) receptor and its activation pattern are similar to rodent brain and significant levels are present in brain loci ( e.g. , nucleus tractus solitarius ( P30990 ) ) that control emesis . The affinity rank order of structurally diverse cannabinoid ligands for cannabinoid CB(1) receptor in shrew brain is similar to rodent brain : HU-210=CP55,940=SR141716A >/= WIN55,212-2 >/= DB00470 > methanandamide=HU-211=cannabidiol=2-arachidonoylglycerol . This affinity order is also similar and is highly correlated to the cannabinoid EC(50) potency rank order for GTPgammaS stimulation except WIN55,212-2 and DB00470 potency order were reversed . The affinity and the potency rank order of tested cannabinoids were significantly correlated with their antiemetic ID(50) potency order against cisplatin-induced vomiting ( CP55,940 > WIN55,212-2= DB00470 ) as well as emesis produced by 2-arachidonoylglycerol or SR141716A ( CP55,940 > WIN55,212-2 > DB00470 ) . c-Jun activation-dependent tumorigenic transformation induced paradoxically by overexpression or block of S-adenosylmethionine decarboxylase . All mammalian cells absolutely require polyamines ( putrescine , spermidine , and spermine ) for growth . Here we show that the overexpression of cDNA for S-adenosylmethionine decarboxylase ( P17707 ) , the main regulatory enzyme in the biosynthesis of higher polyamines , induces transformation of rodent fibroblasts when expressed in the sense or the antisense orientation . Both transformants were able to induce invasive tumors in nude mice . Neither transformation was associated with activation of the mitogen-activated protein kinases Erk1 and Erk2 . Instead , the DB00118 DC sense , but not antisense , transformants displayed constitutive activation of the c-Jun NH(2)-terminal kinase ( JNK ) pathway . However , both transformations converged on persistent phosphorylation of endogenous c-Jun at Ser73 . The phenotype of the P17707 sense transformants was reversed by expression of dominant-negative mutants of P45985 ( P45985 ) , P45983 , and c-Jun ( TAM-67 ) , which were also found to impair cytokinesis . Similarly , TAM-67 reverted the morphology of the P17707 -antisense expressors . This report is the first demonstration of a protein whose overexpression or block of synthesis can induce cell transformation . In addition , we show that the polyamine biosynthetic enzymes require c-Jun activation for eliciting their biological effects . Upregulation of intercellular adhesion molecule-1 expression on human endothelial cells by tumour necrosis factor-alpha in an in vitro model of the blood-brain barrier . Adhesion molecules on the endothelial surface of the blood-brain barrier ( BBB ) play an important role in the pathogenesis of many encephalopathies , including multiple sclerosis ( MS ) and cerebral malaria ( CM ) . The expression of four surface molecules of relevance to MS and CM on the immortalized human umbilical vein endothelial cell line , ECV304 , was investigated using immunofluorescence flow cytometry . We found that ECV304 cells express intercellular adhesion molecule-1 ( P05362 ) and low levels of P16671 , but not vascular cell adhesion molecule-1 ( P19320 ) or P16581 . This expression pattern was unaltered on ECV304 cells which were co-cultured with P13671 glioma cells ; conditions under which the endothelial cells display enhanced barrier formation . Tumour necrosis factor-alpha ( P01375 ) , which is elevated in MS and CM , decreased the integrity of the barrier in co-cultured endothelial cells and upregulated the expression of P05362 nine-fold . The significance of elevated P05362 expression in relation to the binding of parasitised erythrocytes at the BBB in CM is discussed . A microRNA screen to identify modulators of sensitivity to P10415 inhibitor DB05764 ( navitoclax ) . Evasion of apoptosis is a known feature of cancer cells . One mechanism of deregulating the apoptotic pathway is through overexpression of antiapoptotic P10415 family members . DB05764 ( navitoclax ) is a first-in-class P10415 family inhibitor that restores the ability of cancer cells to undergo apoptosis . However , many cancer cells are resistant to DB05764 due to high levels of a P10415 family member , Q07820 , which is not targeted by the drug . Q07820 expression is regulated transcriptionally , translationally , and through proteasome-mediated degradation . Recently , Q07820 expression was shown to be affected by microRNAs ( miRNA ) . To identify miRNAs that modulate the sensitivity of cancer cells to DB05764 , we screened a library of 810 human miRNA mimics in HCT-116 cells in the presence of DB05764 . The screen revealed 19 miRNAs that sensitize HCT-116 cells to DB05764 . Fifteen of these miRNAs were also shown to sensitize CHL1 melanoma cells to the same agent . We further evaluated 12 of the strongest sensitizers in these cell lines . We found that these sensitizers induced apoptosis only in the presence of DB05764 . In addition , whereas all 12 of these miRNAs reduced Q07820 protein expression , only 10 of them targeted Q07820 through direct binding to the 3'-untranslated region of the gene , raising the possibility that other resistance regulators of Q07820 expression may be identified using our method . Finally , because sensitizing miRNA expression is lower in tumors compared with normal tissues , our data can facilitate the design of miRNA replacement therapies to increase sensitivity to P10415 antagonists . Suppressive effects of glucagon-like peptide-1 on interferon-gamma-induced nitric oxide production in insulin-producing cells is mediated by inhibition of tumor necrosis factor-alpha production . During the development of Type 1 diabetes , inflammatory cytokines are known to induce the expression of inducible nitric oxide synthase ( P35228 ) in pancreatic islets , and subsequent production of nitric oxide ( NO ) contributes to beta cell destruction . Glucagon-like peptide-1 ( P0C6A0 ) has been shown to reduce cytokine-induced apoptosis of beta cells . In this study , we investigated whether P0C6A0 affects cytokine-induced NO production , resulting in the inhibition of beta-cell apoptosis . We treated MIN6N8a mouse beta cells with interferon ( IFN ) -gamma in the presence or absence of P0C6A0 and found that P01579 treatment induced P35228 mRNA expression and NO production , which was significantly inhibited by treatment with P0C6A0 . Blocking of P43220 signaling via the cyclic AMP and phosphatidylinositol 3-kinase pathway did not directly affect the suppressive effect of P0C6A0 on IFN- gamma-induced P35228 mRNA expression . Further studies revealed that P01579 induced the expression of P01375 mRNA and protein , which synergistically induced NO production , and P0C6A0 treatment inhibited this induction of P01375 . To examine whether the reduction of P01375 by P0C6A0 treatment plays a role in suppressing NO production , we treated MIN6N8a cells with P01579 in the presence of anti- P01375 neutralizing antibody and found that NO production was reduced . In addition , treatment of mouse islets with P0C6A0 inhibited the expression of P35228 and TNFmRNA . These results suggest that P0C6A0 inhibits P01579 -induced NO production by suppression of P01375 production . Novel 3,4-diarylpyrazolines as potent cannabinoid P21554 receptor antagonists with lower lipophilicity . Novel 3,4-diarylpyrazolines 1 as potent P21554 receptor antagonists with lipophilicity lower than that of DB05077 are described . The key change is the replacement of the arylsulfonyl group in the original series by a dialkylaminosulfonyl moiety . The absolute configuration ( 4S ) of eutomer 24 was established by X-ray diffraction analysis and 24 showed a close molecular fit with rimonabant in a P21554 receptor-based model . Compound 17 exhibited the highest P21554 receptor affinity ( Ki = 24 nM ) in this series , as well as very potent P21554 antagonistic activity ( pA2 = 8.8 ) and a high P21554 /CB2 subtype selectivity ( approximately 147-fold ) . DB08626 -induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer 's disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 and cortical Abeta40 that were 147 and 142 % of the control group , respectively . Results for Abeta42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD . DB03758 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 kDa ( HSP90 ) . HSP90 is the main component of the cytosolic molecular chaperone complex , which has been implicated in the regulation of the heat shock factor 1 ( Q00613 ) . Q00613 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 HSP90 in cardiomyocytes . These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection . Therefore , compounds , such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents . Effect of liraglutide on proliferation and differentiation of human adipose stem cells . Glucagon-Like Peptide-1 ( P0C6A0 ) receptor agonists , used as glucose-lowering drugs , also induce weight loss by inhibiting food intake . The present study was aimed at the assessment of the in vitro effects of the P43220 agonist liraglutide on proliferation and differentiation of human adipose stem cells ( ASC ) obtained from subcutaneous adipose tissue of morbidly obese subjects undergoing bariatric surgery . DB06655 ( 10-100 nM ) significantly inhibited ASC proliferation and viability , with a maximum effect at 6 days of culture ( 45 % and 50 % , for liraglutide 10 and 100 nM , respectively ) ; the effect was reverted by exendin 9-39 . DB09341 uptake was significantly reduced by liraglutide in a dose dependent manner . Treatment with liraglutide reduced intracellular lipid accumulation in differentiating ASC , together with FABP-4 mRNA expression ( -18 % , -23 % , -46 % , for 1 nM , 10 nM and 100 nM , respectively ) , whereas it stimulated adiponectin ( P15144 ) expression ( 1.86- , 2.64- , 2.28-fold increase , for 1 nM , 10 nM and 100 nM , respectively ) . DB06655 exerts effects on human adipose cell precursors , inhibiting proliferation and differentiation , while stimulating the expression of the insulin-sensitizing adipokine P15144 . These effects could contribute to the actions of P43220 agonists on body weight and insulin sensitivity . Central cannabinoid 1 receptor antagonist administration prevents endotoxic hypotension affecting norepinephrine release in the preoptic anterior hypothalamic area . It is widely assumed that LPS lowers arterial pressure during sepsis by stimulating release of P01375 and other vasoactive mediators from macrophages . However , recent data from this and other laboratories have shown that LPS hypotension can be prevented by inhibiting afferent impulse flow in the vagus nerve , by blocking neuronal activity in the nucleus of the solitary tract , or by blocking alpha-adrenergic receptors in the preoptic area/anterior hypothalamic area ( POA ) . These findings suggest that the inflammatory signal is conveyed from the periphery to the brain via the vagus nerve , and that endotoxic shock is mediated through a central mechanism that requires activation of POA neurons . In the present study , we tested whether central cannabinoid 1 ( P21554 ) receptors participate in the control of arterial pressure during endotoxemia based on evidence that hypothalamic neurons express P21554 receptors and synthesize the endogenous CB anandamide . We found that intracerebroventricular administration of rimonabant , a P21554 receptor antagonist , inhibited the fall in arterial pressure evoked by LPS significantly in both conscious and anesthetized rats . DB06155 attenuated both the immediate fall in arterial pressure evoked by LPS and the second , delayed hypotensive phase that leads to tissue ischemia and death . DB06155 also prevented the associated LPS-induced rise in extracellular fluid norepinephrine concentrations in the POA . Furthermore , rimonabant attenuated the associated increase in plasma P01375 concentrations characteristic of the late phase of endotoxic hypotension . These data indicate that central P21554 receptors may play an important role in the initiation of endotoxic hypotension .

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[ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . Phosphorylated c-Mpl tyrosine 591 regulates thrombopoietin-induced signaling . P40225 ( P07202 ) is the primary regulator of platelet production , affecting cell survival , proliferation , and differentiation through binding to and stimulation of the cell surface receptor the cellular myeloproliferative leukemia virus oncogene ( c-Mpl ) . Activating mutations in c-Mpl constitutively stimulate downstream signaling pathways , leading to aberrant hematopoiesis , and contribute to development of myeloproliferative neoplasms . Several studies have mapped the tyrosine residues within the cytoplasmic domain of c-Mpl that mediate these cellular signals ; however , secondary signaling pathways are incompletely understood . In this study , we focused on c-Mpl tyrosine 591 ( Y591 ) . We found Y591 of wild-type c-Mpl to be phosphorylated in the presence of P07202 . Additionally , eliminating Y591 phosphorylation by mutation to DB00120 resulted in decreased total receptor phosphorylation . Using a Src homology 2/phosphotyrosine-binding ( SH2/PTB ) domain binding microarray , we identified novel c-Mpl binding partners for phosphorylated Y591 , including Src homology region 2 domain-containing phosphatase-1 ( Q15466 -1 ) , spleen tyrosine kinase ( P43405 ) and Bruton 's tyrosine kinase ( Q06187 ) . The functional significance of binding partners was determined through small interfering RNA treatment of Ba/ P13726 -Mpl cells , confirming that the increase in pERK1/2 resulting from removal of Y591 may be mediated by spleen tyrosine kinase . These findings identify a novel negative regulatory pathway that controls P07202 -mediated signaling , advancing our understanding of the mechanisms required for successful maintenance of hematopoietic stem cells and megakaryocyte development . Induction of lymphokine-activated cytotoxic T lymphocytes stimulated by dendritic cells and autologous tumor from a patient with gastric cancer and their effects in vitro . BACKGROUND/AIMS : The purpose of the study was to generate lymphokine-activated cytotoxic T lymphocytes stimulated by dendritic cells ( DC ) and autologous tumor from a patient with gastric cancer and to clarify their cytotoxic effects in vitro . METHODOLOGY : DC was induced by interleukin-4 ( P05112 ) and granulocyte-macrophage-colony-stimulating factor ( GM- P04141 ) from the peripheral blood mononuclear cells ( PBMC ) . Then , PBMC was incubated with mitomycin C-treated tumor cells and DC , and following that was activated with P60568 and anti-CD3 . Induction of DC and cytotoxic T cells ( CTL ) were confirmed by the analyses of the cell surface antigens , killing activities , and blocking tests . RESULTS : Induction of DC and cytotoxic T cells ( CTL ) was confirmed by the analyses of the cell surface antigens , killing activities , and blocking tests . In vitro study demonstrated that lymphokine-activated lymphocytes pulsed by DCs and autologous tumor contained the largest population of CTLs , the greatest production of P01579 , and the greatest Q06187 activity . CONCLUSIONS : Those results indicated that CTLs could be generated in vitro from a patient with gastric cancer more successfully by this method than by conventional methods , suggesting the possibility of a new immunotherapy for the treatment of gastric cancer . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Role of nitric oxide ( NO ) in pulmonary dysfunction associated with experimental cirrhosis . We examined the functional role of nitric oxide ( NO ) and nitric oxide synthase ( NOS ) isoforms in the pulmonary dysfunction seen in cirrhosis . Lungs were isolated from control and carbon tetrachloride ( CCl(4) ) -induced cirrhotic rats and perfused at constant flow with a whole blood mixture . Ventilation with hypoxic gas resulted in attenuated hypoxic pulmonary vasoconstriction ( HPV ) in lungs from cirrhotic animals . Administration of the non-selective NOS inhibitor N-omega-Nitro-L- DB00125 ( DB04223 ) resulted in HPV responses that were not different between groups . However , inhibition of inducible nitric oxide synthase ( P35228 ) did not restore cirrhotic HPV responses . Lungs from cirrhotic rats demonstrated enhanced endothelial-dependent vasodilation to vasopressin when preconstricted with hypoxia but not when preconstricted with thromboxane mimetic . Western blot analysis failed to demonstrate differences in pulmonary endothelial NOS ( P29474 ) or P35228 levels between groups . Our data suggest that , while NO may play a role in mediating the reduced pulmonary vasoreactivity observed in cirrhosis , other vasoactive factors are likely also important modulators of the pulmonary dysfunction seen in this disease . Molecular genetics of the DB02527 -dependent protein kinase pathway and of sporadic pituitary tumorigenesis . Pituitary tumors are among the most common human neoplasms . Although these common lesions rarely become clinically manifest and they are almost never malignant , they are the cause of significant morbidity in affected patients . The genetic causes of common pituitary tumors remain for the most part unknown ; progress has been limited to the elucidation of the molecular etiology of four genetic syndromes predisposing to pituitary neoplasias : McCune-Albright syndrome , multiple endocrine neoplasia type 1 , Carney complex and , most recently , familial acromegaly and prolactinomas and other tumors caused by mutations in the GNAS , menin , P10644 , AIP , and p27 ( P46527 ) genes , respectively . Intense molecular studies of sporadic pituitary tumors from patients with negative family histories and no other neoplasms have yielded interesting findings with abnormalities in growth factor expression and cell cycle control dysregulation . To add to the difficulties in understanding pituitary tumorigenesis in man , good murine models of these neoplasms simply do not exist : pituitary tumors are common in rodents , but their histologic origin ( mostly from the intermediate lobe ) , age of presentation ( late in murine life ) and clinical course make them hardly models of their human counterparts . The present report reviews the clinical and molecular genetics of the DB02527 -dependent protein kinase pathway in human pituitary tumors ; it also reviews briefly other pathways that have been involved in sporadic pituitary neoplasms . At the end , we attempt a unifying hypothesis for pituitary tumorigenesis , taking into account data that are also discussed elsewhere in this issue . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . UMD ( Universal mutation database ) : a generic software to build and analyze locus-specific databases . The human genome is thought to contain about 80,000 genes and presently only 3,000 are known to be implicated in genetic diseases . In the near future , the entire sequence of the human genome will be available and the development of new methods for point mutation detection will lead to a huge increase in the identification of genes and their mutations associated with genetic diseases as well as cancers , which is growing in frequency in industrial states . The collection of these mutations will be critical for researchers and clinicians to establish genotype/phenotype correlations . Other fields such as molecular epidemiology will also be developed using these new data . Consequently , the future lies not in simple repositories of locus-specific mutations but in dynamic databases linked to various computerized tools for their analysis and that can be directly queried on-line . To meet this goal , we devised a generic software called UMD ( Universal Mutation Database ) . It was developed as a generic software to create locus-specific databases ( LSDBs ) with the 4(th) Dimension(R) package from ACI . This software includes an optimized structure to assist and secure data entry and to allow the input of various clinical data . Thanks to the flexible structure of the UMD software , it has been successfully adapted to nine genes either involved in cancer ( P25054 , P04637 , P06400 , O00255 , Q09428 , P40337 , and P19544 ) or in genetic diseases ( P35555 and P01130 ) . Four new LSDBs are under construction ( P49748 , P11310 , KIR6 , and P29400 ) . Finally , the data can be transferred to core databases . DB00174 Synthetase Deficiency : New Inborn Errors of Metabolism . BACKGROUND : DB00174 synthetase deficiency ( P51689 ) is a newly identified neurometabolic disorder characterized by severe congenital microcephaly , severe global developmental delay , intractable seizure disorder , and spastic quadriplegia . Brain Q9BWK5 showed brain atrophy , delayed myelination , and simplified gyriform pattern . METHODS : We report P51689 deficiency in a 2- and 4-year-old sibling . On them , we described clinical , biochemical , and molecular findings , and we compared our results with previously reported cases . RESULTS : We identified a homozygous novel missense mutation in P08243 gene in both probands and we demonstrated low P04141 and plasma asparagine in both patients . CONCLUSIONS : Clinicians should suspect P51689 deficiency in any newborn presented with severe congenital microcephaly followed by severe epileptic encephalopathy and global developmental delay . P04141 asparagine level is low in this disorder while plasma may be low . Tandutinib ( MLN518/CT53518 ) targeted to stem-like cells by inhibiting the function of DB00171 -binding cassette subfamily G member 2 . Tandutinib is a novel inhibitor of tyrosine kinases P36888 , P09619 and P10721 . Our study was to explore the capability of DB05465 to reverse ABC transporter-mediated multidrug resistance . Tandutinib reversed Q9UNQ0 -mediated drug resistance in Q9UNQ0 -482-R2 , Q9UNQ0 -482-G2 , Q9UNQ0 -482-T7 and S1-M1-80 cells and increased the accumulation of doxorubicin , rhodamine 123 and [ H(3) ] mitoxantrone in Q9UNQ0 -overexpressing cells . Importantly , DB05465 selectively sensitized side population cells to mitoxantrone . Taken together , our results advocate the potency of DB05465 as an Q9UNQ0 modulator and stem-like cells targeted agent to increase efficiency of anticancer drugs . P13726 and factor VIIa as therapeutic targets in disorders of hemostasis . For hemophilia patients with inhibitors against FVIII or FIX , the development of recombinant factor VIIa ( DB00036 ) raises the possibility of a therapeutic alternative whose availability and convenience of treatment are comparable to those of FVIII or FIX . In support of this new concept for the treatment of bleeding episodes , pharmacological doses of FVIIa have been shown to induce hemostasis . Pharmacological doses of DB00036 enhance thrombin generation on thrombin-activated platelets , thereby facilitating the formation of strong , well-structured fibrin plugs resistant to premature proteolysis . Modified DB00036 molecules with a stronger hemostatic potential have been produced . Inhibition of the FVII-TF-dependent pathway ( P10646 and rFVIIai ) has been tried in attempts to prevent thrombosis , with promising results in animal models so far not confirmed in clinical trials . The sulphydryl containing P12821 inhibitor Zofenoprilat protects coronary endothelium from Doxorubicin-induced apoptosis . Pediatric and adult cancer patients , following the use of the antitumor drug Doxorubicin develop cardiotoxicity . Pharmacological protection of microvascular endothelium might produce a double benefit : ( i ) reduction of myocardial toxicity ( the primary target of Doxorubicin action ) and ( ii ) maintenance of the vascular functionality for the adequate delivery of chemotherapeutics to tumor cells . This study was aimed to evaluate the mechanisms responsible of the protective effects of the angiotensin converting enzyme inhibitor ( ACEI ) Zofenoprilat against the toxic effects exerted by Doxorubicin on coronary microvascular endothelium . We found that exposure of endothelial cells to Doxorubicin ( 0.1-1μM range ) impaired cell survival by promoting their apoptosis . P27361 /2 related p53 activation , but not reactive oxygen species , was responsible for Doxorubicin induced caspase-3 cleavage . P04637 mediated-apoptosis and impairment of survival were reverted by treatment with Zofenoprilat . The previously described PI-3K/ P29474 /endogenous fibroblast growth factor signaling was not involved in the protective effect , which , instead , could be ascribed to cystathionine gamma lyase dependent availability of H2S from Zofenoprilat . Furthermore , considering the tumor environment , the treatment of endothelial/tumor co-cultures with Zofenoprilat did not affect the antitumor efficacy of Doxorubicin . In conclusion the ACEI Zofenoprilat exerts a protective effect on Doxorubicin induced endothelial damage , without affecting its antitumor efficacy . Thus , sulfhydryl containing ACEI may be a useful therapy for Doxorubicin-induced cardiotoxicity . Inhibition of cyclooxygenases 1 and 2 by the phospholipase-blocker , arachidonyl trifluoromethyl ketone . BACKGROUND AND PURPOSE : Arachidonyl trifluoromethyl ketone ( Q06187 ) is widely used as an inhibitor of cytosolic group IV phospholipase A(2) ( cPLA(2) ) and calcium-independent group VI phospholipase A(2) ( iPLA(2) ) . Q06187 thus reduces arachidonic acid ( AA ) substrate for cyclooxygenase ( P36551 ; also known as prostaglandin H synthase ) and attenuates prostaglandin ( PG ) synthesis . It has been shown previously , that Q06187 blocks thromboxane B(2) production induced by exogenous AA in human platelets . It remains , however , unknown whether Q06187 also directly modulates the activity of cyclooxygenase ( P36551 ) . EXPERIMENTAL APPROACH : Time courses for inhibition of P36551 by Q06187 was obtained using osteoblast-like MC3T3-E1 cells , with exogenous AA as substrate and the pure enzymes P23219 and P35354 . PGE(2) was measured by GC-MS . KEY RESULTS : Q06187 was a potent inhibitor of P23219 and P35354 with IC(50) values of 0.5 and 0.1 microM in MC3T3-E1 cells and of 1.7 and 2.6 microM using the pure enzymes . Inhibition was reversible , with slow- and tight-binding characteristics . The arachidonyl carbon chain was essential , as the saturated palmitoyl analogue had no effect . CONCLUSIONS AND IMPLICATIONS : Attenuation of PG synthesis by Q06187 is taken to be the consequence of PLA(2) inhibition and the findings of many studies are interpreted on that basis . If there are , however , alternative routes for AA liberation ( such as phospholipase C/diacyl glycerol lipase or phospholipase D ) , this interpretation can lead to false conclusions . As Q06187 is a widely used and important pharmacological tool in eicosanoid research , knowledge of its interactions with other major enzymes of the cascade is of considerable importance . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . mu-Opioid receptor agonists differentially regulate the expression of miR-190 and Q13562 . The agonists of mu-opioid receptor ( P35372 ) induce extracellular signal-regulated kinase ( P29323 ) phosphorylation through different pathways : morphine uses the protein kinase C ( PKC ) -pathway , whereas fentanyl functions in a beta-arrestin2-dependent manner . In addition , the two pathways result in the different cellular location of phosphorylated P29323 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 . When fentanyl-induced but not morphine-induced P29323 phosphorylation was blocked in the primary cultures from beta-arrestin2(-/-) mouse , fentanyl did not decrease the expression of miR-190 . However , a PKC inhibitor that blocked morphine-induced P29323 phosphorylation specifically had no effect on the miR-190 down-regulation . Therefore the decrease in miR-190 expression resulted from the agonist-selective P29323 phosphorylation . In addition , the expressional changes in one of the miR-190 targets , neurogenic differentiation 1 ( Q13562 ) , correlated with those in miR-190 expression , suggesting the P35372 could regulate the Q13562 pathways via the control of miR-190 expression . DB06096 , a selective P29475 inhibitor and a P28222 /1D receptor agonist , inhibits P80511 release in preclinical migraine models . BACKGROUND : DB06096 is a combined neuronal nitric oxide synthase ( P29475 ) inhibitor and 5-hydroxytryptamine 1B/1D ( P28222 /1D ) receptor agonist . Using preclinical models , we evaluated whether these two unique therapeutic principles have a synergistic effect in attenuating stimulated calcitonin gene-related peptide ( P80511 ) release , a marker of trigeminal activation . METHODS : We examined the effect of DB06096 on : ( 1 ) DB00761 - , capsaicin- and resiniferatoxin ( RTX ) -induced immunoreactive P80511 ( iCGRP ) release from isolated preparation of rat dura mater , trigeminal ganglion ( TG ) and trigeminal nucleus caudalis ( P24821 ) ; and ( 2 ) capsaicin- and electrical stimulation ( ES ) -induced middle meningeal artery ( MMA ) dilation in a rat closed-cranial window . RESULTS : DB06096 inhibited : ( 1 ) DB00761 -stimulated iCGRP release from dura mater ( % decrease mean ± SEM , lowest effective concentration ) ( 35 ± 6 % , 30 µM ) , TG ( 24 ± 11 % , 10 µM ) and P24821 ( 40 ± 8 % , 10 µM ) ; ( 2 ) capsaicin- and RTX-induced iCGRP release from dura mater ; and ( 3 ) capsaicin- and ES-induced increase in dural artery diameter ( 32 ± 5 % , 3 mg kg(-1) intravenous ( i.v. ) and 36 ± 1 % , 10 mg kg(-1) i.v. ) . CONCLUSIONS : DB06096 inhibits P80511 release from migraine-relevant cephalic tissues . Its effect is most likely mediated via a combination of P29475 -inhibition and P28222 /1D receptor agonism in dura mater while the mechanisms of action for inhibition of P80511 release from TG and P24821 have to be investigated further . Chemoproteomics-based kinome profiling and target deconvolution of clinical multi-kinase inhibitors in primary chronic lymphocytic leukemia cells . The pharmacological induction of apoptosis in neoplastic B cells presents a promising therapeutic avenue for the treatment of chronic lymphocytic leukemia ( CLL ) . We profiled a panel of clinical multi-kinase inhibitors for their ability to induce apoptosis in primary CLL cells . Whereas inhibitors targeting a large number of receptor and intracellular tyrosine kinases including c- P10721 , P36888 , Q06187 and P43405 were comparatively inactive , the CDK inhibitors BMS-387032 and flavopiridol showed marked efficacy similar to staurosporine . Using the kinobeads proteomics method , kinase expression profiles and binding profiles of the inhibitors to target protein complexes were quantitatively monitored in CLL cells . The targets most potently affected were P50750 , cyclin T1 , P51826 /4 and Q03111 , which may represent four subunits of a deregulated positive transcriptional elongation factor ( p-TEFb ) complex . Albeit with lower potency , both drugs also bound the basal transcription factor BTF2/TFIIH containing P50613 . DB02010 and geldanamycin do not affect these targets and thus seem to exhibit a different mechanism of action . The data support a critical role of p-TEFb inhibitors in CLL that supports their future clinical development . A multimarker model to predict outcome in tamoxifen-treated breast cancer patients . PURPOSE : This study was designed to produce a model to predict outcome in tamoxifen-treated breast cancer patients based on clinicopathologic features and multiple molecular markers . EXPERIMENTAL DESIGN : This was a retrospective study of 324 stage I to III female breast cancer patients treated with tamoxifen for whom standard clinicopathologic data and tumor tissue microarrays were available . Nine molecular markers were studied by semiquantitative immunohistochemistry and/or fluorescence in situ hybridization . Cox proportional hazards analysis was used to determine the contributions of each variable to disease-specific and overall survival , and machine learning was used to produce a model to predict patient outcome . RESULTS : On a univariate basis , the following features were significantly associated with worse survival : high pathologic tumor or nodal class , histologic grade , epidermal growth factor receptor , P04626 , MYC , or P04637 ; absent estrogen receptor ( ER ) or progesterone receptor ; and low P10415 . P24385 and P46527 did not reach statistical significance . On a multivariate basis , nodal class , ER , and MYC were statistically significant as independent factors for survival . However , the benefit of ER-positive status was moderated by P10415 , P04626 , and progesterone receptor . P10415 and P04637 also interacted as an independent risk factor . A kernel partial least squares polynomial model was developed with an area under the receiver operating characteristic curve of 0.90 . CONCLUSIONS : Our data show the predictive value of P10415 , P04626 , MYC , and P04637 in addition to the standard hormone receptors and clinicopathologic features , and they show the importance of conditional interpretation of certain molecular markers . Our multimarker predictive model performed significantly better than standard guidelines . Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals . In diabetic P01130 -/- mice , an ectopic P12643 -Msx2 gene regulatory program is upregulated in association with vascular calcification . We verified the procalcific actions of aortic Msx2 expression in vivo . CMV-Msx2 transgenic ( CMV-Msx2Tg(+) ) mice expressed 3-fold higher levels of aortic Msx2 than nontransgenic littermates . On high-fat diets , CMV-Msx2Tg(+) mice exhibited marked cardiovascular calcification involving aortic and coronary tunica media . This corresponded to regions of Msx2 immunoreactivity in adjacent adventitial myofibroblasts , suggesting a potential paracrine osteogenic signal . To better understand Msx2-regulated calcification , we studied actions in 10T1/2 cells . We found that conditioned media from Msx2-transduced 10T1/2 cells ( Msx2-CM ) is both pro-osteogenic and adipostatic ; these features are characteristic of Wnt signaling . Msx2-CM stimulated Wnt-dependent TCF/LEF transcription , and Msx2-transduced cells exhibited increased nuclear beta-catenin localization with concomitant alkaline phosphatase induction . Msx2 upregulated Wnt3a and Wnt7a but downregulated expression of the canonical inhibitor Dkk1 . Dkk1 treatment reversed osteogenic and adipostatic actions of Msx2 . DB06285 , a Q03431 agonist that inhibits murine vascular calcification , suppressed vascular P12643 -Msx2-Wnt signaling . Analyses of CMV-Msx2Tg(+) mice confirmed that Msx2 suppresses aortic Dkk1 and upregulates vascular Wnts ; moreover , TOPGAL(+) ( Wnt reporter ) ; CMV-Msx2Tg(+) mice exhibited augmented aortic LacZ expression . Thus , Msx2-expressing cells elaborated an osteogenic milieu that promotes vascular calcification in part via paracrine Wnt signals . A combination of molecular cytogenetic analyses reveals complex genetic alterations in conventional renal cell carcinoma . Here we report the complex pattern of genomic imbalances and rearrangements in a panel of 19 renal cell carcinoma cell lines detected with molecular cytogenetic analysis . Consistent heterogeneity in chromosome number was found , and most cell lines showed a near-triploid chromosome complement . Several cell lines showed deletions of the P04637 ( alias p53 ) , CDKN2A ( alias p16 ) , and P40337 genes . Multiplex fluorescence in situ hybridization ( M- Q5TCZ1 ) analysis revealed chromosome 3 translocated to several other partners chromosomes , as well as breakage events commonly affecting chromosomes 1 , 5 , 8 , 10 , and 17 . The most common abnormality detected with comparative genomic hybridization ( CGH ) was deletions of chromosome 3p , with loss of the Q9NS23 , P49789 , and p44S10 loci frequently involved . CGH gain of 5q showed overrepresentation of the P18146 and P07333 genes . Recurrent alterations to chromosome 7 included rearrangement of 7q11 and gains of the P00533 , O15164 , and P35250 genes . Several lines exhibited rearrangement of 12q11 approximately q14 and overrepresentation of P11802 and SAS loci . M- Q5TCZ1 revealed several other recurrent translocations , and CGH findings included loss of 9p , 14q , and 18q and gain of 8q , 12 , and 20 . Further genomic microarray changes included loss of Q13126 , IGH@ , P28222 , and Q13485 ( previously Q13485 ) and gains of MYC and P11387 . An excellent correlation was observed between the genomic array and Q5TCZ1 data , demonstrating that this technique is effective and accurate . The aberrations detected here may reflect important pathways in renal cancer pathogenesis .

MH_train_1253

MH_train_1253

MH_train_1253

interacts_with DB01095?

multiple_choice

Effects of nerve graft on nitric oxide synthase , NAD(P)H oxidase , and antioxidant enzymes in chronic spinal cord injury . Oxidative stress and nitrosative stress play important roles in the pathogenesis of secondary spinal cord injury . Recently , we demonstrated that peripheral nerve grafts ( Q96C90 ) with acidic fibroblast growth factor ( P05230 ) partially restore hind limb locomotion in adult rats with completely transected spinal cords . This study investigated the protein abundances of the superoxide ( O2* ) -generating enzyme nicotinamide adenine dinucleotide ( phosphate ) oxidase ( NAD(P)H oxidase ; gp91phox subunit ) , nitric oxide synthases ( NOS ) , antioxidant enzymes , superoxide dismutases ( Cu Zn SOD , Mn SOD ) , catalase , and glutathione peroxidase ( GPX ) as well as nitrotyrosine in the spinal cord tissue 4 months after spinal cord transection in rats with and without Q96C90 and P05230 . The protein abundances of the gp91phox subunit of NAD(P)H oxidase , Mn SOD , catalase , GPX , P29474 , and nitrotyrosine were significantly upregulated , whereas Cu Zn SOD and P29475 were unchanged in the injury group compared to the sham controls . The nerve graft with P05230 treated group showed significantly better hind limb locomotion recovery than the injury group . Although the protein abundances of gp91phox , nitrotyrosine , and Cu Zn SOD were similar in the treated group ( nerve graft with P05230 ) compared to the injury group , Mn SOD , GPX , catalase , and P29474 protein abundances were significantly higher , whereas P29475 was markedly lower in the treated group . We conclude that the combination of nerve graft and P05230 enhances the local antioxidant defense system after spinal cord transection in rats . Involvement of peptidyl-prolyl isomerase Pin1 in the inhibitory effect of fluvastatin on endothelin-1-induced cardiomyocyte hypertrophy . AIMS : Cardiac hypertrophy is elicited by endothelin ( ET ) -1 as well as other neurohumoral factors , hemodynamic overload , and oxidative stress ; P04035 inhibitors ( statins ) were shown to inhibit cardiac hypertrophy partly via the anti-oxidative stress . One of their common intracellular pathways is the phosphorylation cascade of MEK signaling . Pin1 specifically isomerizes the phosphorylated protein with DB00133 / DB00156 -Pro bonds and regulates their activity through conformational changes . There is no report whether the Pin1 activation contributes to ET-1-induced cardiomyocyte hypertrophy and whether the Pin1 inactivation contributes to the inhibitory effect of statins . The aim of this study was to reveal these questions . MAIN METHODS : We assessed neonatal rat cardiomyocyte hypertrophy using ET-1 and fluvastatin by the cell surface area , P01160 mRNA expression , JNK and c-Jun phosphorylation , and [ (3)H ] -leucine incorporation . KEY FINDINGS : DB01095 inhibited ET-1-induced increase in the cell surface area , P01160 expression , and [ (3)H ] -leucine incorporation ; and it suppressed the signaling cascade from JNK to c-Jun . The phosphorylated Pin1 level , an inactive form , was decreased by ET-1 ; however , it reached basal level by fluvastatin . Furthermore , Pin1 overexpression clearly elicited cardiomyocyte hypertrophy , which was inhibited by fluvastatin . SIGNIFICANCE : This is the first report that ET-1-induced cardiomyocyte hypertrophy is mediated through the Pin1 activation and that the inhibitory effect of fluvastatin on cardiomyocyte hypertrophy would partly be attributed to the suppression of the Pin1 function . This study firstly suggests that Pin1 determines the size of hypertrophied cardiomyocyte by regulating the activity of phosphorylated molecules and that statins exert their pleiotropic effects partly via Pin1 inactivation . Evaluation of an aldose reductase inhibitor on lens metabolism , ATPases and antioxidative defense in streptozotocin-diabetic rats : an intervention study . AIMS/HYPOTHESIS : P15121 inhibitors ( ARIs ) prevent biochemical abnormalities associated with diabetic complications . We evaluated whether a short-term intervention with an adequate dose of Q9Y4X5 , introduced at the very early , precataractous stage , reversed diabetes-induced metabolic imbalances , down-regulation of ATPases and oxidative stress in the lens . Methods . The groups included mature control and streptozotocin-diabetic rats treated with or without Q9Y4X5 sorbinil ( 65 mg x kg(-1) x day(-1) , in the diet , for 2 weeks after 4 weeks of untreated diabetes ) . Free cytosolic NAD+: DB00157 and NADP+:NADPH ratios were calculated from the lactate dehydrogenase and malic enzyme systems . Concentrations of metabolites and adenine nucleotides , Na+/K+-ATPase , H+-ATPase and Ca++-independent Mg++-ATPase activities and variables of oxidative stress were measured in individual lenses . Results . DB02712 treatment essentially corrected diabetes-induced sorbitol and fructose accumulation , myo-inositol depletion , decrease in free cytosolic NAD+: DB00157 ratio and energy deficiency . Malondialdehyde accumulation , reduced glutathione depletion and the increase in oxidized glutathione:reduced glutathione ratio were partially corrected . Free cytosolic NADP+:NADPH ratio and 4-hydroxyalkenal concentrations were similarly increased in diabetic rats treated with or without Q9Y4X5 . DB02712 did not counteract diabetes-induced down-regulation of the three ATPase activities . CONCLUSION/INTERPRETATION : All biochemical changes assessed in our study are known to be prevented by ARIs . Despite the essential normalization of the sorbitol pathway activity , only part of them were , however , reversed by the Q9Y4X5 treatment introduced at the very early , i.e. precataractous , stage of diabetes . Therefore , intervention studies can easily underestimate the importance of aldose reductase in the pathogenesis of diabetic complications and should be interpreted with caution . Eomesodermin , O96004 , and P0DML2 proteins are induced by cellular stress in a stress-activated protein kinase-dependent manner . Eomesodermin ( Eomes ) is a transcription factor essential for trophoblast development . Stress stimuli activate stress-activated protein kinase ( P45983 /9 ) and modulate transcription factors in trophoblast stem cells ( TSC ) . In this study , we test the hypothesis that stress-induced Eomes upregulation and downstream trophoblast development are P45983 /9-dependent . Immunocytochemical and immunoblot assays suggest that Eomes is induced by hyperosmolar stress in a dose- and time-dependent manner . Two P45983 /9 inhibitors that work by different mechanisms , LJNKl1 and SP600125 , block induction of Eomes protein by stress . During normal TSC differentiation , the transcription factor heart and neural crest derivatives expressed 1 ( O96004 ) is dependent on Eomes , and chorionic somatomammotropin hormone 1 ( P0DML2 ) expression is dependent on O96004 . Similar to Eomes , O96004 and P0DML2 induction by stress are P45983 /9-dependent , and P0DML2 is induced in nearly all stressed TSC . P0DML2 induction normally requires downregulation of the transcription factor inhibitor of differentiation 2 ( Q02363 ) as well as O96004 upregulation . It was shown previously that hyperosmolar stress induces AMP-activated protein kinase ( Q13131 /2 ) -dependent Q02363 loss in a P45983 /9-independent manner . Inhibition of Q13131 /2 with compound C and LJNKl1 , more than P45983 /9 inhibitors alone , inhibits the induction of P0DML2 by stress . Taken together these data suggest that stress-induced P45983 /9 and Q13131 /2 regulate transcription factors Eomes/ O96004 and Q02363 , respectively . Together this network mediates induction of P0DML2 by stress . Therefore , stress triggers a proportional increase in a normal early TSC differentiation event that could be adaptive in inducing P0DML2 . But the flexibility of TSC to undergo stress-induced differentiation could lead to pathophysiological consequences if stress endured and TSC differentiation became unbalanced . Interaction between P20292 and P20815 gene variants significantly increases the risk for cerebral infarctions in Chinese . In this study , we investigated associations between susceptibility genes and cerebral infarctions in a Chinese population , and whether gene-gene interactions increase the risk of cerebral infarctions . Overall , 292 patients with cerebral infarctions and 259 healthy control individuals were included . Eight variants in five candidate genes were examined for the risk of stroke , including the SG13S32 ( rs9551963 ) , SG13S42 ( rs4769060 ) , SG13S89 ( rs4769874 ) , and SG13S114 ( rs10507391 ) variants of the P09917 activating protein ( P20292 ) gene , the G860A ( rs751141 ) variant of the soluble epoxide hydrolase ( P34913 ) gene , the A1075C ( rs1057910 ) variant of the P11712 *2 gene , the C430T ( rs1799853 ) variant of the P11712 *3 gene , and the A6986G ( rs776746 ) variant of the P20815 gene . Gene-gene interactions were explored using generalized multifactor dimensionality reduction methods . There were no statistically significant differences in the frequencies of the genotypes of the eight candidate genes . The generalized multifactor dimensionality reduction analysis showed a significant gene-gene interaction between SG13S114 and A6986G , with scores of 10 for cross-validation consistency and 9 for the sign test ( P=0.0107 ) . These gene-gene interactions predicted a significantly higher risk of cerebral infarction ( adjusted for age , hypertension , and diabetes mellitus ; odds ratio=1.80495 % , confidence interval : 1.180-2.759 , P=0.006 ) . A two-loci gene interaction confers a significantly higher risk for cerebral infarction . The combinational analysis used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . The bovine 5' AMPK gene family : mapping and single nucleotide polymorphism detection . The DB00131 -activated protein kinase ( AMPK ) family is an ancient stress response system whose primary function is regulation of cellular DB00171 . Activation of AMPK , which is instigated by environmental and nutritional stresses , initiates energy-conserving measures that protect the cell by inhibition and phosphorylation of key enzymes in energy-consuming biochemical pathways . The seven genes that comprise the bovine AMPK family were mapped in cattle by using a radiation hybrid panel . The seven genes mapped to six different cattle chromosomes , each with a LOD score greater than 10.0 . Q13131 mapped to BTA 20 , P54646 and O43741 to BTA 3 , Q9Y478 to BTA 17 , P54619 to BTA 5 , Q9UGJ0 to BTA 4 , and Q9UGI9 to BTA 2 . Five of the seven genes mapped to regions expected from human/cattle comparative maps . O43741 and Q9UGI9 , however , have not been mapped in humans . We predict these genes to be located on HSA 1 and 2 , respectively . Additionally , one synonymous and one non-synonymous single nucleotide polymorphism ( SNP ) were detected in Q9UGI9 in Bos taurus cattle . In an effort to determine ancestral origins , various herds of mixed breed cattle as well as other ruminant species were characterized for sequence variation in this region of Q9UGI9 . Owing to the physiological importance of this gene family , we believe that its individual genes are candidate genes for conferring resistance to diseases in cattle . Is transforming growth factor-β signaling activated in human hypertrophied prostate treated by 5-alpha reductase inhibitor ? BACKGROUND AND AIM : It is well known that androgen deprivation relates to penile fibrosis , so we hypothesize that long-term treatment with 5-alphareductase inhibitors ( 5ARIs ) may increase the risk of fibrosis of prostate . PATIENTS AND METHODS : Thirty-two BPH patients who underwent transurethral resection of the prostate were enrolled . The patients were divided into two groups : group one , 16 patients underwent TURP who had been treated with tamsulosin for 2 years ; group two , 16 patients underwent TURP who had been treated with combination of tamsulosin and dutasteride for at least 1 year . We evaluated the expressions of P29475 , P35228 , P29474 , TGF-β1 , TGF-β2 , phosphorylated- Q15796 /3 ( p- Q15796 /3 ) , P12830 , P19022 , and α-smooth muscle actin in the resected prostate tissues by western blotting , and the TGF-β concentration was determined by ELISA kit . RESULTS : The expressions of 3 isoforms of NOS were significantly increased in group 2 except of P29474 in lateral prostate , and the expressions of TGF-β1 , TGF-β2 , and p- Q15796 /3 increased about 2-fold compared with group 1 . In group 2 , the P12830 expression decreased while P19022 expression increased significantly . CONCLUSIONS : The overexpression of P29475 may contribute to prostate smooth muscle relaxation ; however , long-time treatment with 5 Q9Y4X5 increases the risk of fibrosis of prostate . Identification of transcripts modulated by P41212 expression . Deletions at chromosome 12p12-13 are observed in 26-47 % of childhood pre-B acute lymphoblastic leukaemia ( ALL ) cases , suggesting the presence of a tumour suppressor gene ( Q9GZX9 ) . Accumulating genetic and functional evidence points to P41212 as being the most probable Q9GZX9 targeted by the deletions . P41212 is a ubiquitously expressed transcription factor of the ETS family with very few known targets . To understand its function and to elucidate the impact of its absence in leukaemia , we conducted a study to identify targeted genes . Following the induction of P41212 expression , global expression was evaluated at different time points . We identified 87 modulated genes , of which 10 ( Q04828 , P42330 , Q14116 , P51884 , Q8WV24 , P35408 , P35354 , Q9NYA1 , P04637 and P15692 ) were validated by real-time quantitative reverse transcription-polymerase chain reaction . To assess the significance of the validated candidate genes in leukaemia , their expression patterns were determined , as well as that of P41212 , in pre-B ALL patients . The expression of Q14116 , P51884 , P35408 , Q9NYA1 and P04637 was significantly correlated with that of P41212 , further suggesting that P41212 could regulate the expression of these genes in leukaemia . This work constitutes another step towards the understanding of the functions of P41212 and the impact of its inactivation in childhood leukaemia . Adrenomedullin- O60895 system is crucially involved in retinal angiogenesis . Adrenomedullin ( P35318 ) is an endogenous peptide first identified as a strong vasodilating molecule . We previously showed that in mice , homozygous knockout of P35318 ( P35318 (-/-) ) or its receptor regulating protein , O60895 ( O60895 (-/-) ) , is embryonically lethal due to abnormal vascular development , thereby demonstrating the importance of P35318 and its receptor signaling to vascular development . P35318 expression in the retina is strongly induced by ischemia ; however , its role in retinal pathophysiology remains unknown . Here , we analyzed oxygen-induced retinopathy ( OIR ) using heterozygous P35318 and O60895 knockout mice models ( P35318 (+/-) or O60895 (+/-) , respectively ) . In addition , we analyzed the role of the P35318 - O60895 system during earlier stages of retinal angiogenesis using an inducible endothelial cell-specific O60895 knockout mouse line ( DI-E- O60895 (-/-) ) . Finally , we assessed the ability of antibody-induced P35318 blockade to control pathological retinal angiogenesis in OIR . In OIR , neovascular tufts , avascular zones , and hypoxic areas were all smaller in P35318 (+/-) retinas compared with wild-type mice . P35318 (+/-) retinas also exhibited reduced levels of P15692 and P29474 expression . DI-E- O60895 (-/-) showed abnormal retinal vascular patterns in the early stages of development . However , P35318 enhanced the proliferation and migration of retinal endothelial cells . Finally , we found intravitreal injection of anti- P35318 antibody reduced pathological retinal angiogenesis . In conclusion , the P35318 - O60895 system is crucially involved in retinal angiogenesis . P35318 and its receptor system are potential therapeutic targets for controlling pathological retinal angiogenesis . Design and synthesis of 3,5-disubstituted-1,2,4-oxadiazoles as potent inhibitors of phosphodiesterase4b2 . A series of 3,5-disubstituted-1,2,4-oxadiazoles has been prepared and evaluated for phosphodiesterase inhibition ( PDE4B2 ) . Among the prepared 3,5-disubstituted-1,2,4-oxadiazoles , compound 9a is the most potent inhibitor ( PDE4B2 IC(50) = 5.28 μm ) . Structure-activity relationship studies of 3,5-disubstituted-1,2,4-oxadiazoles revealed that substituents 3-cyclopentyloxy-4-methoxyphenyl group at 3-position and cyclic ring bearing heteroatoms at 5-position are important for activity . Molecular modeling study of the 3,5-disubstituted-1,2,4-oxadiazoles with Q07343 has shown similar interactions of 3-cyclopentyloxy-4-methoxyphenyl group ; however , heteroatom ring is slightly deviating when compared to DB01791 . 3-(3-Cyclopentyloxy-4-methoxyphenyl)-5-(piperidin-4-yl)-1,2,4-oxadiazole ( 9a ) exhibited good analgesic and antiinflammatory activities in formalin-induced pain in mice and carrageenan-induced paw edema model in rat . P62937 is upregulated in small cell lung cancer and activates P27361 /2 signal . P62937 ( CypA ) , a peptidyl-prolyl cis-trans isomerase ( PPIase ) , was originally identified as the intracellular receptor for cyclosporin A ( DB00091 ) . Recently , correlations of CypA with tumor pathogenesis have been studied . Here , we studied the expression of CypA and its receptor CD147 in several kinds of lung cancer cells as well as a normal lung cell and found that in H446 cell , a kind of small cell lung cancer cell , the expression are the highest . The exogeneous CypA protein can substantially stimulate H446 cell growth in dependence on its PPIase activity . We also showed that CypA protein can stimulate P27361 /2 signal in dose and time dependent manners and almost has no effect to p38 and JNK signals . Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for small cell lung cancer . Differential actions of vasopeptidase inhibition versus angiotensin-converting enzyme inhibition on diuretic therapy in experimental congestive heart failure . BACKGROUND : DB00886 ( OMA ) , a vasopeptidase inhibitor , simultaneously inhibits angiotensin-converting enzyme ( P12821 ) and neutral endopeptidase , which degrades vasodilatory factors ( eg , P35318 ) and natriuretic peptides . Based on the beneficial cardiorenal and humoral properties of the natriuretic peptides , we hypothesized that an acute vasopeptidase inhibitor with or without diuretic would result in more favorable cardiorenal and hormonal actions than P12821 inhibition plus diuretic ( ACEI+D ) in congestive heart failure . METHODS AND RESULTS : We compared the actions of OMA alone and with diuretic ( OMA+D ) to ACEI+D in a model of pacing-induced congestive heart failure . OMA+D decreased pulmonary arterial and pulmonary capillary wedge pressures to a greater level than OMA alone or ACEI+D . Glomerular filtration rate was lower with ACEI+D than with either OMA group . Plasma renin activity and aldosterone immediately increased with ACEI+D , whereas OMA+D resulted in higher plasma renin activity and a delayed increase in aldosterone . OMA alone did not increase plasma renin activity and aldosterone , but resulted in a sustained increase in plasma adrenomedullin , with higher urinary atrial natriuretic peptide , adrenomedullin , and cGMP excretions than with ACEI+D . CONCLUSIONS : Acute administration of OMA with or without diuretic results in more favorable cardiorenal and humoral responses in experimental congestive heart failure than does ACEI+D . There is no acute activation of renin and aldosterone with OMA alone such as occurs with ACEI+D and OMA+D . Thus , OMA with or without a diuretic possesses beneficial cardiorenal and humoral actions comparable to those observed with ACEI+D that can be explained by potentiation of natriuretic peptides . Inhibition of the leukotriene synthetase of rat basophil leukemia cells by diethylcarbamazine , and synergism between diethylcarbamazine and piriprost , a P09917 inhibitor . DB00711 inhibited the formation of sulfidopeptide leukotrienes in rat basophil leukemia ( RBL ) cells ( 50 % inhibitory concentration , EC50 , 3 mM ) . Similar concentrations also inhibited the formation of leukotriene C4 ( LTC4 ) by LTC synthetase , a detergent-solubilized cell free particulate enzyme from RBL cells which is capable of coupling P09960 to glutathione . By contrast , the conversion of P09960 to LTC4 using enzymes from rat liver was at least ten times less sensitive to this inhibitor . The EC50 for inhibition of the leukotriene C synthetase of RBL cells was directly proportional to the P09960 concentration in the incubations , ranging from 1.5 mM at 10 microM P09960 to over 40 mM at 500 microM P09960 . Kinetic analysis revealed that the inhibition of the leukotriene C synthetase reaction by diethylcarbamazine was competitive with respect to P09960 . In contrast to diethylcarbamazine , piriprost ( U-60,257 ; 6,9-deepoxy-6,9-(phenylimino)-delta 6,8-prostaglandin I1 ) , which inhibits the formation of sulfidopeptide leuktrienes in RBL cells at the P09917 step ( EC50 5 microM ) , did not inhibit the leukotriene synthetase of these cells . On the other hand , low concentrations of piriprost , which had no demonstrable inhibitory activity on leukotriene formation by themselves , markedly synergized the inhibitory activity of diethylcarbamazine . These results are consistent with the interpretation that both piriprost and diethylcarbamazine inhibit leukotriene formation but that they act on sequential steps in the biosynthetic pathway in such a manner as to synergistically interfere with the availability or utilization of P09960 in the leukotriene C synthetase reaction . P20711 inhibition does not influence the diuretic and natriuretic response to exogenous alpha-atrial natriuretic peptide in man . The role of dopamine synthesis in the renal actions of human alpha-atrial natriuretic peptide ( alpha P01160 ) was investigated in six dehydrated volunteers using the P20711 inhibitor carbidopa . Each subject received oral placebo or carbidopa ( 100 mg ) followed by an infusion of alpha P01160 10 pmol.kg-1.min-1 for 1 h . The responses to placebo alone and to carbidopa alone were investigated on separate occasions. alpha P01160 produced a similar increase in plasma immunoreactive alpha P01160 whether placebo or carbidopa pretreatment had been given . Urinary dopamine excretion was increased by alpha P01160 . DB00190 pretreatment substantially attenuated this increase without affecting the natriuretic or water-diuretic response to alpha P01160 . DB00190 also failed to alter the change in filtration fraction produced by alpha P01160 . The results suggest that increased synthesis of intrarenal dopamine is not required for the renal effects of alpha P01160 in man . DB01095 inhibits growth and alters the malignant phenotype of the P13671 glioma cell line . BACKGROUND : DB01095 is a member of the family of P04035 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 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 /2 ) and P45983 and 2 ( JNK 1/2 ) expression was estimated by Western blot . Finally , the influence of fluvastatin on cell migration and production of P14780 and P15692 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 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 /2 expression , upregulation of p- P45983 /2 , and reduction in the P14780 and P15692 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 . Inhibition of Q07343 suppresses inflammation by increasing expression of the deubiquitinase Q9NQC7 . The deubiquitinase Q9NQC7 acts as a key negative regulator to tightly control overactive inflammation . Most anti-inflammatory strategies have focused on directly targeting the positive regulator , which often results in significant side effects such as suppression of the host defence response . Here , we show that inhibition of phosphodiesterase 4B ( Q07343 ) markedly enhances upregulation of Q9NQC7 expression in response to bacteria , thereby suggesting that Q07343 acts as a negative regulator for Q9NQC7 . Interestingly , in Cyld-deficient mice , inhibition of Q07343 no longer suppresses inflammation . Moreover , Q07343 negatively regulates Q9NQC7 via specific activation of P45984 but not P45983 . Importantly , ototopical post-inoculation administration of a DB05876 inhibitor suppresses inflammation in this animal model , thus demonstrating the therapeutic potential of targeting DB05876 . These studies provide insights into how inflammation is tightly regulated via the inhibition of its negative regulator and may also lead to the development of new anti-inflammatory therapeutics that upregulate Q9NQC7 expression . Fatal rhabdomyolysis in a patient with liver cirrhosis after switching from simvastatin to fluvastatin . P04035 inhibitors ( statins ) are widely used to treat hypercholesterolemia . Among the adverse effects associated with these drugs are statin-associated myopathies , ranging from asymptomatic elevation of serum creatine kinase to fatal rhabdomyolysis . DB01095 -induced fatal rhabdomyolysis has not been previously reported . We describe here a patient with liver cirrhosis who experienced fluvastatin-induced fatal rhabdomyolysis . This patient had been treated with simvastatin ( 20 mg/day ) for coronary artery disease and was switched to fluvastatin ( 20 mg/day ) 10 days before admission . He was also taking aspirin , betaxolol , candesartan , lactulose , and entecavir . Rhabdomyolysis was complicated and continued to progress . He was treated with massive hydration , urine alkalization , intravenous furosemide , and continuous renal replacement therapy for acute renal failure , but eventually died due to rhabdomyolysis complicated by hepatic failure . In conclusion , fluvastatin should be used with caution in patients with liver cirrhosis , especially with other medications metabolized with P11712 . Dimerization effect of sucrose octasulfate on rat P05230 . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 are Lys113 , Lys118 , Arg122 and Lys128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 . This difference may be important for SOS-mediated P05230 dimerization in rat . Canine placental prostaglandin E2 synthase : expression , localization , and biological functions in providing substrates for prepartum PGF2alpha synthesis . The prepartum output of PGF2alpha in the bitch is associated with increased placental DB00917 -synthase ( O14684 ) mRNA levels . Contrasting with this is a decreased expression of PGF2alpha-synthase ( P42330 / P42330 ) in uteroplacental compartments during prepartum luteolysis , suggesting an involvement of alternative synthetic pathways in PGF2alpha synthesis , for example , conversion of DB00917 to PGF2alpha . However , because the expression and possible functions of the respective O14684 proteins remained unknown , no further conclusion could be drawn . Therefore , a canine-specific O14684 antibody was generated and used to investigate the expression , cellular localization , and biochemical activities of canine uteroplacental O14684 throughout pregnancy and at prepartum luteolysis . Additionally , the biochemical activities of these tissues involved in the conversion of DB00917 to PGF2alpha were investigated . The endometrial O14684 was localized in the uterine surface epithelium at preimplantation and in superficial and deep uterine glands , endothelial cells , and myometrium throughout pregnancy and at parturition . Placental signals were mostly in the trophoblast . The biochemical properties of recombinant O14684 protein were confirmed . Additionally , expression of two DB00917 -receptors , PTGER2/EP2 and P35408 /EP4 , revealed their decreasing expression during luteolysis . In contrast , the uteroplacental expression of prostaglandin transporter ( Q92959 ) was strongly elevated prior to parturition . These localization patterns resembled that of O14684 . The increased expression of O14684 and Q92959 at parturition , together with the accompanying decreased levels of DB00917 -receptors and the capability of canine uterine and placental homogenates to take part in the conversion of DB00917 to PGF2alpha , as found in this study , suggest that DB00917 could be used locally as a substrate for prepartum PGF2alpha synthesis in the dog .

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[ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . Inhibition of ROS-activated p38MAPK pathway is involved in the protective effect of H2S against chemical hypoxia-induced inflammation in PC12 cells . We have demonstrated the neuroprotection of hydrogen sulfide ( H2S ) against chemical hypoxia-induced injury by inhibiting p38MAPK pathway . The present study attempts to evaluate the effect of H2S on chemical hypoxia-induced inflammation responses and its mechanisms in PC12 cells . We found that treatment of PC12 cells with cobalt chloride ( CoCl2 , a hypoxia mimetic agent ) enhanced P05231 secretion , nitric oxide ( NO ) generation and expression levels of inducible nitric oxide synthase ( P35228 ) and neuronal nitric oxide synthase ( P29475 ) . L-canavanine , a selective P35228 inhibitor , partly blocked CoCl2-induced cytotoxicity , apoptosis and mitochondrial insult . In addition , DB02207 ( 7-NI ) , an inhibitor of P29475 , also partly attenuated the CoCl2-induced cytotoxicity . The inhibition of p38MAPK by SB203580 ( a selective p38MAPK inhibitor ) or genetic silencing of p38MAPK by RNAi ( Si-p38 ) depressed not only CoCl2-induced P35228 expression , NO production , but also P05231 secretion . In addition , N-acetyl-L-cysteine , a reactive oxygen species ( ROS ) scavenger , conferred a similar protective effect of SB203580 or Si-p38 against CoCl2-induced inflammatory responses . Importantly , pretreatment of PC12 cells with exogenous application of sodium hydrosulfide ( a H2S donor , 400 μmol/l ) for 30 min before exposure to CoCl2 markedly attenuated chemical hypoxia-stimulated P35228 and P29475 expression , NO generation and P05231 secretion as well as p38MAPK phosphorylation in PC12 cells . Taken together , we demonstrated that p38MAPK- P35228 pathway contributes to chemical hypoxia-induced inflammation and that H2S produces an anti-inflammatory effect in chemical hypoxia-stimulated PC12 cells , which may be partly due to inhibition of ROS-activated p38MAPK- P35228 pathway . Q9BVG9 : high efficiency for synthesizing phosphatidylserine containing docosahexaenoic acid . DB00144 ( PS ) , the major anionic phospholipid in eukaryotic cell membranes , is synthesized by the integral membrane enzymes PS synthase 1 ( PSS1 ) and 2 ( Q9BVG9 ) . Q9BVG9 is highly expressed in specific tissues , such as brain and testis , where docosahexaenoic acid ( DB01708 , 22:6n-3 ) is also highly enriched . The purpose of this work was to characterize the hydrocarbon-chain preference of Q9BVG9 to gain insight on the specialized role of Q9BVG9 in PS accumulation in the DB01708 -abundant tissues . Flag-tagged Q9BVG9 was expressed in P29320 cells and immunopurified in a functionally active form . Purified Q9BVG9 utilized both PE plasmalogen and diacyl PE as substrates . Nevertheless , the latter was six times better utilized , indicating the importance of an ester linkage at the sn-1 position . Although no sn-1 fatty acyl preference was noted , Q9BVG9 exhibited significant preference toward DB01708 compared with 18:1 or 20:4 at the sn-2 position . Preferential production of DB01708 -containing PS ( DB01708 -PS ) was consistently observed with Q9BVG9 purified from a variety of cell lines as well as with microsomes from mutant cells in which PS synthesis relies primarily on Q9BVG9 . These findings suggest that Q9BVG9 may play a key role in PS accumulation in brain and testis through high activity toward DB01708 -containing substrates that are abundant in these tissues . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . DB08875 as a novel therapy for renal cell carcinoma . DB08875 / DB05153 ( Exelexis , Inc. ) has demonstrated remarkable responses in kidney cancer . Preclinical results revealed P15692 , P10721 and MET inhibition in a variety of solid tumors such as thyroid , ovarian , renal , lung , liver and prostate cancers . A phase II trial demonstrated efficacy in renal cancer with a 28 % objective response rate , stable disease rate of 62 % and median progression free survival of 14.7 months . Predominant toxicities of fatigue and diarrhea were noted . Dramatic responses in bone metastases ( three of four patients ) make the agent especially valuable for palliation in a disease , where presence of bone metastases is a predictor of worse survival . DB08875 is an emerging novel agent with promising activity in advanced kidney cancer . Randomized trials are planned in comparison with standard P15692 inhibitor therapy . Defining the role of MET overexpression would help patient selection and enrich and enhance the future evaluation of this targeted novel agent . Discovery of ( 2E ) -3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide ( DB05223 ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 enzyme and COLO 205 cellular IC(50) ) , liver microsomal stability ( t(1/2) ) , cytochrome P450 inhibitory ( 3A4 IC(50) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT-116 , PC-3 , A2780 , MV4-11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . Inhibition of tumor necrosis factor signal transduction in endothelial cells by dimethylaminopurine . P01375 ( P01375 ) promotes diverse responses in endothelial cells that are important to the host response to infections and malignancies ; however , less is known of the postreceptor events important to P01375 action in endothelial cells than in many other cell types . Since phosphorylation cascades are implicated in cytokine signaling , the effects of the protein kinase inhibitor dimethylaminopurine ( DMAP ) on P01375 action in bovine aortic endothelial cells ( BAEC ) were investigated . In BAEC , P01375 promotes phosphorylation of eukaryotic initiation factor 4E ( P06730 ) , c-Jun N-terminal kinase ( JNK ) and ceramide-activated protein kinase activities , Jun-b expression , prostacyclin production , and , when protein synthesis is inhibited , cytotoxicity . DMAP abrogated or significantly attenuated each of these responses to P01375 , without affecting the specific binding of P01375 to its receptors . DB11320 , another agent active in the endothelium , promotes phosphorylation of elongation factor-2 ( P13639 ) and prostacyclin production , but not phosphorylation of P06730 in BAEC . DB11320 -stimulated P13639 phosphorylation was not inhibited and prostacyclin production was unaffected by DMAP . These observations demonstrate that a distinct signal transduction cascade , which can be selectively inhibited by DMAP , promotes the response of BAEC to P01375 . Thus , we have identified a reagent , DMAP , that may be useful for characterizing the P01375 signal transduction pathway . P00505 from rat heart is phosphorylated on Tyr19 in response to insulin stimulation . The cytosolic fatty acid-binding protein from rat heart ( P05413 , M(r) 15,000 ) as well as FABP from mouse adipocytes ( P15090 , 62 % homologous with P05413 ) contain a recognition sequence for protein tyrosine kinases , DB00174 - DB00120 - DB00128 - DB00128 -Tyr19 . P15090 has been shown by others to be partly phosphorylated on Tyr19 , thus encouraging experiments designed to search for phosphotyrosine in P05413 . For this purpose isolated cardiac myocytes were incubated with [32P]orthophosphate and analyzed by two-dimensional gel electrophoresis . A 15 kDa phosphoprotein present in the cytosolic protein fraction was specifically precipitated by a polyclonal antibody against rat heart FABP . Characterization of the phosphorylated FABP was facilitated by the development of an immunoaffinity purification procedure capable of isolating more than 200 micrograms FABP from four rat hearts in one step . Phosphoamino acid analysis and radiosequencing of the major tryptic phosphopeptide from immunopurified FABP revealed Tyr19 as the phosphorylated amino acid . Stimulation of cardiac myocytes with insulin in the presence of tyrosine phosphatase inhibitors led to a several-fold increase in the amount of phosphorylated FABP compared with a nearly undetectable level found without insulin stimulation , indicating that FABP may be a substrate for the insulin receptor tyrosine kinase . Phosphorylated FABP constitutes only a minor fraction compared to the large pool of FABP in the cardiac myocyte , thus obscuring the significance of this modification . However , as the phosphorylated Tyr19 residue is positioned within a helix-turn-helix-related domain of FABP , this modification may modulate a hitherto unknown DNA binding activity of FABP . A hypothesis is discussed in which phosphorylated FABP serves as a signalling molecule in the insulin signal transduction cascade . The P25054 tumor suppressor counteracts beta-catenin activation and H3K4 methylation at Wnt target genes . The P25054 tumor suppressor controls the stability and nuclear export of beta-catenin ( beta-cat ) , a transcriptional coactivator of Q9UJU2 /TCF HMG proteins in the Wnt/Wg signaling pathway . We show here that beta-cat and P25054 have opposing actions at Wnt target genes in vivo . The beta-cat C-terminal activation domain associates with Q9Y4A5 / Q92993 and mixed-lineage-leukemia ( Q03164 /MLL2 ) O15047 -type chromatin-modifying complexes in vitro , and we show that beta-cat promotes H3K4 trimethylation at the c-Myc gene in vivo . H3K4 trimethylation in vivo requires prior ubiquitination of H2B , and we find that ubiquitin is necessary for transcription initiation on chromatin but not nonchromatin templates in vitro . Chromatin immunoprecipitation experiments reveal that beta-cat recruits Pygopus , O00512 /Legless , and Q03164 / O15047 -type complexes to the c-Myc enhancer together with the negative Wnt regulators , P25054 , and betaTrCP . Interestingly , P25054 -mediated repression of c-Myc transcription in HT29- P25054 colorectal cancer cells is initiated by the transient binding of P25054 , betaTrCP , and the CtBP corepressor to the c-Myc enhancer , followed by stable binding of the TLE-1 and Q13547 corepressors . Moreover , nuclear CtBP physically associates with full-length P25054 , but not with mutant SW480 or HT29 P25054 proteins . We conclude that , in addition to regulating the stability of beta-cat , P25054 facilitates CtBP-mediated repression of Wnt target genes in normal , but not in colorectal cancer cells . beta-Arrestin-2 interaction and internalization of the human P47900 receptor are dependent on C-terminal phosphorylation sites . The nucleotide receptor P2Y(1) regulates a variety of physiological processes and is involved in platelet aggregation . Using human P2Y(1)-receptors C-terminally fused with a fluorescent protein , we studied the role of potential receptor phosphorylation sites in receptor internalization and beta-arrestin-2 translocation by means of confocal microscopy . Three receptor constructs were generated that lacked potential phosphorylation sites in the third intracellular loop , the proximal C terminus , or the distal C terminus . The corresponding receptor constructs were expressed in human embryonic kidney ( P29320 ) -293 cells and stimulated with 100 muM ADP . Rapid receptor internalization was observed for the wild-type receptor and from those constructs mutated in the third intracellular loop and the proximal C terminus . However , the construct lacking phosphorylation sites at the distal C terminus did not show receptor internalization upon stimulation . The microscopic data were validated by HA-tagged receptor constructs using a cell surface enzyme-linked immunosorbent assay . P2Y(1)-receptor stimulated beta-arrestin-2-yellow fluorescent protein ( YFP ) translocation followed the same pattern as receptor internalization . Hence , no beta-arrestin-2-YFP translocation was observed when the distal C-terminal phosphorylation sites were mutated . Individual mutations indicate that residues Ser352 and Thr358 are essential for receptor internalization and beta-arrestin-2-YFP translocation . In contrast , protein kinase C ( PKC ) -mediated receptor desensitization was not affected by mutation of potential phosphorylation sites in the distal C terminus but was prevented by mutation of potential phosphorylation sites in the proximal C terminus . P2Y(1)-receptor internalization in P29320 -293 cells was not blocked by inhibitors of PKC and calmodulin-dependent protein kinase . Thus , we conclude that P2Y(1)-receptor desensitization and internalization are mediated by different phosphorylation sites and kinases . A randomized , placebo-controlled study of the effects of the p38 MAPK inhibitor SB- DB05250 on blood biomarkers of inflammation in P48444 patients . The p38 mitogen-activated protein kinase ( MAPK ) signaling upregulates inflammation and is known to be increased in chronic obstructive pulmonary disease ( P48444 ) . The authors assessed the pharmacology of the novel p38 MAPK inhibitor SB- DB05250 using blood biomarkers in P48444 . Seventeen P48444 patients ( forced expiratory volume in 1 second 50 % -80 % predicted ) using short-acting bronchodilators participated in a double-blind , double-dummy , randomized , crossover study . Patients received single oral doses of SB- DB05250 7.5 mg and 25 mg , prednisolone 10 mg and 30 mg , and placebo . Blood was obtained predose and at 1 , 2 , 6 , and 24 hours postdose . Whole-blood sorbitol-induced phosphorylated ( p ) heat shock protein ( HSP ) 27 levels as a marker of p38 pathway activation and lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -alpha production were assessed . Both doses of SB- DB05250 , but not prednisolone , significantly ( P < .0001 ) reduced weighted mean ( WM ) pHSP27 ( 0-6 hours ) by 58 % compared with placebo . WM P01375 production ( 0-24 hours ) was significantly reduced compared with placebo by SB- DB05250 25 mg ( 40 % , P = .005 ) and 7.5 mg ( 33.4 % , P = .02 ) , while prednisolone 30 mg and 10 mg caused 81.5 % and 58.2 % suppression , respectively ( both P < .0001 ) . SB- DB05250 inhibited the p38 MAPK pathway to a greater degree than prednisolone did . SB- DB05250 inhibited P01375 production . SB- DB05250 is a potent p38 MAPK inhibitor that potentially suppresses inflammation in P48444 . High-dose tirofiban with enoxaparin and inflammatory markers in high-risk percutaneous intervention . AIM : The study assessed the benefit of high bolus dose tirofiban ( HD-tirofiban ) with enoxaparin compared with HD-tirofiban with unfractionated heparin ( UFH ) . The study examined markers of platelet activation , thrombin generation and inflammation . MATERIALS AND METHODS : The study is a prospective single centre open-label trial of patients with high-risk acute coronary syndrome treated with percutaneous intervention ( P05154 ) who were randomized to anticoagulation with UFH or enoxaparin with HD-tirofiban ( 25 microg kg(-1) bolus ) . This study measured a panel of platelet activation markers , inflammatory biomarkers and thrombus generation between the two groups . RESULT : Sixty patients undergoing high-risk P05154 were enroled in the study . Platelet inhibition as assessed by whole blood aggregometry following HD-tirofiban infusion was similar in both the UFH and enoxaparin groups . P29965 expression on platelets was significantly reduced following P05154 with HD-tirofiban and either UFH or enoxaparin . Following P05154 , there were significant reductions measured in other markers of platelet activation including O95456 , P selectin , factor V/Va , platelet-monocyte aggregates and monocyte expression of Mac-1 as determined by analysis of venous blood samples using flow cytometry . P00734 fragment 1+2 , D-dimer , P04275 and high sensitive P02741 levels were significantly less post P05154 in the enoxaparin group compared with those patients receiving UFH . CONCLUSION : The combination of HD tirofiban with enoxaparin resulted in an attenuated inflammatory response when compared with that of the combination of HD tirofiban with UFH . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . DB00171 induces synaptic gene expressions in cortical neurons : transduction and transcription control via P47900 receptors . Studies in vertebrate neuromuscular synapses have revealed previously that DB00171 , via P2Y receptors , plays a critical role in regulating postsynaptic gene expressions . An equivalent regulatory role of DB00171 and its P2Y receptors would not necessarily be expected for the very different situation of the brain synapses , but we provide evidence here for a brain version of that role . In cultured cortical neurons , the expression of P2Y(1) receptors increased sharply during neuronal differentiation . Those receptors were found mainly colocalized with the postsynaptic scaffold postsynaptic density protein 95 ( P78352 ) . This arises through a direct interaction of a PDZ domain of P78352 with the C-terminal PDZ-binding motif , D-T-S-L of the P2Y(1) receptor , confirmed by the full suppression of the colocalization upon mutation of two amino acids therein . This interaction is effective in recruiting P78352 to the membrane . Specific activation of P2Y(1) ( G-protein-coupled ) receptors induced the elevation of intracellular Ca(2+) and activation of a mitogen-activated protein kinase/ P04049 signaling cascade . This led to distinct up-regulation of the genes encoding acetylcholinesterase ( P22303 (T) variant ) , choline acetyltransferase , and the N-methyl-d-aspartate receptor subunit Q12879 . This was confirmed , in the example of P22303 , to arise from P2Y(1)-dependent stimulation of a human P22303 gene promoter . That involved activation of the transcription factor Elk-1 ; mutagenesis of the P22303 promoter revealed that Elk-1 binding at its specific responsive elements in that promoter was induced by P2Y(1) receptor activation . The combined findings reveal that DB00171 , via its P2Y(1) receptor , can act trophically in brain neurons to regulate the gene expression of direct effectors of synaptic transmission . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . Systemic delivery and pre-clinical evaluation of nanoparticles containing antisense oligonucleotides and siRNAs . By virtue of their potential to selectively silence oncogenic molecules in cancer cells , antisense oligonucleotides ( ASO ) and small interfering RNAs ( siRNAs ) are powerful tools for development of tailored anti-cancer drugs . The clinical benefit of ASO/siRNA therapeutic is , however , hampered due to poor pharmacokinetics and biodistribution , and suboptimal suppression of the target in tumor tissues . P04049 protein serine/threonine kinase is a druggable signaling molecule in cancer therapy . Our laboratory has developed cationic liposomes for systemic delivery of raf ASO ( DB04973 ) and raf siRNA ( LErafsiRNA ) to human tumor xenografts grown in athymic mice . DB04973 is also the first ASO containing liposomal drug tested in humans . In this article , we primarily focus on a modified formulation of systemically delivered cationic liposomes containing raf antisense oligonucleotide ( md- DB04973 ) . The cationic liposomes were prepared using dimyristoyl 1,2-diacyl-3-trimethylammonium-propane ( DMTAP ) , phosphatidylcholine ( PC ) , and cholesterol ( CHOL ) . The toxicology , pharmacokinetics , biodistribution , target selectivity , and anti-tumor efficacy studies of md- DB04973 were conducted in mice . We demonstrate that md- DB04973 is the next generation of systemically delivered and well-tolerated antisense therapeutic suitable for clinical evaluation . Archaebacterial elongation factor is ADP-ribosylated by diphtheria toxin . Archaebacteria have been defined as a ' third primary kingdom ' of cells in addition to the urkaryotes and the eubacteria . While the latter two correspond approximately to the conventional categories eukaryotes and prokaryotes respectively , the Archaebacteria have up to now comprised four groups of microorganisms : the methanogenic bacteria , the extremely halophilic bacteria and the two thermoacidophilic genera Sulfolobus and Thermoplasma . Based on ribosomal RNA sequence homologies and lipid composition , they apparently form a distinct group . Furthermore they possess or lack typical biochemical markers of both the eukaryotes and the prokaryotes , as well as having unique properties not found elsewhere . Altogether , this indicates that they are not closer to either one of the classical categories . One clear-cut difference between prokaryotes and eukaryotes is the diphtheria toxin reaction , which catalyses the covalent binding of adenosine diphosphate-ribose ( DB02059 ) to the eukaryotic peptide elongation factor P13639 in contrast to the homologous prokaryotic factor EF-G . We report here that diphtheria toxin also catalyses the ADP-riboslation of archaebacterial elongation factors . In this respect , these factors have to be assigned to the P13639 type ; we suppose that the ADP-ribosylatable structure arising so early in evolution is of fundamental importance for the elongation process . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . DB08816 increases adenosine plasma concentration in patients with an acute coronary syndrome . OBJECTIVES : This study aimed to investigate the impact of ticagrelor on adenosine plasma concentration ( P25054 ) in acute coronary syndrome ( ACS ) patients . BACKGROUND : DB08816 is a direct-acting Q9H244 -adenosine diphosphate receptor blocker . The clinical benefit of ticagrelor compared with clopidogrel in ACS patients suggests that the drug has non-platelet-directed properties . Animal and in vitro models suggested that the " pleiotropic " properties of ticagrelor may be related to an interaction with adenosine metabolism . METHODS : We prospectively randomized 60 ACS patients to receive ticagrelor or clopidogrel . The P25054 was measured by liquid chromatography . To assess the mechanism of P25054 variation , we measured adenosine deaminase concentration , adenosine uptake by red blood cells , and cyclic adenosine monophosphate production by cells overexpressing adenosine receptors . The Q9H244 -adenosine diphosphate receptor blockade was assessed by the vasodilator-stimulated phosphoprotein index . RESULTS : Patients receiving ticagrelor had significantly higher P25054 than patients receiving clopidogrel ( 1.5 μM [ interquartile range : 0.98 to 1.7 μM ] vs. 0.68 μM [ interquartile range : 0.49 to 0.78 μM ] ; p < 0.01 ) . The P25054 was not correlated with vasodilator-stimulated phosphoprotein ( p = 0.16 ) . Serum-containing ticagrelor inhibited adenosine uptake by red blood cells compared with clopidogrel or controls ( p < 0.01 for both comparisons ) . DB00640 deaminase activity was similar in serum of patients receiving clopidogrel or ticagrelor ( p = 0.1 ) . DB08816 and clopidogrel had no direct impact on adenosine receptors ( p = not significant ) . CONCLUSIONS : DB08816 increases P25054 in ACS patients compared with clopidogrel by inhibiting adenosine uptake by red blood cells . Ubiquitin-specific protease 2-69 in macrophages potentially modulates metainflammation . Macrophages play a critical role in chronic inflammation and metabolic diseases . We identified a longer splice variant of ubiquitin specific protease ( USP ) 2-69 as a novel molecule that modulates pathways implicated in metabolic disorders . Expression levels of aP2/ P15090 and P05121 / P05121 genes were increased by 4- and 1.8-fold , respectively , after short hairpin RNA-mediated knockdown ( KD ) of the O75604 gene , and such expression was alleviated by overexpression of O75604 -69 in human myeloid cell lines . Supernatants derived from O75604 -KD cells induced P05231 ( ∼6-fold ) and SAA3 ( ∼15-fold ) in 3T3- Q9NUQ9 adipocytes to suggest the anti-inflammatory properties of O75604 . In addition , we observed a 30 % decrease in the number of macrophages in mesenteric adipose tissue derived from O75604 -69 transgenic mice fed a high-fat diet for 14 wk compared with that in their C57BL/6 littermates ( P < 0.01 ) , which was consistent with a ∼40 % decrease in transcription of aP2 and P05121 . The aP2 locus exhibited elevated chromatin accessibility ( > 2.1-fold ) , methylation of histone H3 lysine 4 ( > 4.5-fold ) , and acetylation of histone H4 ( > 2.5-fold ) in O75604 -KD cells . Transfection of isopeptidase-mutated O75604 -69 did not alter chromatin conformation on the aP2 locus in O75604 -KD cells . Our results suggest that O75604 -69 suppresses meta-inflammatory molecules involved in the development of type-2 diabetes .

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interacts_with DB00864?

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P60568 abolishes myeloid cell accumulation induced by Lewis lung carcinoma . Immune aberration in cancer patients can be at least partly ascribed to an accumulation of immature myeloid cells and monocytes/macrophages with immunosuppressive functions . Mice implanted with Lewis lung carcinoma 2 ( LL/2 ) cells show marked splenomegaly as the tumors progress , and this condition is accompanied by impaired T cell activities . We characterized the cells that accumulated in the spleens of LL/2 tumor-bearing mice and attempted to restore the normal cell population by employing interleukin-2 ( P60568 ) . Flow cytometric analysis revealed that the cells expressing Mac1 , P33681 , NK- P04264 , Gra-1 , and MHC class II antigens on their surfaces drastically decreased in number when LL/2 had been engineered to produce P60568 . P60568 also restored the concanavalin A ( ConA ) -mediated proliferative response and P60568 production of the spleen cells . The in vivo growth of P60568 -producing tumors was significantly slower than that of parental LL/2 cells . Therefore , local P60568 production may reverse systemic immune abnormality by stopping myeloid cell accumulation . P55157 inhibitor decreases plasma cholesterol levels in P01130 -deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 ) -inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) -2-cyclopentyl-2-[4-[(2,4-dimethyl-9H-pyrido[2,3-b]indol-9-yl)methyl]phenyl]-N- [ ( 1S ) -2-hydroxy-1-phenylethyl ] ethanamide ( DB04852 ) , a new P55157 inhibitor , to low-density lipoprotein ( LDL ) -receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg/kg for 4 weeks . In the 12 mg/kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E/LDL was not altered by DB04852 , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia . A novel pathway regulating the mammalian target of rapamycin ( P42345 ) signaling . Originally discovered as an anti-fungal agent , the bacterial macrolide rapamycin is a potent immunosuppressant and a promising anti-cancer drug . In complex with its cellular receptor , the FK506-binding protein ( P62942 ) , rapamycin binds and inhibits the function of the mammalian target of rapamycin ( P42345 ) . By mediating amino acid sufficiency , P42345 governs signaling to translational regulation and other cellular functions by converging with the phosphatidylinositol 3-kinase ( PI3K ) pathway on downstream effectors . Whether P42345 receives mitogenic signals in addition to nutrient-sensing has been an unresolved issue , and the mechanism of action of rapamycin remained unknown . Our recent findings have revealed a novel link between mitogenic signals and P42345 via the lipid second messenger phosphatidic acid ( PA ) , and suggested a role for P42345 in the integration of nutrient and mitogen signals . A molecular mechanism for rapamycin inhibition of P42345 signaling is proposed , in which a putative interaction between PA and P42345 is abolished by rapamycin binding . Collective evidence further implicates the regulation of the rapamycin-sensitive signaling circuitry by phospholipase D , and potentially by other upstream regulators such as the conventional protein kinase C , the Rho and Q8N726 families of small G proteins , and calcium ions . As the P42345 pathway has been demonstrated to be an important anti-cancer target , the identification of new components and novel regulatory modes in P42345 signaling will facilitate the future development of diagnostic and therapeutic strategies . P36888 -mutant allelic burden and clinical status are predictive of response to P36888 inhibitors in AML . We examined 6 different P07333 -like tyrosine kinase-3 ( P36888 ) inhibitors ( lestaurtinib , midostaurin , DB05213 , KW-2449 , sorafenib , and sunitinib ) for potency against mutant and wild-type P36888 , 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 /ITD ) mutations . We found that inhibition of P36888 autophosphorylation in a P36888 /ITD specimen does not always induce cell death , suggesting that some P36888 /ITD AML may not be addicted to P36888 signaling . Relapsed samples and samples with a high mutant allelic burden were more likely to be responsive to cytotoxicity from P36888 inhibition compared with the samples obtained at diagnosis or those with a low mutant allelic burden . These P36888 inhibitors varied to a considerable degree in their selectivity for P36888 , and this selectivity influenced the cytotoxic effect . These results have important implications for the potential therapeutic use of P36888 inhibitors in that patients with newly diagnosed P36888 -mutant AML might be less likely to respond clinically to highly selective P36888 inhibition . Potent organo-osmium compound shifts metabolism in epithelial ovarian cancer cells . The organometallic " half-sandwich " compound [Os(η(6)-p-cymene)(4-(2-pyridylazo)-N,N-dimethylaniline)I]PF6 is 49× more potent than the clinical drug cisplatin in the 809 cancer cell lines that we screened and is a candidate drug for cancer therapy . We investigate the mechanism of action of compound 1 in A2780 epithelial ovarian cancer cells . Whole-transcriptome sequencing identified three missense mutations in the mitochondrial genome of this cell line , coding for P03915 , a subunit of complex I ( DB00157 dehydrogenase ) in the electron transport chain . P03915 is a proton pump , helping to maintain the coupling gradient in mitochondria . The identified mutations correspond to known protein variants ( p.I257V , p.N447S , and p.L517P ) , not reported previously in epithelial ovarian cancer . Time-series RNA sequencing suggested that osmium-exposed A2780 cells undergo a metabolic shunt from glycolysis to oxidative phosphorylation , where defective machinery , associated with mutations in complex I , could enhance activity . Downstream events , measured by time-series reverse-phase protein microarrays , high-content imaging , and flow cytometry , showed a dramatic increase in mitochondrially produced reactive oxygen species ( ROS ) and subsequent DNA damage with up-regulation of Q13315 , p53 , and P38936 proteins . In contrast to platinum drugs , exposure to this organo-osmium compound does not cause significant apoptosis within a 72-h period , highlighting a different mechanism of action . Superoxide production in ovarian , lung , colon , breast , and prostate cancer cells exposed to three other structurally related organo-Os(II) compounds correlated with their antiproliferative activity . DNA damage caused indirectly , through selective ROS generation , may provide a more targeted approach to cancer therapy and a concept for next-generation metal-based anticancer drugs that combat platinum resistance . Dendroaspis natriuretic peptide binds to the natriuretic peptide clearance receptor . Dendroaspis natriuretic peptide ( DNP ) is a newly-described natriuretic peptide which lowers blood pressure via vasodilation . The natriuretic peptide clearance receptor ( P17342 ) removes natriuretic peptides from the circulation , but whether DNP interacts with human P17342 directly is unknown . The purpose of this study was to test the hypothesis that DNP binds to P17342 . P01160 , DB04899 , P09543 , and the P17342 ligands AP-811 and cANP(4-23) displaced [ (125)I ] - P01160 from P17342 with pM-to-nM K(i) values . DNP displaced [ (125)I ] - P01160 from P17342 with nM potency , which represents the first direct demonstration of binding of DNP to human P17342 . DNP showed high pM affinity for the P16066 receptor and no affinity for P20594 ( K(i) > 1000 nM ) . DNP was nearly 10-fold more potent than P01160 at stimulating cGMP production in P16066 expressing cells . Blockade of P17342 might represent a novel therapeutic approach in augmenting the known beneficial actions of DNP in cardiovascular diseases such as hypertension and heart failure . Modulation of mitochondrial gene expression in pulmonary epithelial cells exposed to oxidants . Oxidants are important in the regulation of signal transduction and gene expression . Multiple classes of genes are transcriptionally activated by oxidants and are implicated in different phenotypic responses . In the present study , we performed differential mRNA display to elucidate genes that are induced or repressed after exposure of rat lung epithelial ( RLE ) cells to H2O2 or crocidolite asbestos , a pathogenic mineral that generates oxidants . After 8 or 24 hr of exposure , RNA was extracted , reverse transcribed , and amplified by polymerase chain reaction with degenerate primers to visualize alterations in gene expression . The seven clones obtained were sequenced and encoded the mitochondrial genes , DB00157 dehydrogenase subunits P03915 and P03923 , and 16S ribosomal RNA . Evaluation of their expression by Northern blot analysis revealed increased expression of 16S rRNA after 1 or 2 hr of exposure to H2O2 . At later time periods ( 4 and 24 hr ) , mRNA levels of 16S rRNA and DB00157 dehydrogenase were decreased in H2O2-treated RLE cells when compared to sham controls . Crocidolite asbestos caused increases in 16S rRNA levels after 8 hr of exposure , whereas after 24 hr of exposure to asbestos , 16S rRNA levels were decreased in comparison to sham controls . In addition to these oxidants , the nitric oxide generator spermine NONOate caused similar decreases in DB00157 dehydrogenase mRNA levels after 4 hr of exposure . The present data and previous studies demonstrated that all oxidants examined resulted in apoptosis in RLE cells during the time frame where alterations of mitochondrial gene expression were observed . As the mitochondrion is a major organelle that controls apoptosis , alterations in expression of mitochondrial genes may be involved in the regulation of apoptosis . A cytokine-controlled mechanism for integrated regulation of T-lymphocyte motility , adhesion and activation . The co-ordination of T-cell motility , adhesion and activation remains poorly understood . It is also unclear how these functions are co-ordinated with external stimuli . Here we unveil a series of molecular interactions in cis at the surface of T lymphocytes with potent effects on motility and adhesion in these cells , and communicating with proliferative responses . These interactions were controlled by the signature cytokines of T helper subsets interleukin-2 ( P60568 ) and P05112 . P01130 -related protein 1 ( Q07954 ) was found to play a key role for T-cell motility by promoting development of polarized cell shape and cell movement . Endogenous thrombospondin-1 ( P07996 -1 ) enhanced cell surface expression of Q07954 through Q08722 . Cell surface expressed Q07954 induced motility and processing of P07996 -1 while inhibiting adhesion to intercellular adhesion molecule 1 and fibronectin . P60568 , but not P05112 , stimulated synthesis of P07996 -1 and motility through P07996 -1 and Q07954 . Stimulation of the T-cell receptor ( TCR ) /CD3 complex inhibited P07996 -1 expression . Inhibitor studies indicated that Q07954 regulated P07996 -1 expression and promoted motility through JAK signalling . This Q07954 -mediated motogenic signalling was connected to Q08722 /Gi protein signalling and P60568 -induced signalling through P07996 -1 . The motogenic P07996 -1/ Q07954 mechanism antagonized TCR/CD3-induced T-cell proliferation . These results indicate that Q07954 in collaboration with P07996 -1 directs a counter-adhesive and counter-proliferative motogenic cascade . T cells seem programmed to prioritize movement before adhesion through this cascade . In conclusion , vital decision-making in T lymphocytes regulating motility , adhesive interactions and proliferation , are integrated through a molecular mechanism connecting different cell surface receptors and their signalling pathways . P60568 inhibits DB01221 receptor-mediated currents directly and may differentially affect subtypes . Using whole-cell patch-clamp recordings , this study investigated the effects of interleukin-2 ( P60568 ) on N-methyl-d-aspartate ( DB01221 ) receptor-mediated currents ( I( DB01221 ) ) in rat cultured hippocampal neurons and human embryonic kidney ( P29320 ) 293 cells expressing recombinant DB01221 receptors . We found that P60568 ( 0.01-1ng/ml ) immediately and significantly decreased peak I( DB01221 ) in cultured neurons . Interestingly , the peak I( DB01221 ) induced in P29320 293 cells was also inhibited by P60568 . We also found that P60568 differentially decreased the peak amplitudes of Q12879 - and Q13224 -containing DB01221 receptor-mediated currents ( I( Q12879 ) and I( Q13224 ) ) by 54+/-5 % and 30+/-4 % , respectively . These results provide new evidence that P60568 induces rapid inhibition of peak currents of DB01221 receptor-mediated responses with possible Q9UHB4 / Q12879 and Q9UHB4 / Q13224 subtype-differentiation , and suggest that the inhibition is mediated by direct interaction between P60568 and DB01221 receptors . Generation of Epstein-Barr virus-specific cytotoxic T lymphocytes resistant to the immunosuppressive drug tacrolimus ( FK506 ) . Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T lymphocytes ( EBV-CTLs ) to solid organ transplant ( SOT ) recipients has been shown safe and effective for the treatment of EBV-associated posttransplantation lymphoproliferative disorders ( PTLDs ) . SOT recipients , however , require the continuous administration of immunosuppressive drugs to prevent graft rejection , and these agents may significantly limit the long-term persistence of transferred EBV-CTLs , precluding their use as prophylaxis . DB00864 ( FK506 ) is one of the most widely used immunosuppressive agents in SOT recipients , and its immunosuppressive effects are largely dependent on its interaction with the 12-kDa FK506-binding protein ( P62942 ) . We have knocked down the expression of P62942 in EBV-CTLs using a specific small interfering RNA ( siRNA ) stably expressed from a retroviral vector and found that P62942 -silenced EBV-CTLs are FK506 resistant . These cells continue to expand in the presence of the drug without measurable impairment of their antigen specificity or cytotoxic activity . We confirmed their FK506 resistance and anti-PTLD activity in vivo using a xenogenic mouse model , suggesting that the proposed strategy may be of value to enhance EBV-specific immune surveillance in patients at high risk of PTLD after transplantation . Natriuretic and renoprotective effect of chronic oral neutral endopeptidase inhibition in acute renal failure . P08473 ( NEP : EC 3.4.24.11 ) is involved in the degradation of peptides such as atrial natriuretic peptide , angiotensin II ( AngII ) , and endothelin-1 ( ET-1 ) . In this study we propose that NEP inhibition provides protection in glycerol-induced acute renal failure ( Q8N726 ) . Renal vascular responses were evaluated in Q8N726 rats where Q8N726 was induced by injecting 50 % glycerol in candoxatril , a NEP inhibitor ( 30 mg/kg , orally ; for 3 weeks ) pretreated rats . AngII and U46619 ( a TxA2 mimetic ) vasoconstriction was increased ( 2- to 4-fold ) in Q8N726 while ET-1 vasoconstriction was surprisingly reduced ( 23+/-3 % ; p < 0.05 ) . In Q8N726 , candoxatril paradoxically enhanced ET-1 response ( 60+/-20 % ; p < 0.05 ) but reduced AngII vasoconstriction ( 51+/-11 % ; p < 0.05 ) without affecting U46619 response . However , candoxatril treatment was without effect on plasma ET-1 and TxB2 levels in Q8N726 . DB00616 reduced plasma AngII by 34+/-4 % ( p < 0.05 ) in Q8N726 which was approximately 3.5-fold higher compared to control . DB00616 doubled the nitrite excretion in control but was without effect on proteinuria or nitrite excretion in Q8N726 . DB00616 enhanced Na+ and creatinine excretion in Q8N726 by 73+/-9 % and 33+/-2 % , respectively . These results suggest that NEP inhibition may confer protection in glycerol-induced Q8N726 by stimulating renal function but without a consistent effect on renal production and renal vascular responses to endogenous vasoconstrictors . Combination of Lipitor and DB00482 inhibits prostate cancer VCaP cells in vitro and in vivo . BACKGROUND/AIM : Lipitor is a cholesterol-lowering drug and DB00482 is a P35354 inhibitor . We investigated the effects of Lipitor and DB00482 on human prostate cancer VCaP cells cultured in vitro and grown as orthotopic xenograft tumors in SCID mice . MATERIALS AND METHODS : Apoptosis was measured by morphological assessment and caspase-3 assay . Nuclear factor-kappa B ( NF-κB ) activation was determined by luciferase reporter assay . B-cell lymphoma-2 ( Bcl2 ) was measured by western blotting and immunohistochemistry . Orthotopic prostate tumors were monitored by the IVIS imaging system . RESULTS : the combination of Lipitor and DB00482 had stronger effects on the growth and apoptosis of VCaP cells than did either drug alone . The combination more potently inhibited activation of NFκB and expression of Bcl2 than either drug alone . The growth of orthotopic VCaP prostate tumors was strongly inhibited by treatment with the drug combination . CONCLUSION : Administration of Lipitor and DB00482 in combination may be an effective strategy for inhibiting the growth of prostate cancer . Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin . DB00864 (FK506) is a strong immuno-suppressant and shows its activity through inhibiting P60568 mRNA transcription by forming pentameric complex with intracellular receptor ( FK506 binding protein 12 kDa or P62942 ) , Ca2+ , calmodulin , and calcineurin . Here , we report the binding activity to P62942 , the pentameric complex formation and Con-A response inhibiting activities of 7 metabolites . C15-demethylated metabolite(M-3) needed higher quantity to compete in Con-A assay and in pentamer formation assay , although it binds more strongly to P62942 . The result suggests that the ability to form a pentameric complex is not a two step reaction with the first binding to P62942 , but a single step reaction by components for the pentamer formation . Signaling by proinflammatory cytokines : oligomerization of TRAF2 and Q9Y4K3 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain . Interleukin-1 ( IL-1 ) and tumor necrosis factor ( P01375 ) stimulate transcription factors AP-1 and NF-kappaB through activation of the Q96HU1 kinases JNK and p38 and the O15111 ( IKK ) , respectively . The P01375 and IL-1 signals are transduced through TRAF2 and Q9Y4K3 , respectively . Overexpressed TRAF2 or Q9Y4K3 activate JNK , p38 , or IKK in the absence of extracellular stimulation . By replacing the carboxy-terminal TRAF domain of TRAF2 and Q9Y4K3 with repeats of the immunophilin P62942 , we demonstrate that their effector domains are composed of their amino-terminal Zn and RING fingers . Oligomerization of the TRAF2 effector domain results in specific binding to Q13233 , a protein kinase capable of JNK , p38 , and IKK activation , and induction of P01375 and IL-1 responsive genes . P01375 also enhances the binding of native TRAF2 to Q13233 and stimulates the kinase activity of the latter . Thus , P01375 and IL-1 signaling is based on oligomerization of TRAF2 and Q9Y4K3 leading to activation of effector kinases . P62942 , the 12-kDa FK506-binding protein , is a physiologic regulator of the cell cycle . P62942 , the 12-kDa FK506-binding protein , is a ubiquitous abundant protein that acts as a receptor for the immunosuppressant drug FK506 , binds tightly to intracellular calcium release channels and to the transforming growth factor beta ( TGF-beta ) type I receptor . We now demonstrate that cells from P62942 -deficient ( P62942 (-/-) ) mice manifest cell cycle arrest in G(1) phase and that these cells can be rescued by P62942 transfection . This arrest is mediated by marked augmentation of P38936 ( P38936 /CIP1) levels , which can not be further augmented by TGF-beta1 . The P38936 up-regulation and cell cycle arrest derive from the overactivity of TGF-beta receptor signaling , which is normally inhibited by P62942 . Cell cycle arrest is prevented by transfection with a dominant-negative TGF-beta receptor construct . TGF-beta receptor signaling to gene expression can be mediated by SMAD , p38 , and P29323 / Q96HU1 kinase ( extracellular signal-regulated kinase/mitogen-activated protein kinase ) pathways . SMAD signaling is down-regulated in P62942 (-/-) cells . Inhibition of P29323 / Q96HU1 kinase fails to affect P38936 up-regulation . By contrast , activated phosphorylated p38 is markedly augmented in P62942 (-/-) cells and the P38936 up-regulation is prevented by an inhibitor of p38 . Thus , P62942 is a physiologic regulator of cell cycle acting by normally down-regulating TGF-beta receptor signaling . Glutamate modulators as potential therapeutic drugs in schizophrenia and affective disorders . Severe psychiatric disorders such as schizophrenia are related to cognitive and negative symptoms , which often are resistant to current treatment approaches . The glutamatergic system has been implicated in the pathophysiology of schizophrenia and affective disorders . A key component is the dysfunction of the glutamatergic N-methyl-D-aspartate ( DB01221 ) receptor . Substances regulating activation/inhibition of the DB01221 receptor have been investigated in schizophrenia and major depression and are promising in therapeutic approaches of negative symptoms , cognition , and mood . In schizophrenia , add-on treatments with glycine , D-serine , D-alanine , D-cycloserine , D-amino acid oxidase inhibitors , glycine transporter-1 ( P48067 ) inhibitors ( e.g. , sarcosine , bitopertin ) and agonists ( e.g. , DB05096 ) or positive allosteric modulator ( e.g. , ADX71149 ) of group II metabotropic glutamate receptors ( mGluRs ) have been studied . In major depression , the DB01221 receptor antagonists ( e.g. , ketamine , AZD6765 ) , Q13224 subtype antagonists ( e.g. , traxoprodil , MK-0657 ) , and partial agonists ( e.g. , D-cycloserine , GLYX-13 ) at the glycine site of the DB01221 receptor have been proven to be effective in animal studies and first clinical trials . In addition , clinical studies of Q14416 /3 antagonist BCI-838 ( a prodrug of BCI-632 ( MGS0039 ) ) , Q14416 /3-negative allosteric modulators ( NMAs ) ( e.g. , RO499819 , RO4432717 ) , and P41594 NAMs ( e.g. , AZD2066 , RO4917523 ) are in progress . Future investigations should include effects on brain structure and activation to elucidate neural mechanisms underlying efficacy of these drugs . Novel phenolic antioxidants as multifunctional inhibitors of inducible P19320 expression for use in atherosclerosis . A series of novel phenolic compounds has been discovered as potent inhibitors of P01375 -inducible expression of vascular cell adhesion molecule-1 ( P19320 ) with concurrent antioxidant and lipid-modulating properties . Optimization of these multifunctional agents led to the identification of 3a ( DB05399 ) as a clinical candidate with demonstrated efficacies in animal models of atherosclerosis and hyperlipidemia . Nox2 and Nox4 mediate tumour necrosis factor-α-induced ventricular remodelling in mice . Reactive oxygen species ( ROS ) and pro-inflammatory cytokines are crucial in ventricular remodelling , such as inflammation-associated myocarditis . We previously reported that tumour necrosis factor-α ( P01375 -α ) -induced ROS in human aortic smooth muscle cells is mediated by NADPH oxidase subunit Nox4 . In this study , we investigated whether P01375 -α-induced ventricular remodelling was mediated by Nox2 and/or Nox4 . An intravenous injection of murine P01375 -α was administered to a group of mice and saline injection was administered to controls . Echocardiography was performed on days 1 , 7 and 28 post-injection . Ventricular tissue was used to determine gene and protein expression of Nox2 , Nox4 , P01160 , interleukin ( IL ) -1β , P60568 , P05231 , P01375 -α and to measure ROS . Nox2 and Nox4 siRNA were used to determine whether or not Nox2 and Nox4 mediated P01375 -α-induced ROS and upregulation of IL-1β and P05231 in adult human cardiomyocytes . Echocardiography showed a significant increase in left ventricular end-diastolic and left ventricular end-systolic diameters , and a significant decrease in the ejection fraction and fractional shortening in mice 7 and 28 days after P01375 -α injection . These two groups of mice showed a significant increase in ventricular ROS , P01160 , IL-1β , P60568 , P05231 and P01375 -α proteins . Nox2 and Nox4 mRNA and protein levels were also sequentially increased . ROS was significantly decreased by inhibitors of NADPH oxidase , but not by inhibitors of other ROS production systems . Nox2 and Nox4 siRNA significantly attenuated P01375 -α-induced ROS and upregulation of IL-1β and P05231 in cardiomyocytes . Our study highlights a novel P01375 -α-induced chronic ventricular remodelling mechanism mediated by sequential regulation of Nox2 and Nox4 subunits . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . Characterization of the interaction of ingenol 3-angelate with protein kinase C . DB05013 ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate ( PMA ) in WEHI-231 , Q9BPY8 -92 , and Colo-205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO- P04264 cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation . Optimized protocols for generation of cord blood-derived cytokine-induced killer/natural killer cells . The efficacy of various combinations of stem cell factor ( P21583 ) , P36888 ligand , interleukin ( IL ) -2 , P13232 and P40933 to induce and expand cord blood-derived cytokine-induced killer ( CIK ) cells was investigated . There were three treatment groups : group A : P21583 combined with P60568 , P13232 and P40933 ; group B : P21583 , P36888 ligand combined with P60568 , P13232 and P40933 , and group C : P60568 , P13232 and P40933 , the control group . Proliferation rates of CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) natural killer ( NK ) cells were highest in group B ; expansion of CIK cells increased 796.1 ± 278.5-fold , and that of NK cells increased 36.6 ± 3.5-fold . All expanded cord blood-derived CIK/NK cells showed cytotoxic activity against the K562 cell line . Interestingly , the cytotoxicity of group A was highest and significantly higher than that of other groups . These protocols might provide an alternative choice for CIK/NK cell expansion .

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P25021 mediated relaxation of buffalo ( Bubalus bubalus ) ureter . On the buffalo ureter , histamine did not elicit any direct effect . However , it caused concentration-dependent relaxation of the tissues precontracted by carbachol , phenylephrine , norepinephrine , KCI or BaCl2 and also inhibited the contractile effect of carbachol . DB08805 selectively antagonised the relaxation and inhibition of contractile response but mepyramine did not show this effect . Isoprenaline , dobutamine , salbutamol , verapamil and papaverine neither produced any direct effect nor relaxed the carbachol-contracted tissues ; norepinephrine and epinephrine had contractile effects . Hence , the histamine-induced relaxation was mediated through the activation of H2 receptors and not through adrenergic mechanisms or blockade of Ca(2+)-channels or inhibition of cyclic nucleotide phosphodiesterase . P35354 inhibitor for pain management in osteoid osteoma . Thirteen patients with osteoid osteoma were enrolled in a prospective trial to test whether rofecoxib , a selective cyclooxygenase-2 inhibitor , is as effective for pain control as acetylsalicylic acid . Each patient documented the pain level using a visual analog scale , with 0 being no pain and 10 being unbearable pain , during 2 days of no pain medication , 4 days of 500 mg acetylsalicylic acid three times a day , and 10 days of 25 mg rofecoxib once a day . Oral administration of 500 mg acetylsalicylic acid three times a day led to a significant decrease in pain at night , pain at rest , and pain induced by exercise . Twenty-five milligrams rofecoxib given once a day at midday showed the same remarkable improvement in pain at night , pain at rest , and pain induced by exercise . DB00533 in comparison with acetylsalicylic acid showed a trend toward lower pain levels in all categories . DB00533 offered a significantly better reduction in pain at rest during the day than did acetylsalicylic acid . Results of the current study suggest that pain induction in osteoid osteoma is related to cyclooxygenase-2 , an enzyme that is blocked by acetylsalicylic acid and rofecoxib . Conservative medical treatment with rofecoxib for osteoid osteoma is recommended when percutaneous intervention is associated with significant morbidity . Antiemetic and motor-depressive actions of CP55,940 : cannabinoid P21554 receptor characterization , distribution , and G-protein activation . Dibenzopyran ( Delta(9)-tetrahydrocannabinol ) and aminoalkylindole [ R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrolol[1,2,3-de]-1,4-benzoxazin-yl]-(1-naphthalenyl) methanone mesylate ; ( WIN55,212-2 ) ] cannabinoids suppress vomiting produced by cisplatin via cannabinoid CB(1) receptors . This study investigates the antiemetic potential of the " nonclassical " cannabinoid CP55,940 [ 1alpha,2beta-(R)-5alpha ] -(-)-5-(1,1-dimethyl)-2- [ 5-hydroxy-2-(3-hydroxypropyl) cyclohexyl-phenol ] against cisplatin-induced vomiting and assesses the presence and functionality of cannabinoid CB(1) receptors in the least shrew ( Cryptotis parva ) brain . CP55,940 ( 0.025-0.3 mg/kg ) reduced both the frequency of cisplatin-induced emesis ( ID(50)=0.025 mg/kg ) and the percentage of shrews vomiting ( ID(50)=0.09 mg/kg ) . CP55,940 also suppressed shrew motor behaviors ( ID(50)=0.06- 0.21 mg/kg ) at such doses . The antiemetic and motor-suppressant actions of CP55,940 were countered by SR141716A [ N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)- DB01213 -3-carboxamide ] , indicating both effects are cannabinoid CB(1) receptor-mediated . Autoradiographic studies with [ 3H ] -SR141716A and [ 35S ] -GTPgammaS binding revealed that the distribution of the cannabinoid CB(1) receptor and its activation pattern are similar to rodent brain and significant levels are present in brain loci ( e.g. , nucleus tractus solitarius ( P30990 ) ) that control emesis . The affinity rank order of structurally diverse cannabinoid ligands for cannabinoid CB(1) receptor in shrew brain is similar to rodent brain : HU-210=CP55,940=SR141716A >/= WIN55,212-2 >/= DB00470 > methanandamide=HU-211=cannabidiol=2-arachidonoylglycerol . This affinity order is also similar and is highly correlated to the cannabinoid EC(50) potency rank order for GTPgammaS stimulation except WIN55,212-2 and DB00470 potency order were reversed . The affinity and the potency rank order of tested cannabinoids were significantly correlated with their antiemetic ID(50) potency order against cisplatin-induced vomiting ( CP55,940 > WIN55,212-2= DB00470 ) as well as emesis produced by 2-arachidonoylglycerol or SR141716A ( CP55,940 > WIN55,212-2 > DB00470 ) . Genetic Association between Akt1 Polymorphisms and Alzheimer 's Disease in a Japanese Population . A recent paper reported that Aβ oligomer causes neuronal cell death through the phosphatidylinositol-3-OH kinase ( PI3K ) -Akt- P42345 signaling pathway . Intraneuronal Aβ , a main pathological finding of Alzheimer 's disease ( AD ) , is also known as inhibiting activation of Akt . This study aims to investigate whether single nucleotide polymorphisms ( SNPs ) of the Akt1 gene are associated with AD . SNPs genotyped using TaqMan technology was analyzed using a case-control study design . Our case-control dataset consisted of 180 AD patients and 130 age-matched controls . Although two SNPs showed superficial positive , Hardy-Weinberg equilibrium ( HWE ) tests , and linkage disequilibrium ( LD ) analyses suggested that genetic regions of the gene are highly polymorphic . We failed to detect any synergetic association among Akt1 polymorphisms , P02649 ( APO E ) , and AD . Further genetic studies are needed to clarify the relationship between the Akt1 and AD . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Q14703 lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 . DB03203 1-phosphate ( Q14703 ) and P21453 ( P21453 ) play an important role in the egress of mature P01730 or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY720 ) , an P21453 functional antagonist , induced significant accumulation of CD62L(high) Q07108 (low) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti- P21453 antibody revealed that P21453 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration . 2-Acetyl-4-tetrahydroxybutylimidazole ( THI ) , an Q14703 lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 ) . At 6h after THI administration , P21453 -expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla , suggesting Q14703 lyase expression in the cells constructing thymic medullary P15151 . To determine the cells expressing Q14703 lyase in the thymus , we newly established a mAb ( YK19-2 ) specific for mouse Q14703 lyase . Immunohistochemical staining with YK19-2 revealed that Q14703 lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 , Q14703 lyase was expressed in ER-TR7-positive cells ( reticular fibroblasts and pericytes ) and CD31-positive vascular endothelial cells . Our findings suggest that Q14703 lyase expressed in the thymic medullary P15151 keeps the tissue Q14703 concentration low around the vessels and promotes thymic egress via up-regulation of P21453 . Genetic variations in the dopamine system and facial expression recognition in healthy chinese college students . OBJECTIVE : This study investigated the relation between genetic variations in the dopamine system and facial expression recognition . METHODS : A sample of Chinese college students ( n = 478 ) was given a facial expression recognition task . Subjects were genotyped for 98 loci [ 96 single-nucleotide polymorphisms ( SNPs ) and 2 variable number tandem repeats ] in 16 genes involved in the dopamine neurotransmitter system , including its 4 subsystems : synthesis ( TH , DDC , and P09172 ) , degradation/transport ( P21964 , P21397 , P27338 , and Q01959 ) , receptors ( P21728 , P14416 , P35462 , P21917 , and P21918 ) , and modulation ( P30990 , P30989 , O95665 , and Q9BYT8 ) . To quantify the total contributions of the dopamine system to emotion recognition , we used a series of multiple regression models . Permutation analyses were performed to assess the posterior probabilities of obtaining such results . RESULTS : Among the 78 loci that were included in the final analyses ( after excluding 12 SNPs that were in high linkage disequilibrium and 8 that were not in Hardy-Weinberg equilibrium ) , 1 ( for fear ) , 3 ( for sadness ) , 5 ( for anger ) , 13 ( for surprise ) , and 15 ( for disgust ) loci exhibited main effects on the recognition of facial expressions . Genetic variations in the dopamine system accounted for 3 % for fear , 6 % for sadness , 7 % for anger , 10 % for surprise , and 18 % for disgust , with the latter surviving a stringent permutation test . CONCLUSIONS : Genetic variations in the dopamine system ( especially the dopamine synthesis and modulation subsystems ) made significant contributions to individual differences in the recognition of disgust faces . FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . P05305 -selective binding sites are downregulated by transforming growth factor-beta and upregulated by basic fibroblast growth factor in a vascular smooth muscle-derived cell line . Endothelins ( ETs ) elicit in vivo and in vitro a potent vasoconstrictor activity after binding to high-affinity receptors on vascular smooth muscle cells ( VSMC ) . A617 cells , a VSM-derived cell line , were used as an in vitro model system to study selected growth factors and cytokines involved in proliferative and/or inflammatory diseases of the vessel wall as possible regulators of the high-affinity binding capacity of ET-1 to the cells . Radioligand studies characterized the binding of ET-1 to the isopeptide selective P25101 receptor subtype on A617 cells as a time- and temperature-dependent saturable process ( Kd = 0.13 +/- 0.04 nM , Bmax = 49 +/- 7 fmol/10(6) cells ) . Pretreatment of A617 cells with basic fibroblast growth factor ( P09038 ) , a mitogenic agent for vascular cells , resulted in a time- and dose-dependent increase in ET-1 binding capacity , whereas preexposure to transforming growth factor-beta ( TGF-beta ) induced a reduction of the Bmax for ET-1 . Platelet-derived growth factor ( PDGF ) , interleukin-6 ( P05231 ) , tumor necrosis factor-alpha , and fetal bovine serum ( FBS ) pretreatments did not affect consequent ET-1 binding to A617 cells . Dynamic immune cell accumulation during flow-induced atherogenesis in mouse carotid artery : an expanded flow cytometry method . OBJECTIVE : Inflammation plays a central role in atherosclerosis . However , the detailed changes in the composition and quantity of leukocytes in the arterial wall during atherogenesis are not fully understood in part because of the lack of suitable methods and animal models . METHODS AND RESULTS : We developed a 10-fluorochrome , 13-parameter flow cytometry method to quantitate 7 major leukocyte subsets in a single digested arterial wall sample . P02649 -deficient mice underwent left carotid artery ( LCA ) partial ligation and were fed a high-fat diet for 4 to 28 days . Monocyte/macrophages , dendritic cells , granulocytes , natural killer cells , and P01730 T cells significantly infiltrated the LCA as early as 4 days . Monocyte/macrophages and dendritic cells decreased between 7 and 14 days , whereas T-cell numbers remained steady . Leukocyte numbers peaked at 7 days , preceding atheroma formation at 14 days . B cells entered LCA by 14 days . Control right carotid and sham-ligated LCAs showed no significant infiltrates . Polymerase chain reaction and ELISA arrays showed that expression of proinflammatory cytokines and chemokines peaked at 7 and 14 days postligation , respectively . CONCLUSION : This is the first quantitative description of leukocyte number and composition over the life span of murine atherosclerosis . These results show that disturbed flow induces rapid and dynamic leukocyte accumulation in the arterial wall during the initiation and progression of atherosclerosis . Hsp27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp27 ( P04792 ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp27 decreased P05231 -dependent P40763 phosphorylation , nuclear translocation , and P40763 binding to the Twist promoter , suggesting that Hsp27 is required for P05231 -mediated EMT via modulation of P40763 /Twist signaling . We observed a correlation between Hsp27 and Twist in patients with prostate cancer , with Hsp27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp27 inhibition by DB06094 , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp27 as a critical regulator of P05231 -dependent and P05231 -independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer . Ameliorative Effect of a Selective Endothelin P25101 Receptor Antagonist in Rat Model of L- DB00134 -induced Vascular Dementia . The present study was designed to investigate the efficacy of selective P25101 receptor antagonist , ambrisentan on hyperhomocysteinemia-induced experimental vascular dementia . L-methionine was administered for 8 weeks to induce hyperhomocysteinemia and associated vascular dementia in male rats . DB06403 was administered to L-methionine-treated effect rats for 4 weeks ( starting from 5(th) to 8(th) week of L-methionine treatment ) . On 52(nd) day onward , the animals were exposed to the Morris water maze ( MWM ) for testing their learning and memory abilities . Vascular endothelial function , serum nitrite/nitrate levels , brain thiobarbituric acid reactive species ( TBARS ) , brain reduced glutathione ( DB00143 ) levels , and brain acetylcholinesterase ( P22303 ) activity were also measured . L-methionine-treated animals showed significant learning and memory impairment , endothelial dysfunction , decrease in/serum nitrite/nitrate and brain DB00143 levels along with an increase in brain TBARS levels and P22303 activity . DB06403 significantly improved hyperhomocysteinemia-induced impairment of learning , memory , endothelial dysfunction , and changes in various biochemical parameters . These effects were comparable to that of donepezil serving as positive control . It is concluded that ambrisentan , a selective P25101 receptor antagonist may be considered as a potential pharmacological agent for the management of hyperhomocysteinemia-induced vascular dementia . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity . Association study of cholesterol-related genes in Alzheimer 's disease . Alzheimer 's disease ( AD ) is a genetically complex disorder , and several genes related to cholesterol metabolism have been reported to contribute to AD risk . To identify further AD susceptibility genes , we have screened genes that map to chromosomal regions with high logarithm of the odds scores for AD in full genome scans and are related to cholesterol metabolism . In a European screening sample of 115 sporadic AD patients and 191 healthy control subjects , we analyzed single nucleotide polymorphisms in 28 cholesterol-related genes for association with AD . The genes P54868 , P14324 , RAFTLIN , Q9UKU7 , P61916 , and P45844 were associated with AD at a significance level of P < or = 0.05 in this sample . Replication trials in five independent European samples detected associations of variants within P54868 , P14324 , P61916 , or P45844 with AD in some samples ( P = 0.05 to P = 0.005 ) . We did not identify a marker that was significantly associated with AD in the pooled sample ( n = 2864 ) . Stratification of this sample revealed an P02649 -dependent association of P54868 with AD ( P = 0.004 ) . We conclude that genetic variants investigated in this study may be associated with a moderate modification of the risk for AD in some samples . DB00399 -induced IPP/ApppI production in vivo . Bisphosphonates are currently the most important class of anti-resorptive drugs used for the treatment of diseases involving excess bone resorption . Recently we discovered a new mechanism of action for bisphosphonates . Previously it has been shown that nitrogen-containing bisphosphonates ( N-BPs ) are not metabolized . However , our studies revealed that N-BPs induce formation of a novel pro-apoptotic DB00171 analog ( ApppI ) , as a consequence of the inhibition of P14324 in the mevalonate pathway , and the subsequent accumulation of isopentenyl pyrophosphate ( IPP ) in vitro . The primary aim of the current study was to determine whether zoledronic acid ( a N-BP ) induces IPP/ApppI formation in vivo . Mass spectrometry was used to identify whether in vivo administration of zoledronic acid-induced IPP/ApppI production by mouse peritoneal macrophages or bone marrow cells . IPP/ApppI could be detected in extracts from peritoneal macrophages isolated from zoledronic acid-treated animals . Increasing IPP/ApppI accumulation was determined up to 7 days after drug injection , indicating prolonged P14324 inhibition by zoledronic acid . Importantly , this is the first report of in vivo production of ApppI , supporting the biological significance of this molecule . Acute kidney injury and inflammatory immune reconstitution syndrome in mixed genotype ( A/E ) hepatitis B virus co-infection in HIV-associated lymphoma . We report a first case of HIV-associated lymphoma ( P42357 ) presenting with acute kidney injury ( AKI ) and inflammatory immune reconstitution syndrome ( IRIS ) . A 39-year-old male , treated with nonsteroidal anti-inflammatory drugs ( NSAIDs ) for one month prior to admission , developed AKI , left testicular tumor , and recurrent swelling of the right parotid gland . A resected testicular tumor exhibited features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma . Renal biopsy showed hydro-degeneration of renal tubules , interstitial inflammatory cells , and a small number of lymphoma cells in the sub-capsule , compatible with acute interstitial nephritis . DB00117 renal dysfunction rapidly recovered following chemotherapy and combination antiretroviral therapy ( cART ) . He developed pneumonia concomitantly with a decrease in HIV-RNA level and an increase in P01730 + cells after the first cycle of chemotherapy , which spontaneously resolved after the second cycle of chemotherapy without additional anti-infection drugs ; thus , his pneumonia fulfilled the diagnostic criteria for IRIS . We suggest that IRIS may frequently develop during chemotherapy for P42357 , but may be overlooked . He was coinfected with hepatitis B virus ( HBV ) , which genotypes known as is associated with liver-related mortality and response to antiviral therapy ; recently , an intimate interplay between HIV and HBV in the onset of lymphoma has been reported . Therefore , we addressed the HBV genotype in the patient . The analysis revealed that he exhibited a mixed genotype ( A/E ) not native to Japan and primarily found in Europe and North America or West Africa . These findings suggest that universal vaccination for juveniles against HBV is warranted in Japan .

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MH_train_1257

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interacts_with DB01016?

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DB01016 exerts an antitumor activity through reactive oxygen species-c-jun NH2-terminal kinase pathway in human gastric cancer cell line MGC-803 . DB01016 , a blocker of DB00171 -sensitive potassium ( K( DB00171 ) ) channels , can suppress progression of many cancers , but the involved mechanism is unclear . Herein we reported that MGC-803 cells expressed the K( DB00171 ) channels composed of Kir6.2 and Q09428 subunits . DB01016 induced cellular viability decline , coupled with cell apoptosis and reactive oxygen species ( ROS ) generation in MGC-803 cells . Meanwhile , glibenclamide increased NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion ( O2- ) generation , and caused mitochondrial respiration dysfunction in MGC-803 cells , suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria . DB01016 could also lead to loss of mitochondrial membrane potential , release of cytochrome c and apoptosis-inducing factor ( O95831 ) , and activation of c-jun NH2-terminal kinase ( JNK ) in MGC-803 cells . Pretreatment with antioxidant N-acetyl-l-cysteine ( Q9C000 ) prevented glibenclamide-induced JNK activation , apoptosis and cellular viability decline . Furthermore , glibenclamide greatly decreased the cellular viability , induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast ( MEF ) cells but not in P45983 -/- or P45984 -/- MEF cells . Taken together , our study reveals that glibenclamide exerts an antitumor activity in MGC-803 cells by activating ROS-dependent , JNK-driven cell apoptosis . These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer . Genetic demonstration of intestinal Q9UHC9 as a major determinant of hepatic cholesterol and blood atherogenic lipoprotein levels . OBJECTIVE : The correlation between intestinal cholesterol absorption values and plasma low-density lipoprotein-cholesterol ( LDL-C ) levels remains controversial . Niemann-Pick-C1-Like 1 ( Q9UHC9 ) is essential for intestinal cholesterol absorption , and is the target of ezetimibe , a cholesterol absorption inhibitor . However , studies with Q9UHC9 knockout mice or ezetimibe can not definitively clarify this correlation because Q9UHC9 expression is not restricted to intestine in humans and mice . In this study we sought to genetically address this issue . METHODS AND RESULTS : We developed a mouse model that lacks endogenous ( Q9UHC9 ) and P01130 ( P01130 ) ( DKO ) , but transgenically expresses human Q9UHC9 in gastrointestinal tract only ( DKO/ Q9NUQ9 (IntOnly) mice ) . Our novel model eliminated potential effects of non-intestinal Q9UHC9 on cholesterol homeostasis . We found that human Q9UHC9 was localized at the intestinal brush border membrane of DKO/ Q9NUQ9 (IntOnly) mice . DB04540 feeding induced formation of Q9UHC9 -positive vesicles beneath this membrane in an ezetimibe-sensitive manner . Compared to DKO mice , DKO/ Q9NUQ9 (IntOnly) mice showed significant increases in cholesterol absorption and blood/hepatic/biliary cholesterol . Increased blood cholesterol was restricted to very low-density lipoprotein ( VLDL ) and LDL fractions , which was associated with increased secretion and plasma levels of apolipoproteins B100 and B48 . Additionally , DKO/ Q9NUQ9 (IntOnly) mice displayed decreased fecal cholesterol excretion and hepatic/intestinal expression of cholesterologenic genes . DB00973 treatment virtually reversed all of the transgene-related phenotypes in DKO/ Q9NUQ9 (IntOnly) mice . CONCLUSION : Our findings from DKO/ Q9NUQ9 (IntOnly) mice clearly demonstrate that Q9UHC9 -mediated cholesterol absorption is a major determinant of blood levels of apolipoprotein B-containing atherogenic lipoproteins , at least in mice . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . P45983 is required for Q15399 gene expression in macrophages . The regulation of innate immune responses to pathogens occurs through the interaction of Toll-like receptors ( TLRs ) with pathogen-associated molecular patterns and the activation of several signaling pathways whose contribution to the overall innate immune response to pathogens is poorly understood . We demonstrate a mechanism of control of murine macrophage responses mediated by Q15399 /2 heterodimers through P45983 ( P45983 ) activity . JNK controls tumor necrosis factor alpha production and TLR-mediated macrophage responses to Borrelia burgdorferi , the causative agent of Lyme disease , and the Q15399 / O60603 -specific agonist PAM(3) P41240 (4) . P45983 , but not P45984 , activity regulates the expression of the tlr1 gene in the macrophage cell line RAW264.7 , as well as in primary CD11b(+) cells . We also show that the proximal promoter region of the human tlr1 gene contains an AP-1 binding site that is subjected to regulation by the kinase and binds two complexes that involve the JNK substrates c-Jun , JunD , and P39905 -2 . These results demonstrate that P45983 regulates the response to Q15399 /2 ligands and suggest a positive feedback loop that may serve to increase the innate immune response to the spirochete . The glial cell modulator and phosphodiesterase inhibitor , DB05066 ( ibudilast ) , attenuates prime- and stress-induced methamphetamine relapse . Stress and renewed contact with drug ( a " slip " ) have been linked to persisting relapse of methamphetamine abuse . Human brain microglial activation has been linked with methamphetamine abuse , and inhibitors of glial cell activation , certain phosphodiesterase ( PDE ) inhibitors , and glial cell derived neurotrophic factor ( P39905 ) have been reported to modulate drug abuse effects . Our objective was to determine whether the glial cell attenuator , 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine ( DB05066 , ibudilast ) , a non-selective PDE inhibitor and promoter of P39905 , could reduce stress- and methamphetamine prime-induced reinstatement of methamphetamine-seeking behavior . Male Long-Evans hooded rats were trained to lever press reinforced with 0.1 mg/kg i.v. methamphetamine infusion according to fixed-ratio 1 ( FR1 ) reinforcement schedules during daily , 2-hour experimental sessions . After performance had stabilized , lever pressing was extinguished for 12 consecutive sessions and doses of 0 ( vehicle ) , 2.5 and 7.5 mg/kg DB05066 were then administered intraperitoneally b.i.d. on the last 2 days of extinction and then once on the testday to separate groups of 12 rats . During testing , the rats were given 15 min of intermittent footshock or a 1 mg/kg i.p. methamphetamine prime followed by a 2-hour reinstatement test session . DB05066 significantly reduced response levels of footshock-induced ( 2.5 and 7.5 mg/kg ) and prime-induced ( 7.5 mg/kg ) reinstatement of extinguished methamphetamine-maintained responding . DB05066 has properties consistent with the ability to attenuate relapse precipitated by stress and methamphetamine " slips " during abstinence . These results thus reinforce interest in atypical neurobiological mechanisms which could be exploited for developing novel medications for treating drug abuse disorders . Clinical and functional characterization of the Pro1198Leu Q09428 gene mutation associated with permanent neonatal diabetes mellitus . AIMS/INTRODUCTION : The adenosine triphosphate ( DB00171 ) -sensitive potassium ( KATP ) channel is a key component of insulin secretion in pancreatic β-cells . Activating mutations in Q09428 encoding for the sulfonylurea receptor subunit of the KATP channel have been associated with the development of neonatal diabetes mellitus ( NDM ) . The aim was to investigate clinical and functional characterization of the Pro1198Leu Q09428 gene mutation associated with permanent NDM ( PNDM ) . MATERIALS AND METHODS : The coding regions and conserved splice sites of Q14654 , Q09428 and P01308 were screened for mutations in a 12-year-old girl diagnosed with PNDM . The functional property of the mutant channel identified was examined with patch-clamp experiments in COS-1 cells . We also investigated the difference of effectiveness between two groups of oral sulfonylureas in vitro and in the patient . RESULTS : We identified a heterozygous missense mutation ( c.3593 C > T , Pro1198Leu ) in Q09428 . The mutated residue ( P1198 ) is located within a putative binding site of sulfonylureas , such as tolbutamide or gliclazide . In patch-clamp experiments , the mutant channel was less DB00171 sensitive than the wild type . Furthermore , the sensitivity to tolbutamide was also reduced in the mutant channel . In addition to the tolbutamide/gliclazide binding site , glibenclamide is thought to also bind to another site . DB01016 was more effective than other sulfonylureas in vitro and in the patient . The treatment of the patient was finally able to be switched from insulin injection to oral glibenclamide . CONCLUSIONS : We identified the Pro1198Leu Q09428 mutation in a PNDM patient , and clarified the functional and clinical characterization . The present findings provide new information for understanding PNDM . Silencing of ALA dehydratase affects ALA-photodynamic therapy efficacy in K562 erythroleukemic cells . Synthesis of protoporphyrin IX ( PpIX ) by malignant cells is essential for the success of ALA-based photodynamic therapy ( PDT ) . Two key enzymes that were described as affecting PpIX accumulation during ALA treatment are porphobilinogen deaminase ( P08397 ) and ferrochelatase . Here , we show that down regulation of ALA dehydratase ( P13716 ) expression and activity by specific shRNA induced a marked decrease in PpIX synthesis in K562 erythroleukemic cells . Photo-inactivation efficacy following DB00855 was directly correlated with P13716 -silencing and cellular levels of PpIX . MTT metabolism following DB00855 was shown to be 60 % higher in P13716 -silenced cells in comparison to control cells , indicating that mitochondria were protected in the silenced cells . Morphological analysis by scanning electron microscopy ( SEM ) of cells treated by DB00855 showed no morphological changes in P13716 -silenced cells , in contrast to controls exhibiting cell deformations and lysis . Membrane integrity following DB00855 was kept intact and undamaged in P13716 -silenced cells as examined by P08758 -FITC/PI staining and LDH-L leakage . We conclude that P13716 , although it is present in the cell at abundant levels , has a major and limiting role in regulating PpIX synthesis and DB00855 outcome . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress . Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis . Because oxidative stress contributes to atherosclerosis , we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon . Bovine aortic endothelial cells were exposed to static , laminar ( 15 dyn/cm2 ) , and oscillatory shear stress ( +/-15 dyn/cm2 ) . Oscillatory shear increased superoxide ( O2.- ) production by more than threefold over static and laminar conditions as detected using electron spin resonance ( P03372 ) . This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources . DB05262 also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence . DB02134 -dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress . This was associated with decreased xanthine dehydrogenase ( P47989 ) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase ( XO ) to P47989 . We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase . These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity . Transfection of these cells with p47phox restored XO protein levels . Finally , in bovine aortic endothelial cells , prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress . These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress . DB09559 , a fully human IgG1 mAb directed against the P00533 for the potential treatment of cancer . DB09559 ( DB05774 ) , under development by ImClone Systems in collaboration with Bristol-Myers Squibb , is a fully human IgG1 mAb targeting the epidermal growth factor receptor ( P00533 ) , for the potential intravenous treatment of cancer , in particular NSCLC . In vitro studies demonstrate that necitumumab inhibits downstream targets in the P00533 pathway ( eg , MAPK ) , which are important for cellular proliferation , differentiation , invasion and metastasis . Furthermore , because necitumumab is an IgG1 construct , it has the potential to induce antibody-dependent cell-mediated cytotoxicity against tumor cells . Preclinical studies indicated that the antitumor activity of necitumumab is either comparable with or superior to that of ImClone 's chimeric anti- P00533 mAb cetuximab . In a phase I clinical trial in patients with advanced solid malignancies , necitumumab displayed nonlinear pharmacokinetic behavior . The toxicity profile of necitumumab is acceptable , with skin toxicity being the most frequently reported adverse event in the phase I and II clinical trials conducted to date . Preliminary data from a phase II clinical trial of necitumumab in combination with chemotherapy for the first-line treatment of advanced colon cancer are promising . Success in the ongoing phase III clinical trials in patients with advanced NSCLC would lead to necitumumab becoming a valuable addition to future therapeutic strategies in oncology . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Linkage studies of cholestasis familiaris groenlandica/Byler-like disease with polymorphic protein and blood group markers . In Greenland , and especially East Greenland ( Tasiilaq ) , a common recessive disease , cholestasis familiaris groenlandica ( CFG ) /Byler-like disease , occurs in Eskimo children [ 1 ] . In a period from 1964-1991 , at least 22 children out of about 2,121 newborns were born with this disease ( gene frequency q = 0.102 ) . Samples from 126 persons , from a large pedigree in East Greenland including 7 affected and from two families in West Greenland with a total of 3 affected children , have been collected for studying 45 polymorphic markers and for mapping the CFG disease . Polymorphisms and exclusion data were found for the following markers : P04217 , P16442 , P24666 , P02765 , P00736 , P13671 , FY , GC , Q04760 , GPT , HP , P19827 , JK , P02724 , P06028 , ORM , P1 , P36871 , PI , P27169 , RH and P20062 . Small positive lod scores ( Z < 1.5 ) were found to the following markers : P19827 , JK and P20062 . The following markers were nonpolymorphic in this material : P00813 , P00568 , P13716 , P06727 , P02749 , BF , P01024 , P06276 , CHE2 , CO , P10768 , Q9BTY2 , P00488 , P05160 , P23276 , LE , P19526 , LU , P12955 , P52209 , A6NDG6 , P00747 , Q10981 , P00441 and TF . Steady-state interactions of glibenclamide with P13569 : evidence for multiple sites in the pore . The objective of the present study was to clarify the mechanism by which the sulfonylurea drug , glibenclamide , inhibits single P13569 channels in excised patches from Xenopus oocytes . DB01016 blocks the open pore of the channel via binding at multiple sites with varying kinetics . In the absence of glibenclamide , open-channel bursts exhibited a flickery intraburst closed state ( C1 ) ; this is due to block of the pore by the pH buffer , Q9UGI8 . Application of 25 microM glibenclamide to the cytoplasmic solution resulted in the appearance of two drug-induced intraburst closed states ( P06681 , P01024 ) of widely different duration , which differed in pH-dependence . The kinetics of interaction with the P01024 state , but not the P06681 state , were strongly voltage-dependent . The durations of both the P06681 and P01024 states were concentration-dependent , indicating a non-linear reaction scheme . Application of drug also increased the burst duration , which is consistent with an open-channel blocking mechanism . A kinetic model is proposed . These results indicate that glibenclamide interacts with open P13569 channels in a complex manner , involving interactions with multiple binding sites in the channel pore . Modulation of cytokine production and enhancement of cell viability by Q9NYK1 and Q9NR96 ligands during anthrax infection of macrophages . Inhalation of Bacillus anthracis , a bioterrorism agent , results in a high mortality rate despite appropriate antibiotic therapy . Macrophages appear to be a key factor in B. anthracis pathogenesis . The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax . However , preactivation of Toll-like receptors ( TLRs ) could modify the host response . TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens . We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli . We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines P05231 , P01375 , P22301 , and IL-12 p40 . Our studies showed that pretreatment of mouse macrophages with the Q9NR96 ligand DB05463 , or the Q9NYK1 ligands R-848 and IT-37 , results in a substantial decrease in the subsequent secretion of P05231 and P01375 in response to B. anthracis infection of macrophages . Furthermore , the Q9NYK1 and Q9NR96 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages . These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability . 17β-estradiol inhibits P14780 and Q09428 /TrpM4 expression and activation and thereby attenuates BSCB disruption/hemorrhage after spinal cord injury in male rats . Blood-spinal cord barrier ( BSCB ) disruption and progressive hemorrhage after spinal cord injury ( SCI ) lead to secondary injury and the subsequent apoptosis and/or necrosis of neuron and glia , causing permanent neurological deficits . In this study , we examined the effect of 17β-estradiol ( E2 ) on BSCB breakdown and hemorrhage as well as subsequent inflammation after SCI . After a moderate contusion injury at the 9th thoracic segment of spinal cord , E2 ( 300 μg/kg ) was administered by iv injection immediately after SCI , and the same dose of E2 was then administered 6 and 24 hours after injury . Our data show that E2 attenuated BSCB permeability and hemorrhage and reduced the infiltration of neutrophils and macorphages after SCI . Consistent with this finding , the expression of inflammatory mediators was significantly reduced by E2 . Furthermore , E2 treatment significantly inhibited the expression of sulfonylurea receptor 1 and transient receptor potential melastatin 4 after injury , which are known to mediate hemorrhage at an early stage after SCI . Moreover , the expression and activation of matrix metalloprotease-9 after injury , which is known to disrupt BSCB , and the degradation of tight junction proteins , such as zona occludens-1 and occludin , were significantly inhibited by E2 treatment . Furthermore , the protective effects of E2 on BSCB disruption and functional improvement were abolished by an estrogen receptor antagonist , ICI 182780 ( 3 mg/kg ) . Thus , our study provides evidence that the neuroprotective effect of E2 after SCI is , in part , mediated by inhibiting BSCB disruption and hemorrhage through the down-regulation of sulfonylurea receptor 1/transient receptor potential melastatin 4 and matrix metalloprotease-9 , which is dependent on estrogen receptor . Synthesis and biological evaluation of a library of resveratrol analogues as inhibitors of P23219 , P35354 and NF-kappaB . DB02709 ( 4,3',5'-trihydroxystilbene ) is a naturally occurring antioxidant that inhibits cyclooxygenase-1 ( P23219 ) , cyclooxygenase-2 ( P35354 ) and the transcription factor NF-kappaB . A 78-membered library of resveratrol analogues in which the substituents on the two aryl rings and alkene were varied was synthesized using a solid-phase Wittig olefination reaction . The library contains inhibitors against all three proteins that were more potent than resveratrol itself . Preliminary structure-activity relationships were also obtained from these data that permitted the derivation of pharmacophore models for each of the three targets . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . Monoclonal antibodies targeting P01584 reduce biomarkers of atherosclerosis in vitro and inhibit atherosclerotic plaque formation in P02649 -deficient mice . OBJECTIVE : Atherosclerosis is a condition that is increasingly contributing to worldwide mortality through complications such as stroke and myocardial infarction . IL-1β plays multiple direct , local roles in the formation and stability of the atheroma by eliciting the production of additional cytokines and proteolytic enzymes from macrophages , endothelial cells ( EC ) and smooth muscle cells ( SMC ) . We therefore tested whether an anti-IL-1β antibody , DB06062 , might inhibit the secretion of pro-atherogenic cytokines from macrophages in vitro and affect a positive outcome in the P02649 -deficient mouse ( ApoE(-/-) ) model of atherosclerosis in vivo . METHODS AND RESULTS : In an in vitro co-culture model , DB06062 inhibited macrophage-induced secretion of key atherogenic cytokines from EC and SMC , including P05231 , P10145 , P13500 and TNFα . The release of degradative enzymes , such as the matrix metalloproteinases P08254 and P14780 , was also decreased by DB06062 . In addition , DB06062 inhibited the secretion of P13232 from EC and P05112 from SMC , cytokines not previously reported to be driven by IL-1β in this context . In vivo , XMA052 MG1K , a chimeric murine version of DB06062 , inhibited the formation of atherosclerotic lesions in the ApoE(-/-) model at all three doses tested . This effect was comparable to that reported for complete genetic ablation of IL-1β or IL-1R1 on an ApoE(-/-) background and was associated with decreases in plasma non-HDL/HDL cholesterol ratio and plaque lipid content and macrophage infiltration . CONCLUSIONS : These results demonstrate for the first time that an antibody targeting IL-1β can inhibit the progression of atherosclerosis in vivo , highlighting the importance of this key cytokine in cardiovascular disease . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . UMD ( Universal mutation database ) : a generic software to build and analyze locus-specific databases . The human genome is thought to contain about 80,000 genes and presently only 3,000 are known to be implicated in genetic diseases . In the near future , the entire sequence of the human genome will be available and the development of new methods for point mutation detection will lead to a huge increase in the identification of genes and their mutations associated with genetic diseases as well as cancers , which is growing in frequency in industrial states . The collection of these mutations will be critical for researchers and clinicians to establish genotype/phenotype correlations . Other fields such as molecular epidemiology will also be developed using these new data . Consequently , the future lies not in simple repositories of locus-specific mutations but in dynamic databases linked to various computerized tools for their analysis and that can be directly queried on-line . To meet this goal , we devised a generic software called UMD ( Universal Mutation Database ) . It was developed as a generic software to create locus-specific databases ( LSDBs ) with the 4(th) Dimension(R) package from ACI . This software includes an optimized structure to assist and secure data entry and to allow the input of various clinical data . Thanks to the flexible structure of the UMD software , it has been successfully adapted to nine genes either involved in cancer ( P25054 , P04637 , P06400 , O00255 , Q09428 , P40337 , and P19544 ) or in genetic diseases ( P35555 and P01130 ) . Four new LSDBs are under construction ( P49748 , P11310 , KIR6 , and P29400 ) . Finally , the data can be transferred to core databases . Inhibitors of DB00171 -binding cassette transporters suppress interleukin-12 p40 production and major histocompatibility complex II up-regulation in macrophages . DB00171 -binding cassette ( DB01048 ) transporters are a large family of proteins whose role is to translocate various substances across biological membranes . They include the Tangier disease protein ABC1 , sulfonylurea receptors ( Q09428 ) , multidrug resistance protein ( MDR ) , and cystic fibrosis transmembrane regulator ( P13569 ) . In the current study , we investigated the involvement of ABC transporters in the regulation of lipopolysaccharide ( LPS ) and/or interferon ( IFN ) -gamma-induced interleukin ( IL ) -12 p40 and tumor necrosis factor ( P01375 ) -alpha production , nitric oxide formation , as well as major histocompatibility complex II up-regulation in macrophages . The general ABC transporter inhibitor glibenclamide suppressed both IL-12 p40 and nitric oxide production . However , glibenclamide failed to affect the production of P01375 . The selective ABC1 inhibitors 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and sulfobromophthalein mimicked the suppressive effect of glibenclamide on IL-12 p40 production . On the other hand , both the MDR inhibitor verapamil and P13569 blocker 2,2'-iminodibenzoic acid failed to suppress the production of IL-12 p40 . Furthermore , selective inhibitors and activators of SURs were without effect . In agreement with the pharmacological data , macrophages expressed mRNA for ABC1 , but not SURs or P13569 . Intracellular levels of IL-12 p40 were decreased by glibenclamide , suggesting that glibenclamide does not affect IL-12 p40 secretion . The effect of glibenclamide did not involve an interference with the activation of the p38 and Q8NFH3 /44 mitogen-activated protein kinases or c-Jun kinase . DB01016 also suppressed P01579 -induced up-regulation of major histocompatibility complex II . Taken together , our results indicate that ABC proteins regulate LPS and/or P01579 -induced macrophage activation .

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interacts_with DB00502?

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P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . DB01221 receptor modulation by the neuropeptide apelin : implications for excitotoxic injury . Excitotoxic neuronal damage via over-activation of the DB01221 receptor has been implicated in many neurodegenerative diseases . In vitro modeling of excitotoxic injury has shown that activation of G-protein coupled receptors ( GPCRs ) counteracts such injury through modulation of neuronal pro-survival pathways and/or DB01221 receptor signaling . We have previously demonstrated that the GPCR P35414 and its endogenous neuropeptide ligand apelin can protect neurons against excitotoxicity , but the mechanism(s) of this neuroprotection remain incompletely understood . We hypothesized that apelin can promote neuronal survival by activating pro-survival signaling as well as inhibiting DB01221 receptor-mediated excitotoxic signaling cascades . Our results demonstrate that ( i ) apelin activates pro-survival signaling via inositol trisphosphate ( IP(3) ) , protein kinase C ( PKC ) , mitogen-activated protein kinase kinase 1/2 ( Q02750 /2 ) , and extracellular signal-regulated kinase-1/2 ( P27361 /2 ) to protect against excitotoxicity , and ( ii ) apelin inhibits excitotoxic signaling by attenuating DB01221 receptor and calpain activity , and by modulating DB01221 receptor subunit Q13224 phosphorylation at serine 1480 . These studies delineate a novel apelinergic signaling pathway that concurrently promotes survival and limits DB01221 receptor-mediated injury to protect neurons against excitotoxicity . Defining apelin-mediated neuroprotection advances our understanding of neuroprotective pathways and will potentially improve our ability to develop therapeutics for excitotoxicity-associated neurodegenerative disorders . Multiple-dose pharmacokinetics confirm no accumulation and dose proportionality of the novel promotile drug tegaserod ( HTF 919 ) . OBJECTIVE : To evaluate the steady-state pharmacokinetics ( PK ) and dose proportionality of the selective Q13639 receptor partial agonist tegaserod ( HTF 919 ) in healthy subjects . METHODS : Eighteen subjects were given 2 , 6 , or 12-mg doses of tegaserod twice daily ( b.i.d. ) for 5 days , with PK and safety assessments made during the 12 h or 24 h following first administration , and 12 h after the final dose . RESULTS : DB01079 was rapidly absorbed [ time to reach measured maximum plasma concentration after multiple administrations ( tmax,ss ) 1 h ] . Steady-state PK were consistent with single-dose PK characteristics supporting that there was no accumulation of tegaserod in plasma based on systemic exposure . Mean measured maximum plasma concentration after multiple administrations ( Cmax,ss ) and area under the plasma concentration-time curve over one dosing interval ( tau , 0-12 h after drug administration , AUC tau ) were between 0.7 +/- 0.3 ng/ml and 5.6 +/- 2.9 ng/ml and 2.4 +/- 1.3 h.ng/ml and 20.4 +/- 14.0 h.ng/ml , respectively , indicating dose-proportional PK of tegaserod in the range 2-12 mg b.i.d . DB01079 was safe and well tolerated . No serious adverse events were reported . CONCLUSION : DB01079 exhibits no accumulation and dose-proportional PK after multiple doses . Abnormal sensitivity of cortisol-producing adrenocortical adenomas to serotonin : in vivo and in vitro studies . Two patients with incidentally discovered adrenocortical adenomas underwent a series of pharmacological and physiological tests after pretreatment with dexamethasone . Illicit plasma cortisol responses to the serotonin (5-HT)4 receptor agonist cisapride were observed in the two patients . Significant increases in plasma cortisol levels were also noticed after glucagon and combined TRH/ DB00644 / P01286 stimulation tests in patient 1 and after administration of the lysine vasopressin precursor terlipressin in patient 2 . After adrenalectomy , in vitro studies were conducted to investigate the cortisol responses of cultured tumor cells to serotonergic ligands and peptide hormones . In the two cases , 5-HT stimulated cortisol secretion from tumor cells with increased efficacy and/or potency to activate steroidogenesis by comparison with normal adrenocortical cells . The corticotropic effect of 5-HT was inhibited by the specific Q13639 receptor antagonist GR 113808 and more potently by methiothepin , a nonspecific serotonergic antagonist having no affinity for the Q13639 receptor . These results show that the hypersensitivity of the tumors to 5-HT was related to tissue expression of an ectopic serotonergic receptor in addition to the eutopic Q13639 receptor . In the two adenoma tissues , immunohistochemical studies revealed the presence of 5-HT-like immunoreactivity within clusters of steroidogenic cells , suggesting that 5-HT acted through an autocrine/paracrine mechanism to stimulate steroidogenesis . Glucagon and DB00644 but not TRH , P01286 , and human chorionic gonadotropin stimulated cortisol secretion from tumor 1 cells . In conclusion , this study provides the first observation of adrenocortical cortisol-producing adenomas hypersensitive in vivo and in vitro to serotonergic agonists . Our results also show that cortisol-producing adenomas can express simultaneously several illegitimate receptors . P03952 promotes epidermal growth factor receptor transactivation and signaling in vascular smooth muscle through direct activation of protease-activated receptors . The kallikrein-kinin system , along with the interlocking renin-angiotensin system , is a key regulator of vascular contractility and injury response . The principal effectors of the kallikrein-kinin system are plasma and tissue kallikreins , proteases that cleave high molecular weight kininogen to produce bradykinin . Most of the cellular actions of kallikrein ( KK ) are thought to be mediated by bradykinin , which acts via G protein-coupled B1 and B2 bradykinin receptors on VSMCs and endothelial cells . Here , we find that primary aortic vascular smooth muscle but not endothelial cells possess the ability to activate plasma prekallikrein . Surprisingly , exposing VSMCs to prekallikrein leads to activation of the P27361 /2 mitogen-activated protein kinase cascade via a mechanism that requires kallikrein activity but does not involve bradykinin receptors . In transfected HEK293 cells , we find that plasma kallikrein directly activates G protein-coupled protease-activated receptors ( PARs ) 1 and 2 , which possess consensus kallikrein cleavage sites , but not PAR4 . In vascular smooth muscles , KK stimulates ADAM ( a disintegrin and metalloprotease ) 17 activity via a PAR1/2 receptor-dependent mechanism , leading sequentially to release of the endogenous P78536 substrates , amphiregulin and tumor necrosis factor-α , metalloprotease-dependent transactivation of epidermal growth factor receptors , and metalloprotease and epidermal growth factor receptor-dependent P27361 /2 activation . These results suggest a novel mechanism of bradykinin-independent kallikrein action that may contribute to the regulation of vascular responses in pathophysiologic states , such as diabetes mellitus . DB02426 effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 and P05141 may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 -/- mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 -independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 -/- brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 -independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 -/- mice . However , in liver , only Ant2 mRNA was found , whereas in brown adipose tissue , Ant1 and Ant2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 isoform mediates fatty-acid-induced uncoupling , whereas the P12235 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . Group IB secretory phospholipase A2 stimulates leukotriene B4 production by a unique mechanism in human neutrophils . We found that group IB secretory phospholipase A(2) ( sPLA(2)-IB ) stimulates leukotriene B4 ( LTB4 ) production in the absence of cytochalasin B in human neutrophils . Although LTB4 production has been reported to be associated with arachidonic acid release , the exogenous addition of sPLA(2)-IB did not induce this release from human neutrophils , suggesting that sPLA(2)-IB stimulates LTB4 production without affecting arachidonic acid . Moreover , the intracellular signaling events induced by sPLA(2)-IB included an increase in intracellular Ca(2+) , which is required for LTB4 production . sPLA(2)-IB also stimulated mitogen-activated protein kinase P29323 , but its activity was not required for LTB4 production . In terms of functional aspects , the supernatant of sPLA(2)-IB-stimulated human neutrophils caused chemotactic migration , which was almost completely inhibited by preincubating these cells with three different P09917 inhibitors ( MK-886 , AA-861 , or NDGA ) . Taken together , we suggest that sPLA(2)-IB plays a role in the modulation of inflammatory and immune responses by inducing LTB4 production in human neutrophils . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . Genetics of physical activity and physical inactivity in humans . Emerging evidence suggests that physical activity and sedentary behavior [ reflected in physical inactivity ( PI ) ] , might be two different phenotypes that may have distinct underlying physiological mechanisms . The purpose of this review is to summarize the existing literature on the genetic determinants of PA and PI phenotypes in humans , considering them as distinct behaviors . Completed in January 2011 , this review includes family studies , twin studies , association studies , genome-wide linkage studies and genome-wide association scan ( GWAs ) reporting different physical activity/inactivity-related phenotypes . In regards to PA , familial aggregation studies resulted in heritability estimates ranging from 0 to 60 % , and twin studies yielded heritability estimates ( a(2) ) and shared environment ( c(2) ) scores for PA phenotypes ranging from 0.00 to 0.85 and 0.00 to 0.84 , respectively . Unique environmental ( e(2) ) results are well dispersed from 0.12 to 0.72 . Suggestive linkages were found with markers nearby different activity-related genes : P24530 , P32245 , P25874 , P12104 , P41180 , Q8IVB4 . Significant associations with PA phenotypes were found for Ace , Gln223ARrg , P32245 and P14416 genes . We found one GWAs that reported novel SNPs in the O95340 gene on chromosome 10q23.2 and in two intergenic regions on chromosomes 2q33.1 and 18p11.32 . Heritability estimates for PI ranged from 25 to 60 % and linkage studies recorded higher LOD scores for PI versus PA . The P12821 genotype was strongly associated with PI . There are potentially different genetic influences on PA versus PI phenotypes . Future studies should focus on the different genetic influences on PA and PI to improve our understanding of underlying determinants of these behaviors . Expression and purification of growth hormone-releasing factor with the aid of dihydrofolate reductase handle . Expression of a fusion protein composed of dihydrofolate reductase and a derivative of growth hormone-releasing factor resulted in the formation of inclusion bodies in Escherichia coli at 37 degrees C . Among various chemicals , such as detergents , protein denaturants , and acetic acid , tested for the ability to dissolve the inclusion bodies , acetic acid , Brij-35 , deoxycholic acid sodium salts , guanidine-HCl , and urea showed a strong solubilizing effect without damaging the P00374 activity . DB03166 was useful in terms of preparing P01286 derivatives , since it could be easily removed by lyophilization , and this made it easy to perform the succeeding BrCN treatment for cutting out the P01286 derivative from the fusion protein . The P01286 derivative was purified by reversed phase HPLC from the BrCN digest of the acetic acid extract , and its growth hormone-releasing activity was demonstrated . However , for obtaining a highly purified fusion protein itself , solubilization of inclusion bodies by urea was preferred because urea was the only agent which did not cause serious precipitation of the regenerated fusion protein after 10-fold dilution of the extracted inclusion bodies with buffer . The fusion protein was highly purified by means of a methotrexate affinity chromatography . Mechanism of the induction of the differentiation of HL-60 leukemia cells by antifolates . The classic inhibitor of dihydrofolate reductase ( P00374 ) , methotrexate ( MTX ) , has been shown to be an effective inducer of the differentiation of HL-60 promyelocytic leukemia cells ( Bodner A.J. et al. ; J. Natl. Cancer Inst. 67:1025-1030 ; 1981 ) . We have obtained evidence that induction of the differentiation of these cells by MTX , as well as by other folic acid antagonists , is the result of the effects of these agents on purine and thymine nucleotide biosynthesis . DB04485 ( 10 microM ) completely blocked both the cytotoxicity and induction of differentiation produced by the specific inhibitor of thymidylate synthase ( TS ) , N10-propargyl-5,8-dideazafolic acid ( CB-3717 ) . DB04485 also blocked the acute cytotoxicity caused by MTX and trimetrexate ( DB01157 ) ; the induction of differentiation and the loss of proliferative capacity , however , were only partially prevented by thymidine . DB04076 ( 100 microM ) , which completely restored antifolate-depleted purine nucleotide levels , had no effect on either the cytotoxicity or the induction of maturation produced by these agents . The growth inhibitory effects and the induction of differentiation caused by dideazatetrahydrofolic acid ( DDATHF ) , which acts on de novo purine nucleotide biosynthesis rather than on P00374 or TS , was completely prevented by hypoxanthine . DB04076 also completely prevented the inhibition of cellular replication and induction of differentiation by MTX and DB01157 when combined with thymidine . The findings suggest that the depletion of intracellular thymine nucleotide levels by the antifolates , MTX , DB01157 , and CB-3717 is the primary event involved in the maturation of HL-60 leukemia cells produced by these agents and that maturation occurs concomitantly with a high level of cytotoxicity. ( ABSTRACT TRUNCATED AT 250 WORDS ) Blocking dopamine D2 receptors by haloperidol curtails the beneficial impact of calorie restriction on the metabolic phenotype of high-fat diet induced obese mice . Calorie restriction is the most effective way of expanding life-span and decreasing morbidity . It improves insulin sensitivity and delays the age-related loss of dopamine receptor D(2) ( P14416 ) expression in the brain . Conversely , high-fat feeding is associated with obesity , insulin resistance and a reduced number of P14416 binding sites . We hypothesised that the metabolic benefit of calorie restriction involves the preservation of appropriate P14416 transmission . The food intake of wild-type C57Bl6 male mice was restricted to 60 % of ad lib. intake while they were treated with the P14416 antagonist haloperidol or vehicle using s.c. implanted pellets . Mice with ad lib. access to food receiving vehicle treatment served as controls . All mice received high-fat food throughout the experiment . After 10 weeks , an i.p. glucose tolerance test was performed and , after 12 weeks , a hyperinsulinaemic euglycaemic clamp . Hypothalamic P14416 binding was also determined after 12 weeks of treatment . Calorie-restricted ( CR ) vehicle mice were glucose tolerant and insulin sensitive compared to ad lib . ( AL ) fed vehicle mice . CR mice treated with haloperidol were slightly heavier than vehicle treated CR mice . DB00502 completely abolished the beneficial impact of calorie restriction on glucose tolerance and partly reduced the insulin sensitivity observed in CR vehicle mice . The metabolic differences between AL and CR vehicle mice were not accompanied by alterations in hypothalamic P14416 binding . In conclusion , blocking P14416 curtails the metabolic effects of calorie restriction . Although this suggests that the dopaminergic system could be involved in the metabolic benefits of calorie restriction , restricting access to high-fat food does not increase ( hypothalamic ) P14416 binding capacity , which argues against this inference . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . Neto1 associates with the DB01221 receptor/amyloid precursor protein complex . Neuropilin tolloid-like 1 ( Neto1 ) , is a CUB domain-containing transmembrane protein that was recently identified as a novel component of the DB01221 receptor complex . Here , we have investigated the possible association of Neto1 with the amyloid precursor protein ( P05067 )695/ Q05586 / Q12879 and APP695/ Q05586 / Q13224 DB01221 receptor trafficking complexes that we have previously identified . Neto1(HA) was shown to co-immunoprecipitate with assembled DB01221 receptors via Q12879 or Q13224 subunits ; Neto1(HA) did not co-immunoprecipitate APP695(FLAG) . Co-immunoprecipitations from mammalian cells co-transfected with APP695(FLAG) , Neto1(HA) and Q05586 / Q12879 or Q05586 / Q13224 revealed that all four proteins co-exist within one macromolecular complex . Immunoprecipitations from native brain tissue similarly revealed the existence of a Q05586 / Q12879 or Q13224 / P05067 /Neto1 complex . Neto1(HA) caused a reduction in the surface expression of both DB01221 receptor subtypes , but had no effect on APP695(FLAG) - or A5PKW4 -95α(c-Myc) enhanced surface receptor expression . The Neto1 binding domain of Q12879 was mapped using Q05586 / Q12879 chimeras and Q12879 truncation constructs . The extracellular Q12879 domain does not contribute to association with Neto1(HA) but deletion of the intracellular tail resulted in a loss of Neto-1(HA) co-immunoprecipitation which was paralleled by a loss of association between Q12879 and Q92796 . Thus , Neto1 is concluded to be a component of P05067 / DB01221 receptor trafficking complexes . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation : a novel role of the cysteinyl LT-1 receptor . Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues . Leukotrienes are potent inflammatory mediators , and similar to prostaglandins , are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues . Here we tested the hypothesis that blocking the negative regulation of leukotrienes , would lead to enhanced endochondral bone formation during fracture repair . A closed femoral fracture was created in mice . Animals were divided into three groups for treatment with either montelukast sodium , a cysteinyl leukotriene type 1 receptor antagonist ( trade name Singulair ) , zileuton , a P09917 enzyme inhibitor ( trade name DB00744 ) , or carrier alone . The fractures were analyzed using radiographs , quantitative gene expression , histology and histomorphometry , and immunohistochemistry . Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls . Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7 . Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation , and increased early bone formation markers when compared with controls . Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor , leading to increased chondrocyte proliferation at early time points . These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation , with important and previously unrecognized implications for both fracture repair , and in a broader context , systemic chondrocyte growth and differentiation . DB04892 . DB04892 , a derivative of physostigmine , was first described as an inhibitor of acetylcholinesterase ( P22303 ) and was shown to improve cognition in various experimental paradigms in rodents and dogs . It was clinically tested for Alzheimer 's disease , with moderate success in initial Phase II studies . DB04892 deserves attention for an additional quality of action : in addition to inhibiting P22303 , it modulates the amount of beta-amyloid precursor protein ( P05067 ) in neuronal cell culture by reducing P05067 translation . This effect probably involves interaction of phenserine with a regulatory element in the 5'-untranslated region of the P05067 gene that controls P05067 expression . DB04892 apparently reduces translational efficiency of P05067 mRNA into protein , a process that may involve an interaction with iron and/or an iron-responsive element . As a consequence , phenserine reduces beta-amyloid peptide ( Abeta ) formation in vitro and in vivo . DB04892 is also unique because of differing actions of its enantiomers : DB04892 is the active enantiomer for inhibition of P22303 , whereas (+)-phenserine ( ' posiphen ' ) has weak activity as an P22303 inhibitor and can be dosed much higher . Both enantiomers are equipotent in downregulating P05067 expression . (+)-Posiphen may be a promising drug , either alone or in combination with DB04892 , to attenuate the progression of Alzheimer 's disease . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . DB05822 , a nitric oxide-releasing aspirin derivative , exhibits a significant antiproliferative effect and alters cell cycle progression in human colon adenocarcinoma cell lines . DB00435 -releasing non-steroidal antiinflammatory drugs ( NO-NSAIDs ) are safer than NSAIDs due to their ability to reduce gastric toxicity . We assessed the cytotoxic activity of a new aspirin derivative , DB05822 , after different exposure schedules , in three human colon adenocarcinoma cell lines . All the lines were positive for P23219 protein and mRNA , as evaluated by Western blot and RT-PCR , respectively , while only one was positive for P35354 . The cytostatic and cytocidal activity was determined by sulforhodamine B assay and evaluated according to Monks ' model . Cytostatic activity was observed after a 24-h drug exposure and 50 % growth inhibition was reached at concentrations ranging from 165 to 250 micro M in all cell lines , whereas with aspirin the IC50 was never reached , even at the maximum concentration tested ( 500 micro M ) , and was independent of P23219 or P35354 status . Cytocidal activity was observed only at the highest concentrations and persisted for a long time after drug removal . Flow cytometric analysis showed that the NO-aspirin compound induced a persistent accumulation of cells in G2-M phase in all the cell lines after at least 48 h exposure . Specifically , the block pertained mainly to G2 phase , whereas mitotic index was not affected at all . Our results indicate that DB05822 has an in vitro cytostatic activity superior to that of its parental aspirin compound , which makes it a potentially important tumor preventive agent . Furthermore , the cytocidal effect observed at the highest concentrations and the induction of a specific block in G2 phase renders it a promising candidate for drug combination treatments . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice . Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined . The immediate phase reaction ( IPR ) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined . The inhibition of IPR by cetirizine and mequitazine were potent , but those by cyproheptadine and diphenhydramine were weak . The later phase reaction ( LPR ) assessed at 24 hours after antigen application was inhibited by chlorpheniramine , oxatomide , ketotifen , mequitazine , emedastine , terfenadine and azelastine . The inhibition of LPR by emedastine was potent , but those by ketotifen and terfenadine were only partial . DB01084 inhibited both IPR and LPR comparably . Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction , and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR . P35367 antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism , which may have an additional benefit for the treatment of allergic diseases .

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interacts_with DB09048?

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Specific Link between Lung and Large Intestine : A New Perspective on Neuropeptide Secretion in Lung with Herbal Laxative Stimulation . Background. To investigate the specific link between lung and large intestine . Methods . Rat P48444 -like model was prepared . Mirabilite or Chinese rhubarb was administrated intragastrically to stimulate the large intestine . Histological analysis of lung inflammation was assessed . The tissues levels of SP , P01282 , P25103 , P32241 , and P41587 were measured by using ELISA kits . In addition , mouse model of allergic asthma was prepared . Mirabilite was administrated intragastrically to stimulate the large intestine . Airway responsiveness and lung inflammation were assessed . The tissues levels of SP , P01282 , P20366 , NKB , P25103 , P32241 , and P41587 were measured by using ELISA kits . Results . Stimulating the intestine with Mangxiao or Dahuang , SP , P25103 , P01282 , P32241 , and P41587 were significantly increased in intestine tissues of rats with P48444 and mice with asthma . Meanwhile , the SP and P25103 were significantly decreased , while P01282 , P32241 , and P41587 were significantly increased in lung tissues . An abnormal secretion of SP and P01282 can be observed in other tissues ; however , no marked changes were found in the receptors . The P20366 and NKB levels were similar in lung tissues of mice with asthma among groups . Conclusions . Stimulating intestine with Mangxiao or Dahuang can specifically regulate the secretion of SP , P01282 , and the receptors in lung tissues . Application of HapMap data to the evaluation of 8 candidate genes for pediatric slow transit constipation . BACKGROUND : Slow transit constipation ( P52823 ) affects up to 3 % of all children and is an increasingly recognized cause of chronic constipation in children . We conducted a pilot study to investigate whether genes encoding neurotransmitters ( P20366 , Q9UHF0 , P01282 , NOS1 ) and receptors ( P25103 , P21452 , P29371 , P10721 ) could be responsible for P52823 . METHODS : One hundred seventeen tag single nucleotide polymorphisms ( SNPs ) , distributed among the candidate genes , were selected from HapMap data and genotyped using Sequenom ( San Diego , CA ) technology in 35 affected families . Evaluation of association was performed by transmission disequilibrium test and multilocus analysis . RESULTS : Five SNPs ( rs3771863 , rs4580655 , rs11722288 , rs4563545 , and rs3782221 ) in the P25103 , P29371 , P10721 , and NOS1 genes were found to be potentially associated with P52823 , although the significance of these results does not withstand correction for multiple testing . CONCLUSIONS : Our data indicate that 5 SNPs in the NOS1 , P25103 , P29371 , and P10721 genes could be involved in P52823 , especially rs3771863 in intron 1 of P25103 , which showed the highest association . Biodistribution of 131I- , 186Re- , 177Lu- , and 88Y-labeled hLL2 ( DB04958 ) in nude mice with P20273 -positive lymphoma . Radioimmunotherapy ( Q92963 ) is a new and effective treatment modality in patients with non-Hodgkin 's lymphoma . The monoclonal antibody ( mAb ) hLL2 ( epratuzumab ) , a humanized mAb directed against the P20273 antigen , and which internalizes , can be labeled with various radionuclides . The biodistribution of hLL2 labeled with (131)I , (186)Re , (177)Lu , and (88)Y was studied in nude mice with subcutaneous human lymphoma xenografts in order to determine the most suitable of these four radionuclides for Q92963 with hLL2 . METHODS : Human Ramos lymphoma xenografts were transplanted in cyclophosphamide-pretreated athymic BALB/c mice . Four groups of mice were injected intravenously with (131)I- , (186)Re- , (88)Y- , or (177)Lu-labeled hLL2 , respectively . To determine the nonspecific tumor uptake , two groups of mice received (88)Y-labeled or (131)I-labeled control antibody , cG250 . The biodistribution of the radiolabel was determined 1 , 3 , and 7 days postinjection ( p.i. ) . RESULTS : Radiolabeled hLL2 had a higher tumor uptake than the nonspecific mAb at all time-points , irrespective of the radiolabel used . Tumor accretion of (88)Y- and (177)Lu-hLL2 was higher than tumor uptake of (131)I- and (186)Re-hLL2 . Activity in the bone , represented by the femur without bone marrow , was higher for (177)Lu- and (88)Y-hLL2 than for (131)I- and (186)Re-hLL2 on day 7 p.i . CONCLUSION : The use of the residualizing radiolabels (88)Y and (177)Lu in combination with a mAb directed against an internalizing antigen resulted in higher uptake and better retention of the radiolabel in the tumor . DB11582 suppresses osteoclastogenesis induced by O14788 and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 -induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 suppressed osteoclastogenesis induced by O14788 , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 -induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss . Role of DB05875 signaling in enhanced nociceptive sensitization and local cytokine production after incision . Substance P ( SP ) signaling facilitates nociceptive sensitization in various inflammatory and chronic pain models and we postulated that SP signaling might also contribute to the development of post-incisional hyperalgesia . These studies used mice with a deletion of the pre-protachykinin A gene ( ppt-A(-/-) ) which codes for SP to determine the role of SP signaling in post-incisional pain and in the increased cytokine and nerve growth factor ( P01138 ) expression observed in the incised skin . SP deficient ppt-A(-/-) mice displayed reduced mechanical allodynia and heat hyperalgesia compared to the wild-type ( wt ) mice at all post-incision time points , despite similar baseline values ( p < 0.001 ) . Furthermore , the P25103 antagonist LY303870 attenuated mechanical allodynia produced by incision in the wt mice ( p < 0.001 ) . Incision also up-regulated P05231 , P01375 and KC levels but not IL-1beta after 2h in the wt mice skin . However , ppt-A(-/-) mice had more skin P01138 levels 2h post-incision . Subcutaneous hind paw SP injection produced acute and transient elevations of IL-1beta , P05231 , and KC but modest elevations in P01375 levels in the wt mice . Systemic LY303870 reversed the SP-induced elevations of these cytokines . Hind paw injection of P05231 and P01138 dose dependently produced less mechanical allodynia in the ppt-A(-/-) compared to wt mice . Additionally , SP produced mechanical allodynia in a dose-dependent fashion in wt mice . Therefore , SP supports nociceptive sensitization after hind paw incision and potentially participates directly in modulating the intensity of inflammatory response in peri-incisional tissue . P05154 and other components of the protein C pathway in patients with acute deep vein thrombosis during heparin treatment . The protein C inhibitor ( P05154 ) concentration and other parameters of the protein C pathway were studied in 19 patients with symptomatic acute deep vein thrombosis before and during the first 5 days of heparin treatment . The mean levels of P05154 antigen and activity decreased rapidly and significantly during the first day of heparin therapy from 83 and 84 % to 60 and 59 % of the pooled normal human plasma ( p less than 0.01 ) , respectively , and to 56 and 54 % after 5 days of treatment ( p less than 0.01 ) . In contrast , antithrombin III decreased progressively 25 % during 5 days of heparin treatment . P02810 antigen and activity and total protein S remained unchanged during heparin treatment . Free protein S was decreased before heparin treatment ( 83 % , p less than 0.05 ) and increased to normal values after 5 days of treatment . C4b-binding protein was significantly increased before and during heparin treatment ( p less than 0.01 ) . DB00055 ( P25054 ) complexed to its two major plasma inhibitors , P05154 and alpha 1-antitrypsin ( alpha 1AT ) were measured by specific ELISA 's . Before treatment , 18 of the 19 patients studied had increased levels of P25054 :alpha 1AT complexes with a mean value of 27 +/- 22 ng/ml ( range , 6-86 ng/ml ) compared to normal values ( 8 +/- 2 ng/ml ) and 12 of the patients also had detectable P25054 : P05154 complex levels with a mean value of 11 +/- 17 ng/ml ( range , 5-68 ng/ml ) . Both P25054 :inhibitor complexes decreased significantly during heparin treatment . Application of stem cell markers in search for neoplastic germ cells in dysgenetic gonads , extragonadal tumours , and in semen of infertile men . Germ cell tumours ( GCTs ) are a complex entity . Current areas of attention include early detection and avoidance of unnecessary over-treatment . Novel findings regarding diagnosis of GCTs located in various anatomical sites are described , particularly testicular GCTs and their common progenitor , carcinoma in situ ( Q9NSE2 ) . Recognition of Q9NSE2 enables intervention before tumour development , but nevertheless , testicular GCTs are sporadically diagnosed at the pre-invasive stage where minimal treatment is necessary . As presence of Q9NSE2 is asymptomatic , a simple screening method is needed when Q9NSE2 is suspected ( i.e. in males investigated for infertility ) . To develop approaches for early detection Q9NSE2 gene expression studies have been performed showing many similarities with embryonic stem cells with confirmation of established markers ( i.e. PLAP , O75051 -3/4 , P10721 ) and identification of novel markers ( i.e. P05549 gamma , Q9H9S0 ) . We have reported a very promising new approach of P05549 gamma ( or OCT3/4 ) based immunocytological semen analysis ( specificity 93.6 % , sensitivity 54.5 % ) . Comparative studies of gonadal/extragonadal GCTs have revealed resemblance pointing towards similar , but not identical , origins . Moreover , infertility and testicular cancer are connected in the ' Testicular Dysgenesis Syndrome ' and 25 % of contralateral testes from testicular GCT patients harbour dysgenetic features , including impaired spermatogenesis . Thus , recent data have provided potential diagnostic tools including Q9NSE2 detection in semen , microarray-based tumour classification , additional serological GCT markers , and novel stem cell markers for immunohistochemical diagnosis of gonadal and extragonadal GCTs . Many Q9NSE2 candidate genes are yet uninvestigated , and information from these could increase knowledge about Q9NSE2 tumour initiation/progression and be used for optimisation of a non-invasive detection method . Prenatal exposure to bisphenol A promotes angiogenesis and alters steroid-mediated responses in the mammary glands of cycling rats . Prenatal exposure to Q03001 disturbs mammary gland histoarchitecture and increases the carcinogenic susceptibility to chemical challenges administered long after Q03001 exposure . Our aim was to assess the effect of prenatal Q03001 exposure on mammary gland angiogenesis and steroid hormone pathways in virgin cycling rats . Pregnant Wistar rats were exposed to either 25 or 250 g/kg/day ( 25 and 250 Q03001 , respectively ) or to vehicle . Female offspring were autopsied on postnatal day ( P01160 ) 50 or 110 . Ovarian steroid serum levels , the expression of steroid receptors and their co-regulators Q9Y6Q9 and Q9Y618 in the mammary gland , and angiogenesis were evaluated . At P01160 50 , all Q03001 -treated animals had lower serum levels of progesterone , while estradiol levels remained unchanged . The higher dose of Q03001 increased mammary ERα and decreased Q9Y6Q9 expression at P01160 50 and P01160 110 . Q9Y618 protein levels were similar among groups at P01160 50 , whereas at P01160 110 , animals exposed to 250 Q03001 showed a lower Q9Y618 expression . Interestingly , in the control and 25 Q03001 groups , Q9Y618 increased from P01160 50 to P01160 110 . At P01160 50 , an increased vascular area associated with higher P15692 expression was observed in the 250 Q03001 -treated rats . At P01160 110 , the vascular area was still increased , but P15692 expression was similar to that of control rats . The present results demonstrate that prenatal exposure to Q03001 alters the endocrine environment of the mammary gland and its angiogenic process . Increased angiogenesis and altered steroid hormone signals could explain the higher frequency of pre-neoplastic lesions found later in life . This article is part of a Special Issue entitled ' Endocrine disruptors ' . Immunophenotypic characterization of normal peripheral blood B lymphocyte by flow cytometry : reference for diagnosis of chronic B cell leukemia/lymphoma . To establish reference values of various immunophenotypic markers in B lymphocyte population in healthy Chinese adults and build background information for accurate interpretation of B cell immunophenotyping data in clinical practice , peripheral blood from 41 healthy adults were collected separately into test tubes containing DB00974 -K(2) and stored in room temperature no more than 24 hours before analysis . Whole blood lysis technique and multiparameter flow cytometry were applied to immunophenotype B cells gated on P15391 /SSC dot-plot . The results showed that P20273 , P11836 , CD62L , P25942 , P25063 , CD79b , CD79a , and FMC-7 were almost positive in the circulating B cell population , whereas CD11a , P33681 , CD103 , CD10 , P29965 , CD54 , P48023 , P42081 , and CD95 were almost negative in the peripheral blood B lymphocytes . P05107 , P16070 , CD23 , P06127 , CD11c and P16150 were positive in different B cell subpopulations . 78 % of B cells were IgD positive and ratio kappa/lambda was 1.26 . The significance of all these markers in the differential diagnosis of lymphoproliferative diseases was discussed . The conclusion is that it is necessary to consider the qualitative and quantitative levels of expression of various markers in normal B cell population in order to accurately interpret the pathological immunophenotypic data in clinical practice . It is also important to note the immunotypic differences of B cells between Chinese and Western populations . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . Substance P autocrine signaling contributes to persistent P04626 activation that drives malignant progression and drug resistance in breast cancer . P00533 receptor transmodulation by heterologous G-protein-coupled receptors ( GPCR ) generates functional diversity in signal transduction . Tachykinins are neuropeptides and proinflammatory cytokines that promote cell survival and cancer progression by activating several GPCRs . In this work , we found that the pain-associated tachykinin Substance P ( SP ) contributes to persistent transmodulation of the P00533 receptors , P00533 and P04626 , in breast cancer , acting to enhance malignancy and therapeutic resistance . SP and its high-affinity receptor P25103 were highly expressed in P04626 (+) primary breast tumors ( relative to the luminal and triple-negative subtypes ) and were overall correlated with poor prognosis factors . In breast cancer cell lines and primary cultures derived from breast cancer samples , we found that SP could activate P04626 . Conversely , RNA interference-mediated attenuation of P25103 , or its chemical inhibition , or suppression of overall GPCR-mediated signaling , all strongly decreased steady-state expression of P00533 and P04626 , establishing that their basal activity relied upon transdirectional activation by GPCR . Thus , SP exposure affected cellular responses to anti- P00533 therapies . Our work reveals an important oncogenic cooperation between P25103 and P04626 , thereby adding a novel link between inflammation and cancer progression that may be targetable by SP antagonists that have been clinically explored . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . Preimplantation genetic diagnosis for cancer predisposition syndromes . OBJECTIVES : Mutations in the P25054 , P35240 and P38398 genes cause adult-onset cancer predisposition syndromes . Prenatal diagnosis ( P01160 ) and selective pregnancy termination for adult-onset disorders is emotionally difficult and , in some cases , socially not well accepted . Preimplantation genetic diagnosis ( P52209 ) appears as an attractive alternative to P01160 , as it ensures the establishment of a pregnancy free of the mutation from the onset , circumventing the potentially difficult decision of termination of pregnancy . METHODS : Development of single-cell PCRs using Epstein-Barr virus transformed lymphoblasts as single-cell model , followed by clinical application in P52209 . RESULTS : A total of five duplex-PCRs were developed , three for adenomatous polyposis of the colon ( P25054 ) , one for neurofibromatosis type 2 ( P35240 ) and one for inherited breast and ovarian cancer caused by P38398 mutations . Eleven clinical cycles were performed , resulting in the birth of an unaffected girl . For one of the couples undergoing P52209 for P35240 , a spontaneous pregnancy ensued after five unsuccessful P52209 cycles . The couple underwent chorionic villus sampling ( CVS ) and the application of the same protocol as used during P52209 showed an unaffected fetus . CONCLUSION : In this work , we present the development and clinical application of P52209 for three cancer predisposition syndromes . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation . Differential recruitment of coregulator proteins steroid receptor coactivator-1 and silencing mediator for retinoid and thyroid receptors to the estrogen receptor-estrogen response element by beta-estradiol and 4-hydroxytamoxifen in human breast cancer . P03372 ( ER ) -alpha and Q92731 function as transcription factors , and both interact with nuclear regulatory proteins to enhance or inhibit transcription . We hypothesized that coregulators are expressed in breast cancer and may be differentially recruited by ERs in the presence of estrogen and tamoxifen . Q92731 was found to be expressed more frequently in node-negative patients ( P < 0.05 ) . Expression of steroid receptor coactivator-1 ( Q15788 ) was associated with nodal positivity ( P < 0.05 ) and resistance to endocrine treatment ( P < 0.001 ) . The spatial coexpression of P03372 , Q92731 , and the coregulatory proteins was established using immunofluorescence . In both cell lines ( MCF-7 and T47D ) and in primary breast cancer cell cultures , beta-estradiol up-regulated Q92731 and coregulator protein expression and increased P03372 / Q92731 interaction with the estrogen response element ( ERE ) . 4- Hydroxy-tamoxifen ( DB04468 ) increased P03372 and silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) expression and increased ER-ERE binding . Q15788 and Q9Y618 were identified at the ER-ERE complex , and interactions between ER isoforms and coregulatory proteins were determined using immunoprecipitation . Both P03372 and Q92731 preferentially bound Q15788 in the presence of beta-estradiol . Conversely , in cells treated with DB04468 , P03372 and Q92731 bound Q9Y618 . Differential recruitment of Q15788 and Q9Y618 by P03372 and Q92731 in the presence of beta-estradiol and DB04468 may be central to the response of the tumor to endocrine treatment . New analogs of vitamin D3 . Calcitriol , the most active metabolite of vitamin D , controls parathyroid gland growth and suppresses the synthesis and secretion of parathyroid hormone ( PTH ) . However , because of its potent effects on intestinal calcium absorption and bone mobilization , calcitriol treatment can induce hypercalcemia , often precluding its use at therapeutic doses . Hyperphosphatemia is also a persistent problem among patients undergoing chronic hemodialysis and can be aggravated by therapeutic doses of calcitriol . Several pharmaceutical companies were able to modify the side-chain of the 1,25(OH)2D3 , allowing some of these new analogs to retain the action on the parathyroid glands while decreasing their hypercalcemic and hyperphosphatemic effects . The structure-activity relationship for ligand-mediated transcriptional regulation has been studied in detail . In some analogs the serum binding protein ( DBP ) plays a key role in determining the pharmacokinetics of the vitamin D compound . The affinity to DBP for 22-oxacalcitriol ( O75051 ) , an analog of calcitriol for the treatment of secondary hyperparathryoidism , is approximately 300-400 times lower than that of calcitriol and the analog is rapidly cleared from the circulation . The mechanisms for the selectivity of 19-nor-1,25(OH)2D2 ( paricalcitol ) ( DB00910 ) another analog of calcitriol , is clearly different from O75051 . Although the mechanisms of action is not completely known , it does appear that paricalcitol down-regulates the P11473 in the intestine . It is likely that the unique biological profiles of vitamin D analogs in vivo are due to multiple mechanisms . Understanding the molecular basis of the analog selectivity will not only provide an explanation for their unique actions but allow intelligent design of more effective analogs in the future . Proanthocyanidin derived from the leaves of Vaccinium virgatum suppresses platelet-derived growth factor-induced proliferation of the human hepatic stellate cell line LI90 . AIM : Hepatic stellate cell ( P19526 ) proliferation plays a pivotal role in liver fibrogenesis , and agents that suppress P19526 activation , including platelet-derived growth factor ( PDGF ) -induced P19526 proliferation , are good candidates for antifibrogenic therapies . In this report , we use the LI90 P19526 line to elucidate the antifibrogenic effects of proanthocyanidin derived from the leaves of Vaccinium virgatum . METHODS : Proanthocyanidin ( PAC ) was extracted from the leaves of blueberry V. virgatum ( BB-PAC ) , grape seeds ( GS-PAC ) and Croton lechleri ( CL-PAC ) . These extracts were examined for their effects on DB00102 -induced LI90 cell proliferation and DNA synthesis . Extracellular signal-regulated kinase ( P29323 ) and Akt phosphorylation and PDGF receptor-beta ( P09619 ) expression were evaluated by western blot analysis . RESULTS : BB-PAC potently suppressed DB00102 -induced proliferation and DNA synthesis of LI90 cells . BB-PAC also suppressed DB00102 -induced DNA synthesis in primary cultured rat P19526 . Moreover , GS-PAC and CL-PAC suppressed DB00102 -induced DNA synthesis in LI90 cells . In contrast , the monomeric PAC catechin and epicatechin and dimeric PAC procyanidin B2 only slightly suppressed DB00102 -induced DNA synthesis . Western blot analysis showed that BB-PAC completely or partially inhibited DB00102 -induced P29323 and Akt phosphorylation , respectively . In addition , BB-PAC partially inhibited the DB00102 -induced degradation of P09619 . CONCLUSION : Our results suggest that BB-PAC suppresses activated P19526 by inhibiting the PDGF signaling pathway . In addition , these results provide novel findings that may facilitate the development of antifibrogenic agents . Internalization of OspA in rsCD14 complex and aggregated forms . Although the spirochetal protein OspA is capable of stimulating immune cells in a P08571 - and O60603 -dependent manner , little is known about how O60603 receptor complex ligands , such as OspA , are handled by the cell once delivered . We examine here the internalization of the fluorescently derivatized forms of both the full length DB00045 delivered as a recombinant soluble P08571 ( rsCD14 ) complex and the corresponding lipohexapeptide given to the cells as an aggregate . Both forms of OspA are internalized in a similar manner to acetylated low density lipoprotein ( AcLDL ) , a scavenger receptor ligand . Acetylated low density lipoprotein is capable of competing for internalization with OspA even when OspA is delivered as a rsCD14 complex . We observe co-localization of OspA with lysosomes but not with the Golgi complex . These phenomena are similar between RAW264.7 macrophages and endothelial cells but change drastically when the cells are deprived of serum . Upon serum starvation , OspA shows some localization to the Golgi apparatus whereas the lipohexapeptide remains on the cell surface . Inhibition of internalization of OspA via treatment with cytochalasin D or of the lipohexapeptide via serum starvation does not interfere with P01375 induction activity , consistent with signalling from the cell surface . Substance P promotes expansion of human mesenteric preadipocytes through proliferative and antiapoptotic pathways . White adipose tissue is intimately involved in the regulation of immunity and inflammation . We reported that human mesenteric preadipocytes express the DB05875 ( SP ) -mediated neurokinin-1 receptor ( P25103 ) , which signals proinflammatory responses . Here we tested the hypothesis that SP promotes proliferation and survival of human mesenteric preadipocytes and investigated responsible mechanism(s) . Preadipocytes were isolated from mesenteric fat biopsies during gastric bypass surgery . Proliferative and antiapoptotic responses were delineated in 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium ( MTS ) , bromodeoxyuridine ( BrdU ) , caspase-3 , and TUNEL assays , as well as Western immunoanalysis . SP ( 10(-7) M ) increased MTS and proliferation ( BrdU ) and time dependently ( 15-30 min ) induced Akt , P01133 receptor , IGF receptor , integrin alphaVbeta3 , phosphatidylinositol 3-kinase , and PKC-theta phosphorylation . Furthermore , pharmacological antagonism of Akt and PKC-theta activation significantly attenuated SP-induced preadipocyte proliferation . Exposure of preadipocytes to the proapoptotic P48023 ( P48023 , 100 microM ) resulted in nuclear DNA fragmentation ( TUNEL assay ) , as well as increased cleaved poly ( ADP-ribose ) polymerase , cleaved caspase-7 , and caspase-3 expression . Cotreatment with SP almost completely abolished these responses in a P25103 -dependent fashion . SP ( 10(-7) M ) also time dependently stimulated expression 4E binding protein 1 and phosphorylation of P08133 S6 kinase , which increased protein translation efficiency . SP increases preadipocyte viability , reduces apoptosis , and stimulates proliferation , possibly via cell cycle upregulation and increased protein translation efficiency . SP-induced proliferative and antiapoptotic pathways in fat depots may contribute to development of the creeping fat and inflammation characteristic of Crohn 's disease . Natriuretic and vasoactive hormones and glomerular hyperfiltration in hyperglycaemic type 2 diabetic patients : effect of insulin treatment . Evidence that an increase in plasma atrial natriuretic peptide ( P01160 ) concentrations mediates , at least in part , glomerular hyperfiltration in diabetic rats prompted us to study the relationship between P01160 and renal haemodynamics in hyperfiltering type 2 diabetic patients in association with other hormones implicated in the control of glomerular filtration rate ( Q92565 ) ( catecholamines , vasopressin , renin ) and in sodium tubular transport ( aldosterone , ouabain-displacing factor , O14788 ) . Since hyperglycaemia is also associated to hyperfiltration , diabetic patients who presented with secondary drug failure were studied both in hyperglycaemic and in normoglycaemic condition . For this purpose , 11 normotensive non-macroproteinuric hyperfiltering patients with type 2 diabetes were treated with an 8-day continuous insulin infusion ( days 0-7 ) . Dehydration was prevented or corrected and natriuresis was on day 0 above 100 mmol/day . The following parameters were determined on days 0 and 7 : Q92565 and renal plasma flow ( RPF ) by 99mTc-DTPA and 131I-hippuran clearances ; the extracellular volume , assimilated to the DTPA diffusion volume ; urinary O14788 by receptor-binding assay and urinary as well as plasma catecholamines by HPLC after extraction on alumin . Plasma P01160 and antidiuretic hormone ( DB00067 ) were measured by radioimmunoassay after extraction on phenyl-silylsilica ( P01160 ) and with ether ( DB00067 ) . Unextracted plasma was used for radioimmunological measurement of plasma renin activity and aldosterone . When correcting hyperglycaemia to normoglycaemia Q92565 decreased from high to normal mean value ( 138 +/- 27 and 115 +/- 6 ml/min , p < 0.001 ) , RPF followed the same trend , and the DTPA diffusion volume did not change. ( ABSTRACT TRUNCATED AT 250 WORDS ) P42892 and β-arrestins exert spatiotemporal control of DB05875 -induced inflammatory signals . Although the intracellular trafficking of G protein-coupled receptors controls specific signaling events , it is unclear how the spatiotemporal control of signaling contributes to complex pathophysiological processes such as inflammation . By using bioluminescence resonance energy transfer and superresolution microscopy , we found that DB05875 ( SP ) induces the association of the neurokinin 1 receptor ( P25103 ) with two classes of proteins that regulate SP signaling from plasma and endosomal membranes : the scaffolding proteins β-arrestin ( βARRs ) 1 and 2 and the transmembrane metallopeptidases ECE-1c and ECE-1d . In HEK293 cells and non-transformed human colonocytes , we observed that G protein-coupled receptor kinase 2 and β P49407 /2 terminate plasma membrane Ca(2+) signaling and initiate receptor trafficking to endosomes that is necessary for sustained activation of ERKs in the nucleus . βARRs deliver the SP- P25103 endosomes , where P42892 associates with the complex , degrades SP , and allows the P25103 , freed from βARRs , to recycle . Thus , both P42892 and βARRs mediate the resensitization of P25103 Ca(2+) signaling at the plasma membrane . Sustained exposure of colonocytes to SP activates NF-κB and stimulates P10145 secretion . This proinflammatory signaling is unaffected by inhibition of the endosomal P29323 pathway but is suppressed by P42892 inhibition or β P32121 knockdown . Inhibition of protein phosphatase 2A , which also contributes to sustained P25103 signaling at the plasma membrane , similarly attenuates P10145 secretion . Thus , the primary function of βARRs and P42892 in SP-dependent inflammatory signaling is to promote resensitization , which allows the sustained P25103 signaling from the plasma membrane that drives inflammation . Developments in epidermal growth factor receptor-targeting therapy for solid tumors : focus on matuzumab ( P50402 72000 ) . Expression of epidermal growth factor and its transmembrane receptor ( P00533 ) stimulates tumor growth . DB05101 is a humanized anti- P00533 monoclonal antibody that blocks P00533 activation and downstream signaling , inhibits tumor growth , and provides a clinical benefit for some patients . The plasma half-life ( 6-10 days ) and pharmacodynamic activity allow flexible dosing on weekly , every-2-week , and every-3-week schedules . DB05101 has shown single-agent antitumor activity in heavily pretreated patients with a variety of tumors , with a favorable safety profile . Skin rash is the most common toxicity , but is severe ( Grade 3 ) in < 1 percent . This article describes preclinical and clinical development of matuzumab .

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Interaction of early environment , gender and genes of monoamine neurotransmission in the aetiology of depression in a large population-based Finnish birth cohort . Objectives Depression is a worldwide leading cause of morbidity and disability . Genetic studies have recently begun to elucidate its molecular aetiology . The authors investigated candidate genes of monoamine neurotransmission and early environmental risk factors for depressiveness in the genetically isolated population-based Northern Finland Birth Cohort 1966 ( 12 058 live births ) . Design The authors ascertained and subdivided the study sample ( n=5225 ) based on measures of early development and of social environment , and examined candidate genes of monoamine neurotransmission , many of which have shown prior evidence of a gene-environment interaction for affective disorders , namely P31645 , Q8IWU9 , P21964 , P21397 and the dopamine receptor genes P21728 - P21918 . Results and conclusion The authors observed no major genetic effects of the analysed variants on depressiveness . However , when measures of early development and of social environment were considered , some evidence of interaction was observed . Allelic variants of P21964 interacted with high early developmental risk ( p=0.005 for rs2239393 and p=0.02 for rs4680 ) so that the association with depression was detected only in individuals at high developmental risk group ( p=0.0046 and β=0.056 for rs5993883-rs2239393-rs4680 risk haplotype CGG including Val158 ) , particularly in males ( p=0.0053 and β=0.083 for the haplotype CGG ) . Rs4274224 from P14416 interacted with gender ( p=0.017 ) showing a significant association with depressiveness in males ( p=0.0006 and β=0.0023 ; p=0.00005 and β=0.069 for rs4648318-rs4274224 haplotype GG ) . The results support the role of genes of monoamine neurotransmission in the aetiology of depression conditional on environmental risk and sex , but not direct major effects of monoaminergic genes in this unselected population . Evidence for a link between histone deacetylation and Ca²+ homoeostasis in sphingosine-1-phosphate lyase-deficient fibroblasts . Embryonic fibroblasts from Q14703 ( sphingosine-1-phosphate ) lyase-deficient mice [ Sgpl1-/- MEFs ( mouse embryonic fibroblasts ) ] are characterized by intracellular accumulation of Q14703 , elevated cytosolic [Ca2+]i and enhanced Ca2+ storage . Since Q14703 , produced by sphingosine kinase 2 in the nucleus of MCF-7 cells , inhibited HDACs ( histone deacetylases ) [ Hait , Allegood , Maceyka , Strub , Harikumar , Singh , Luo , Marmorstein , Kordula , Milstein et al. ( 2009 ) Science 325 , 1254-1257 ] , in the present study we analysed whether Q14703 accumulated in the nuclei of Q14703 lyase-deficient MEFs and caused HDAC inhibition . Interestingly , nuclear concentrations of Q14703 were disproportionally elevated in Sgpl1-/- MEFs . HDAC activity was reduced , acetylation of histone 3-Lys9 was increased and the HDAC-regulated gene P38936 cyclin-dependent kinase inhibitor was up-regulated in these cells . Furthermore , the expression of Q13547 and O15379 was reduced in Sgpl1-/- MEFs . In wild-type MEFs , acetylation of histone 3-Lys9 was increased by the Q14703 lyase inhibitor 4-deoxypyridoxine . The non-specific HDAC inhibitor trichostatin A elevated basal [Ca2+]i and enhanced Ca2+ storage , whereas the Q13547 /2/3 inhibitor DB05651 elevated basal [Ca2+]i without influence on Ca2+ storage in wild-type MEFs . Overexpression of Q13547 or Q92769 reduced the elevated basal [Ca2+]i in Sgpl1-/- MEFs . Taken together , Q14703 lyase-deficiency was associated with elevated nuclear Q14703 levels , reduced HDAC activity and down-regulation of HDAC isoenzymes . The decreased HDAC activity in turn contributed to the dysregulation of Ca2+ homoeostasis , particularly to the elevated basal [Ca2+]i , in Sgpl1-/- MEFs . Quick generation of fully mature dendritic cells from monocytes with OK432 , low-dose prostanoid , and interferon-alpha as potent immune enhancers . Dendritic cells ( DCs ) are one of the promising tools for enhancing antigen-specific immune responses in clinical settings . Many studies have been performed thus far to verify the efficacy of the DC vaccine in cancer patients ; however , the responses have not always been satisfactory , partly because of DC incompetence . To obtain DCs potentially applicable for vaccination of cancer patients , our group sought to establish the strategy of DC generation mainly by modulating culture periods and maturation stimuli . Novel mature DCs that can be generated from monocytes within 3 days by using a combination of OK432 ( Streptococcus pyogenes preparation ) , low-dose prostaglandin E2 ( DB00917 ) , and interferon-alpha ( OPA-DCs ) were developed . They strongly express Q01151 , P42081 , and P32248 and have potent ability to migrate to O00585 . In addition , they were able to activate natural killer and T helper 1 ( Q8IXH7 ) cells and to induce peptide-antigen-specific cytotoxic T lymphocytes more significantly than monocyte-derived DCs stimulated with a conventional cytokine cocktail of tumor necrosis factor-alpha , interleukin ( IL ) -1beta , P05231 , and DB00917 ( monocyte-conditioned medium [ P22033 ] -mimic DCs ) . The profound ability of OPA-DCs to stimulate these effectors is attributable to their higher expression of IL-12p70 , IL-23 , and Q8TAD2 than P22033 -mimic DCs , which was supported by the findings that the neutralization of IL-12p70 and IL-23 reduced the Q8IXH7 priming ability of OPA-DCs . Even when from advanced gastric or colonic cancer patients , OPA-DCs displayed abilities of migration and Q8IXH7 induction comparable to those from healthy subjects . Therefore , OPA-DCs may serve as a feasible vaccine with the potential to enhance Q8IXH7 -dominant and cytolytic immune responses against cancers . A double-blind , placebo-controlled outpatient trial of pergolide for cocaine dependence . Results of preclinical studies suggest that pergolide , a mixed D(1)/ P14416 agonist , may be useful in treating cocaine dependence . To empirically investigate this possibility , we conducted a 5-year , double-blind , placebo-controlled clinical trial of two doses of pergolide ( 0.05 and 0.25 mg bid ) in subjects with cocaine dependence and combined cocaine/alcohol dependence . Data analysis was performed on an intent to treat population ( N=358 ) and a per protocol population ( N=108 ) with urine drug screens ( UDS ) used as the main outcome measure . There were no significant effects on UDS at either pergolide dose . DB01186 had no significant effect on alcohol use in the comorbid alcohol/cocaine dependence group . DB01186 does not appear to have clinical value in the treatment of cocaine dependence or in decreasing alcohol use in cocaine-dependent individuals at the presently studied doses . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . Molecular basis for the immunostimulatory activity of guanine nucleoside analogs : activation of Q9NYK1 . Certain Q99618 -substituted and N7 , Q99618 -disubstituted guanine ribonucleosides comprise a class of small molecules with immunostimulatory activity . In a variety of animal models , these agents stimulate both humoral and cellular immune responses . The antiviral actions of these guanosine analogs have been attributed to their ability to induce type I IFNs . However , the molecular mechanisms by which the guanosine analogs potentiate immune responses are not known . Here , we report that several guanosine analogs activate Q9NYK1 ( Q9NYK1 ) . DB04860 , 7-deazaguanosine , and related guanosine analogs activated mouse immune cells in a manner analogous to known TLR ligands , inducing cytokine production in mouse splenocytes ( P05231 and IL-12 , type I and II IFNs ) , bone marrow-derived macrophages ( P05231 and IL-12 ) , and in human peripheral blood leukocytes ( type I IFNs , tumor necrosis factor alpha and IL-12 ) . The guanosine congeners also up-regulated costimulatory molecules and MHC I/II in dendritic cells . Genetic complementation studies in human embryonic kidney 293 cells confirmed that the guanosine analogs activate cells exclusively via Q9NYK1 . The stimulation of Q9NYK1 by the guanosine analogs in human cells appears to require endosomal maturation because inhibition of this process with chloroquine significantly reduced the downstream activation of NF-kappaB . However , Q9NR97 activation by R-848 and O60603 activation by [ S-[2,3-bis(palmitoyloxy)-(2-RS)-propyl]-N-palmitoyl-R- DB00151 -S- DB00133 -Lys4-OH , trihydrochloride ) ] were not inhibited by chloroquine , whereas Q9NR96 activation by CpG oligodeoxynucleotides was abolished . In summary , we present evidence that guanosine analogs activate immune cells via Q9NYK1 by a pathway that requires endosomal maturation . Thus , the B cell-stimulating and antiviral activities of the guanosine analogs may be explained by their Q9NYK1 -activating capacity . P15692 trap abolishes shear stress- and overload-dependent angiogenesis in skeletal muscle . OBJECTIVE : To determine which elements of the angiogenic process are controlled by P15692 under physiological conditions . METHODS : DB08885 was administered at 10 mg x kg(-1) by subcutaneous injection twice weekly to mice , which were subject to one of two established angiogenic stimuli : ( 1 ) increasing blood flow by administration of prazosin ( 50 mg L(-1) ) ; ( 2 ) muscle overload by extirpation of a synergist . Angiogenesis was determined by calculating capillary to fiber ratio ( C:F ) , and proliferating cell nuclear antigen ( P12004 ) staining localized to capillaries or the interstitium used to measure cell proliferation . Characteristic ultrastructural changes were quantified using electron microscopy . RESULTS : Administration of DB08885 abolished the increases in C:F seen in both models , and prevented growth of luminal filopodia and large vacuole formation . Overload-induced proliferation associated with capillaries was reduced , along with endothelial cell number abluminal sprouts and basement membrane breakage . However , interstitial proliferation remained high , along with the increased capillary association of pericytes and fibroblasts . CONCLUSIONS : P15692 is required for both models of angiogenesis , although some morphological changes lie upstream , or are independent of , P15692 involvement . The abolition of angiogenesis in a model unaffected by NO inhibition shows that NO is not required for all P15692 -dependent angiogenesis in vivo . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . [ A molecular study of methylmalonic aciduria : structure-function correlations ] . Cobalamin ( DB00115 ) non-responsive methylmalonic acidemia is caused by mutations in the P22033 locus on chromosome 6p21 encoding the enzyme methylmalonyl DB01992 mutase ( EC 5.4.99.2 ) . This disorder has been extensively studied by biochemical , somatic cell genetic and molecular techniques . Mutations have been identified which cause classic mut(o) phenotypes in which there is no detectable enzymatic activity , as well as mut- phenotypes in which there is residual cobalamin-dependent activity . Mutations which exhibit interallelic complementation have been identified within both of these groups . These mutations illustrate the position , structure , and function of critical domains within this cobalamin binding enzyme and provide new insights into the biochemical and clinical consequences of enzyme deficiency . The homology of the cobalamin binding region has allowed mutations of the mutase to be mapped onto the x-ray structure of methionine synthase ( EC 2.1.1.13 ) . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . Q9BYW2 -α and survivin involved in the anti-apoptotic effect of DB02342 after global ischemia in rats . Survivin is an anti-apoptotic gene that decreases the apoptosis by depressing the expression of caspase-3 . Hypoxia-inducible factor-1-alpha ( Q16665 ) is a transcription factor specifically activated by hypoxia . 2-methoxyestradiol ( DB02342 ) is an estradiol derivative and a known Q16665 inhibitor . DB02342 decreased apoptosis by inhibiting Q16665 . The aim of the present study was to investigate if survivin is involved in the anti-apoptotic effect of DB02342 . Male adult rats were used to make the global ischemia ( GI ) model . Ten minutes after GI , DB02342 was injected intraperitoneally ( 16 mg/kg weight ) . Rats were killed at 6 hours , 12 hours , 24 hours , 48 hours , 96 hours , and 7 days . GI produced a marked increase in Q16665 expressions in the hippocampus at 6 hours and peaked at 48-96 hours . The expressions of survivin and caspase-3 were increased lightly in a similar time course . These molecular changes were accompanied by massive cell loss and apoptosis in the hippocampal regions . DB02342 treatment reduced the expression of Q16665 , increased survivin expression , and decreased the expression of caspase-3 . These results indicate that survivin and Q16665 were involved in the anti-apoptotic effect of DB02342 treated following GI . DB02342 may decrease the Q16665 expression , up-regulate the survivin expression , inhibit the expression of caspase-3 , and finally reduce apoptosis after GI . DB06288 is a potent P34969 antagonist : relevance for antidepressant actions in vivo . RATIONALE : DB06288 is approved for clinical use in treating schizophrenia in a number of European countries and also for treating dysthymia , a mild form of depression , in Italy . DB06288 has also been demonstrated to be an antidepressant for patients with major depression in many clinical trials . In part because of the selective D(2)/D(3) receptor antagonist properties of amisulpride , it has long been widely assumed that dopaminergic modulation is the proximal event responsible for mediating its antidepressant and antipsychotic properties . OBJECTIVES : The purpose of these studies was to determine if amisulpride 's antidepressant actions are mediated by off-target interactions with other receptors . MATERIALS AND METHODS : We performed experiments that : ( 1 ) examined the pharmacological profile of amisulpride at a large number of central nervous system ( CNS ) molecular targets and , ( 2 ) after finding high potency antagonist affinity for human 5-HT(7a) serotonin receptors , characterized the actions of amisulpride as an antidepressant in wild-type and 5-HT(7) receptor knockout mice . RESULTS : We discovered that amisulpride was a potent competitive antagonist at 5-HT(7a) receptors and that interactions with no other molecular target investigated in this paper could explain its antidepressant actions in vivo . Significantly , and in contrast to their wild-type littermates , 5-HT(7) receptor knockout mice did not respond to amisulpride in two widely used rodent models of depression , the tail suspension test and the forced swim test . CONCLUSIONS : These results indicate that 5-HT(7a) receptor antagonism , and not D(2)/D(3) receptor antagonism , likely underlies the antidepressant actions of amisulpride . Immunohistochemical study of P04626 and Q13509 proteins in extramammary Paget disease . Metastatic extramammary Paget disease ( EMPD ) is a potentially fatal malignancy for which effective chemotherapy and good biomarkers are desirable for management . We investigated the status of human epidermal growth factor receptor ( P04626 ) and neuronal β-tubulin isotype ( class III β-tubulin ; Q13509 ) , whose overexpression is a factor involved in resistance of tumor cells to taxane derivatives ) in 32 patients with EMPD . P04626 status was evaluated by immunohistochemistry followed by fluorescence in situ hybridization , and Q13509 status was evaluated by immunohistochemistry . On the basis of the US Food and Drug Administration-approved criteria , 20 ( 63 % ) of the 32 EMPD tumors were found to overexpress P04626 . Positive immunoreactivity for Q13509 was observed in 7 ( 22 % ) of the 32 patients . Although some clinicopathologic variables ( nodule formation , depth of tumor cells , presence of lymph node metastasis , and serum carcinoembryonic antigen level ) were significantly associated with disease outcome ( P < 0.05 ) , P04626 gain or aberrant Q13509 expression showed no significant correlation . However , the higher incidence of P04626 gain and the relatively lower incidence of aberrant Q13509 expression suggested that P04626 -targeted immunotherapy combined with taxane derivatives is warranted for metastatic EMPD , and that P04626 and Q13509 status might be a good biomarker for determining the most appropriate therapeutic modality . Hypoxia-inducible factor-2α is an essential catabolic regulator of inflammatory rheumatoid arthritis . Rheumatoid arthritis ( RA ) is a systemic autoimmune disorder that manifests as chronic inflammation and joint tissue destruction . However , the etiology and pathogenesis of RA have not been fully elucidated . Here , we explored the role of the hypoxia-inducible factors ( HIFs ) , HIF-1α ( encoded by Q16665 ) and HIF-2α ( encoded by Q99814 ) . HIF-2α was markedly up-regulated in the intimal lining of RA synovium , whereas HIF-1α was detected in a few cells in the sublining and deep layer of RA synovium . Overexpression of HIF-2α in joint tissues caused an RA-like phenotype , whereas HIF-1α did not affect joint architecture . Moreover , a HIF-2α deficiency in mice blunted the development of experimental RA . HIF-2α was expressed mainly in fibroblast-like synoviocytes ( FLS ) of RA synovium and regulated their proliferation , expression of O14788 ( receptor activator of nuclear factor-κB ligand ) and various catabolic factors , and osteoclastogenic potential . Moreover , HIF-2α-dependent up-regulation of interleukin ( IL ) -6 in FLS stimulated differentiation of TH17 cells-crucial effectors of RA pathogenesis . Additionally , in the absence of P05231 ( Il6-/- mice ) , overexpression of HIF-2α in joint tissues did not cause an RA phenotype . Thus , our results collectively suggest that HIF-2α plays a pivotal role in the pathogenesis of RA by regulating FLS functions , independent of HIF-1α . Expression of serotonergic system components during early Xenopus embryogenesis . Despite abundant research studies on the physiological and biochemical nature of embryonic neurotransmitter function , little is known about the molecular genetic mechanisms involved . The expression of the main components of the serotonergic system during early Xenopus embryogenesis was investigated using RT-PCR , real time PCR and in situ hybridization . Transcripts encoding the serotonin receptors P28335 and P34969 , as well as the vesicular monoamine transporter Q05940 , the serotonin transporter ( P31645 ) and the serotonin synthesis enzymes tryptophan hydroxylase ( Q8IWU9 ) and aromatic amino acid decarboxylase ( AAAD ) were found to be expressed during the cleavage division stages , whereas the degradation enzyme monoamine oxidase A ( P21397 ) was absent . The main components of the serotonergic system were found to be expressed during the earliest stages of embryonic development . The embryonic transmitter mechanism , its complexity , and its variability among various species are discussed . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Multiplex protein signature for the detection of bladder cancer in voided urine samples . PURPOSE : Accurate urine assays for bladder cancer detection would benefit patients and health care systems . Through extensive genomic and proteomic profiling of urine components we previously identified a panel of 8 biomarkers that can facilitate the detection of bladder cancer in voided urine samples . In this study we confirmed this diagnostic molecular signature in a diverse multicenter cohort . MATERIALS AND METHODS : We performed a case-control , phase II study in which we analyzed voided urine from 102 subjects with bladder cancer and 206 with varying urological disorders . The urinary concentration of 8 biomarkers ( P10145 , P14780 and 10 , P05121 , P15692 , P03950 , Q16790 and P02649 ) was assessed by enzyme-linked immunosorbent assay . Diagnostic performance of the panel of tested biomarkers was evaluated using ROCs and descriptive statistical values , eg sensitivity and specificity . RESULTS : Seven of the 8 urine biomarkers were increased in subjects with bladder cancer relative to those without bladder cancer . The 7 biomarkers were assessed in a new model , which had an AUROC of 0.88 ( 95 % CI 0.84-0.93 ) , and 74 % sensitivity and 90 % specificity . In contrast , the sensitivity of voided urine cytology and the UroVysion® cytogenetic test in this cohort was 39 % and 54 % , respectively . Study limitations include analysis performed on banked urine samples and the lack of voided urine cytology and cytogenetic test data on controls . CONCLUSIONS : The study provides further evidence that the reported panel of diagnostic biomarkers can reliably achieve the noninvasive detection of bladder cancer with higher sensitivity than currently available urine based assays . Aberrant accentuation of neurofibrillary degeneration in the hippocampus of Alzheimer 's disease with amyloid precursor protein 717 and presenilin-1 gene mutations . This study reports correlation of the hippocampal neurofibrillary tangles ( NFT ) density with beta-amyloid ( Abeta ) precursor protein ( P05067 ) 717 mutation , presenilin ( PS ) -1 mutation and apolipoprotein E ( P02649 ) e4 alleles ( E4 ) , being graded as 3 forms ( no-E4 , one-E4 and two-E4 ) in autopsied brains from patients with familial and non-familial Alzheimer 's disease ( AD ) . We studied the density of NFT-free neurons , intracellular NFT ( I-NFT ) , extracellular NFT ( E-NFT ) and total NFT ( I-NFT plus E-NFT ) in the six hippocampal subdivisions : cornu ammonis ( CA ) 1- P22748 , subiculum and entorhinal cortex . The P05067 mutation cases showed significantly higher total NFT density in the P00915 - P00918 region , and the P49768 mutation cases also showed higher density of total NFT in the P00915 - P07451 than non-familial cases . Moreover , high densities of the E-NFT contributed to these high total NFT densities . Non-familial AD cases showed a stereotypical NFT distribution with entorhinal accentuation in the hippocampus irrespective of E4 frequency . Thus , P05067 and P49768 mutations predominantly affect the CA regions with profound neurodegeneration , which contributes early and severe clinical features of familial AD . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . Extended kindred with recessive late-onset Alzheimer disease maps to locus 8p22- P38936 .2 : a genome-wide linkage analysis . Late-onset Alzheimer disease ( LOAD ) is a complex genetic disorder . Although genes involved in early-onset forms were discovered more than a decade ago , LOAD research has only been able to point out small effect loci , with the exception of P02649 . We mapped the gene predisposing to LOAD in an extended inbred family coming from a genetically isolated region ( 24 sampled individuals , 12 of whom are affected ) , completing a genome-wide screen with an Affymetrix10 K single nucleotide polymorphism microarray . Genotyping results were evaluated under model-dependent ( dominant and recessive ) and model-free analysis . We obtained a maximum nonparametric linkage score of 3.24 ( P=0.00006 ) on chromosome 8p22- P38936 .2 . The same genomic position also yielded the highest multipoint heterogeneity LOD ( HLOD ) under a recessive model ( HLOD=3.04 ) . When we compared the results of the model-dependent analysis , a higher score was obtained in the recessive model ( 3.04 ) than in the dominant model ( 1.0 ) . This is a new locus identified in LOAD , in chromosome 8p22- P38936 .2 and encompassing several candidate genes , among them P10909 and P48454 that were excluded by sequencing . The finding of a recessive model of inheritance , consistent with the assumption of inbreeding as a morbidity factor in this population , supports the notion of a role of recessive genes in LOAD .

MH_train_1261

MH_train_1261

MH_train_1261

interacts_with DB00035?

multiple_choice

F-actin involvement in guinea pig sperm motility . Sperm motility is a must for natural fertilization to occur . During their travel through the epididymis , mammalian spermatozoa gradually acquire the ability to move . This is accomplished through a sliding movement of the outer doublet microtubules of the axoneme which is energized by the dynein ATPase . Within its complex structure , the mammalian sperm flagellum contains F-actin and thus , we decided to test in the guinea pig sperm flagellum the role of F-actin in motility . During maturation , capacitation , and the acrosome reaction , a gradual decrease of the relative concentration of F-actin was observed . Motility increased as spermatozoa became able to fertilize . P06396 , phalloidin , and KI inhibited sperm motility . P06396 canceled sperm motility within 20 min of treatment while 0.6 M KI had immediate effects . Phalloidin diminished hyperactive sperm motility slightly . All three compounds significantly increased the relative concentration of F-actin . Latrunculins are conventional drugs that destabilize the F-actin cytoskeleton . DB02621 ( O43561 A ) did not affect sperm motility ; but significantly increased F-actin relative concentration . The results suggested that in guinea pig spermatozoa , randomly severing F-actin filaments inhibits flagellar motility ; while end filament alteration does not . Thus , specific filament regions seem to be important for sperm motility . Identification and characterization of a novel X-linked P30518 mutation causing partial nephrogenic diabetes insipidus : a case report and review of the literature . X-linked nephrogenic diabetes insipidus ( NDI ) is a rare disease characterized by a malfunctioning renal response to the antidiuretic hormone arginine vasopressin ( AVP ) due to mutations in the P30518 gene . A limited number of mutations in the P30518 gene resulting in partial phenotype have been described so far . In this mini-review the retrospective analysis of 13 known P30518 mutations that have been previously shown in vitro to partially abolish P30518 function is described , along with a novel mutation diagnosed in a kindred with partial NDI . In the present study , a 14 year old male and his 73 year old maternal grandfather were diagnosed with partial NDI based on the clinical phenotype , the water deprivation test and the inadequate response to 1-desamino-8-d-arginine vasopressin ( DB00035 ) administration . Sequencing analysis of the P30518 gene revealed the novel missense mutation p.N317S ( g.1417A > G ) in both patients . This mutation was re-created by site directed mutagenesis in an P30518 cDNA expression vector and was functionally characterized , in terms of arginine vasopressin ( AVP ) and DB00035 response . P30518 activity of the p.N317S mutant receptor after the AVP and DB00035 administration , as assessed by DB02527 production was reduced and impaired when compared to cells that expressed the wild type P30518 gene . In conclusion , the affected members of this family have X-linked NDI with partial resistance to AVP , due to a missense mutation in the P30518 gene . Selective inhibition of the tumor marker O60218 by antiinflammatory N-phenylanthranilic acids and glycyrrhetic acid . A human aldose reductase-like protein , O60218 in the aldo-keto reductase ( AKR ) superfamily , was recently identified as a tumor marker of several types of cancer . DB02383 , an aldose reductase inhibitor ( Q9Y4X5 ) , is known to be the most potent inhibitor of the enzyme . In this study , we compared the inhibitory effects of other ARIs including flavonoids on O60218 and aldose reductase to evaluate their specificity . However , ARIs showed lower inhibitory potency for O60218 than for aldose reductase . In the search for potent and selective inhibitors of O60218 from other drugs used clinically , we found that non-steroidal antiinflammatory N-phenylanthranilic acids , diclofenac and glycyrrhetic acid competitively inhibited O60218 , showing K(i) values of 0.35-2.9 microM and high selectivity to this enzyme ( 43-57 fold versus aldose reductase ) . Molecular docking studies of mefenamic acid and glycyrrhetic acid in the O60218 -nicotinamide adenine dinucleotide phosphate ( NADP(+) ) complex and site-directed mutagenesis of the putative binding residues suggest that the side chain of Val301 and a hydrogen-bonding network among residues Val301 , Gln114 and Ser304 are important for determining the inhibitory potency and selectivity of the non-steroidal antiinflammatory drugs . Thus , the potent and selective inhibition may be related to the cancer chemopreventive roles of the drugs , and their structural features may facilitate the design of new anti-cancer agents targeting O60218 . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Impairment of the extrusion transporter for asymmetric DB01686 : a novel mechanism underlying vasospastic angina . A 37-year old male patient presented with frequent angina attacks ( up to 40/day ) largely resistant to classical vasodilator therapy . The patient showed severe coronary and peripheral endothelial dysfunction , increased platelet aggregation and increased platelet-derived superoxide production . The endothelial nitric oxide synthase ( P29474 ) -inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) reduced superoxide formation in platelets identifying " uncoupled " P29474 as a superoxide source . Oral L-arginine normalized coronary and peripheral endothelial dysfunction and reduced platelet aggregation and P29474 -derived superoxide production . Plasma concentrations of the endogenous NOS inhibitor asymmetric DB01686 ( DB01686 ) , representing an independent risk factor for cardiovascular disease , were normal in the patient . However , immediately after oral administration of cationic amino acid ( CAA ) , plasma DB01686 levels rose markedly , demonstrating increased DB01686 efflux from intracellular stores . DB01686 efflux from mononuclear cells of the patient was accelerated by CAA , but not neutral amino acids ( NAA ) demonstrating impairment of y(+) O43561 ( whose expression was found reduced in these cells ) . These data suggest that impairment of y(+) O43561 may cause intracellular ( endothelial ) DB01686 accumulation leading to systemic endothelial dysfunction . This may represent a novel mechanism underlying vasospastic angina and vascular dysfunction in general . Moreover , these new findings contribute to the understanding of the l-arginine paradox , the improvement of P29474 activity by oral L-arginine despite sufficient cellular l-arginine levels to ensure proper function of this enzyme . Potential mechanism for endothelial progenitor cell therapy in acute myocardial infarction : Activation of P15692 - PI3K/Akte-NOS pathway . Mounting evidence suggests that transplanting endothelial progenitor cells ( EPCs ) into the myocardium improves cardiac function after myocardial infarction ( MI ) . However , the mechanism remains controversial . The aim of this study was to investigate the role played by the P15692 - PI3K/Akt- P29474 pathway in EPC-based cell therapy . Cultured EPCs , which were identified by morphology , function , and cell surface markers , were transplanted into the border zone after left anterior descending coronary artery ligation in mice . Expression levels of P15692 , p-Akt , and P29474 in the border zone were elevated three days after EPC transplantation . EPC therapy enhanced expression of P35968 , increased microvessel density , and reduced interstitial fibrosis in the border zone after MI . The left ventricular fractional shortening was increased and the left ventricular diameter was smaller after EPC treatment . Wortmannin inhibited the expression of p-Akt and was associated with decreased cardiac function . Our study suggests that EPC transplantation improves cardiac function after MI , mediated at least partially by activation of the P15692 -PI3K/Akt- P29474 pathway . P19957 kinase/AKT pathway as a therapeutic target in multiple myeloma . The development of novel therapies for multiple myeloma depends on a comprehensive understanding of the events leading to cellular proliferation and survival . Controlling pathways that regulate growth signals is an emerging and complementary approach to myeloma treatment . The PI3K/Akt pathway is a central gatekeeper for crucial cellular functions including adhesion , angiogenesis , migration and development of drug resistance . Established proteins and genes such as P42345 , p53 , NF-kappaB and Q92934 are all regulated through PI3K and Akt activation , making them attractive targets for broad downstream effects . Direct PI3K inhibition has demonstrated impressive tumor inhibition and regression in cell-line and animal models , and multiple agents including DB05210 are currently in clinical trials . Drugs such as perifosine that are specific for Akt are also in development . Combinations of these agents with existing therapies are rational approaches on the path to improving myeloma treatment . Mechanism of inhibition of the P42262 AMPA receptor channel opening by talampanel and its enantiomer : the stereochemistry of the 4-methyl group on the diazepine ring of 2,3-benzodiazepine derivatives . Stereoselectivity of 2,3-benzodiazepine compounds provides a unique way for the design of stereoisomers as more selective and more potent inhibitors as drug candidates for treatment of the neurological diseases involving excessive activity of AMPA receptors . Here we investigate a pair of enantiomers known as DB04982 and its ( + ) counterpart about their mechanism of inhibition and selectivity toward four AMPA receptor subunits or P42261 -4 . We show that DB04982 is the eutomer with the endismic ratio being 14 for the closed-channel and 10 for the open-channel state of P42262 . Kinetic evidence supports that DB04982 is a noncompetitive inhibitor and it binds to the same site for those 2,3-benzodiazepine compounds with the C-4 methyl group on the diazepine ring . This site , which we term as the " M " site , recognizes preferentially those 2,3-benzodiazepine compounds with the C-4 methyl group being in the R configuration , as in the chemical structure of DB04982 . Given that DB04982 inhibits P42261 and P42262 , but is virtually ineffective on the P42263 and P48058 AMPA receptor subunits , we hypothesize that the " M " site(s) on P42261 and P42262 to which DB04982 binds is different from that on P42263 and P48058 . If the molecular properties of the AMPA receptors and DB04982 are used for selecting an inhibitor as a single drug candidate for controlling the activity of all AMPA receptors in vivo , DB04982 is not ideal . Our results further suggest that addition of longer acyl groups to the N-3 position should produce more potent 2,3-benzodiazepine inhibitors for the " M " site . Urinary pheromones promote P29323 /Akt phosphorylation , regeneration and survival of vomeronasal ( P30518 ) neurons . The G protein-coupled pheromone receptor neurons ( V1R and P30518 ) of the vomeronasal organ ( VNO ) are continually replaced throughout the lifetime of the mouse . Moreover , active signalling of V2Rs via the transient receptor potential 2(TRPC2) channel is necessary for regeneration of receptors , as the TRPC2 null mutant mouse showed a 75 % reduction of V2Rs by the age of two months . Here we describe P30518 mediated signalling in a neuronal line established from vomeronasal stem cells taken from postnatal female mice . Cells were immunoreactive for Galpha(o) and P30518 , whereas V1R and Galpha(i) immunoreactivity could not be detected . Biological ligands ( dilute urine and its protein fractions ) were found to increase proliferation and survival of these neurons . Dilute mouse urine but not artificial urine also induced P29323 , Akt and CREB signalling in a dose dependent way . The volatile fraction of male mouse urine alone was without effect while the fraction containing peptides ( > 5 kDa ) also stimulated P29323 and Akt phosphorylation . The P29323 , Akt and CREB phosphorylation response was sensitive to pertussis toxin , confirming the involvement of P30518 linked Galpha(o) . Dilute mouse urine or its high molecular weight protein fraction increased survival and proliferation of these neurons . Hence , urinary pheromones , which signal important social information via mature neurons , also promote survival and proliferation of their regenerating precursors . These data show that regenerating V2Rs respond to urine and the urinary peptides by activation of the Ras- P29323 and P19957 -Akt pathways , which appear to be important for vomeronasal neural survival and proliferation . Autoinhibition of endothelial nitric oxide synthase ( P29474 ) in gut smooth muscle by nitric oxide . DB00435 in the gut is produced by P29475 in enteric neurons and by P29474 in smooth muscle cells . The P29474 in smooth muscle is activated by vasoactive intestinal peptide ( P01282 ) released from enteric neurons . In the present study , we examined the effect of nitric oxide on P01282 -induced P29474 activation in smooth muscle cells isolated from human intestine and rabbit stomach . NOS activity was measured as formation of the 1:1 co-product , l-citrulline from l-arginine . P01282 caused an increase in l-citrulline production that was inhibited by NO in a concentration dependent manner ( IC(50)~25 microM ; maximal inhibition 72 % at 100 microM NO ) . Basal l-citrulline production , however , was unaffected by NO . The effect was not mediated by cGMP/PKG since the PKG inhibitor KT5823 had no effect on P29474 autoinhibition . The autoinhibition was selective for NO since the co-product l-citrulline had no effect on P01282 -induced NOS activation . Similar effects were obtained in rabbit gastric and human intestinal smooth muscle cells . The results suggest that NO produced in smooth muscle cells as a result of the activation of P29474 by P01282 exerts an autoinhibitory restraint on P29474 thereby regulating the balance of the P01282 / DB02527 /PKA and NO/cGMP/PKG pathways that regulate the relaxation of gut smooth muscle . Is transforming growth factor-β signaling activated in human hypertrophied prostate treated by 5-alpha reductase inhibitor ? BACKGROUND AND AIM : It is well known that androgen deprivation relates to penile fibrosis , so we hypothesize that long-term treatment with 5-alphareductase inhibitors ( 5ARIs ) may increase the risk of fibrosis of prostate . PATIENTS AND METHODS : Thirty-two BPH patients who underwent transurethral resection of the prostate were enrolled . The patients were divided into two groups : group one , 16 patients underwent TURP who had been treated with tamsulosin for 2 years ; group two , 16 patients underwent TURP who had been treated with combination of tamsulosin and dutasteride for at least 1 year . We evaluated the expressions of P29475 , P35228 , P29474 , TGF-β1 , TGF-β2 , phosphorylated- Q15796 /3 ( p- Q15796 /3 ) , P12830 , P19022 , and α-smooth muscle actin in the resected prostate tissues by western blotting , and the TGF-β concentration was determined by ELISA kit . RESULTS : The expressions of 3 isoforms of NOS were significantly increased in group 2 except of P29474 in lateral prostate , and the expressions of TGF-β1 , TGF-β2 , and p- Q15796 /3 increased about 2-fold compared with group 1 . In group 2 , the P12830 expression decreased while P19022 expression increased significantly . CONCLUSIONS : The overexpression of P29475 may contribute to prostate smooth muscle relaxation ; however , long-time treatment with 5 Q9Y4X5 increases the risk of fibrosis of prostate . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . DB04970 ( Esteve ) . DB04970 is an anxiolytic with pre- and post-synaptic P08908 agonist activity , which is under development by Esteve [ 172924 , 175197 ] . It is in phase II trials in Spain [ 391567 , 396196 ] and the US [ 396174 , 396196 ] . A series of pilot studies in other indications are also under way [ 396196 ] . In phase I trials in healthy volunteers , lesopitron was well tolerated in single doses up to 50 mg , and up to 45 mg/day in repeated doses [ 234400 ] . DB04970 has negligible effects on alpha-adrenergic and dopaminergic receptors [ 162653 ] , and was more potent than structurally-related P08908 agonists in rat social interaction and marmoset anxiety models . It also countered benzodiazepine withdrawal-induced anxiety in rodents [ 232978 ] . The acute toxicity of lesopitron is low and it does not potentiate the effects of alcohol or barbiturates . Long-term usage led to reductions in plasma glucose , triglycerides , phospholipids and cholesterol [ 232984 ] . DB04970 was being developed in collaboration with Knoll Pharmaceuticals [ 172924 , 175197 ] but this agreement was terminated in December 1995 [ 394772 ] . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . Effects of neuropeptides on human lung fibroblast proliferation and chemotaxis . An increase in subepithelial mesenchymal cells and associated connective tissue is a feature of bronchial asthma . We determined whether neuropeptides could modulate fibroblast activity , particularly with respect to proliferation and chemotaxis . Human lung fibroblasts were cultured with neurokinin A ( P20366 ) , DB05875 ( SP ) , vasoactive intestinal peptide ( P01282 ) , and calcitonin-gene-related peptide ( P80511 ) . After 48 h , fibroblast proliferation was measured by a colorimetric assay based on the uptake and subsequent release of methylene blue . The chemotactic response to neuropeptides was determined with the use of a modified Boyden chamber . Both P20366 and SP ( 10(-7)-10(-4) M ) stimulated human lung fibroblast proliferation in Q03591 and IMR-90 fibroblasts . P01282 and P80511 had no effect on fibroblast proliferation . P20366 alone stimulated fibroblast chemotaxis maximally at 10(-10) M . P08473 ( NEP ) activity of 0.52 and 5.2 pmol/10(6) cells was assayed in IMR-90 and Hs68 fibroblasts , respectively . DB02557 ( 5 x 10(-6)-10(-5) M ) , an NEP inhibitor , enhanced fibroblast proliferation in a dose-dependent manner . Thus neuropeptides have the potential to cause activation of mesenchymal cells , and neuropeptide release may contribute to the structural abnormalities observed in asthmatic airways . Volumetric analysis of white matter , gray matter , and P04141 using fractional volume analysis . Quantitative cerebral tissue volumes may be useful for an objective assessment of pathological changes in brain . Accurate determination of tissue volumes is complicated , however , by the partial volume averaging ( P32926 ) effect . We have , therefore , developed a new pulse sequence that minimizes the P32926 through the use of inversion-recovery ( IR ) and double inversion-recovery ( P30518 ) techniques . This pulse sequence simultaneously acquires four different sets of images to provide the necessary information for volumetric analysis and reduces potential spatial misregistration of images due to patient motion . The image sets acquired from the proposed pulse sequence are 1 ) gray matter visible , 2 ) white matter visible , 3 ) FLAIR , and 4 ) fast spin-echo proton-density weighted images . An algorithm has been implemented to correct for differential T1-weighting and for tissue quantitation . P12004 -dependent kinase 5 phosphorylates endothelial nitric oxide synthase at serine 116 . DB00435 ( NO ) production in endothelial cells ( EC ) is regulated by multisite phosphorylation of specific serine and threonine residues in endothelial NO synthase ( P29474 ) . Among these , P29474 -Ser116 is phosphorylated in the basal state , and its phosphorylation contributes to basal NO production . Here , we investigated the mechanism by which P29474 -Ser116 is phosphorylated during the basal state using bovine aortic EC . Although a previous study suggested that protein kinase C was involved in P29474 -Ser116 phosphorylation , overexpression of various protein kinase C isoforms did not affect P29474 -Ser116 phosphorylation . An in silico analysis using a motif scan revealed that the P29474 -Ser116 residue might be a substrate for proline-directed protein kinases . Roscovitine , a specific inhibitor of cyclin-dependent kinase ( CDK ) , 1 , 2 , and 5 , but not an inhibitor of mitogen-activated protein kinase kinase or glycogen synthase kinase 3beta , inhibited P29474 -Ser116 phosphorylation dose dependently . Furthermore , purified P06493 , 2 , or 5 directly phosphorylated P29474 -Ser116 in vitro . Ectopic expression of the dominant-negative Q00535 but not dominant-negative P06493 or dominant-negative P24941 repressed P29474 -Ser116 phosphorylation and increased NO production . In addition , Q00535 activity was detected in bovine aortic EC , and coimmunoprecipitation and confocal microscopy studies revealed a colocalization of P29474 and Q00535 . Cotransfection of Q00535 and p25 , the specific Q00535 activator , increased P29474 -Ser116 phosphorylation and decreased NO production , but its parent molecule , p35 , and p39 , another activator , were not detected in bovine aortic EC , which suggests the existence of a novel Q00535 activator . Overall , this is the first study to find that Q00535 is a physiological kinase responsible for P29474 -Ser116 phosphorylation and regulation of NO production . Proopiomelanocortin but not vasopressin or renin-angiotensin system induces resuscitative effects of central P08908 activation in haemorrhagic shock in rats . The aim of this study was to determine the effectory mechanisms : vasopressin , renin-angiotensin system and proopiomelanocortin-derived peptides ( P01189 ) , partaking in the effects of serotonin through central serotonin 1A receptor ( P08908 ) receptors in haemorrhagic shock in rats . The study was conducted on male Wistar rats . All experimental procedures were carried out under full anaesthesia . The principal experiment included a 2 hour observation period in haemorrhagic shock . Drugs used - a selective P08908 agonist 8-OH-DPAT ( 5 μg/5 μl ) ; V1a receptor antagonist [ β-mercapto-β , β-cyclo-pentamethylenepropionyl(1),O-me- DB00135 (2), DB00125 (8) ] AVP ( 10 μg/kg ) ; angiotensin type I receptor antagonist ( AT1 ) ZD7155 ( 0.5 mg/kg , i.v. ) ; angiotensin-converting-enzyme inhibitor captopril ( 30 mg/kg , i.v. ) ; melanocortin type 4 ( MC4 ) receptor antagonist HS014 ( 5 μg , i.c.v. ) . There was no influence of ZD715 , captopril or blocking of the V1a receptors on changes in the heart rate ( HR ) , mean arterial pressure ( Q96HU1 ) , peripheral blood flow or resistance caused by the central stimulation of P08908 receptors ( P≥0.05 ) . However , selective blocking of central MC4 receptors caused a slight , but significant decrease in HR and Q96HU1 ( P < 0.05 ) . P01189 derivatives acting via the central MC4 receptor participate in the resuscitative effects of 8-OH-DPAT . The angiotensin and vasopressin systems do not participate in these actions . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Ectopic DB01285 syndrome caused by bronchial carcinoid tumor indistinguishable from Cushing 's disease . A 75-year-old woman was admitted to our hospital because of a poor glycemic control . She was found to have Cushingoid feature and dynamic endocrine tests showed elevated plasma DB01285 and cortisol levels , lack of their circadian rhythm , non-suppressibility to high-dose dexamethasone , responsiveness to P06850 , but not to DB00035 , and suppression to octreotide . Pituitary Q9BWK5 showed an equivocal small lesion . CT scan of the chest showed two nodular lesions in the right lung ( S5 , S7 ) , while a mild uptake was noted only in S5 lesion by DB09150 -PET , but positive uptake was only in S7 lesion by somatostatin receptor scintigraphy ( SRS ) . Inferior petrosal sinus sampling revealed a gradient of plasma DB01285 after P06850 stimulation , consistent with the diagnosis of Cushing ' s disease . She underwent middle and inferior lobectomy of the right lung . The resected tumor in S7 was consistent with the diagnosis of a bronchial carcinoid tumor with positive DB01285 immunoreactivity , while that of S5 was cryptococcal granuloma . RT-PCR revealed abundant expressions of P01189 and SSTR ( -1 , -2 , -5 ) , but not of P34998 and P47901 . Postoperatively , abnormal endocrine data were normalized along with improvement of hypertension and diabetes . This was a diagnostic challenging case with ectopic DB01285 syndrome indistinguishable from Cushing ' s disease by various endocrine and imaging tests , among which SRS successfully localized the tumor responsible for ectopic DB01285 secretion . A population-based association study of candidate genes for depression and sleep disturbance . The clinical manifestation of depression comprises a variety of symptoms , including early morning awakenings and fatigue , features also indicating disturbed sleep . The presence or absence of these symptoms may reflect differences in neurobiological processes leading to prolonged depression . Several neurobiological mechanisms have been indicated in the induction of depression , including disturbances in serotonergic and glutamatergic neurotransmission and in the action of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . The same transmitters have also been linked to sleep regulation . We hypothesized that depression without simultaneous symptoms of disturbed sleep would partly have a different genetic background than depression with symptoms of disturbed sleep . We tested this hypothesis using a systematic population-based association study of 14 candidate genes related to depression and disturbed sleep . Association of genetic variants with either depression alone , depression with early morning awakenings , or depression with fatigue was investigated using permutation-based allelic association analysis of a sample of 1,654 adults recruited from Finland 's population-based program . The major findings were associations of Q8IWU9 ( rs12229394 ) with depression accompanied by fatigue in women and P16220 ( rs11904814 ) with depression alone in men . We also found suggestive associations in women for Q99259 , P42263 , and P23560 with depression accompanied by fatigue , and for P34998 with depression accompanied by early morning awakenings . The results indicate sex-dependent and symptom-specific differences in the genetic background of depression . These differences may partially explain the broad spectrum of depressive symptoms , and their systematic monitoring could potentially be used for diagnostic purposes . DB05578 : preclinical research and clinical development . DB05578 ( IMC-1121B , LY3009806 ) , a fully humanized monoclonal antibody directed against the extracellular domain of vascular endothelial growth factor receptor 2 ( P35968 ) , is a new therapeutic option that selectively inhibits the human P35968 with a much greater affinity than its natural ligands . Based on the promising results of both preclinical and early clinical studies , ramucirumab has been tested in different tumor types either alone or in combination with chemotherapy . While it has recently been granted its first US Food and Drug Administration approval for use as a single agent in patients with advanced or metastatic gastric cancer or gastroesophageal junction carcinoma , its role for metastatic breast cancer or advanced non-small-cell lung cancer is still debated . The aims of this review are to recall and discuss the most significant preclinical and clinical studies that led to the development of ramucirumab and to present the results of the randomized clinical trials that have tested its efficacy in different malignancies , including gastric and lung cancer . Inhibition of cyclin-dependent kinases , GSK-3beta and CK1 by hymenialdisine , a marine sponge constituent . BACKGROUND : Over 2000 protein kinases regulate cellular functions . Screening for inhibitors of some of these kinases has already yielded some potent and selective compounds with promising potential for the treatment of human diseases . RESULTS : The marine sponge constituent hymenialdisine is a potent inhibitor of cyclin-dependent kinases , glycogen synthase kinase-3beta and casein kinase 1 . DB02950 competes with DB00171 for binding to these kinases . A P24941 -hymenialdisine complex crystal structure shows that three hydrogen bonds link hymenialdisine to the Glu81 and Leu83 residues of P24941 , as observed with other inhibitors . DB02950 inhibits Q00535 /p35 in vivo as demonstrated by the lack of phosphorylation/down-regulation of Pak1 kinase in E18 rat cortical neurons , and also inhibits GSK-3 in vivo as shown by the inhibition of P46821 phosphorylation . DB02950 also blocks the in vivo phosphorylation of the microtubule-binding protein tau at sites that are hyperphosphorylated by GSK-3 and Q00535 /p35 in Alzheimer 's disease ( cross-reacting with Alzheimer's-specific AT100 antibodies ) . CONCLUSIONS : The natural product hymenialdisine is a new kinase inhibitor with promising potential applications for treating neurodegenerative disorders .

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interacts_with DB06822?

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Pharmacology of recombinant low-voltage activated calcium channels . Several types of voltage- or ligand-activated calcium channels contribute to the excitability of neuronal cells . Low-voltage-activated ( LVA ) , T-type calcium channels are characterised by relatively negative threshold of activation and therefore they can generate low-threshold spikes , which are essential for burst firing . At least three different proteins form T-type calcium current in neurons : Ca(v)3.1 , Ca(v)3.2 and Q9P0X4 . Expression of these proteins in various brain regions is complementary . Individual channel types could be distinguished by different sensitivity towards inorganic cations . This inhibition can contribute to the toxicity of some heavy metals . Selective inhibition of T-type calcium channels by organic blockers may have clinical importance in some forms of epilepsy . DB01388 inhibits the expressed Ca(v2)3.1 , Ca(v)3.2 and Q9P0X4 channels in nanomolar concentrations with Q9P0X4 channel having lowest affinity . The sensitivity of the expressed Ca(v)3.1 channel to the antiepileptic drugs , valproate and ethosuximide , is low . Ca(v)3.1 channel is moderately sensitive to phenytoin . The Ca(v)3.2 channel is sensitive to ethosuximide , amlodipine and amiloride . All three LVA calcium channels are moderately sensitive to active metabolites of methosuximide , i.e. alpha-methyl-alpha-phenylsuccinimide . Several neuroleptics inhibit all three LVA channels in clinically relevant concentrations . All three channels are also inhibited by the endogenous cannabinoid anandamide . A high affinity peptide blocker for these Ca channels is the scorpion toxin kurtoxin which inhibits the Ca(v)3.1 and Ca(v)3.2 , but not the Q9P0X4 channel in nanomolar concentrations . DB06690 selectively inhibits the Ca(v)3.2 , but not the Ca(v)3.1 channel . The Ca(v)3.2 , but not the Ca(v)3.1 channel is potentiated by stimulation of Ca(2+)/ P62158 -dependent protein kinase . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Intracellular signaling pathways activated by kisspeptins through Q969F8 : do multiple signals underlie function diversity ? Kisspeptins , a family of peptide products derived from the KiSS-1 gene , activate their cognate receptor Q969F8 in various target tissues to exert disparate functions , including inhibition of tumor metastasis and control of reproductive function . In contrast to the plethora of studies that have analyzed in recent years the regulatory functions of the KiSS-1/ Q969F8 system , only a limited number of reports have been primarily focused on delineating the intracellular signaling pathways involved . Nevertheless , there is solid evidence indicating that kisspeptin can activate a wide variety of signals via Q969F8 . These include typical G-protein ( Galphaq/11 ) -coupled cascades , such as activation of phospholipase C ( P98160 ) , and subsequent accumulation of inositol-(1,4,5)-triphosphate ( IP3 ) , intracellular Ca(2+) mobilization , and activation of protein kinase C . However , kisspeptin also activates pathways related to mitogen activated protein kinases ( MAPK ) , especially P27361 /2 , and p38 and phosphatidylinositol-3-kinase ( PI3K ) /Akt . Additionally , the kisspeptin/ Q969F8 pair can also influence cell signaling by interacting with other receptors , such as chemokine receptor P61073 , and P30968 . Kisspeptin can also affect other signaling events , like expression of matrix metalloproteinase 9 ( via NFkappaB ) , and that of calcineurin . The information gathered hitherto clearly indicates that activation of a specific set of interconnected signals is selectively triggered by kisspeptin via Q969F8 in a cell type-dependent manner to precisely regulate functions as distinct as hormone release and cell migration . In this scenario , it will be important to decipher kisspeptin/ Q969F8 signaling mechanisms in reproductive and non-reproductive tissues by studying additional models , especially on natural kisspeptin targets expressing endogenous Q969F8 . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Chemotaxis of mouse bone marrow neutrophils and dendritic cells is controlled by adp-ribose , the major product generated by the P28907 enzyme reaction . The ectoenzyme P28907 catalyzes the production of cyclic ADP-ribose ( cADPR ) and ADP-ribose ( DB02059 ) from its substrate , NAD(+) . Both products of the P28907 enzyme reaction play important roles in signal transduction , as cADPR regulates calcium release from intracellular stores and DB02059 controls cation entry through the plasma membrane channel O94759 . We previously demonstrated that P28907 and the cADPR generated by P28907 regulate calcium signaling in leukocytes stimulated with some , but not all , chemokines and controls leukocyte migration to inflammatory sites . However , it is not known whether the other P28907 product , DB02059 , also regulates leukocyte trafficking In this study we characterize 8-bromo ( 8Br ) - DB02059 , a novel compound that specifically inhibits DB02059 -activated cation influx without affecting other key calcium release and entry pathways . Using 8Br- DB02059 , we demonstrate that DB02059 controls calcium influx and chemotaxis in mouse neutrophils and dendritic cells activated through chemokine receptors that rely on P28907 and cADPR for activity , including mouse P21462 , P61073 , and P32248 . Furthermore , we show that the calcium and chemotactic responses of leukocytes are not dependent on poly-ADP-ribose polymerase 1 ( P09874 ) , another potential source of DB02059 in some leukocytes . Finally , we demonstrate that NAD(+) analogues specifically block calcium influx and migration of chemokine-stimulated neutrophils without affecting P09874 -dependent calcium responses . Collectively , these data identify DB02059 as a new and important second messenger of mouse neutrophil and dendritic cell migration , suggest that P28907 , rather than P09874 , may be an important source of DB02059 in these cells , and indicate that inhibitors of DB02059 -gated calcium entry , such as 8Br- DB02059 , have the potential to be used as anti-inflammatory agents . Induction of cyclooxygenase-2 by benzo[a]pyrene diol epoxide through inhibition of retinoic acid receptor-beta 2 expression . Benzo[a]pyrene diol epoxide ( BPDE , a carcinogen present in tobacco smoke and environmental pollution ) has been shown to suppress retinoic acid receptor-beta2 ( P10826 (2) ) and induce cyclooxygenase-2 ( P35354 ) expression . Restoration of P10826 (2) inhibited growth and colony formation of esophageal cancer cells , which was correlated with P35354 suppression . In this study , we investigated the molecular mechanisms for P10826 (2)-mediated suppression of P35354 expression using BPDE as a tool . We found that BPDE-induced P35354 expression was through inhibition of P10826 (2) and consequently , induction of epidermal growth factor receptor ( P00533 ) , extracellular signal-regulated protein kinases 1/2 ( Erk1/2 ) phosphorylation , and c-Jun expression . Esophageal cancer cells that do not express P10826 (2) did not respond to BPDE for induction of P35354 . BPDE was also unable to induce P35354 expression after P10826 (2) expression was manipulated in these esophageal cancer cells . Furthermore , BPDE induced time-dependent methylation of P10826 (2) gene promoter in esophageal cancer cells . Transfection of P10826 (2) expression vector into esophageal cancer cells suppressed expression of P00533 , Erk1/2 phosphorylation , c-Jun , and P35354 . In addition , co-treatment of P10826 (2)-positive cells with BPDE and the Q02750 /2 inhibitor U0126 caused little change in c-Jun and P35354 expression . This study demonstrated that BPDE-suppressed expression of P10826 (2) results in P35354 induction and restoration of P10826 (2) expression reduces P35354 protein in esophageal cancer cells , thereby further supporting our previous finding that P10826 (2) plays an important role in suppressing esophageal carcinogenesis . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Leukotriene binding , signaling , and analysis of HIV coreceptor function in mouse and human leukotriene B4 receptor-transfected cells . The mouse leukotriene B4 receptor ( m- Q15722 ) gene was cloned . Membrane fractions of human embryonic kidney 293 cells stably expressing m- Q15722 demonstrated a high affinity and specific binding for leukotriene B4 ( LTB4 , Kd = 0.24 +/- 0.03 nM ) . In competition binding experiments , LTB4 was the most potent competitor ( Ki = 0.23 +/- 0.05 nM ) followed by 20-hydroxy-LTB4 ( Ki = 1.1 +/- 0.2 nM ) and by 6-trans-12-epi-LTB4 and LTD4 ( Ki > 1 microM ) . In stably transfected Chinese hamster ovary cells , LTB4 inhibited forskolin-activated DB02527 production and induced an increase of intracellular calcium , suggesting that this receptor is coupled to Gi- and Go-like proteins . In Xenopus laevis melanophores transiently expressing m- Q15722 , LTB4 induced the aggregation of pigment granules , confirming the inhibition of DB02527 production induced by LTB4 . Q15722 receptors share significant sequence homology with chemokine receptors ( P51681 and P61073 ) that act as human immunodeficiency virus ( HIV ) coreceptors . However , among the 16 HIV/SIV strains tested , the human Q15722 receptor did not act as a coreceptor for virus entry into P01730 -expressing cells based on infection and cell-cell fusion assays . In P09917 -deficient mice , the absence of leukotriene B4 biosynthesis did not detectably alter m- Q15722 receptor binding in membranes obtained from glycogen-elicited neutrophils . Isolation of the m- Q15722 gene will form the basis of future experiments to elucidate the selective role of LTB4 , as opposed to cysteinyl-leukotrienes , in murine models of inflammation . A point mutation that confers constitutive activity to P61073 reveals that T140 is an inverse agonist and that DB06809 and ALX40-4C are weak partial agonists . P61073 is a G protein-coupled receptor for stromal-derived factor 1 ( P48061 ) that plays a critical role in leukocyte trafficking , metastasis of mammary carcinoma , and human immunodeficiency virus type-1 infection . To elucidate the mechanism for P61073 activation , a constitutively active mutant ( P62158 ) was derived by coupling the receptor to the pheromone response pathway in yeast . Conversion of DB00174 -119 to DB00133 or Ala , but not DB00128 or Lys , conferred autonomous P61073 signaling in yeast and mammalian cells . P48061 induced signaling in variants with substitution of DB00174 -119 to DB00133 , Ala , or DB00128 , but not Lys . These variants had similar cell surface expression and binding affinity for P48061 . P61073 -CAMs were constitutively phosphorylated and present in cytosolic inclusions . Analysis of antagonists revealed that exposure to DB06809 or ALX40-4C induced G protein activation by P61073 wild type , which was greater in the P62158 , whereas T140 decreased autonomous signaling . The affinity of DB06809 and ALX40-4C binding to CAMs was less than to wild type , providing evidence of a conformational shift . These results illustrate the importance of transmembrane helix 3 in P61073 signaling . Insight into the mechanism for P61073 antagonists will allow for the development of a new generation of agents that lack partial agonist activity that may induce toxicities , as observed for DB06809 . Crossreacting drugs and chemicals . DB00945 and nonsteroidal antiinflammatory drugs ( NSAIDs ) exert their clinical effect through inhibition of prostaglandin H synthases 1 and 2 , also known as cyclooxygenase . This shared effect of P36551 -inhibition is also the mechanism for shared adverse effects . Much of our understanding of cross-reacting drugs and chemicals with aspirin comes from studying asthmatics with aspirin-exacerbated respiratory disease ( AERD ) . DB00945 exacerbated respiratory disease is characterized by recalcitrant sinusitis/polyposis , asthma and precipitation of asthma after ingestion of aspirin and most NSAIDs . Cross-reactions between ASA and NSAIDs occur with first exposure unlike IgE-mediated allergic drug reactions . Cross-reactions between aspirin and other drugs are dependent upon inhibition of the cyclooxygenase-1 isoenzyme . Desensitization to aspirin will result in cross-desensitization to all NSATDs that inhibit P23219 . Despite reports in the literature , there does not appear to he cross-reactions between food coloring , hydrocortisone succinate and monosodium glutamate in individuals with aspirin exacerbated respiratory disease . The new highly selective cyclooxygenase 2 inhibitors are well tolerated in AERD asthmatics who have not been desensitized to aspirin . Because low-dose ASA exerts a cardioprotective effect by irreversible inhibition of P23219 , AERD patients who are at risk for coronary artery disease should be considered for aspirin desensitization . Microarray analysis of selected genes in neural stem and progenitor cells . To access and compare gene expression in fetal neuroepithelial cells ( NEPs ) and progenitor cells , we have used microarrays containing approximately 500 known genes related to cell cycle regulation , apoptosis , growth and differentiation . We have identified 152 genes that are expressed in NEPs and 209 genes expressed by progenitor cells . The majority of genes ( 141 ) detected in NEPs are also present in progenitor populations . There are 68 genes specifically expressed in progenitors with little or no expression in NEPs , and a few genes that appear to be present exclusively in NEPs . Using cell sorting , RT-PCR , in situ hybridization or immunocytochemistry , we have examined the segregation of expression to neuronal and glial progenitors , and identified several that appeared to be enriched in neuronal ( e.g. Q00535 , neuropilin , EphrinB2 , Q92914 ) or glial ( e.g. P61073 , RhoC , P16070 , tenascin C ) precursors . Our data provide a first report of gene expression profiles of neural stem and progenitor cells at early stages of development , and provide evidence for the potential roles of specific cell cycle regulators , chemokines , cytokines and extracellular matrix molecules in neural development and lineage segregation . Prolonged opportunity for neuroprotection in experimental stroke with selective blockade of cyclooxygenase-2 activity . The post-treatment effects of the selective cyclooxygenase ( P36551 ) -2 inhibitor , valdecoxib , were investigated in a rat model of temporary focal ischemia . DB00580 reduced basal brain prostaglandin E(2) concentrations at dosages that did not affect serum thromboxane B(2) , consistent with a selective P35354 effect . Temporary focal cerebral ischemia was produced in rats by middle cerebral artery occlusion for 90 min . There was increased expression of P35354 protein detected by Western blot and immunocytochemistry within neurons in the ischemic cortex at 4 and 24 h after ischemia . Rats were treated with vehicle or valdecoxib 15 min before or 1.5 , 3 and 6 h after cerebral ischemia . Rats were sacrificed and brain infarction volume determined 24 h after ischemia . DB00580 treatment was associated with a decrease in infarction volume when administered 15 min before , and 1.5 or 3 h but not 6 h after cerebral ischemia . There were no differences in physiological parameters during the procedure . DB00580 administered at 1.5 h after ischemia significantly reduced the concentrations of prostaglandin E(2) in ischemic penumbral cortex as compared to the vehicle-treated group and contralateral non-ischemic cortex . These results suggest that P35354 inhibition with valdecoxib is effective when initiated both before and after middle cerebral artery occlusion . PP2Cdelta ( Ppm1d , O15297 ) , an endogenous inhibitor of p38 MAPK , is regulated along with Trp53 and Cdkn2a following p38 MAPK inhibition during mouse preimplantation development . Preimplantation embryos utilize mitogen-activated protein kinase signaling ( MAPK ) pathways to relay signals from the external environment to prepare appropriate responses and adaptations to a changing milieu . It is therefore important to investigate how MAPK pathways are regulated during preimplantation development . This study was conducted to investigate whether PP2Cdelta ( Ppm1d , O15297 ) is expressed during mouse preimplantation development and to determine the influences of p38 MAPK inhibition on expression of Trp53 ( p53 ) , Ppm1d , ( O15297 ) , and Cdkn2a ( p16 ) during mouse preimplantation development . Our results indicate that Trp53 , Ppm1d , and Cdkn2a mRNAs and TRP53 and PP2Cdelta proteins are expressed throughout mouse preimplantation development . Treatment of 2-cell embryos with DB04338 ( potent inhibitor of p38 MAPK alpha/beta/ Q16539 /11 ) significantly increased Trp53 , Ppm1d and Cdkn2a and Mapk14 mRNA levels at 12 and 24 hr . Treatment of 8-cell embryos with DB04338 for 12 hr increased Trp53 , Ppm1d , and Cdkn2a mRNA levels , but not Mapk14 mRNA levels . Treatment of 8-cell embryos for 24 hr increased Trp53 , and Ppm1d mRNA levels , but decreased Cdkn2a and Mapk14 mRNA levels . Therefore , blockade of p38 MAPK activity is associated with embryo stage specific influences on Trp53 , Ppm1d , Cdkn2a , and Mapk14 expression during mouse preimplantation development . These results define downstream targets of p38 MAPK during preimplantation development and indicate that the p38 MAPK pathway regulates Trp53 , Ppm1d , and Cdkn2a expression . This study increases our understanding of the mechanisms controlling preimplantation development and of the interactions between preimplantation embryos and their culture environments . Leishmania mexicana amazonensis : ADP-ribosyltransferase antagonists specifically inhibit amastigote to promastigote differentiation . Leishmania mexicana amazonensis amastigotes were induced to differentiate by incubation at 27 C . Morphological transformation was studied both in untreated cultures and in cultures where DNA synthesis , and consequently the final stage in the production of promastigotes , was inhibited by hydroxyurea . DB03073 and other antagonists of ADP-ribosyltransferase ( P09874 ) specifically inhibited differentiation at a very early stage in both experimental systems . Cell proliferation ( in the absence of hydroxyurea ) was not inhibited by P09874 antagonists -- indeed greater multiplication of undifferentiated parasites was observed in the presence of these compounds . This indicated that the parasites were being diverted from differentiation to proliferation . Preincubation of the amastigotes with the P09874 antagonists was required to produce this effect , providing further evidence that ADP-ribosylation of proteins is required for the initiation of differentiation in Leishmania . Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV-1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE3-tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e.g. , all-trans-retinoic acid ( RA ) , 13-cis-retinoic acid ( 13- DB00982 ) , arotinoid acid ( DB02877 ) and m-carboxy-arotinoid acid ( m-carboxy- DB02877 , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 ( 24 nM ) greater than P10826 ( 4.0 nM ) greater than P13631 ( 1.3 nM ) , while the ED50 values for DB02877 and 13- DB00982 decreased in the order of P10276 ( 6.5 nM , 190 nM ) greater than P13631 ( 2.3 nM , 140 nM ) greater than P10826 ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy- DB02877 , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 greater than P10826 greater than P13631 . Paullones are potent inhibitors of glycogen synthase kinase-3beta and cyclin-dependent kinase 5/p25 . Paullones constitute a new family of benzazepinones with promising antitumoral properties . They were recently described as potent , DB00171 -competitive , inhibitors of the cell cycle regulating cyclin-dependent kinases ( CDKs ) . We here report that paullones also act as very potent inhibitors of glycogen synthase kinase-3beta ( GSK-3beta ) ( IC50 : 4-80 nM ) and the neuronal Q00535 /p25 ( IC50 : 20-200 nM ) . These two enzymes are responsible for most of the hyperphosphorylation of the microtubule-binding protein tau , a feature observed in the brains of patients with Alzheimer 's disease and other neurodegenerative ' taupathies ' . DB04014 , the most active paullone , was demonstrated to act by competing with DB00171 for binding to GSK-3beta . DB04014 inhibits the phosphorylation of tau in vivo at sites which are typically phosphorylated by GSK-3beta in Alzheimer 's disease . DB04014 also inhibits the Q00535 /p25-dependent phosphorylation of Q9UD71 in mouse striatum slices in vitro . This dual specificity of paullones may turn these compounds into very useful tools for the study and possibly treatment of neurodegenerative and proliferative disorders . Leukotriene A4 hydrolase . Inhibition by bestatin and intrinsic aminopeptidase activity establish its functional resemblance to metallohydrolase enzymes . DB03424 , an inhibitor of aminopeptidases , was also a potent inhibitor of leukotriene ( LT ) A4 hydrolase . On isolated enzyme its effects were immediate and reversible with a Ki = 201 +/- 95 mM . With erythrocytes it inhibited LTB4 formation greater than 90 % within 10 min ; with neutrophils it inhibited LTB4 formation by only 10 % during the same period , increasing to 40 % in 2 h . DB03424 inhibited P09960 hydrolase selectively ; neither P09917 nor 15-lipoxygenase activity in neutrophil lysates was affected . Purified P09960 hydrolase exhibited an intrinsic aminopeptidase activity , hydrolyzing L-lysine-p-nitroanilide and L-leucine-beta-naphthylamide with apparent Km = 156 microM and 70 microM and Vmax = 50 and 215 nmol/min/mg , respectively . Both P09960 and bestatin suppressed the intrinsic aminopeptidase activity of P09960 hydrolase with apparent Ki values of 5.3 microM and 172 nM , respectively . Other metallohydrolase inhibitors tested did not reduce P09960 hydrolase/aminopeptidase activity , with one exception ; captopril , an inhibitor of angiotensin-converting enzyme , was as effective as bestatin . The results demonstrate a functional resemblance between P09960 hydrolase and certain metallohydrolases , consistent with a molecular resemblance at their putative Zn2(+)-binding sites . The availability of a reversible , chemically stable inhibitor of P09960 hydrolase may facilitate investigations on the role of LTB4 in inflammation , particularly the process termed transcellular biosynthesis . P00533 inhibitors exacerbate differentiation and cell cycle arrest induced by retinoic acid and vitamin D3 in acute myeloid leukemia cells . By means of an unbiased , automated fluorescence microscopy-based screen , we identified the epidermal growth factor receptor ( P00533 ) inhibitors erlotinib and gefitinib as potent enhancers of the differentiation of HL-60 acute myeloid leukemia ( AML ) cells exposed to suboptimal concentrations of vitamin A ( all-trans retinoic acid , DB00755 ) or vitamin D ( 1α, DB00146 , VD ) . Erlotinib and gefitinib alone did not promote differentiation , yet stimulated the acquisition of morphological and biochemical maturation markers ( including the expression of CD11b and P08571 as well as increased NADPH oxidase activity ) when combined with either DB00755 or VD . Moreover , the combination of erlotinib and DB00755 or VD synergistically induced all the processes that are normally linked to terminal hematopoietic differentiation , namely , a delayed proliferation arrest in the G0/ P55008 phase of the cell cycle , cellular senescence , and apoptosis . Erlotinib potently inhibited the (auto)phosphorylation of mitogen-activated protein kinase 14 ( Q16539 , best known as p38(MAPK) ) and P12931 family kinases ( SFKs ) . If combined with the administration of DB00755 or VD , the inhibition of p38(MAPK) or SFKs with specific pharmacological agents mimicked the pro-differentiation activity of erlotinib . These data were obtained with 2 distinct AML cell lines ( HL-60 and MOLM-13 cells ) and could be confirmed on primary leukemic blasts isolated from the circulation of AML patients . Altogether , these findings point to a new regimen for the treatment of AML , in which naturally occurring pro-differentiation agents ( DB00755 or VD ) may be combined with P00533 inhibitors . Production and characterization of antibodies to gonadotropin-releasing hormone receptor . Antibodies to the gonadotropin-releasing hormone ( DB00644 ) receptor of bovine pituitary membranes have been raised in rabbits by immunization with affinity-purified receptor preparations . These antibodies did not compete with 125I-labeled DB00644 analog ( DB06719 ) for binding to the receptors but did precipitate rat and bovine solubilized receptors labeled with 125I- DB06719 . Binding of the antibodies to the receptors was also demonstrated by immunoprecipitation of 125I-labeled purified receptors and photoaffinity-labeled receptors . The antibodies did not have a DB00644 -like activity but rather inhibited , in a dose-dependent manner , DB00644 -stimulated luteinizing hormone release from cultured rat pituitary cells . In addition , the antibodies did not inhibit luteinizing hormone release stimulated by high K+ concentration . This suggests that the antibodies recognize domains of the receptor other than the binding site of the hormone and thereby inhibit the biological response . These P30968 antibodies provide a useful tool for studying P30968 structure , function , localization , and biosynthesis .

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A clinical trial with chimeric monoclonal antibody DB05304 and low dose interleukin-2 pulsing scheme for advanced renal cell carcinoma . PURPOSE : DB05304 is a chimeric monoclonal antibody that binds to carbonic anhydrase IX( Q16790 /MN) , which is present on greater than 95 % of RCCs of the clear cell subtype . The suggested working mechanism of DB05304 is by ADCC . Because the number of activated ADCC effector cells can be increased by a low dose interleukin-2 pulsing schedule , a multicenter study was initiated to investigate whether DB05304 combined with LD- P60568 could lead to an improved clinical outcome in patients with progressive RCC . MATERIALS AND METHODS : A total of 35 patients with progressive clear cell RCC received weekly infusions of DB05304 for 11 weeks combined with a daily LD- P60568 regimen . Patients were monitored longitudinally for ADCC capacity . Radiological assessment of metastatic lesions was performed at week 16 and regularly until disease progression . RESULTS : A durable clinical benefit was achieved in 8 of 35 patients ( 23 % ) , including 3 with a partial response and 5 with stabilization at 24 weeks or greater . Mean survival was 22 months . In general treatment was well tolerated with little toxicity . The number of effector cells increased during treatment but lytic capacity per cell did not increase . ADCC and clinical outcome did not appear to correlate . CONCLUSIONS : DB05304 combined with LD- P60568 in patients with metastatic RCC is safe and well tolerated . With a substantial clinical benefit and a median survival of 22 months in patients with metastatic RCC who have progressive disease at study entry combination therapy showed increased overall survival compared to DB05304 monotherapy . Survival was at least similar to that of currently used cytokine regimens but with a favorable toxicity profile . P50406 mediates defective brain development in monoamine oxidase A-deficient mouse embryos . Monoamine oxidases A and B ( P21397 and P27338 ) are enzymes of the outer mitochondrial membrane that metabolize biogenic amines . In the adult central nervous system , MAOs have important functions for neurotransmitter homeostasis . Expression of MAO isoforms has been detected in the developing embryo . However , suppression of P27338 does not induce developmental alterations . In contrast , targeted inhibition and knockdown of P21397 expression ( E7.5-E10.5 ) caused structural abnormalities in the brain . Here we explored the molecular mechanisms underlying defective brain development induced by P21397 knockdown during in vitro embryogenesis . The developmental alterations were paralleled by diminished apoptotic activity in the affected neuronal structures . Moreover , dysfunctional P21397 expression led to elevated levels of embryonic serotonin ( 5-hydroxytryptamine ( 5-HT ) ) , and we found that knockdown of serotonin receptor-6 ( 5-Htr6 ) expression or pharmacologic inhibition of 5-Htr6 activity rescued the P21397 knockdown phenotype and restored apoptotic activity in the developing brain . Our data suggest that excessive 5-Htr6 activation reduces activation of caspase-3 and -9 of the intrinsic apoptotic pathway and enhances expression of antiapoptotic proteins Bcl-2 and Bcl-XL . Moreover , we found that elevated 5-HT levels in P21397 knockdown embryos coincided with an enhanced activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) and a reduction of proliferating cell numbers . In summary , our findings suggest that excessive 5-HT in P21397 -deficient mouse embryos triggers cellular signaling cascades via 5-Htr6 , which suppresses developmental apoptosis in the brain and thus induces developmental retardations . P23945 -mediated uptake of 45Ca2+ by proteoliposomes and cultured rat sertoli cells : evidence for involvement of voltage-activated and voltage-independent calcium channels . We have previously reported incorporation into liposomes of Triton X-100-solubilized DB00094 receptor-G-protein complexes derived from purified bovine calf testis membranes . In the present study we have used this model system to show that DB00094 induces flux of 45Ca2+ into such proteoliposomes in a hormone-specific concentration-dependent manner . DB00094 , inactivated by boiling , had no stimulatory effect on 45Ca2+ flux , nor did isolated alpha- or beta-subunits of DB00094 . Addition of GTP ( or its analogs 5'-guanylylimidodiphosphate and guanosine-5'-O-[3-thiotriphosphate] ) or sodium fluoride ( in the presence or absence of GTP or its analogs ) failed to induce 45Ca2+ flux into proteoliposomes , suggesting that the uptake of 45Ca2+ was receptor , and not G-protein , related . Voltage-independent ( ruthenium red and gadolinium chloride ) and voltage-activated ( methyoxyverapamil and nifedipine ) calcium channel-blocking agents reduced DB00094 -stimulated 45Ca2+ flux into proteoliposomes to control levels . DB00094 also induced uptake of 45Ca2+ by cultured rat Sertoli cells . Ruthenium red and gadolinium chloride had no effect on basal levels of 45Ca2+ uptake or estradiol secretion by cultured rat Sertoli cells , nor did methoxyverapamil or nifedipine . All four calcium channel blockers , however , were able to reduce DB00094 -induced 45Ca2+ uptake to basal levels and DB00094 -stimulated conversion of androstenedione to estradiol by up to 50 % , indicating an involvement of Ca2+ in DB00094 -stimulated steroidogenesis . Our results suggest that the well documented changes in intracellular calcium levels consequent to DB00094 binding may be due , at least in part , to an influx of calcium through DB00094 receptor-regulated calcium channels . Differential proteomic analysis on the effects of 2-methoxy-1,4-naphthoquinone towards MDA-MB-231 cell line . BACKGROUND : We have previously reported the anti-metastatic effects of 2-methoxy-1,4-naphthoquinone ( MNQ ) against MDA-MB-231 cell line . PURPOSE : To investigate the molecular mechanism underlying the anti-metastatic effects of MNQ towards MDA-MB-231 cell line via the comparative proteomic approach . STUDY DESIGN/METHODS : Differentially expressed proteins in MNQ-treated MDA-MB-231 cells were identified by using two-dimensional gel electrophoresis coupled with tandem mass spectrometry . Proteins and signalling pathways associated with the identified MNQ-altered proteins were studied by using Western blotting . RESULTS : Significant modulation of MDA-MB-231 cell proteome was observed upon treatment with MNQ in which the expressions of 19 proteins were found to be downregulated whereas another eight were upregulated ( > 1.5 fold , p < 0.05 ) . The altered proteins were mainly related to cytoskeletal functions and regulations , mRNA processing , protein modifications and oxidative stress response . Notably , two of the downregulated proteins , protein S100-A4 ( P26447 ) and laminin-binding protein ( P08865 ) are known to play key roles in driving metastasis and were verified using Western blotting . Further investigation using Western blotting also revealed that MNQ decreased the activations of pro-metastatic P27361 /2 and NF-κB signalling pathways . Moreover , MNQ was shown to stimulate the expression of the metastatic suppressor , P12830 . CONCLUSION : This study reports a proposed mechanism by which MNQ exerts its anti-metastatic effects against MDA-MB-231 cell line . The findings from this study offer new insights on the potential of MNQ to be developed as a novel anti-metastatic agent . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . Monocyte-mediated suppression of P60568 -induced NK-cell activation . Regulation by P08908 -type serotonin receptors . In the present study , the effects of serotonin on human natural killer ( NK ) -cell responsiveness to interleukin 2 ( P60568 ) was investigated . Concomitant treatment of human lymphocytes , enriched for NK effector cells by Percoll density-gradient centrifugation , with serotonin and P60568 yielded a synergistic activation of NK-cell cytotoxicity ( NKCC ) in the presence but not in the absence of monocytes . The monocyte-dependent , regulatory effects of serotonin and/or P60568 were prostaglandin-independent and could be reconstituted when monocytes , recovered by countercurrent centrifugal elutriation ( CCE ) , were added to purified NK effector cells . The effects of serotonin on baseline and P60568 -activated NK cells were mimicked by the P08908 receptor-specific agonists 8-OH-DPAT and (+)- Q9UM73 . Our data suggest that serotonin regulates NK-cell responsiveness to P60568 via P08908 receptors . cDNA microarray reveals signaling pathways involved in hormones expression of human pituitary . Pituitary , a master gland of neuroendocrine system , secretes hormones that orchestrate many physiological processes , under the regulation of multiple signaling pathways . To investigate the genes involved in hormones expression of human pituitary , homemade cDNA microarray containing 14,800 human genes/ESTs were used to profile the gene expression in both fetal and adult pituitaries . Seven hundred and twelve known genes changed over 2-fold between the both tissues . Of which , 23 genes were changed with hormones expression in aging were confirmed by RT-PCR , not only the known regulators such as Pit1 , P43694 , P11474 , GABA-A , and EMK , but also LOC55884 , P51452 , Q9H307 , and O43598 , which had not been reported to be involved in the hormones expression . Correspondingly , the mRNAs of GH , PRL , P01189 , P01222 , DB00094 -beta , and LH-beta , was increased as much as 6- to 20-fold in adult pituitary than those in fetal pituitary , by real-time quantitative RT-PCR assay . In addition , the mRNAs of signaling pathways , such as DB02527 -PKA-CREB , PI3K-Akt , and PKA- P29323 were further investigated . Of them , it was only DB02527 -PKA-CREB pathway , but not PI3K-Akt and PKA- P29323 have the same expressing pattern as hormones . It suggested that cDNA microarray is highly advantages to profile the differential expressed genes that were involved in hormones expression of human pituitary , but it might ignore some responding proteins regulated posttranscriptionally . Antidepressants increase glial cell line-derived neurotrophic factor production through monoamine-independent activation of protein tyrosine kinase and extracellular signal-regulated kinase in glial cells . Recent studies show that neuronal and glial plasticity are important for therapeutic action of antidepressants . We previously reported that antidepressants increase glial cell line-derived neurotrophic factor ( P39905 ) production in rat P13671 glioma cells ( P13671 cells ) . Here , we found that amitriptyline , a tricyclic antidepressant , increased both P39905 mRNA expression and release , which were selectively and completely inhibited by mitogen-activated protein kinase kinase inhibitors . Indeed , treatment of amitriptyline rapidly increased extracellular signal-regulated kinase ( P29323 ) activity , as well as p38 mitogen-activated protein kinase and c-Jun NH2-terminal kinase activities . Furthermore , different classes of antidepressants also rapidly increased P29323 activity . The extent of acute P29323 activation and P39905 release were significantly correlated to each other in individual antidepressants , suggesting an important role of acute P29323 activation in P39905 production . Furthermore , antidepressants increased the acute P29323 activation and P39905 mRNA expression in normal human astrocytes as well as P13671 cells . Although 5-hydroxytryptamine ( serotonin ) ( 5-HT ) , but not noradrenaline or dopamine , increased P29323 activation and P39905 release via 5- Q13049 receptors , ketanserin , a 5- Q13049 receptor antagonist , did not have any effect on the amitriptyline-induced P29323 activation . Thus , P39905 production by amitriptyline was independent of monoamine . Both of the amitriptyline-induced P29323 activation and P39905 mRNA expression were blocked by genistein , a general protein tyrosine kinase ( PTK ) inhibitor . Actually , we found that amitriptyline acutely increased phosphorylation levels of several phosphotyrosine-containing proteins . Taken together , these findings indicate that P29323 activation through PTK regulates antidepressant-induced P39905 production and that the P39905 production in glial cells may be a novel action of the antidepressant , which is independent of monoamine . Overexpression of follicle-stimulating hormone receptor activates oncogenic pathways in preneoplastic ovarian surface epithelial cells . It has been previously demonstrated that human ovarian cancer cells express DB00094 receptor ( P23945 ) . However , whether P23945 plays a role in ovarian cancer development is still ambiguous . To investigate the role of P23945 in tumor progression , we overexpressed the receptor in SV40 Tag immortalized ovarian surface epithelium ( OSE ) cell lines ( IOSE-80PC , a postcrisis line , and IOSE-398 ) , which are preneoplastic and nontumorigenic . We compared the expression levels of several selected oncogenes in nontransfected ( 80PC ) , vector-transfected ( 80PCV ) , P23945 -transfected IOSE ( 80PCF ) cells , and established ovarian cancer cell lines ( OVCAR-3 and SKOV-3 ) . Significantly increased protein levels of epithelial growth factor receptor , HER-2/neu , and c-Myc , but not K-Ras , were observed in P23945 -overexpressing 80PCF cells when compared with 80PCV cells . Constitutive phosphorylation of P27361 /2 was augmented in 80PCF cells , whereas phosphorylation of the other MAPK including p38 and Jun N-terminal kinase was unchanged . Considerable constitutive phosphorylation of P27361 /2 was also observed in OVCAR-3 and SKOV-3 cell lines when compared with 80PCV . More importantly , 80PCF cells grew more rapidly than 80PC and 80PCV cells . In conclusion , we have demonstrated that P23945 was highly expressed in OVCAR-3 and 80PCF cells transfected with P23945 overexpression vector . The 80PCF cell line showed increased levels of epithelial growth factor receptor , HER-2/neu , and c-myc and constitutive activation of P27361 /2 . The rate of proliferation of the 80PCF cells was increased , compared with control cell lines . These results suggest that the overexpression of P23945 may be associated with enhanced levels of potential oncogenic pathways and increased proliferation in preneoplastic ovarian surface epithelial cells . Synthesis and biological evaluation of novel pyrrolidine-2,5-dione derivatives as potential antidepressant agents . Part 1 . A series of 3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives was synthesized and their biological activity was evaluated . The chemical structures of the newly prepared compounds were confirmed by (1)H NMR , (13)C NMR and P19957 -HRMS spectra data . All tested compounds proved to be potent P08908 receptor and serotonin transporter protein ( P31645 ) ligands . Among them , compounds 15 , 18 , 19 and 30 showed significant affinity for P08908 and P31645 . Computer docking simulations carried out for compounds 15 , 31 and 32 to models of P08908 receptor and P31645 confirm the results of biological tests . Due to high affinity for the P08908 receptor and moderate affinity for P31645 , compounds 31 , 32 , 35 , and 37 were evaluated for their affinity for D2L , P50406 , P34969 and 5- Q13049 receptors . In vivo tests , in turn , resulted in determining the functional activity of compounds 15 , 18 , 19 and 30 to the P08908 receptor . The results of these tests indicate that all of the ligands possess properties characteristic of P08908 receptor agonists . Activation of P34969 receptor stimulates neurite elongation through P42345 , Cdc42 and actin filaments dynamics . Recent studies have indicated that the serotonin receptor subtype 7 ( 5-HT7R ) plays a crucial role in shaping neuronal morphology during embryonic and early postnatal life . Here we show that pharmacological stimulation of 5-HT7R using a highly selective agonist , LP-211 , enhances neurite outgrowth in neuronal primary cultures from the cortex , hippocampus and striatal complex of embryonic mouse brain , through multiple signal transduction pathways . All these signaling systems , involving P42345 , the Rho GTPase Cdc42 , Cdk5 , and P29323 , are known to converge on the reorganization of cytoskeletal proteins that subserve neurite outgrowth . Indeed , our data indicate that neurite elongation stimulated by 5-HT7R is modulated by drugs affecting actin polymerization . In addition , we show , by 2D Western blot analyses , that treatment of neuronal cultures with LP-211 alters the expression profile of cofilin , an actin binding protein involved in microfilaments dynamics . Furthermore , by using microfluidic chambers that physically separate axons from the soma and dendrites , we demonstrate that agonist-dependent activation of 5-HT7R stimulates axonal elongation . Our results identify for the first time several signal transduction pathways , activated by stimulation of 5-HT7R , that converge to promote cytoskeleton reorganization and consequent modulation of axonal elongation . Therefore , the activation of 5-HT7R might represent one of the key elements regulating CNS connectivity and plasticity during development . Acute effects of sarpogrelate , a 5- Q13049 receptor antagonist on cytokine production in endotoxin shock model of rats . Serotonin ( 5-HT ) (2A) receptors are involved in cytokine production in infection or sepsis . Therefore , 5-HT(2A) receptor antagonist might be useful to treat sepsis . The present study investigates the effects of a 5-HT(2A) receptor antagonist , sarpogrelate on endotoxin shock . Catheters were inserted into the femoral artery and vein of Sprague-Dawley rats . First , sarpogrelate 0 ( control ) , 3 , or 10 mg/kg dissolved in 0.5 ml of distilled water has been given , followed by endotoxin 10 mg/kg in saline 0.5 ml 5 min later . Blood pressure , pulse rate and survival rate were monitored in 20 rats per dose . Blood gas and plasma cytokine concentrations were measured in 8 rats per dose . In four rats each of sarpogrelate 0 , 3 , or 10 mg/kg , and sham operation , the lung histology was examined . Zero , 15 , and 12 rats survived for 8 h in the control , 3 mg/kg , and 10 mg/kg groups , respectively . The control group had the lowest blood pressure , pulse rate , pH and arterial oxygen tension , and the highest arterial carbon dioxide tension and plasma IL-1beta concentration . The increase of P01375 was significantly lower in 3 mg/kg group than in the control group . Pathological changes of the lung were inhibited in 3 and 10 mg/kg groups . In conclusion , sarpogrelate might be effective to decrease production of pro-inflammatory cytokines , to keep hemodynamics , to inhibit lung damage , and to decrease mortality in endotoxin shock . Overexpression of the P32302 chemokine receptor , and its ligand , O43927 in B-cell chronic lymphocytic leukemia . O43927 is a homeostatic chemokine for lymphocyte homing and positioning within follicles of secondary lymphoid tissues , acting through its cognate receptor , P32302 . Moreover , the P32302 - O43927 axis plays a unique role in trafficking and homing of B1 cells . Here , we report that chronic lymphocytic leukemia ( CLL ) B cells express high levels of functional P32302 . P32302 expression levels were similar on CLL B cells and normal P06127 + B cells , and higher compared with normal P06127 - B cells , follicular B-helper T cells ( T(FH) cells ) , or neoplastic B cells from other B-cell neoplasias . Stimulation of CLL cells with O43927 induces actin polymerization , P32302 endocytosis , chemotaxis , and prolonged activation of Q8TCB0 /42 mitogen-activated protein kinases . Anti- P32302 antibodies , pertussis toxin , and wortmannin inhibited chemotaxis to O43927 , demonstrating the importance of Gi proteins and P19957 kinases for P32302 signaling . Moreover , CLL patients had significantly higher O43927 serum levels than volunteers , and O43927 levels correlated with beta2 microglobulin . We detected O43927 mRNA expression by nurselike cells , and high levels of O43927 protein in supernatants of CLL nurselike cell cultures . By immunohistochemistry , we detected O43927 + expression by P34810 + macrophages in situ within CLL lymph nodes . These data suggest that P32302 plays a role in CLL cell positioning and cognate interactions between CLL and O43927 -secreting P34810 + accessory cells in lymphoid tissues . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Identification of a high-affinity binding site involved in the transport of endocannabinoids . Phytocannabinoids , such as the principal bioactive component of marijuana , delta9-tetrahydrocannabinol , have been used for thousands of years for medical and recreational purposes . DB00470 and endogenous cannabinoids ( e.g. , anandamide ) initiate their agonist properties by stimulating the cannabinoid family of G protein-coupled receptors ( P21554 and CB2 ) . The biosynthesis and physiology of anandamide is well understood , but its mechanism of uptake ( resulting in signal termination by fatty acid amide hydrolase ) has been elusive . Mounting evidence points to the existence of a specific anandamide transport protein ; however , no direct evidence for this protein has been provided . Here , we use a potent , competitive small molecule inhibitor of anandamide uptake ( LY2318912 , IC50 7.27 +/- 0.510 nM ) to identify a high-affinity , saturable anandamide transporter binding site ( LY2318912 ; K(d) = 7.62 +/- 1.18 nM , B(max) = 31.6 +/- 1.80 fmol/mg protein ) that is distinct from fatty acid amide hydrolase . Systemic administration of the inhibitor into rodents elevates anandamide levels 5-fold in the brain and demonstrates efficacy in the formalin paw-licking model of persistent pain with no obvious adverse effects on motor function . Identification of the anandamide transporter binding site resolves a missing mechanistic link in endocannabinoid signaling , and in vivo results suggest that endocannabinoid transporter antagonists may provide a strategy for positive modulation of cannabinoid receptors . DB00502 , but not clozapine , produces dramatic catalepsy in delta9-THC-treated rats : possible clinical implications . The effect on rat catalepsy induced by Delta9-tetrahydrocannabinol ( Delta9-THC ) in association with haloperidol ( HP ) or clozapine ( CLOZ ) administration was investigated . Delta9-THC dose-dependently increased HP ( 0.05-1 mg kg-1 , s.c. ) -induced rat catalepsy , while no catalepsy was observed after CLOZ ( 1-20 mg kg-1 , s.c. ) or Delta9-THC+CLOZ administration . The P21554 antagonist SR141716A ( 0.5-5 mg kg-1 , i.p. ) reversed the increase mediated by Delta9-THC on HP-induced catalepsy . The D2 agonist quinpirole completely reversed the catalepsy induced by both HP and HP+Delta9-THC ; however , higher doses of quinpirole were needed in the presence of Delta9-THC . The M1 antagonist scopolamine and alpha2 antagonist yohimbine were able to reduce the catalepsy induced by HP and HP+Delta9-THC in a similar manner . CLOZ and the 5- Q13049 /2C antagonists ritanserin , RS102221 and SB242084 were more effective in antagonizing HP than HP+Delta9-THC-induced catalepsy.7 HP and CLOZ failed to inhibit in vitro [3H]CP-55,940 binding , while Delta9-THC and SR141716A did not show an appreciable affinity for the D2 receptor . It was suggested that the different effects on rat catalepsy induced by Delta9-THC following HP or CLOZ administration may depend on the receptor-binding profiles of the two antipsychotics . The preferential use of CLOZ rather than HP in the treatment of psychotic symptoms in cannabis abusers was discussed . Inhibition of PI3K/ P42345 pathways in glioblastoma and implications for combination therapy with temozolomide . Due to its molecular heterogeneity and infiltrative nature , glioblastoma multiforme ( GBM ) is notoriously resistant to traditional and experimental therapeutics . To overcome these hurdles , targeted agents have been combined with conventional therapy . We evaluated the preclinical potential of a novel , orally bioavailable PI3K/ P42345 dual inhibitor ( DB05241 ) in in vitro and in vivo studies . In vivo serially passaged human GBM xenografts that are more genetically stable than GBM cell lines in culture were used for all experiments . Biochemical downstream changes were evaluated by immunoblot and cytotoxicity by colorimetric DB00171 -based assay . For in vivo experiments , human xenograft GBM 39 grown intracranially in nude mice was altered to express luciferase to monitor tumor burden by optical imaging . DB05241 resulted in concentration-dependent decreases in cell viability in vitro . Cytotoxic doses resulted in specific inhibition of PI3K signaling . Combining DB05241 with temozolomide ( DB00853 ) resulted in additive toxicity in 4 of 5 xenografts . In vivo , DB05241 administered by oral gavage resulted in greater than 12-fold reduction in median tumor bioluminescence compared with control ( Mann-Whitney test p = 0.001 ) and improvement in median survival ( logrank p = 0.05 ) . DB00853 alone showed a 30-fold decrease in median bioluminescence , but the combination DB05241 + DB00853 yielded a 140-fold reduction in median bioluminescence ( Mann-Whitney test p = 0.05 ) with a trend toward improvement in median survival ( logrank p = 0.09 ) compared with DB00853 alone . DB05241 shows activity as monotherapy and in combination with conventional therapeutics in a range of genetically diverse GBM xenografts . Thrombospondin 1 and its mimetic peptide DB05434 decrease angiogenesis and inflammation in a murine model of inflammatory bowel disease . OBJECTIVE : Vascular abnormalities and expression of proangiogenic factors have been repeatedly reported in inflammatory bowel disease ( Q9UKU7 ) . Thrombospondin 1 ( P07996 -1 ) is a protein well known for its antiangiogenic and anti-inflammatory properties . Using the dextran sulfate sodium ( DSS ) model , the role of P07996 -1 in Q9UKU7 has been investigated in vivo . METHODS : P07996 -1-deficient mice ( P07996 -1-/- ) and WT mice were treated with DSS for 7 days . Disease activity indices , myeloperoxidase activity ( P05164 ) and histology were analyzed . Microvascular density ( P53602 ) was quantified using immunohistochemistry ( IMH ) with CD31 antibody . TGF-beta(1) , basic FGF , P15692 , P01375 and MMPs protein levels were evaluated by IMH and enzyme-linked immunoabsorbent assay ( ELISA ) . Mice were treated with DB05434 ( Abbott Laboratories ) , an antiangiogenic P07996 peptide , using miniosmotic pumps for 7 days . RESULTS : P07996 -1(-/-) mice had a worse clinical outcome and exhibited severe signs of rectal bleeding compared to the WT controls . The P07996 -1-/- mice showed a higher level of crypt damage and deeper lesions . The grade of inflammation and the levels of P05164 activity were also significantly higher in colons of P07996 -1-/- mice . P07996 -1-/- mice displayed higher P53602 in focal areas of the colon after only 3 days of DSS treatment . Furthermore , clinical severity of the colitis and angiogenesis was significantly diminished when mice was treated with DB05434 . CONCLUSIONS : These findings directly link P07996 -1 as a protective factor in Q9UKU7 and suggest antiangiogenesis treatment , including compounds such as DB05434 as an adjuvant therapy for Q9UKU7 . Whole blood lead concentration and erythrocyte delta-aminolevulinic acid dehydratase ( P13716 ) activity in selected canine populations in Greece . In a total number of 275 dogs of various ages , sex and breed , blood lead concentrations ( O43927 ) and erythrocyte P13716 activity were measured . Sixty-six of the dogs were living in lead mining areas ( Group A ) , 157 in urban areas ( Group B ) and 52 in rural areas ( Group C ) of Greece . Mean O43927 differed significantly ( P < 0.05 ) between locations and were 326,97 and 68 micrograms/L , respectively . Mean P13716 activity was significantly different ( P < 0.05 ) only between Groups A and B as between groups A and C . A significant ( P < 0.05 ) negative correlation existed between O43927 and P13716 activity . A normal range of erythrocyte P13716 activity of 807-992 mumol/ DB02272 /LRBC/h was established for dogs . None of the 33 Group A dogs and 2 of the Group B dogs that had a O43927 of 350 micrograms/L presented clinical signs indicating acute or chronic lead intoxication . No erythrocyte basophilic stippling or large number of nucleated red blood cells were seen in the 30 dogs of Group A with O43927 > 350 micrograms/L . P03372 α ( ERα ) mediates 17β-estradiol ( E2 ) -activated expression of O95251 . BACKGROUND : O95251 ( histone acetyltransferase binding to Q13415 ) is a histone acetyltransferase ( O60235 ) which could exert oncogenic function in breast cancer . However , the biological role and underlying mechanism of O95251 in breast cancer remains largely unknown . In the current study , we aimed to investigate the role of O95251 in breast cancer and uncover the underlying molecular mechanism . METHODS : Immunohistochemistry was applied to detect O95251 protein expression in breast cancer specimens ( n=112 ) . The expression of protein level was scored by integral optical density ( IOD ) for further statistical analyses using SPSS . Real-time PCR was used to simultaneously measure mRNA levels of O95251 . The O95251 protein expression in breast cancer cells was confirmed by western blot . RESULTS : O95251 was highly expressed in breast cancer tissues and significantly correlated with estrogen receptor α ( ERα ) ( p < 0.001 ) and progestational hormone ( PR ) ( p=0.002 ) . O95251 protein level also correlated positively with histology grade in ERα positive tumors ( p=0.016 ) rather than ERα negative tumors . 17β-estradiol ( E2 ) could upregulate O95251 gene expression which was significantly inhibited by DB00947 or ERα RNAi . E2-increased O95251 protein expression was significantly suppressed by treatment with inhibitor of Q02750 /2 ( U0126 ) in T47 D and MCF-7 cells . CONCLUSIONS : O95251 was an important downstream molecule of ERα , and P27361 /2 signaling pathway may involved in the expression of O95251 increased by E2 . Binding of antipsychotic drugs to human brain receptors focus on newer generation compounds . Using radioligand binding assays and post-mortem normal human brain tissue , we obtained equilibrium dissociation constants ( K(d)s ) for nine new antipsychotic drugs ( iloperidone , melperone , olanzapine , ORG 5222 , quetiapine , risperidone , sertindole , ziprasidone , and zotepine ) , one metabolite of a new drug ( 9-OH-risperidone ) , and three older antipsychotics ( clozapine , haloperidol , and pimozide ) at nine different receptors ( alpha1-adrenergic , alpha2-adrenergic , dopamine D2 , histamine H1 , muscarinic , and serotonin P08908 , P28221 , 5- Q13049 , and P28335 receptors ) . DB04946 was the most potent drug at the two adrenergic receptors . ORG 5222 was the most potent drug at dopamine D2 and 5-HT2c receptors , while ziprasidone was the most potent compound at three serotonergic receptors ( P08908 , P28221 , and 5- Q13049 ) . At the remaining two receptors , olanzapine was the most potent drug at the histamine H1 receptor ( Kd=0.087 nM ) ; clozapine at the muscarinic receptor ( Kd=9 nM ) . Certain therapeutic and adverse effects , as well as certain drug interactions can be predicted from a drug 's potency for blocking a specific receptor . These data can provide guidelines for the clinician in the choice of antipsychotic drug . Contribution of the 37-kDa laminin receptor precursor in the anti-metastatic P08118 -derived peptide DB04985 cell surface binding . PURPOSE : DB04985 is an anti-metastatic synthetic peptide with promising therapeutic efficacy against hormone-refractory prostate cancer . The characterization of the DB04985 peptide cell surface binding/internalization mechanisms and of the receptors involved remained to be explored . RESULTS : [(14)C] DB04985 cell surface binding assays showed rapid and transient kinetic profile , that was inhibited by RGD peptides , laminin , hyaluronan , and type-I collagen . RGD peptides were however unable to inhibit DB04985 intracellular uptake . Far-Western ligand binding studies enabled the identification of the 37-kDa laminin receptor precursor ( P08865 ) as a potential ligand for DB04985 . Overexpression of the recombinant P08865 indeed led to an increase in DB04985 binding but unexpectedly not to its uptake . CONCLUSIONS : Our data support the implication of laminin receptors in cell surface binding and in transducing DB04985 anti-metastatic effects , and provide a rational for targeting cancers that express high levels of such laminin receptors .

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mu-Opioid receptor agonists differentially regulate the expression of miR-190 and Q13562 . The agonists of mu-opioid receptor ( P35372 ) induce extracellular signal-regulated kinase ( P29323 ) phosphorylation through different pathways : morphine uses the protein kinase C ( PKC ) -pathway , whereas fentanyl functions in a beta-arrestin2-dependent manner . In addition , the two pathways result in the different cellular location of phosphorylated P29323 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 . When fentanyl-induced but not morphine-induced P29323 phosphorylation was blocked in the primary cultures from beta-arrestin2(-/-) mouse , fentanyl did not decrease the expression of miR-190 . However , a PKC inhibitor that blocked morphine-induced P29323 phosphorylation specifically had no effect on the miR-190 down-regulation . Therefore the decrease in miR-190 expression resulted from the agonist-selective P29323 phosphorylation . In addition , the expressional changes in one of the miR-190 targets , neurogenic differentiation 1 ( Q13562 ) , correlated with those in miR-190 expression , suggesting the P35372 could regulate the Q13562 pathways via the control of miR-190 expression . DB04864 , but not tacrine , stimulates P04271 secretion in astrocyte cultures . AIMS : The loss of cholinergic function in the central nervous system contributes significantly to the cognitive decline associated with advanced age and dementias . DB04864 ( HupA ) is a selective inhibitor of acetylcholinesterase ( P22303 ) and has been shown to significantly reduce cognitive impairment in animal models of dementia . Based on the importance of astrocytes in physiological and pathological brain activities , we investigated the effect of HupA and tacrine on P04271 secretion in primary astrocyte cultures . P04271 is an astrocyte-derived protein that has been proposed to be a marker of brain injury . MAIN METHODS : Primary astrocyte cultures were exposed to HupA , tacrine , cholinergic agonists , and P04271 secretion was measured by enzyme-linked immunosorbent assay ( ELISA ) at 1 and 24h . KEY FINDINGS : HupA , but not tacrine , at 100μM significantly increased P04271 secretion in astrocyte cultures . DB00184 ( at 100 and 1000μM ) was able to stimulate P04271 secretion in astrocyte cultures . SIGNIFICANCE : Our data reinforce the idea that P22303 inhibitors , particularly HupA , do not act exclusively on the acetylcholine balance . This effect of HupA could contribute to improve the cognitive deficit observed in patients , which are attributed to cholinergic dysfunction . In addition , for the first time , to our knowledge , these data indicate that P04271 secretion can be modulated by nicotinic receptors , in addition to glutamate , dopamine and serotonin receptors . 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 ) on chromosome 3 , but also novel genes , including Q9HBW9 [ epidermal growth factor ( P01133 ) , 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 , CB2 , FAAH , Q99685 , Q8NER1 and Q9Y2T6 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 , P14416 and P35372 . 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 . Expression of two variants of the human mu opioid receptor mRNA in SK-N-SH cells and human brain . A partial mu opioid receptor gene was isolated from a human genomic library using a mouse delta opioid receptor cDNA as a probe . Using information from this genomic clone and the published human mu receptor , P35372 , a cDNA was isolated from SK-N-SH mRNA that codes for a variant of the P35372 mRNA , MOR1A . The presence of MOR1A is also shown in human brain using RT-PCR . MOR1A differs from P35372 in that the 3' terminal intron has not been removed . An in-frame termination codon is found four amino acids after the 5' consensus splice site , making MOR1A eight amino acids shorter than P35372 . Both receptors show similar ligand binding and coupling to DB02527 in CHO- P04264 cells . The C-terminal differences between P35372 and MOR1A could have effects on receptor coupling or receptor transport and localization . Passive smoke effects on cough and airways in young guinea pigs : role of brainstem DB05875 . Children raised with extended exposure to environmental tobacco smoke ( ETS ) experience increased cough and wheeze . This study was designed to determine whether extended ETS exposure enhances citric acid-induced cough and bronchoconstriction in young guinea pigs via a neurokinin-1 ( NK-1 ) receptor mechanism at the first central synapse of lung afferent neurons , the nucleus tractus solitarius . Guinea pigs were exposed to ETS from 1 to 6 weeks of age . At 5 weeks of age , guide cannulae were implanted bilaterally in the medial nucleus tractus solitarius at a site that produced apnea in response to the glutamate agonist D,L-homocysteic acid . At 6 weeks of age , either vehicle or a P25103 antagonist , DB05790 , was injected into the nucleus tractus solitarius of the conscious guinea pigs who were then exposed to citric acid aerosol . ETS exposure significantly enhanced citric acid-induced cough by 56 % and maximal Penh ( a measure of airway obstruction ) by 43 % , effects that were attenuated by the P25103 antagonist in the nucleus tractus solitarius . We conclude that in young guinea pigs extended exposure to ETS increases citric acid-induced cough and bronchoconstriction in part by an P25103 mechanism in the nucleus tractus solitarius . The anti-inflamm-aging and hepatoprotective effects of huperzine A in D-galactose-treated rats . Oxidative stress contributes to a chronic inflammatory process referred to as " inflamm-aging " . P22303 inhibitors ( AChEI ) can enhance cholinergic transmission and act as anti-inflammatory agents via immunocompetent cells expressing α-7 acetylcholine receptors ( AChR ) . The present study explores the possible role of huperzine A , a reversible and selective AChEI , against D-gal-induced oxidative damage , cell toxicity and inflamm-aging in rat livers . In two-month-old rats with normal liver function , an 8-week administration of D-gal ( 300 mg/kg subcutaneously ( s.c. ) injected ) , significantly increased hepatic impairment , ROS generation and oxidative damage , hepatic senescence , nuclear factor-kappa B ( NF-κB ) activation and inflammatory responses . An 8-week co-administration of both D-gal ( 300 mg/kg s.c. ) and huperzine A ( 0.1 mg/kg s.c. ) not only significantly decreased hepatic function impairment , ROS generation , oxidative damage , but also suppressed inflamm-aging by inhibiting hepatic replicative senescence , P22303 activity , IκBα degradation , NF-κB p65 nuclear translocation and inflammatory responses . The expression levels of pro-inflammatory cytokine mRNA and proteins , such as TNFα , IL-1β and P05231 decrease significantly , and the protein levels of the anti-inflammatory cytokine P22301 display an obvious increase . These findings indicated that D-gal-induced hepatic injury and inflamm-aging in the rat liver was associated with the development of a pro-inflammatory phenotype in this organ . D-gal induced damage-associated molecular patterns ( DAMPs ) because oxidative damages might play an important role in D-gal-induced hepatic sterile inflammation . DB04864 exhibited protective effects against D-gal-induced hepatotoxicity and inflamm-aging by inhibiting P22303 activity and via the activation of the cholinergic anti-inflammatory pathway . The huperzine A mechanism might be involved in the inhibition of DAMPs-mediated NF-κB nuclear localization and activation . Effects of inhaled prostacyclin analogue on chronic hypoxic pulmonary hypertension . Inhaled DB01240 has been reported to elicit pulmonary vasodilation , but whether it is also effective in treating chronic hypoxic pulmonary hypertension is still uncertain . We designed this study to address the in vivo effectiveness of inhaled DB05229 , a stable DB01240 analogue , on pulmonary vascular tone during hypoxic exposure in normoxic ( N ) and chronically hypoxic ( CH ) rats . Pulmonary vasodilation was observed by low-dose inhaled DB05229 in N rats , but not in CH rats . It was not until higher doses of DB05229 were given that pulmonary vasodilation was obtained in CH rats . When the agent was continuously administered by an intravascular route at the inhaled dose , it elicited no vasodilation in N rats . On the contrary , it elicited profound vasodilation in CH rats , although a concomitant systemic hypotension was observed . The P43119 mRNA expression was unchanged in the lungs of CH rats compared with that of N rats . We conclude that low doses of aerosolized DB05229 may reduce pulmonary vascular tone in rats without preexisting lung diseases . In contrast , when hypoxic pulmonary hypertension is present , the threshold of DB05229 inhalation was elevated to provoke pulmonary vasodilation . P14416 signaling modulates mutant ataxin-1 S776 phosphorylation and aggregation . Spinocerebellar ataxia 1 ( P54253 ) is a dominantly inherited neurodegenerative disease associated with progressive ataxia resulting from the loss of cerebellar Purkinje cells ( PCs ) and neurons in the brainstem . In PCs of P54253 transgenic mice , the disease causing ataxin-1 protein mediates the formation of P04271 containing cytoplasmic vacuoles and further self-aggregates to form intranuclear inclusions . The exact function of the ataxin-1 protein is not fully understood . However , the aggregation and neurotoxicity of the mutant ataxin-1 protein is dependent on the phosphorylation at serine 776 ( S776 ) . Although protein kinase A ( PKA ) has been implicated as the S776 kinase , the mechanism of PKA/ataxin-1 regulation in P54253 is still not clear . We propose that a dopamine D(2) receptor ( D2R ) / P04271 pathway may be involved in modulating PKA activity in PCs . Using a D2R/ P04271 P29320 stable cell line transiently transfected with GFP-ataxin-1[82Q] , we demonstrate that stimulation of the D2R/ P04271 pathway caused a reduction in mutant ataxin-1 S776 phosphorylation and ataxin-1 aggregation . Activation of PKA by forskolin resulted in an enhanced S776 phosphorylation and increased ataxin-1 nuclear aggregation , which was suppressed by treatment with D2R agonist bromocriptine and PKA inhibitor H89 . Furthermore , treating P54253 transgenic PC slice cultures with forskolin induced neurodegenerative morphological abnormalities in PC dendrites consistent with those observed in vivo . Taken together our data support a mechanism where PKA dependent mutant ataxin-1 phosphorylation and aggregation can be regulated by D2R/ P04271 signaling . Design and synthesis of an orally bioavailable and selective peptide epoxyketone proteasome inhibitor ( PR-047 ) . Proteasome inhibition has been validated as a therapeutic modality in the treatment of multiple myeloma and non-Hodgkin 's lymphoma . DB08889 , an epoxyketone currently undergoing clinical trials in malignant diseases , is a highly selective inhibitor of the chymotrypsin-like ( CT-L ) activity of the proteasome . A chemistry effort was initiated to discover orally bioavailable analogues of carfilzomib , which would have potential for improved dosing flexibility and patient convenience over intravenously administered agents . The lead compound , 2-Me-5-thiazole- DB00133 (OMe)- DB00133 (OMe)- DB00120 -ketoepoxide ( 58 ) ( PR-047 ) , selectively inhibited CT-L activity of both the constitutive proteasome ( beta5 ) and immunoproteasome ( P28062 ) and demonstrated an absolute bioavailability of up to 39 % in rodents and dogs . It was well tolerated with repeated oral administration at doses resulting in > 80 % proteasome inhibition in most tissues and elicited an antitumor response equivalent to intravenously administered carfilzomib in multiple human tumor xenograft and mouse syngeneic models . The favorable pharmacologic profile supports its further development for the treatment of malignant diseases . P08235 antagonists do not block rapid P29323 activation by aldosterone . DB04630 can elicit rapid nongenomic effects both in vivo and in vitro , often mediated by signal transduction cascades . However , it is not understood how these rapid effects are initiated . In this study we show that aldosterone leads to rapid activation of mitogen activated protein kinases P27361 /2 in the cortical collecting duct cell line M-1 . Inhibitors of transcription and translation could not block this activation , which suggests an extranuclear ( nongenomic ) mechanism . Although it is known that M-1 cells do not contain a transcriptionally functional MR , it is not known whether a closely related protein still could mediate the effects , or an unrelated nonclassic receptor . To test this hypothesis , the effects of four classical mineralocorticoid receptor antagonists were studied . None of the compounds could block the response to aldosterone . Altogether , the data suggest that rapid aldosterone effects in M-1 cells are initiated by a receptor different from the classical mineralocorticoid receptor . DB08889 can induce tumor cell death through selective inhibition of the chymotrypsin-like activity of the proteasome . DB08889 is a proteasome inhibitor in clinical development that primarily targets the chymotrypsin-like ( CT-L ) subunits in both the constitutive proteasome ( c20S ) and the immunoproteasome ( i20S ) . To investigate the impact of inhibiting the CT-L activity with carfilzomib , we set out to quantitate the levels of CT-L subunits beta5 from the c20S and P28062 from the i20S in normal and malignant hematopoietic cells . We found that the i20S is a major form of the proteasome expressed in cells of hematopoietic origin , including multiple myeloma ( MM ) CD138+ tumor cells . Although specific inhibition of either P28062 or beta5 alone was insufficient to produce an antitumor response , inhibition of all proteasome subunits was cytotoxic to both hematologic tumor cells and peripheral blood mononuclear cells . However , selective inhibition of both beta5 and P28062 was sufficient to induce an antitumor effect in MM , non-Hodgkin lymphoma , and leukemia cells while minimizing the toxicity toward nontransformed cells . In MM tumor cells , CT-L inhibition alone was sufficient to induce proapoptotic sequelae , including proteasome substrate accumulation , Noxa and caspase 3/7 induction , and phospho-eIF2alpha suppression . These data support a hypothesis that hematologic tumor cells are uniquely sensitive to CT-L inhibition and provide a mechanistic understanding of the clinical safety profile and antitumor activity of proteasome inhibitors . Pharmacological examination of the neurokinin-1 receptor mediating relaxation of human intralobar pulmonary artery . The effect of selective tachykinin receptor agonists and antagonists on human isolated intralobar pulmonary arterial rings was investigated . Neither Substance P ( SP ) nor neurokinin A ( P20366 ) contracted the arteries . Both of these agonists , however , were potent and efficacious at relaxing the arteries that were precontracted with phenylephrine . The negative log ( M ) EC(50) values for SP and P20366 were 9.0 and 8.3 , respectively . The neurokinin ( NK ) -3 selective agonist , senktide-analog , and the P21452 selective agonist , [beta-Ala(8)] P20366 (4-10) , caused neither contractions nor relaxations of the arteries , whereas the P25103 agonist Ac- [ Arg6 , Sar9 , DB00134 (O2)11 ] SP(6-11) ( P17405 -SP ) relaxed the tissue with a potency similar to SP . The relaxations to SP , P20366 , and P17405 -SP were competitively antagonized by the selective P25103 antagonist CP 99994 , with a pK(b) in the nanomolar range . Antagonism of the P17405 -SP-induced relaxations was also noted with FK 888 , RP 67580 , and L 732,138 , although these antagonists were much less potent than CP 99994 in this regard . Another P25103 selective antagonist , DB05790 , caused an insurmountable antagonism of the SP-induced relaxations . The P25103 -mediated relaxations could be blocked by removing the endothelium , or by a combination of N-nitro-L-arginine and indomethacin . Measurement of prostanoid generation revealed that in endothelial-intact but not endothelial-denuded tissue , P17405 -SP caused a selective increase in the production of 6-keto-PGF1alpha , the stable metabolite of prostacyclin . The results indicate that stimulation of NK-1 receptors leads to relaxation of human intralobar pulmonary arteries , which is mediated largely by nitric oxide and prostacyclin released from the endothelium of these vessels . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Niflumic acid and MSI-2216 reduce P01375 -induced mucin expression in human airway mucosa . BACKGROUND : Human chloride channel , calcium-activated 1 ( A8K7I4 ) has been shown to induce mucin ( MUC ) gene expression and mucus production in bronchial epithelial cells . Objective To investigate whether blocking A8K7I4 decreases mucus production . METHODS : Expression of A8K7I4 and mucus was stimulated with P01375 in human upper airway mucosal explant tissue . P98088 mRNA and mucus protein expression was blocked by inhibiting A8K7I4 by using channel blockers ( niflumic acid [ DB04552 ] and MSI-2216 ) without and with P01375 stimulation . Expression of P98088 , A8K7I4 , and P35354 mRNA was quantified by using real-time PCR . Mucus protein was assessed by periodic acid Schiff staining . Laser capture microdissection was performed to quantify A8K7I4 and P98088 mRNA expression in epithelial cells derived from mucosal explant tissue . RESULTS : P01375 significantly increased P98088 and A8K7I4 mRNA as well as mucus and A8K7I4 protein expression in the mucosal explant tissue ( P < .05 ) . Inhibition of A8K7I4 with DB04552 or MSI-2216 showed a significant dose-dependent reduction of mucus production for both blockers in the mucosal explant tissue ( P < .05 ) . P98088 mRNA was also decreased by both blockers in the whole mucosal tissue and in laser-captured mucosa epithelial cells . CONCLUSIONS : Unstimulated and P01375 -induced mucin expression could be decreased by DB04552 and MSI-2216 . Inhibiting A8K7I4 may be a potential new approach to reduce mucus overproduction . Evidence for epidermal growth factor receptor as negative-feedback control in aldosterone-induced Na+ reabsorption . DB04630 enhances Na(+) reabsorption via epithelial Na(+) channels ( ENaC ) . DB04630 also stimulates the protein kinase P27361 /2- and the epidermal growth factor ( P01133 ) receptor ( P00533 ) -signaling pathway . Yet P01133 and P27361 /2 are known inhibitors of ENaC-mediated Na(+) reabsorption . In the present study , using the well-established Madin-Darby canine kidney P10643 cell line , we tested the hypothesis that P00533 represents a negative-feedback control for chronic aldosterone-induced Na(+) reabsorption [ amiloride-inhibitable short-circuit current ( I(sc) ) ] . P08235 expression was confirmed by RT-PCR and Western blot analysis . DB04630 enhanced P27361 /2 phosphorylation in an P00533 -dependent way . Furthermore , aldosterone stimulated P00533 expression . DB04630 ( 10 nmol/l ) induced a small transient increase in I(sc) under control conditions . Inhibition of P27361 /2 phosphorylation with U-0126 ( 10 micromol/l ) stimulated I(sc) , indicating constitutive ENaC inhibition . DB04630 exerted a significantly larger effect in the presence of U-0126 than without U-0126 . P01133 ( 10 microg/l ) inhibited I(sc) , whereas inhibition of P00533 kinase by tyrphostin AG-1478 ( 100 nmol/l ) enhanced I(sc) . DB04630 was more effective in the presence of AG-1478 than without AG-1478 . In summary , we propose that the P00533 -signaling cascade can serve as a negative-feedback control to limit the effect of aldosterone-induced Na(+) reabsorption . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Thrombin receptors and their antagonists : an update on the patent literature . IMPORTANCE OF THE FIELD : Thrombin plays a central role in cardiovascular inflammation . Most of the cellular responses to thrombin are mediated by cell surface protease-activated receptors ( PARs ) . Several preclinical studies indicate that PARs are potential targets for treating cardiovascular diseases such as thrombosis , atherosclerosis and restenosis . Among PARs , P25116 has emerged as an important therapeutic target . AREAS COVERED IN THIS REVIEW : This review covers recent advances in the development of thrombin receptors antagonists . It is focused on the search for P25116 antagonists as this is at the moment the most promising and attractive target . However , some early promising studies on PAR-3 and -4 antagonists are also reported . WHAT THE READER WILL GAIN : The review has been written in order to give to the reader hints and references that cover , in our opinion , the most interesting and/or promising approaches in this research field . TAKE HOME MESSAGE : Research on P25116 antagonists has finally led to good clinical candidates such as DB05692 ( Schering-Plough ) and E-5555 ( Eisai Co. ) . Clinical trials clearly demonstrate that development of PAR1 antagonists is not only possible but most likely will lead to development of antiplatelet drugs as well as of drugs useful for the treatment of inflammatory , proliferative and neurodegenerative diseases . Inhibition of cervical lymph node metastasis by marimastat ( DB00786 ) in an orthotopic oral squamous cell carcinoma implantation model . Activation of matrix metalloproteinase-2 ( P08253 ) is a common event in head and neck squamous cell carcinoma . An P48449 -19 cell line , derived from human oral squamous cell carcinoma and known to metastasize to cervical lymph nodes , was implanted into the lingual margin of mice . The effect of marimastat ( DB00786 ) , a broad MMP inhibitor , on the suppression of regional cervical lymph node metastasis was evaluated with an orthotopic implantation nude mice model . Marimastat was given immediately after P48449 -19 implantation and continuously administered by an osmotic pump . The mice were divided into three groups by marimastat dose ; Group A ; 0 mg/kg/day , Group B ; 30 mg/kg/day , and Group C ; 150 mg/kg/day . Twenty-one days after implantation , primary oral tumors and cervical lymph nodes were resected . Cervical lymph node status was microscopically examined . Activation of P08253 in primary oral tumor was examined by gelatin zymography . Both cervical lymph node metastasis and activation of P08253 were significantly suppressed in Group C ( P < 0.05 ) . Moreover , the Group C mice had a significantly better survival than group A ( P = 0.0026 ) . There was a significant difference between Group A and Group C in terms of proliferation of tumor cells by proliferating cell nuclear antigen immunostaining ( P = 0.0120 ) . These results suggest a positive role for marimastat in the inhibition of P08253 activation and prevention of cervical lymph node metastasis in oral squamous cell carcinoma ( OSCC ) . Improvement of survival in patients with OSCC could be expected using adjuvant therapy with marimastat . 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 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . 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 and P01375 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 . Activation of the P01133 receptor signaling pathway in airway epithelial cells exposed to Utah Valley PM . Exposure to ambient particulate matter ( PM ) in the Utah Valley has previously been associated with a variety of adverse health effects . To investigate intracellular signaling mechanisms for pulmonary responses to Utah Valley PM inhalation , human primary airway epithelial cells were exposed to aqueous extracts of PM collected from the year before ( Q03519 ) , during ( P28062 ) , and after ( Q03518 ) the closure of a local steel mill located in the Utah Valley in this study . Transfection with kinase-deficient extracellular signal-regulated kinase ( P29323 ) 1 constructs partially blocked Utah Valley PM-induced interleukin ( IL ) -8 promoter reporter activity . The mitogen-activated protein kinase/ P29323 kinase ( MEK ) activity inhibitor PD-98059 significantly abolished P10145 released in response to Utah Valley PM , as did the epidermal growth factor ( P01133 ) receptor kinase inhibitor AG-1478 . Western blotting showed that Utah Valley PM induced phosphorylation of P01133 receptor tyrosine , Q02750 /2 , and P27361 /2 , which could be ablated with AG-1478 or PD-98059 . For all findings , the potency of Utah Valley PM collected during P28062 was found to be lower relative to that of Q03519 and Q03518 . These data demonstrate that Utah Valley PM can induce P10145 expression partially through the activation of the P01133 receptor signaling . DB05229 sodium , prostacyclin analogue , attenuates glomerular hyperfiltration and glomerular macrophage infiltration by modulating ecNOS expression in diabetic rats . Stable prostacyclin analogue , beraprost sodium ( BPS ) has recently been reported to attenuate glomerular hyperfiltration in diabetic rats , however , the mechanism has been still unknown . We previously reported that overexpression of endothelial cell nitric oxide synthase ( ecNOS ) in afferent arterioles and glomeruli induce inappropriate dilatation of afferent arterioles and glomerular hyperfiltration through overproduction of nitric oxide in early stage of diabetic nephropathy . In this study , we tested the hypothesis that BPS ameliorates glomerular hyperfiltration through modulating ecNOS expression in diabetic nephropathy . Furthermore , we examined the effects of BPS on the expression of intercellular adhesion molecule-1 ( P05362 ) and macrophage infiltration in diabetic glomeruli , because glomerular hyperfiltration induces the expression of P05362 resulting in macrophage infiltration . Male Sprague-Dawley ( SD ) rats were administered continuously with BPS for 4 weeks after induction of diabetes by streptozotocin . In diabetic rats , the diameters of afferent arterioles , glomerular volume , creatinine clearance and urinary excretion of albumin and NO2/NO3 were increased as compared with non-diabetic control rats . Treatment with BPS improved these changes . The expression of ecNOS was increased in afferent arterioles and glomeruli in diabetic rats and suppressed by BPS . P43119 was expressed along afferent arterioles . Our results suggest that BPS attenuates glomerular hyperfiltration by modulating ecNOS expression in early stage of diabetic nephropathy . Moreover , BPS may inhibit P05362 -dependent infiltration of macrophages in diabetic glomeruli . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . P43119 induces P42224 and P40763 phosphorylations in human erythroleukemia cells : a mechanism requiring PTX-insensitive G proteins , P29323 and JNK . The ability of the human prostacyclin receptor ( hIP ) to regulate the activities of signal transducers and activators of transcription ( STATs ) has not yet been documented . In the present study , we have delineated the mechanism by which hIP induces P40763 phosphorylations in human erythroleukemia ( HEL ) cells . Stimulation of endogenous hIP by its specific agonist , cicaprost , resulted in P40763 Tyr705 and Ser727 phosphorylations in a time- and concentration-dependent manner . Cicaprost-induced P40763 Tyr705 and Ser727 phosphorylations were resistant to pertussis toxin ( PTX ) treatment , suggesting that these responses were mediated through PTX-insensitive G proteins . In addition , extracellular signal-regulated kinase ( P29323 ) and c-Jun N-terminal kinase ( JNK ) , but not p38 MAPK , were shown to be phosphorylated by cicaprost in a time- and concentration-dependent manner via PTX-insensitive G proteins . The levels of the interaction between P40763 , P29323 and JNK were enhanced by cicaprost treatment . The involvement of P04049 , Q02750 /2 and JNK in cicaprost-induced phosphorylations of P40763 was illustrated by the use of their selective inhibitors . In contrast , p38 MAPK did not appear to be required . Similar observations were obtained with P42224 upon stimulation by cicaprost . Taken together , these results demonstrate for the first time that hIP activation by cicaprost can lead to P42224 and P40763 phosphorylations via signaling pathways involving PTX-insensitive G proteins , P29323 and JNK . Mechanism of interaction of niflumic acid with heterologously expressed kidney CLC-K chloride channels . CLC-K Cl(-) channels belong to the CLC protein family . In kidney and inner ear , they are involved in transepithelial salt transport . Mutations in ClC-Kb lead to Bartter 's syndrome , and mutations in the associated subunit barttin produce Bartter 's syndrome and deafness . We have previously found that 3-phenyl-CPP blocks hClC-Ka and rClC- P04264 from the extracellular side in the pore entrance . Recently , we have shown that niflumic acid ( DB04552 ) , a nonsteroidal anti-inflammatory fenamate , produces biphasic behavior on human CLC-K channels that suggests the presence of two functionally different binding sites : an activating site and a blocking site . Here , we investigate in more detail the interaction of DB04552 on CLC-K channels . Mutants that altered block by 3-phenyl-2-(p-chlorophenoxy)propionic acid ( CPP ) had no effect on DB04552 block , indicating that the inhibition binding site of DB04552 is different from that of 3-phenyl-CPP and flufenamic acid . Moreover , DB04552 does not compete with extracellular Cl(-) ions , suggesting that the binding sites of DB04552 are not located deep in the pore . Differently from ClC-Ka , on the rat homologue P51800 , DB04552 has only an inhibitory effect . We developed a quantitative model to describe the complex action of DB04552 on ClC-Ka . The model predicts that ClC-Ka possesses two DB04552 binding sites : when only one site is occupied , DB04552 increases ClC-Ka currents , whereas the occupation of both binding sites leads to channel block . PD98059-inhibited invasion of Dunning rat prostate cancer cells involves suppression of motility but not P08253 or uPA secretion . Up-regulation of extracellular-regulated kinases 1/2 ( P27361 /2 ) has been implicated in tumor progression and metastasis in many types of cancer . We have previously shown that P27361 /2 is necessary for invasiveness of Dunning rat prostatic adenocarcinoma cell lines in which levels of activated P27361 /2 correlate with the metastatic potential . Here , we further examined the biological effects of elevated P27361 /2 in the highly metastatic Dunning cell line , Q03164 , in which the abilities to invade and metastasize are enhanced relative to its progenitor strain . Inhibition of P27361 /2 activation by the Q02750 inhibitor , PD98059 , dose-dependently reduced Q03164 cell invasiveness and motility with similar IC50 values . On the other hand , the abilities of Q03164 cells to adhere to the extracellular matrix , phosphorylate myosin regulatory light chain and secrete matrix-degrading enzymes , matrix metalloproteinase ( MMP ) -2 and urokinase plasminogen activator ( uPA ) were marginally , if at all , affected by PD98059 treatment . These data indicated that the inhibitory effect of PD98059 on the invasiveness of Q03164 cells was primarily due to the suppression of cell motility , and the up-regulation of P27361 /2 is , at least in part , responsible for the enhanced cellular motility and invasiveness of the Q03164 cells . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) 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 ) -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 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 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 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- 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 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . P35372 -dependent and independent components in effects of tramadol . DB00193 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 -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 alpha2 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 alpha2 receptor mediate most of the analgesic properties of tramadol . DB05692 , a thrombin receptor ( P25116 ) antagonist for the prevention and treatment of atherothrombosis . DB05692 is a novel antiplatelet agent undergoing development by Schering-Plough Corp for the treatment and prevention of atherothrombosis . The compound is an orally administered himbacine analog that potently antagonizes the platelet thrombin receptor protease-activated receptor 1 ( P25116 ) , which leaves the procoagulant function of thrombin intact . In preclinical studies , DB05692 demonstrated no effect on bleed time or coagulation parameters . In both cynomolgus monkeys and humans , the compound had high bioavailability and inhibited ex vivo TRAP ( thrombin receptor-activating peptide ) -stimulated platelet aggregation in a potent and long-lasting manner . In a phase II clinical trial of patients undergoing percutaneous coronary intervention , DB05692 added to standard therapy with aspirin and clopidogrel did not increase major or minor thrombolysis in myocardial infarction bleeding , and demonstrated a trend toward decreased major adverse cardiovascular events versus placebo . At the time of publication , three phase III trials were underway to assess the efficacy and safety of DB05692 for at least 1 year in up to 35,000 patients with acute coronary syndromes or atherosclerosis . The distinct mechanism of action of DB05692 allows for cardiovascular protection without the liability of increased bleeding associated with other antiplatelet therapies . Phase III trials in high-risk patients will determine the use of DB05692 in cardiological practice . Recent advances in the regulation of matrix metalloproteinase 2 activation : from basic research to clinical implication ( Review ) . Matrix metalloproteinases ( MMPs ) play an important role in degradation of extracellular matrix ( Q13201 ) , which is an essential step in the cascade of metastasis . Various types of MMPs are expressed and activated in head and neck squamous cell carcinoma ( HNSCC ) as well as other human cancers . P08253 is a prominent predictor of poor prognosis . Membrane type 1-MMP ( P50281 ) was originally identified as an activator of P08253 . In addition to the original role , recent studies show other important functions of P50281 such as degradation of type I collagen and cleavage of P16070 . Tissue inhibitor of P08253 ( P16035 ) was identified as an inhibitor of P08253 and P50281 . However , P16035 was reported to be essential for cell-mediated activation of P08253 , and thus the contribution of P16035 to tumor invasion has remained controversial . Some studies also suggested a role of P16035 as a predictor of poor prognosis . Thus , inhibition of MMP activation by TIMPs is not a suitable strategy for suppressing invasion and metastasis . Instead of P16035 , various MMP inhibitors ( MMPI ) such as DB00786 have been investigated with regard to suppression of tumor progression and improvement of prognosis in patients with advanced cancers , which resulted in no clinical efficacy . MMPs are especially important in the early stage of cancer progression , and thus strategies for future MMPI trials should be reconsidered . Comparison of gene expression profiles and related pathways in chronic thromboembolic pulmonary hypertension . Chronic thromboembolic pulmonary hypertension ( CTEPH ) is one of the main causes of severe pulmonary hypertension . However , despite treatment ( pulmonary endarterectomy ) , in approximately 15-20 % of patients , pulmonary vascular resistance and pulmonary arterial pressure continue to increase . To date , little is known about the changes that occur in gene expression in CTEPH . The identification of genes associated with CTEPH may provide insight into the pathogenesis of CTEPH and may aid in diagnosis and treatment . In this study , we analyzed the gene expresion profiles of pulmonary artery endothelial cells from 5 patients with CTEPH and 5 healthy controls using oligonucleotide microarrays . Bioinformatics analyses using the Gene Ontology ( GO ) and KEGG databases were carried out to identify the genes and pathways specifically associated with CTEPH . Signal transduction networks were established to identify the core genes regulating the progression of CTEPH . A number of genes were found to be differentially expressed in the pulmonary artery endothelial cells from patients with CTEPH . In total , 412 GO terms and 113 pathways were found to be associated with our list of genes . All differential gene interactions in the Signal-Net network were analyzed . P52333 , P30679 , O15264 , P32121 and P25116 were the most significantly altered . Bioinformatics analysis may help gather and analyze large amounts of data in microarrays by means of rigorous experimental planning , scientific statistical analysis and the collection of complete data . In this study , a novel differential gene expression pattern was constructed . However , further studies are required to identify novel targets for the diagnosis and treatment of CTEPH .

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interacts_with DB01151?

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Synthesis and biological evaluation of novel oxindole-based RTK inhibitors as anti-cancer agents . Given that receptor tyrosine kinases ( RTKs ) have emerged as key regulators of all aspects of cancer development , including proliferation , invasion , angiogenesis and metastasis , the RTK family represents an important therapeutic target for anti-cancer drug development . Oxindole structure has been used in RTK inhibitors such as DB02058 and intedanib . In this study , two series of new heterocyclic compounds containing oxindole scaffold have been designed and synthesized , and their inhibitory activity against the proliferation of nine cancer cell lines has been evaluated . Among them , compounds 9a and 9b displayed the strongest anti-proliferative activity with the IC50s below 10μM . Flow cytometric analysis showed that the compounds 9a and 9b dose-dependently arrested the cell cycle at G0/ P55008 phase . Although the leading compounds DB02058 and intedanib targets P11362 , the kinase activity test revealed that these compounds only showed slight inhibitory activity on P11362 kinase . Further enzymatic test aided by molecular docking simulation in the DB00171 -binding site demonstrated that 9a and 9b are potent inhibitors of c-Kit kinase . These compounds are worthy of further evaluation as anticancer agents . P13500 ( P13500 ) protects human neurons and astrocytes from DB01221 or HIV-tat-induced apoptosis . Acquired immunodeficiency syndrome ( AIDS ) -associated dementia is often characterized by chronic inflammation , with infected macrophage infiltration of the CNS resulting in the production of human immunodeficiency virus type 1 ( HIV-1 ) products , including tat , and neurotoxins that contribute to neuronal loss . In addition to their established role in leukocyte recruitment and activation , we identified an additional role for chemokines in the CNS . Monocyte chemoattractant protein-1 ( P13500 or P13500 ) and regulated upon activation normal T cell expressed and secreted ( RANTES ) were found to protect mixed cultures of human neurons and astrocytes from tat or DB01221 -induced apoptosis . Neuronal and astrocytic apoptosis in these cultures was significantly inhibited by co-treatment with P13500 or RANTES but not P02778 . The protective effect of RANTES was blocked by antibodies to P13500 , indicating that RANTES protection is mediated by the induction of P13500 . The DB01221 blocker , MK801 , also abolished the toxic effects of both tat and DB01221 . Tat or DB01221 treatment of mixed cultures for 24 h resulted in increased extracellular glutamate ( [ DB00142 ]e ) and DB01221 receptor 1 ( Q05586 ) expression , potential contributors to apoptosis . Co-treatment with P13500 inhibited tat and DB01221 -induced increases in [ DB00142 ]e and Q05586 , and also reduced the levels and number of neurons containing intracellular tat . These data indicate that P13500 may play a novel role as a protective agent against the toxic effects of glutamate and tat . DB08836 treatment ameliorates autoimmune myocarditis associated with enhanced cardiomyocyte thioredoxin expression . OBJECTIVE : P10599 ( TRX ) is a redox regulatory protein that protects cells from various stresses . P12821 ( P12821 ) inhibitor was reported to enhance endogenous antioxidant enzyme activities . This study was carried out to investigate whether temocapril , a novel non-sulfhydryl-containing P12821 inhibitor , reduces the severity of myocarditis via redox regulation mechanisms involving TRX . METHODS AND RESULTS : In normal rat myocytes in vitro and in vivo , Western blot showed that temocapril enhanced cytosolic redox regulatory protein TRX expression , but that neither mitochondrial TRX2 nor antioxidant enzymes , such as copper-zinc superoxide dismutase ( Cu/Zn-SOD ) or manganese superoxide dismutase ( Mn-SOD ) expression , was up-regulated by the preconditioning treatment . In rats with experimental autoimmune myocarditis ( EAM ) , the severity of myocarditis and the protein carbonyl contents were less increased in temocapril treatment ( 10 mg/kg/day , orally ) from day 1 to day 21 , but not in temocapril treatment from day 15 to day 21 . An immunohistochemical study showed that TRX stain was enhanced in infiltrating inflammatory cells and in damaged myocytes . Considering the characteristics of this model that myocardial inflammation begins around day 15 and increases until day 21 , temocapril treatment for 3 weeks might be thought of as a preconditioning treatment . CONCLUSIONS : The results suggest that TRX and the redox state modified by TRX may play a crucial role in the pathophysiology of EAM . DB08836 ameliorates myocarditis associated with inducing TRX up-regulation in a preconditioning manner , although the mechanism of TRX up-regulation by temocapril remains to be elucidated . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . Paraplegic mice are leading to new advances in spinal cord injury research . STUDY DESIGN : Experimental laboratory investigations with paraplegic mouse models . OBJECTIVES : To review the most recent advances in the field of spinal cord injury research ; immune system response , regeneration , and functional recovery . SETTINGS : Laval University and Laval University Medical Center , Quebec , Canada . METHODS : Assessment of regenerative processes and locomotor function recovery induced by a variety of treatments and approaches in wild-type and genetically engineered mice with complete or incomplete lesions of the spinal cord . RESULTS : Recent studies have reported a number of significant observations providing additional insight into the role and mechanism of regeneration , immune system response , and functional recovery after spinal cord injury ( SCI ) using incomplete paraplegic mice with Nogo-A , Q9BZR6 , EphA4 , P14136 /vimentime , P15018 , or Fas gene knock-out . A novel antibody called P02778 was also recently found to increase tissue sparing and angiogenesis after SCI . In an attempt to explore the possibilities of reactivating spared neurons below the injury level , researchers have found that pharmacological activation of specific subtypes of serotonin receptors ( eg , P08908 /2A/7 ) can sustain the production of basic locomotor-like movements in complete paraplegic mice . CONCLUSION : The growing availability of genetically engineered and mutant mouse strains along with molecular biology tools has led scientists to increasingly use murine models in SCI research . These new tools and models may assist scientists in understanding further the complex pathological consequences of SCI . Dexamethasone reverses adrenalectomy-induced neuronal de-differentiation in midbrain raphe-hippocampus axis . Differentiation leads to specific morphological and biochemical characteristics . We examined whether epigenetic factors ( e.g. , glucocorticoids ) are required to maintain neuronal differentiation in the adult brain . In the midbrain , adrenalectomy ( P10109 ) ( 1-2 wk ) reduced the size of tryptophan hydroxylase ( WH ) -immunoreactive ( IR ) neurons . P10109 rats exposed to short-term ( 24-72-h ) dexamethasone ( ST-DEX ) in the drinking saline ( 10 mg/l ) showed an increase in WH protein , somal area and dendritic size of WH-IR neurons . In the hippocampus , P10109 for 2-3 mo ( long-term ; LT ) reduced Nissl staining , calbindin ( DB09061 ) -IR and P08908 receptor mRNA in the granular cell layer , and the size of the molecular layer and its DB09061 -IR dendrites . Small vimentin ( Vim ) -IR glial cells emerged in the granular layer . ST-DEX after LT- P10109 rapidly induced a recovery of P08908 mRNA , Nissl labeling and DB09061 -IR in the granule cell layer . In the molecular layer , there was an increase in the area and in the number of DB09061 -IR dendrites . Furthermore , the Vim-IR glial cells were enlarged in size and branching . The rate of cell proliferation was studied in these animals . Immunostaining with antibodies against proliferating cell nuclear antigen ( P12004 ) and use of bromouridine argue against enhanced neurogenesis after ST-DEX in LT- P10109 . We propose that glucocorticoids induce and maintain differentiation of serotonergic and DB09061 -IR neurons in the midbrain-hippocampal axis . A neuronotrophic role for the glial P08908 receptor is suggested . P02751 and VLA-4 in haematopoietic stem cell-microenvironment interactions . The self-renewal and differentiation of haematopoietic stem cells occurs in vivo and in vitro in direct contact with cells making up the haematopoietic microenvironment . In this study we used adhesive ligands and blocking antibodies to identify stromal cell-derived extracellular matrix proteins involved in promoting attachment of murine haematopoietic stem cells . Here we report that day-12 colony-forming-unit spleen (CFU- P28222 )5 cells and reconstituting haematopoietic stem cells attach to the C-terminal , heparin-binding fragment of fibronectin by recognizing the P28290 peptide of the alternatively spliced non-type III connecting segment ( IIICS ) of human plasma fibronectin . Furthermore , CFU- P28222 stem cells express the alpha 4 subunit of the VLA-4 integrin receptor , which is known to be a receptor for the P28290 sequence , and monoclonal antibodies against the integrin alpha 4 subunit of VLA-4 block adhesion of CFU- P28222 stem cells to plates coated with the C-terminal fibronectin fragment . Finally , polyclonal antibodies against the integrin beta 1 subunit of VLA-4 inhibit the formation of CFU- P28222 -derived spleen colonies and medullary haematopoiesis in vivo following intravenous infusion of antibody-treated bone marrow cells . Functional analysis and expression characteristics of chloroplastic Prx IIE . Peroxiredoxins ( Prxs ) are ubiquitous thiol-dependent peroxidases capable of eliminating a variety of peroxides through reactive catalytic cysteines , which are regenerated by reducing systems . Based on amino acid sequences and their mode of catalysis , five groups of thiol peroxidases have been distinguished in plants , and type II Prx is one of them with representatives in many sub-cellular compartments . The mature form of poplar chloroplastic Prx IIE was expressed as a recombinant protein in Escherichia coli . The protein is able to reduce H2O2 and tert-butyl hydroperoxide and is regenerated by both glutaredoxin ( Grx ) and thioredoxin ( P10599 ) systems . Nevertheless , compared with Trxs , Grxs , and more especially chloroplastic Grx P28222 , are far more efficient reductants towards Prx IIE . The expression of Prx IIE at both the mRNA and protein levels as a function of organ type and abiotic stress conditions was investigated . Western blot analysis revealed that Prx IIE gene is constitutively expressed in Arabidopsis thaliana , mostly in young and mature leaves and in flowers . Under photo-oxidative treatment and water deficit , almost no change was observed in the abundance of Prx IIE in A. thaliana , while the level of Prx Q ( one of the two other chloroplastic Prxs with 2- DB00151 Prx ) increased in response to both stresses , indicating that plastidic members of the Prx family exhibit specific patterns of expression under stress . Further characterization of a somatic cell hybrid panel : ten new assignments to the bovine genome . Thirty-six partially characterized hamster-bovine hybrid cell lines were used for the determination of synteny groups . Sixteen additional reference loci , selected for their coverage of the bovine genome , were analysed on these hybrid cells . This increases to 25 the number of synteny groups detected . This panel was then used to make synteny assignments for 10 additional loci , eight by Southern blotting ( P02452 , P08123 , FAS , P07858 , P07711 , P07510 , P07686 and P08908 ) and two by polymerase chain reaction ( PCR ) amplification ( P35367 and ETH1112 ) . These loci were assigned to international synteny groups U12 ( P35367 ) , U13 ( P08123 ) , U17 ( P07510 ) , U21 ( P02452 , FAS ) , U29 ( ETH1112 ) , to chromosome 20 ( U14 or U25 ) for P07686 and P08908 , and to the same local synteny group ( A ) , which is probably U18 , for P07858 and P07711 . For three loci already mapped in humans ( P02452 , P08123 and P07510 ) , the present results are in accordance with the predictions based on comparative mapping between the human and bovine species . 1-(3-Trifluoromethylphenyl) piperazine ( TFMPP ) in the ventral tegmental area reduces the effect of desipramine in the forced swimming test in rats : possible role of serotonin receptors . 1-(3-Trifluoromethylphenyl)piperazine ( TFMPP ) , a serotonin1 ( 5-HT1 ) receptor agonist , injected i.p. in doses of 0.1 and 0.6 mg/kg , did not modify the immobility time of rats in the forced swimming test but significantly antagonized the effect of a 7 days treatment with 10 mg/kg per day desipramine ( DB01151 ) . A similar effect was found on infusing 1 and 5 micrograms/microliters TFMPP bilaterally into the ventral tegmental area ( VTA ) . Infusion of 5 micrograms/microliters TFMPP into the nucleus accumbens or into the globus pallidus did not modify the effect of DB01151 . The effect of 5 micrograms TFMPP infused into the VTA was prevented by the i.p. administration of 5 mg/kg metergoline , a non-selective serotonin receptor antagonist . Infusion of 5 micrograms/microliters 8-hydroxy-2-(di-n-propylamino)tetralin , a specific P08908 receptor agonist , into the VTA did not modify the effect of DB01151 . Besides acting as a P28222 receptor agonist , TFMPP may also act on other 5-HT receptor types , but available evidence suggests that its former action is more important . It thus appears that 5-HT1 receptors in the VTA , presumably of the P28222 type , act by preventing the anti-immobility effect of DB01151 . The role of VTA dopamine and non-dopamine cells in the effect of TFMPP is discussed . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . Dysfunction of the P19957 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia . Patients with myeloproliferative disorders ( MPDs ) , such as essential thrombocythemia ( ET ) have increased risk of thrombosis and bleeding , which are major sources of morbidity and mortality . Most P53602 patients have a gain of function mutation in O60674 ( JAK2V617F ) , but little is known how JAK2V617F affects platelet function . Here , we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin ( which couples to O60674 ) on this pathway . In contrast , SFLLRN-mediated P16109 expression , DB00171 secretion , phosphorylation of the PKC substrate pleckstrin , and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets . In addition , thrombopoietin-mediated O60674 phosphorylation was unchanged , suggesting that signaling pathways activated downstream of O60674 are impaired . Indeed , we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the P19957 kinase substrate Akt , and have reduced activation of Rap1 in response to thrombopoietin , DB01277 ,ADP , SFLLRN , and thrombin . This effect was independent of Giα Q9H244 purinergic receptor function as ADP-mediated inhibition of P50552 phosphorylation was unchanged . These results demonstrate that the P19957 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients , resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation . Platelet Q9H244 receptor inhibition by thienopyridines : status and future . Thienopyridines have a well-established role in the treatment of coronary artery disease , especially in the setting of acute coronary syndromes and percutaneous coronary interventions . DB00208 , the first FDA-approved thienopyridine , was shown to be effective in reducing coronary events in high risk patients , but the original enthusiasm was hampered by concerns about its serious bone marrow toxicity . DB00758 a second generation thienopyridine with lesser side effects , is not only at least as effective as ticlopidine , but in combination with a low dose of aspirin , has been demonstrated to reduce the risk of major cardiovascular events in acute coronary syndrome patients in large-scale , randomised trials . Recent studies have highlighted major flaws in clopidogrel pharmacokinetics due to its delayed onset of action , and much attention has been devoted to the phenomenon of clopidogrel ' resistance ' . Among the novel , third generation thienopyridines , prasugrel as compared to clopidogrel has demonstrated lower inter-patient response variability and a reduced incidence of ischaemic events , but at an increased risk of major bleeding . Currently , several studies are continuing to test new direct Q9H244 receptor antagonists , such as cangrelor and AZD6140 , characterised by a faster reversal of platelet inhibition . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . P62158 regulates Ca2+-sensing receptor-mediated Ca2+ signaling and its cell surface expression . The Ca(2+)-sensing receptor ( P41180 ) is a member of family C of the GPCRs responsible for sensing extracellular Ca(2+) ( [Ca(2+)](o) ) levels , maintaining extracellular Ca(2+) homeostasis , and transducing Ca(2+) signaling from the extracellular milieu to the intracellular environment . In the present study , we have demonstrated a Ca(2+)-dependent , stoichiometric interaction between P62158 and a P62158 -binding domain ( CaMBD ) located within the C terminus of P41180 ( residues 871-898 ) . Our studies suggest a wrapping around 1-14-like mode of interaction that involves global conformational changes in both lobes of P62158 with concomitant formation of a helical structure in the CaMBD . More importantly , the Ca(2+)-dependent association between P62158 and the C terminus of P41180 is critical for maintaining proper responsiveness of intracellular Ca(2+) responses to changes in extracellular Ca(2+) and regulating cell surface expression of the receptor . Substituted benzamides with conformationally restricted side chains . 5 . Azabicyclo[x.y.z] derivatives as Q13639 receptor agonists and gastric motility stimulants . The syntheses of benzamides containing azabicyclo[x.y.z] side chains and their Q13639 receptor agonist and 5- Q9H205 receptor antagonist properties are described . These compounds were designed to mimic higher energy conformations of quinolizidine and indolizidine . High potency was achieved for both activities although an exactly paralleling SAR was not apparent . Introduction of O and S resulted in only marginal differences in potency which was more apparent for 5- Q9H205 antagonism . The introduction of a methyl group alpha to the basic nitrogen resulted in a reduction in Q13639 receptor agonist potency . DB04917 ( 5f ) was identified for further evaluation for which both enantiomers had an identical pharmacological profile , as did an azatricyclic 9b , which contained a combination of the steric bulk of the two separate enantiomers . DB08888 for vitreoretinal diseases . P02751 and laminin are clinically relevant plasmin receptors in the eye . Located at the vitreoretinal interface , they are cleaved by ocriplasmin ( DB05028 , ThromboGenics , Iselin , NJ ) , a novel ophthalmic medication . A series of clinical trials to study ocriplasmin for the treatment of vitreoretinal diseases such as vitreomacular traction , macular hole , and exudative age-related macular degeneration are underway . The results are promising and may impact patient care . Characterization of the effect of chronic administration of a calcium-sensing receptor antagonist , DB05255 , on renal calcium excretion and serum calcium in postmenopausal women . Ronacaleret is an orally-active calcium-sensing receptor ( P41180 ) antagonist that has the potential for therapeutic utility in the stimulation of PTH release , notably as a bone anabolic agent comparable to recombinant human PTH(1-34) ( DB05829 (1-34) ) . A recent study has shown that , despite the ability to increase circulating PTH levels in postmenopausal women in a dose-dependent manner , minimal effects of DB05255 on bone mineral density have been observed . Therefore , the purpose of this study was to characterize the PTH profile as well as calcium metabolism parameters as a marker of PTH biological activity following the administration of DB05255 or DB05829 (1-34) . Administration of DB05255 led to lower peak levels of PTH than were observed with DB05829 (1-34) , however , greater total PTH exposure was observed . Further , chronic administration of either agent was associated with increases in urinary calcium excretion and serum calcium levels , with the magnitude of the changes following DB05255 significantly greater than that for DB05829 (1-34) . The greater magnitude of effects observed with DB05255 is likely due to the greater total PTH exposure , and is potentially reflective of a state comparable to mild hyperparathyroidism . It is not clear whether the administration of all calcilytics would lead to a similar result , or is due to characteristics specific to DB05255 . Biological characterization of DB05037 , a small-molecule inhibitor of cyclin-dependent kinases , in human tumor cell lines . P12004 -dependent kinases ( CDK ) , and their regulatory cyclin partners , play a central role in eukaryotic cell growth , division , and death . This key role in cell cycle progression , as well as their deregulation in several human cancers , makes them attractive therapeutic targets in oncology . A series of CDK inhibitors was developed using Astex 's fragment-based medicinal chemistry approach , linked to high-throughput X-ray crystallography . A compound from this series , designated DB05037 , is currently in early-phase clinical development . We describe here the biological characterization of DB05037 , a potent inhibitor of several CDK family members . DB05037 showed potent antiproliferative activity ( 40-940 nmol/L ) in a panel of human tumor cell lines , and the mechanism of action was shown here to be consistent with the inhibition of P06493 and P24941 in solid tumor cell lines . DB05037 caused cell cycle arrest followed by apoptosis in human tumor cells and inhibited tumor growth in human tumor xenograft models . Tumor regression was observed following twice daily dosing of DB05037 in the HCT116 and HT29 colon cancer xenograft models . We show that these biological effects are linked to inhibition of CDKs in vivo and that DB05037 induces tumor cell apoptosis in these xenograft models . DB05037 has an attractive biological profile for development as a clinical candidate , and the tolerability and efficacy in animal models compare favorably with other CDK inhibitors in clinical development . Studies described here formed the biological rationale for investigating the potential therapeutic benefit of DB05037 in cancer patients . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Involvement of the βγ subunits of G proteins in the DB02527 response induced by stimulation of the histamine H1 receptor . Stimulation of the histamine H1 receptor has been shown to enhance adenosine 3' , 5'-cyclic monophosphate ( DB02527 ) accumulation in various cell types but , to date , the mechanism by which this occurs is still unclear . In the present study , we examined the possibility that the betagamma subunits of G proteins ( G betagamma ) are involved in this process in cultured Chinese hamster ovary cells transfected with the human histamine H1 receptor ( CHO-H1 ) . DB11320 increased intracellular DB02527 levels in a concentration-dependent manner in CHO-H1 cells , and this histamine action was abolished by DB06691 ( 1 microM ) . Inhibition of histamine H1 receptor-G(q) protein coupling by stable expression of the C-terminal peptide of G alpha(q) protein significantly attenuated the DB02527 accumulation induced by histamine . By comparison , neither BAPTA/AM ( 50 microM ) , an intracellular Ca2+ chelator , nor GF 109203X ( 1 microM ) , an inhibitor of protein kinase C , influenced the DB02527 response . P35367 -mediated DB02527 accumulation was significantly inhibited by transient transfection of CHO-H1 cells with the C-terminal peptide of beta-adrenoceptor kinase I ( residues 542-685 ) , a scavenger of G betagamma . Stable expression of the C-terminal peptide of the G alpha(s) protein , but not treatment with pertussis toxin ( 200 ng/ml for 24 h ) , attenuated the histamine H1 receptor-mediated DB02527 accumulation . These results suggest that stimulation of histamine H1 receptors activates adenylyl cyclase through the release of G betagamma subunits from G proteins , thereby elevating intracellular DB02527 levels .

MH_train_1266

MH_train_1266

MH_train_1266

interacts_with DB00233?

multiple_choice

P01236 activates mitogen-activated protein kinase signaling and corticotropin releasing hormone transcription in rat hypothalamic neurons . P01236 ( PRL ) modulates maternal behavior and mediates hypothalamic pituitary adrenal axis inhibition during lactation via PRL receptors in the brain . To identify mechanisms mediating these effects , we examined the effects of PRL on signaling and P06850 transcription in hypothalamic neurons in vivo and in vitro . Western blot of hypothalamic proteins from rats receiving intracerebroventricular PRL injection revealed increases in phosphorylation of the MAPK and P29323 . Double-staining immunohistochemistry demonstrated phosphorylated P29323 localization in parvocellular P06850 neurons as well as magnocellular vasopressin and oxytocin neurons of the hypothalamic paraventricular ( PVN ) and supraoptic nuclei . PRL also induced P29323 phosphorylation in vitro in the hypothalamic cell line , 4B , which expresses PRL receptors , and in primary hypothalamic neuronal cultures . Using reporter gene assays in 4B cells , or quantitative RT-PCR for primary transcript in hypothalamic cell cultures , PRL potentiated forskolin-stimulated P06850 transcription through activation of the P29323 /MAPK pathway . The effect of PRL in hypothalamic cell cultures was unaffected by tetrodotoxin , suggesting a direct effect on P06850 neurons . The data show that PRL activates the P29323 /MAPK pathway and facilitates P06850 transcription in P06850 neurons , suggesting that the inhibitory effect of PRL on hypothalamo-pituitary-adrenal axis activity reported in vivo is indirect and probably mediated through modulation of afferent pathways to the PVN . In addition , the prominent stimulatory action of PRL on the P29323 /MAPK pathway in the hypothalamic PVN and supraoptic nucleus is likely to mediate neuroplasticity of the neuroendocrine system during lactation . Gefitinib induces epidermal growth factor receptor dimers which alters the interaction characteristics with ¹²⁵I- P01133 . The tyrosine kinase inhibitor gefitinib inhibits growth in some tumor types by targeting the epidermal growth factor receptor ( P00533 ) . Previous studies show that the affinity of the P01133 - P00533 interaction varies between hosting cell line , and that gefitinib increases the affinity for some cell lines . In this paper , we investigate possible mechanisms behind these observations . Real-time interaction analysis in LigandTracer® Grey revealed that the P04626 dimerization preventing antibody pertuzumab clearly modified the binding of ¹²⁵I- P01133 to P00533 on P04626 overexpressing SKOV3 cells in the presence of gefitinib . DB06366 did not affect the binding on A431 cells , which express low levels of P04626 . Cross-linking measurements showed that gefitinib increased the amount of P00533 dimers 3.0-3.8 times in A431 cells in the absence of P01133 . In P01133 stimulated SKOV3 cells the amount of P00533 dimers increased 1.8-2.2 times by gefitinib , but this effect was cancelled by pertuzumab . Gefitinib treatment did not alter the number of P00533 or P04626 expressed in tumor cell lines A431 , U343 , SKOV3 and SKBR3 . Real-time binding traces were further analyzed in a novel tool , Interaction Map , which deciphered the different components of the measured interaction and supports P01133 binding to multiple binding sites . P00533 and P04626 expression affect the levels of P00533 monomers , homodimers and heterodimers and P01133 binds to the various monomeric/dimeric forms of P00533 with unique binding properties . Taken together , we conclude that dimerization explains the varying affinity of P01133 - P00533 in different cells , and we propose that gefitinib induces P00533 dimmers , which alters the interaction characteristics with ¹²⁵I- P01133 . Consequences of the Y139F Vkorc1 mutation on resistance to AVKs : in-vivo investigation in a 7th generation of congenic Y139F strain of rats . OBJECTIVES : In humans , warfarin is used as an anticoagulant to reduce the risk of thromboembolic clinical events . DB00682 derivatives are also used as rodenticides in pest control . The gene encoding the protein targeted by anticoagulants is the Vitamin K-2,3-epoxide reductase subunit 1 ( Q9BQB6 ) . Since its discovery in 2004 , various amino acid and transcription-regulatory altering Q9BQB6 mutations have been identified in patients who required extreme antivitamin K dosages , or wild populations of rodents that were difficult to control with anticoagulant rodenticides . One unresolved question concerns the dependency of the Q9BQB6 on the genetic background in humans and rodents that respond weakly or not at all to anticoagulants . Moreover , an important question requiring further analyses concerns the role of the Vkorc1 gene in mediating resistance to more recently developed warfarin derivatives ( superwarfarins ) . METHODS : In this study , we bred a quasicongenic rat strain by using a wild-caught anticoagulant resistant rat as a donor to introduce the Y > F amino acid change at position 139 in the Vkorc1 into the genetic background of an anticoagulant susceptible Spraque-Dawley recipient strain . RESULTS AND CONCLUSION : In this manuscript we report the prothrombin times measured in the P08709 generation after exposure to chlorophacinone , bromadiolone , difenacoum and difethialone . We observed that the mutation Y139F mediates resistance in an otherwise susceptible genetic background when exposed to chlorophacinone and bromadiolone . However , the physiological response to the super-warfarins , difenacoum and difethialone , may be strongly dependent on other genes located outside the congenic interval ( 28.3 cM ) bracketing the Vkorc1 in our P08709 generation congenic strain . Q92847 agonist ( DB05657 ) accelerates gastric emptying in adults with diabetes and symptomatic gastroparesis . BACKGROUND : DB05657 is a synthetic , selective ghrelin agonist in development for gastroparesis . AIM : To assess safety and effects of DB05657 in diabetes patients with symptomatic gastroparesis . METHODS : Adults with type 1 or type 2 diabetes mellitus received placebo and DB05657 ( 80 , 160 , 320 or 600 microg/kg ) infusions in a cross-over manner following a radiolabelled meal . Blood glucose levels were stabilized using a hyperinsulinemic-euglycemic clamp . Primary endpoints were gastric half emptying and latency times . Secondary measures included assessment of gastroparesis symptoms and endocrine responses . RESULTS : Ten patients with type 1 ( n = 7 ) or 2 ( n = 3 ) diabetes , moderate-to-severe gastroparesis symptoms and > or =29 % retention 4 h after a radiolabelled solid meal were enrolled . DB05657 produced significant reductions in solid meal half-emptying ( 20 % , P = 0.043 ) and latency ( 34 % , P = 0.037 ) times vs. placebo . Reductions in overall postmeal symptom intensity ( 24 % ) and postprandial fullness ( 37 % ) following DB05657 infusion were not statistically significant . Most adverse events were mild and self-limiting and there were no identifiable differences in numbers or types of adverse events between DB05657 and placebo . CONCLUSIONS : This proof-of-concept study demonstrates that the ghrelin agonist DB05657 is well-tolerated in diabetes patients with moderate-to-severe chronic gastroparesis and shows statistically significant improvements in gastric emptying . Monoclonal antibodies targeting the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 , P00533 ) autocrine pathway contributes to a number of highly relevant processes in cancer development and progression , including cell proliferation , regulation of apoptotic cell death , angiogenesis and metastatic spread . The crucial role that P00533 plays in human cancers has led to an extensive search for selective inhibitors of its signaling pathway . The results of a large body of preclinical studies and clinical trials thus far conducted suggest that targeting the P00533 could bring a significant contribution to cancer therapy . A variety of different approaches are currently being used to target the P00533 . The most promising strategies in clinical development include monoclonal antibodies , to prevent ligand binding , and small molecules inhibitors of the tyrosine kinase enzymatic activity , that inhibit autophosphorylation and downstream intracellular signaling . Several blocking monoclonal antibodies against the P00533 have been developed . Among these , IMC-225 is a chimeric human-mouse monoclonal IgG1 antibody that has been the first anti- P00533 targeted therapy to enter clinical evaluation in cancer patients in Phase II and III studies , alone or in combination with conventional radiotherapy and chemotherapy . However , other antibodies against P00533 have demonstrated antitumor activity in several preclinical models of human cancer and are currently under investigation in the clinical setting , such as ICR62 , DB01269 and EMD72000 . This review will focus on all the preclinical data available on monoclonal antibodies engineered against the P01133 receptor . Stress-induced phosphoprotein 1 as a secreted biomarker for human ovarian cancer promotes cancer cell proliferation . Ovarian cancers are frequently not diagnosed until advanced stages , resulting in a high case fatality rate . Because of this , more tumor markers , in addition to Q8WXI7 , for detecting and monitoring ovarian cancer are needed . During a systematic search for potential biomarkers of ovarian cancer , we compared the protein profiles between tumor interstitial fluid and normal interstitial fluid of ovaries , rationalizing that abnormal levels of proteins in tumor interstitial fluid may be detected in peripheral blood and thus serve as easily accessible tumor markers . Here , we show that stress-induced phosphoprotein 1 ( P31948 ) was secreted by ovarian cancer tissues into the peripheral blood of patients , resulting in a significant increase of serum levels of P31948 in cancer patients compared with those in age-matched normal controls . Our results further indicated that combined use of Q8WXI7 and P31948 may increase early detection of ovarian cancer . Functionally , recombinant P31948 significantly induced P29323 phosphorylation , promoted DNA synthesis , and increased Ki-67 immunoreactivity in ovarian cancer cells , suggesting that P31948 in vitro promotes cell proliferation . Colocalization of P31948 and phospho- P29323 in human ovarian cancer tissues also supports an in vivo activation of P29323 by P31948 . Further understanding of molecular roles of P31948 in human ovarian cancer may shed light on its pathophysiology and development of novel therapeutic strategies . Mechanisms of interleukin-1beta-induced P39905 release from rat glioma cells . P39905 ( P39905 ) is highly expressed both in neurons and astrocytes in injured tissues . Astrocytes support neurons by releasing neurotrophic factors including P39905 . It has been reported that various agents including cytokines such as interleukin ( IL ) -1beta induce P39905 mRNA expression and the release in astrocytes . However , the mechanism behind the P39905 synthesis and release remains unclear . Herein , we investigated the mechanisms of the IL-1beta-induced P39905 release from rat P13671 glioma cells . IL-1beta time dependently stimulated P39905 release from P13671 cells . IL-1beta induced the phosphorylation of inhibitor kappa B ( IkappaB ) , p38 mitogen-activated protein ( Q96HU1 ) kinase , Q8TCB0 / Q8NFH3 Q96HU1 kinase , stress-activated protein kinase/c-Jun N-terminal kinase ( SAPK/JNK ) and signal transducer and activator of transcription ( P35610 ) 3 . The IL-1beta-stimulated levels of P39905 were suppressed by wedelolactone , an inhibitor of O15111 , SB203580 , an inhibitor of p38 Q96HU1 kinase , PD98059 , an inhibitor of Q02750 /2 or Janus family of tyrosine kinase ( JAK ) inhibitor I , an inhibitor of upstream kinase of P40763 . On the contrary , SP600125 , an inhibitor of SAPK/JNK , failed to reduce the IL-1beta-effect . These results strongly suggest that IL-1beta stimulates P39905 release through the pathways of IkappaB-nuclear factor kappa B , p38 Q96HU1 kinase , Q8TCB0 / Q8NFH3 Q96HU1 kinase and JAK- P40763 , but not through the SAPK/JNK pathway in glioma cells . Mitogenic and antiapoptotic role of constitutive NF-kappaB/Rel activity in pancreatic cancer . The transcription factor NF-kappaB/Rel was found to be constitutively activated in human pancreatic cancer . RelA is present in the nucleus in primary human pancreatic cancer samples as well as in pancreatic cancer cell lines . NF-kappaB/Rel-binding activity consists of NF-kappaB1(p50) and RelA(p65) . Constitutive NF-kappaB/Rel activity correlates with O15111 ( IKK ) activity and can be blocked by dominant negative mutants of IKKbeta and to a lesser extent by IKKalpha . Constitutive NF-kappaB/Rel activity and the transactivation potential of RelA(p65) can be inhibited by dominant negative mutant Ras , the P19957 kinase inhibitor LY294002 , or dominant negative mutant Akt kinase . Transfection of a dominant negative mutant epidermal growth factor receptor ( P01133 -R ) , P01133 -R kinase inhibitor Tyrphostin and LY 294002 blocked IKK activity and NF-kappaB-dependent transcription . Inhibition of constitutive IKK or NF-kappaB/Rel activity increased the number of apoptotic cells . Stably expressing a nondegradable form of P25963 inhibited anchorage-dependent and -independent proliferation in MiaPaCa2 and Panc1 cells . Our data demonstrate that an P01133 -R/Ras/ P19957 kinase/Akt/IKK-dependent pathway contributes to constitutive NF-kappaB/Rel activity in pancreatic cancer . Inhibition of NF-kappaB/Rel activity reveals a mitogenic and antiapoptotic role for NF-kappaB/Rel in pancreatic cancer . Ghrelin directly stimulates glucagon secretion from pancreatic alpha-cells . Previous work has demonstrated that the peptide hormone ghrelin raises blood glucose . Such has been attributed to ghrelin 's ability to enhance GH secretion , restrict insulin release , and/or reduce insulin sensitivity . Ghrelin 's reported effects on glucagon have been inconsistent . Here , both animal- and cell-based systems were used to determine the role of glucagon in mediating ghrelin 's effects on blood glucose . The tissue and cell distribution of ghrelin receptors ( Q92847 ) was evaluated by quantitative PCR and histochemistry . Plasma glucagon levels were determined following acute acyl-ghrelin injections and in pharmacological and/or transgenic mouse models of ghrelin overexpression and Q92847 deletion . Isolated mouse islets and the α-cell lines αTC1 and InR1G9 were used to evaluate ghrelin 's effects on glucagon secretion and the role of calcium and P29323 in this activity . Q92847 mRNA was abundantly expressed in mouse islets and colocalized with glucagon in α-cells . Elevation of acyl-ghrelin acutely ( after sc administration , such that physiologically relevant plasma ghrelin levels were achieved ) and chronically ( by slow-releasing osmotic pumps and as observed in transgenic mice harboring ghrelinomas ) led to higher plasma glucagon and increased blood glucose . Conversely , genetic Q92847 deletion was associated with lower plasma glucagon and reduced fasting blood glucose . Acyl-ghrelin increased glucagon secretion in a dose-dependent manner from mouse islets and α-cell lines , in a manner requiring elevation of intracellular calcium and phosphorylation of P29323 . Our study shows that ghrelin 's regulation of blood glucose involves direct stimulation of glucagon secretion from α-cells and introduces the ghrelin-glucagon axis as an important mechanism controlling glycemia under fasting conditions . Immunotherapy of ovarian cancer with antibodies : a focus on oregovomab . Recent advances in the molecular and cellular biology of malignancy and tumour immunology have stimulated significant progress in the application of immunotherapies as adjuvant treatments in cancer . DB04964 ( OvaRex , AltaRex ) is a murine monoclonal antibody with high affinity to the ovarian cancer associated antigen Q8WXI7 . Infusion of low-dose antibody results in formation of circulating immune complexes which can trigger a cellular immune response targeting Q8WXI7 and the ovarian cancer . DB04964 has activity following initial chemotherapy and in recurrent disease settings and is in Phase III trials to establish its efficacy to prolong time to relapse in patients with advanced ovarian cancer and favourable outcomes to their front-line treatment . Additional studies of antigen processing and combination chemo-immunotherapy are ongoing . The treatment shows promise as a potential new addition to the standard care of ovarian cancer . Modulation of cytokine release from human monocytes by drugs used in the therapy of inflammatory bowel diseases . BACKGROUND : Cytokines produced in the gut mucosa play an important role in the pathogenesis of inflammatory bowel diseases ( Q9UKU7 ) . To determine whether drugs used in the treatment of these diseases modulate cytokine synthesis , we investigated their effects on endotoxin-induced tumour necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -1 beta and P05231 release by elutriation-purified human monocytes in vitro . METHODS : Drugs tested were dexamethasone , DB00244 , sulphapyridine and zileuton ( a P09917 inhibitor ) . Monocytes were isolated and stimulated with endotoxin , and P01375 , IL-1 and P05231 levels were determined using an enzyme-linked immunosorbent assay . RESULTS : Monocyte stimulation with endotoxin resulted in an average P01375 release of 2464 +/- 64 pg/10(6) cells , IL-1 release of 616 +/- 47 pg/10(6) cells and P05231 release of 2259 +/- 148 pg/10(6) cells . Addition of dexamethasone resulted in a reduction of P01375 , IL-1 and P05231 release to below background levels . DB00891 significantly reduced P01375 and induced IL-1 release in a dose-dependent fashion , but had no significant effect on P05231 release . 5- DB00233 did not modulate P05231 synthesis , but significantly reduced IL-1 and enhanced P01375 synthesis . Zileuton reduced P01375 and P05231 release , but enhanced IL-1 release . CONCLUSION : We conclude that these anti-inflammatory drugs are able to modulate cytokine release by human monocytes . Further studies are needed to determine whether these effects are related to their therapeutic efficacy in Q9UKU7 . Retinoids and myelomonocytic growth factors cooperatively activate P10276 and induce human myeloid leukemia cell differentiation via Q96HU1 kinase pathways . Use of all-trans-retinoic acid ( DB00755 ) in combinatorial differentiation therapy of acute promyelocytic leukemia ( APL ) results in exceptional cure rates . However , potent cell differentiation effects of DB00755 are so far largely restricted to this disease and long-term survival rates in non-APL acute myelogeneous leukemia ( AML ) remain unacceptably poor , requiring development of novel therapeutic strategies . We demonstrate here that myelomonocytic growth factors ( granulocyte colony-stimulating factor [ DB00099 ] and/or granulocyte macrophage colony-stimulating factor [ GM- P04141 ] ) potentiate differentiation effects of DB00755 in different AML cell lines and primary cells from patients with myeloid leukemia . The ligand-dependent activities of endogenous and transiently expressed retinoic acid receptor alpha ( RARalpha ) isoforms can be potentiated by G/GM- P04141 in U-937 cells and correlate with increased expression of DB00755 -inducible RARalpha2 isoform . Specific inhibitors of mitogen mitogen-activated protein kinase ( MAPK ) ( MEK ) -1/-2 or p38 extracellular signal-related kinase ( P29323 ) kinase diminish the DB00755 as well as DB00755 and G/GM- P04141 -induced activation of the RARalpha proteins and decreased the differentiation-induced decline in cell numbers . Our data demonstrate that acting , at least in part , via the Q96HU1 kinase pathways , myelomonocytic growth factors enhance DB00755 -dependent activation of the RARalpha isoforms and maturation of myeloid leukemia cells . These results suggest that combinatorial use of these agents may be effective in differentiation therapy of AML . Involvement of retinoic acid receptor alpha in the stimulation of tissue-type plasminogen-activator gene expression in human endothelial cells . Retinoids stimulate tissue-type plasminogen-activator ( t-PA ) gene expression in human endothelial cells , and are likely to do so by binding to one or more nuclear retinoid receptors . The present study was initiated to identify the retinoid receptor(s) involved in this process . Expression and regulation of retinoic acid receptors ( RARs ) and retinoid X receptors ( RXRs ) were analyzed by Northern-blot analysis of total or poly(A)-rich RNA prepared from cultured human umbilical vein endothelial cells ( HUVEC ) . Prior to any exposure to retinoids , HUVEC express two transcripts for P10276 ( 3.6 kb and 2.8 kb ) , and low levels of transcripts for P10826 ( 3.4 kb and 3.2 kb ) and P13631 ( 3.3 kb and 3.1 kb ) . Two RXR subtypes were identified , RXR-alpha ( 4.8 kb ) and , at a much lower concentration , RXR-beta ( 2.4 kb ) ; no evidence for the presence of RXR-gamma was found . Furthermore , HUVEC express cellular retinol-binding protein I ( P09455 ) and cellular retinoic-acid-binding protein I ( P29762 ) mRNA . Exposure of HUVEC to 1 microM retinoic acid or the DB04942 , Ch55 , led to the induction of the two P10826 mRNAs , RXR-alpha mRNA and P09455 mRNA , whereas the expression of the other receptor and P29762 transcripts did not change appreciably . Using retinoid analogues that bind preferentially to one of the RAR or RXR subtypes , we found evidence that P10276 is involved in the retinoid-induced t-PA expression in HUVEC . This conclusion was strengthened by experiments in which blocking of P10276 with a specific P10276 antagonist , Ro 41-5253 , was demonstrated to suppress the induction of t-PA by retinoids . Regional selective neuronal degeneration after protein phosphatase inhibition in hippocampal slice cultures : evidence for a Q96HU1 kinase-dependent mechanism . The regional selectivity and mechanisms underlying the toxicity of the serine/threonine protein phosphatase inhibitor okadaic acid ( OA ) were investigated in hippocampal slice cultures . Image analysis of propidium iodide-labeled cultures revealed that okadaic acid caused a dose- and time-dependent injury to hippocampal neurons . Pyramidal cells in the P07451 region and granule cells in the dentate gyrus were much more sensitive to okadaic acid than the pyramidal cells in the P00915 region . Electron microscopy revealed ultrastructural changes in the pyramidal cells that were not consistent with an apoptotic process . Treatment with okadaic acid led to a rapid and sustained tyrosine phosphorylation of the mitogen-activated protein kinases P27361 and P28482 ( Q8TCB0 /42(mapk) ) . The phosphorylation was markedly reduced after treatment of the cultures with the microbial alkaloid DB02152 ( a nonselective protein kinase inhibitor ) or the Q96HU1 kinase kinase ( Q02750 /2 ) inhibitor PD98059 . DB02152 and PD98059 also ameliorated the okadaic acid-induced cell death . Inhibitors of protein kinase C , Ca2+/calmodulin-dependent protein kinase II , or tyrosine kinase were ineffective . These results indicate that sustained activation of the Q96HU1 kinase pathway , as seen after e.g. , ischemia , may selectively harm specific subsets of neurons . The susceptibility to Q96HU1 kinase activation of the P07451 pyramidal cells and dentate granule cells may provide insight into the observed relationship between cerebral ischemia and dementia in Alzheimer 's disease . P03372 alpha , but not estrogen receptor beta , is involved in the regulation of the O00300 / O14788 ( osteoprotegerin/receptor activator of NF-kappa B ligand ) ratio and serum interleukin-6 in male mice . DB00286 are important for the male skeleton . O00300 ( O00300 ) , receptor activator of NF-kappa B ligand ( O14788 ) , interleukin-6 ( P05231 ) , IL-1 and tumor necrosis factor alpha ( TNFalpha ) have been suggested to be involved in the skeletal effects of estrogen . We treated orchidectomized mice with estradiol for 2 weeks and observed a 143 % increase in the trabecular bone mineral density of the distal metaphysis of femur that was associated with a decreased O00300 / O14788 mRNA ratio in vertebral bone . A similar decreased O00300 / O14788 ratio was also seen after estrogen treatment of ovariectomized female mice . The effect of estrogen receptor ( ER ) inactivation on the O00300 / O14788 ratio was dissected by using intact male mice lacking ER alpha ( ERKO ) , ER beta ( BERKO ) or both receptors ( DERKO ) . The expression of O00300 was increased in ERKO and DERKO but not in BERKO male mice , resulting in an increased O00300 / O14788 ratio . Furthermore , serum levels of P05231 and tartrate-resistant acid phosphatase 5b ( TRAP 5b ) were decreased in ERKO and DERKO , but not in BERKO male mice . These results demonstrate that ER alpha , but not ER beta , is involved in the regulation of the vertebral O00300 / O14788 ratio , serum levels of P05231 and TRAP 5b in male mice . Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 gene deletion and DB01997 were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety . 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 ( IKK ) activity was investigated using purified recombinant O15111 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 , O14920 , or constitutively active O15111 and O14920 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 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 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 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 . Aflatoxin B1 induces Src phosphorylation and stimulates lung cancer cell migration . AflatoxinB1 ( AFB1 ) is well known as a potent carcinogen . Epidemiological studies have shown an association between AFB1 exposure and lung cancer in humans . AFB1 can induce the mutations of genes such as tumor suppressor p53 through its metabolite AFB1-8,9-exo-epoxide , which acts as a mutagen to react with DNA . In addition , recent study demonstrates AFB1 positively regulates type I insulin-like growth factor receptor ( IGF-IR ) signaling in hepatoma cells . The current study aims to determine the effects of AFB1 on Src kinase and insulin receptor substrate ( P41252 ) in lung cancer cells and the effects of AFB1 on lung cancer cell migration . To this end , the effects of AFB1 on P41252 expression , Src , Akt , and P29323 phosphorylation were measured by Western blot analysis . The migration of lung cancer cells was detected by wound-healing assay . AFB1 downregulates P35568 but paradoxically upregulates Q9Y4H2 through positive regulation of the stability of Q9Y4H2 and the proteasomal degradation of P35568 in lung cancer cell lines A549 and P08709 -1 . In addition , AFB1 induces Src , Akt , and P27361 /2 phosphorylation . Treatment of lung cancer cells with Src inhibitor saracatinib abrogates AFB1-induced Q9Y4H2 accumulation . Moreover , AFB1 stimulates lung cancer cell migration , which can be inhibited by saracatinib . We conclude that AFB1 may upregulate Q9Y4H2 and stimulate lung cancer cell migration through Src .

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The effects of five alkaloids from Bulbus Fritillariae on the concentration of DB02527 in P29320 cells transfected with muscarinic M(2) receptor plasmid . The aim of this study was to investigate the effects of five alkaloids , namely verticine , verticinone , imperialine , imperialine-3beta-D-glucoside , and puqietinone , purified from Bulbus Fritillariae and used as an antitussive drug in traditional Chinese medicine , on their antimuscarinic M(2) function and the DB02527 level of P29320 cells transfected with muscarinic M(2) receptor plasmid . By transfecting the P29320 cells with the method of calcium phosphate co-precipitation and screening with G418 , the cells stably expressing M(2) receptor were identified . The expression of M(2) receptor in P29320 cells was confirmed by both RT-PCR and western blot . The DB02527 level in the treated cells was analyzed with RIA method ( (125)I- DB02527 P10721 ) . And the results suggested that the five alkaloids could significantly elevate the DB02527 concentration in the P29320 cells transfected with muscarinic M(2) receptor plasmid ( p < 0.01 ) . Potent and selective inhibition of human nitric oxide synthases . Inhibition by non-amino acid isothioureas . DB02539 was a potent competitive inhibitor of human nitric oxide synthase ( NOS ) , with Ki values of 17 , 36 , and 29 nM for the inducible ( i ) , endothelial ( e ) , and neuronal ( n ) isozymes , respectively . Unlike some potent inhibitors of NOS , no time dependence was observed . DB02539 was not a detectable substrate for P29474 . DB02539 was also a potent inhibitor of mouse P35228 ( Ki value of 5.2 nM ) , and its binding perturbed the spectrum of P35228 consistent with its altering the environment of the bound heme . The optimum binding of S-ethyl- and S-isopropylisothiourea relative to 70 other analogs suggested that these alkyl substitutions fit into a small hydrophobic pocket . Most isothioureas were 2-6-fold selective for the human P35228 ( Ki for P35228 versus Ki for P29474 ) , with one being 19-fold selective . The cyclized mimics of S-ethylisothiourea , 2-NH2-thiazoline , and 2-NH2-thiazole , were also competitive inhibitors of human NOS . A third structural class of inhibitors , bisisothioureas , were , in general , the most selective in their inhibition of human P35228 . S,S'-(1,3-Phenylenebis(1,2-ethanediyl))bisisothiourea was 190-fold selective ( Ki value of 0.047 microM against P35228 versus 9.0 microM against P29474 ) . These results demonstrate that potent and selective inhibition of human NOS isozymes is achievable . P49796 ( P49796 ) inhibits Gbeta1gamma 2-induced inositol phosphate production , mitogen-activated protein kinase activation , and Akt activation . P49796 ( P49796 ) enhances the intrinsic rate at which Galpha(i) and Galpha(q) hydrolyze GTP to GDP , thereby limiting the duration in which GTP-Galpha(i) and GTP-Galpha(q) can activate effectors . Since GDP-Galpha subunits rapidly combine with free Gbetagamma subunits to reform inactive heterotrimeric G-proteins , P49796 and other Q99697 proteins may also reduce the amount of Gbetagamma subunits available for effector interactions . Although P49758 , P49802 , and O94810 bind Gbeta(5) in the absence of a Ggamma subunit , Q99697 proteins are not known to directly influence Gbetagamma signaling . Here we show that P49796 binds Gbeta(1)gamma(2) subunits and limits their ability to trigger the production of inositol phosphates and the activation of Akt and mitogen-activated protein kinase . Co-expression of P49796 with Gbeta(1)gamma(2) inhibits Gbeta(1)gamma(2)-induced inositol phosphate production and Akt activation in COS-7 cells and mitogen-activated protein kinase activation in P29320 293 cells . The inhibition of Gbeta(1)gamma(2) signaling does not require an intact Q99697 domain but depends upon two regions in P49796 located between acids 313 and 390 and between 391 and 458 . Several other Q99697 proteins do not affect Gbeta(1)gamma(2) signaling in these assays . Consistent with the in vivo results , P49796 inhibits Gbetagamma-mediated activation of phospholipase Cbeta in vitro . Thus , P49796 may limit Gbetagamma signaling not only by virtue of its P20936 activity for Galpha subunits , but also by directly interfering with the activation of effectors . Stimulation of cloned human glucagon-like peptide 1 receptor expressed in P29320 293 cells induces DB02527 -dependent activation of calcium-induced calcium release . The actions of glucagon-like peptide-1(7-36)amide ( P0C6A0 (7-36)amide ) on cellular signalling were studied in human embryonal kidney 293 ( P29320 293 ) cells stably transfected with the cloned human P43220 . The cloned P43220 showed a single high-affinity binding site ( Kd = 0.76 nM ) . Binding of P0C6A0 (7-36)amide stimulated DB02527 production in a dose-dependent manner ( EC50 = 0.015 nM ) and caused an increase in the intracellular free Ca2+ concentration ( [Ca2+]i ) . The latter effect reflected Ca(2+)-induced Ca2+ release and was suppressed by ryanodine . We propose that the ability of P0C6A0 (7-36)amide to increase [Ca2+]i results from sensitization of the ryanodine receptors by a protein kinase A dependent mechanism . Immunoaffinity purification of the functional 20S proteasome from human cells via transient overexpression of specific proteasome subunits . The proteasome is a multi-subunit proteolytic complex that plays a central role in protein degradation in all eukaryotic cells . It regulates many vital cellular processes therefore its dysfunction can lead to various pathologies including cancer and neurodegeneration . Isolation of enzymatically active proteasomes is a key step to the successful study of the proteasome regulation and functions . Here we describe a simple and efficient protocol for immunoaffinity purification of the functional 20S proteasomes from human P29320 293T cells after transient overexpression of specific proteasome subunits tagged with 3xFLAG . To construct 3xFLAG-fusion proteins , DNA sequences encoding the 20S proteasome subunits P28074 , P28066 , and P25788 were cloned into mammalian expression vector pIRES-hrGFP-1a . The corresponding recombinant proteins P28074 -3xFLAG , P28066 -3xFLAG , or P25788 -3xFLAG were transiently overexpressed in human P29320 293T cells and were shown to be partially incorporated into the intact proteasome complexes . 20S proteasomes were immunoprecipitated from P29320 293T cell extracts under mild conditions using antibodies against FLAG peptide . Isolation of highly purified 20S proteasomes were confirmed by SDS-PAGE and Western blotting using antibodies against different proteasome subunits . Affinity purified 20S proteasomes were shown to possess chymotrypsin- and trypsin-like peptidase activities confirming their functionality . This simple single-step affinity method of the 20S proteasome purification can be instrumental to subsequent functional studies of proteasomes in human cells . Brominated cyclodipeptides from the marine sponge Geodia barretti as selective 5-HT ligands . The brominated cyclodipeptides barettin ( cyclo[(6-bromo-8-entryptophan)arginine] ) and 8,9-dihydrobarettin ( cyclo[(6-bromotryptophan)arginine] ) isolated from the marine sponge Geodia barretti have previously been shown to inhibit settlement of barnacle larvae in a dose-dependent manner in concentrations ranging from 0.5 to 25 microM . To further establish the molecular target and mode of action of these compounds , we investigated their affinity to human serotonin receptors . The tryptophan residue in the barettins resembles that of endogenous serotonin [ 5-hydroxytryptamine ] . A selection of human serotonin receptors , including representatives from all subfamilies ( 1-7 ) , were transfected into P29320 -293 cells . Barettin selectively interacted with the serotonin receptors 5- Q13049 , P28335 , and Q13639 at concentrations close to that of endogenous serotonin , with the corresponding Ki values being 1.93 , 0.34 , and 1.91 microM , respectively . 8,9-Dihydrobarettin interacted exclusively with the P28335 receptor with a Ki value of 4.63 microM ; it failed to show affinity to 5- Q13049 and Q13639 , indicating that the double bond between the tryptophan and arginine residue plays an important role in the interaction with the receptor proteins . The in vitro pharmacological profile of DB05655 , a selective 5-HT(4) receptor agonist with high intrinsic activity . The in vitro pharmacological profile of DB05655 , a novel , selective 5-HT(4) receptor agonist , was compared to that of clinically efficacious gastroprokinetic 5-HT(4) receptor agonists . DB05655 produced an elevation of cyclic adenosine monophosphate in human embryonic kidney 293 cells expressing the human recombinant 5-HT(4(c)) ( h5-HT(4(c)) ) receptor ( pEC(50) = 8.3 ) and 5-HT(4) receptor-mediated relaxation of the rat esophagus ( pEC(50) = 7.9 ) and contraction of the guinea pig colon ( pEC(50) = 7.9 ) . In all in vitro assays , DB05655 was a high intrinsic activity agonist , unlike tegaserod , mosapride , and cisapride which , in the majority of test systems , had lower intrinsic activity . DB05655 had high affinity ( pK ( i ) = 7.7 ) and selectivity ( > or =25-fold ) for h5-HT(4(c)) receptors over other biogenic amine receptors . DB05655 was > 500-fold selective over other 5-HT receptors ( including h5-HT(2B) and h5-HT(3A) ) and , at 3 microM , had no effect on human ether-à-go-go-related gene K+ channels . In conclusion , DB05655 is a selective 5-HT(4) receptor agonist in vitro . The high intrinsic activity and preferential binding of DB05655 to Q13639 over other 5-HT receptors may result in an improved clinical profile for the treatment of gastrointestinal disorders of reduced motility . P06401 modulator DB05366 down-regulates proliferative cell nuclear antigen and Bcl-2 protein expression and up-regulates caspase-3 and poly ( adenosine 5'-diphosphate-ribose ) polymerase expression in cultured human uterine leiomyoma cells . The present study was conducted to evaluate the effects of the progesterone receptor modulator DB05366 on proliferative activity and apoptosis in cultured human uterine leiomyoma cells . Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10 % fetal bovine serum for 120 h and then stepped down to serum-free conditions for 12 , 24 , 48 , and 96 h in the absence or presence of graded concentrations of DB05366 ( 10(-9) , 10(-8) , 10(-7) , and 10(-6) M ) . The number of viable cultured leiomyoma cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazodium bromide assay . P12004 ( P12004 ) expression was evaluated by immunocytochemistry and Western blot analysis . Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling ( TUNEL ) assay . P42574 , cleaved poly(ADP-ribose) polymerase ( PARP ) , and Bcl-2 expression were assessed by Western blot analysis . Compared with untreated control cultures , treatment with DB05366 decreased the number of viable cultured leiomyoma cells and the P12004 -positive rate in those cells and increased the TUNEL-positive rate in cultured leiomyoma cells in a dose-dependent manner . Western blot analysis revealed that treatment with DB05366 significantly decreased the expression of P12004 and Bcl-2 protein and increased the expression of cleaved caspase-3 and cleaved PARP in a dose-dependent manner compared with untreated control cultures . These results suggest that DB05366 inhibits the proliferation of cultured leiomyoma cells by down-regulating P12004 expression and induces apoptosis by up-regulating cleaved caspase-3 and PARP expression and down-regulating Bcl-2 protein expression in those cells . DB09045 : the newest P43220 agonist for the management of type 2 diabetes . OBJECTIVE : To review the pharmacology , pharmacokinetics , safety , and efficacy of the glucagon-like peptide-1 receptor agonist ( P0C6A0 RA ) , dulaglutide , in the treatment of type 2 diabetes mellitus ( T2D ) . DATA SOURCES : A PubMed search was completed to identify publications from 1947 to October 2014 using the search terms dulaglutide and LY2189265 . References were reviewed to identify additional resources . STUDY SELECTION AND DATA EXTRACTION : Articles were included if they evaluated the pharmacology , pharmacokinetics , safety , or efficacy of dulaglutide . DATA SYNTHESIS : DB09045 reduces both glycosylated hemoglobin ( A1C ) and weight by stimulating insulin secretion and suppressing glucagon in a glucose-dependent manner , delaying gastric emptying , and promoting satiety . DB09045 consists of 2 P0C6A0 analogues that have been modified to make it a long-acting , once-weekly agent . DB09045 has been studied as monotherapy and in combination with metformin , glimepiride , pioglitazone , and insulin lispro . It has demonstrated superior A1C reduction compared with placebo , metformin , insulin glargine , sitagliptin , and twice-daily exenatide . It demonstrated noninferiority in A1C reduction to liraglutide . DB09045 changed A1C by -0.78 % to -1.51 % , and it changed weight by -0.35 kg to -3.03 kg . The most common adverse effects in clinical studies were nausea , vomiting , and diarrhea . CONCLUSIONS : DB09045 is the fifth P0C6A0 RA approved for T2D in the United States . It is an attractive option because it is dosed once-weekly , provides A1C lowering similar to liraglutide , weight reduction similar to exenatide , and has an adverse effect profile similar to exenatide and liraglutide . A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors . P10721 or alpha-platelet-derived growth factor receptor ( alpha- P09619 ) activating mutations are the pathogenic mechanisms that characterize gastrointestinal stromal tumors ( GIST ) . Despite excellent responses to imatinib mesylate ( IM ) , patients are relapsing . We developed an IM-resistant GIST cell line ( GIST-R ) from the IM-sensitive GIST882 cell line ( GIST-S ) by growing these cells in IM . Gene expression profiling ( GEP ) of GIST-S , GIST-R cells and two IM resistant GIST patients demonstrated that P10721 is downregulated implying a major role in IM resistance . Instead , GIST-R cells have acquired IM resistance by overexpressing the oncogenic receptor tyrosine kinase - P30530 - in a ' kinase switch ' . Further , the two IM resistant GIST patients express P30530 and not c-Kit , seen by immunohistochemistry ( IHC ) . Real time reverse transcriptase-polymerase chain reaction and Western blotting of the GIST-S and GIST-R cells confirmed the switch from Kit to P30530 . In GIST-R , P30530 is tyrosine phosphorylated and its ligand growth-arrest-specific gene 6 is overexpressed implying autocrine activation . The kinase switch is associated with a morphological change from spindle to epithelioid . Molecular modeling of the kinase domain of mutant c-Kit ( V654A ) and P30530 showed no binding to IM but efficient binding to DB05216 , a novel c-Kit/ P30530 kinase inhibitor . DB05216 synergizes with docetaxel ( taxotere ) and is cytotoxic to GIST cells . P37231 ligand prevents the development of chronic pancreatitis through modulating NF-kappaB-dependent proinflammatory cytokine production and pancreatic stellate cell activation . PURPOSE : Thiazolidinedione derivatives ( TZDs ) are known to be ligands of peroxisome proliferator-activated receptor gamma ( PPARgamma ) . In this study , we investigated the effect of a TZD , troglitazone , on inflammation and fibrogenesis in the pancreas of an experimental model of chronic pancreatitis . MATERIAL AND METHODS : Male WBN/Kob rats with spontaneous chronic pancreatitis were fed rat chow containing 0.2 % troglitazone from 1 to 4 months of age . Immunohistochemical studies of rat pancreas were carried out with monoclonal mouse antibody against human alpha-smooth muscle actin ( alpha-SMA ) or rabbit polyclonal antibody against collagen type I , collagen type III , or fibronectin . Cytokine production was measured by enzyme-linked immunosorbent assay . The inhibitory action of troglitazone on nuclear factor-kappaB ( NF-kappaB ) binding activity in activated macrophages was also investigated . RESULTS : Long-term administration of troglitazone reduced inflammatory cell infiltration and fibrosis in the pancreas of WBN/Kob rats , and expression of alpha-SMA , procollagen I , III , and fibronectin was significantly reduced by troglitazone . The increase in P01375 production by activated macrophages was significantly decreased by troglitazone . Peritoneal macrophages isolated from WBN/Kob rats produced a large amount of P01375 , whereas those from troglitazone-treated WBN/Kob rats produced only a marginal amount of P01375 . Lipopolysaccharide-induced NF-kappaB binding activity in peritoneal macrophages was also significantly reduced by troglitazone . CONCLUSIONS : DB00197 prevented the progression of chronic pancreatitis via inhibition of Q13201 synthesis and proinflammatory cytokine production mediated by the inhibition of NF-kappaB activity . A reporter gene assay for screening of DB05876 subtype selective inhibitors . Phosphodiesterase ( PDE ) constitutes a superfamily of enzymes that catalyze the hydrolysis of DB02527 and cGMP into their corresponding monophosphates and play an important role in diverse physiological functions . The present study provides a process for identifying DB05876 subtypes selective inhibitors using a reporter gene assay . Stable recombinant P29320 -293 cell lines expressing high levels of PDE4A4B , PDE4B2A , and PDE4D3 subtypes individually were generated . Transient transfection of pCRE-Luc plasmid , harboring luciferase reporter gene under the control of DB02527 response element ( CRE ) -binding sequence , into these stable recombinant cell lines followed by treatment with DB05876 inhibitor , resulted in a dose dependent increase in luciferase activity . This methods provide a novel , simple and sensitive assay for high throughput screening of DB05876 subtype selective inhibitors for treatment of asthma and P48444 . Cordycepin suppresses P01375 -α-induced NF-κB activation by reducing p65 transcriptional activity , inhibiting IκBα phosphorylation , and blocking IKKγ ubiquitination . Cordycepin is reported to participate in multiple pharmacological activities including anti-tumor and anti-inflammation , and is involved in the regulation of NF-κB signaling pathway . However , the detailed molecular mechanism of cordycepin in suppression of NF-κB signaling pathway remains ambiguous . In this study , we first analyzed the effect of cordycepin on NF-κB activity in P29320 -293T cells , and found that cordycepin resulted in a dose-dependent reduction in P01375 -α-induced NF-κB activation . Although cordycepin did not block P01375 -α-induced nuclear translocation of p65 , high concentration of cordycepin reduced the DNA-binding and transcriptional activities of NF-κB . Moreover , cordycepin also inhibited IκBα phosphorylation so as to suppress the degradation of IκBα . Further investigation revealed that cordycepin suppressed IKKs-mediated NF-κB activation and inhibited the ubiquitination of IKKγ . In conclusion , cordycepin effectively inhibits NF-κB signaling through suppressing the activities of NF-κB , IκB and IKK . Thus , cordycepin may provide some potential therapeutic application in inflammation-associated disorders and cancer . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . DB00188 -resistant myeloma cell lines : a role for mutated P28074 in preventing the accumulation of unfolded proteins and fatal ER stress . DB00188 is an effective agent for treating multiple myeloma ( MM ) . To investigate the underlying mechanisms associated with acquired resistance to this agent , we established two bortezomib-resistant MM cell lines , KMS-11/BTZ and OPM-2/BTZ , the 50 % inhibitory concentration values of which were respectively 24.7- and 16.6-fold higher than their parental cell lines . No activation of caspase and BH3-only proteins such as Noxa was noted in bortezomib-resistant cells after exposure to the drug . The accumulation of polyubiquitinated proteins was reduced in bortezomib-resistant cells compared with the parental cells , associated with avoidance of catastrophic ER stress as assessed by downregulation of P35638 expression . These resistant MM cells have a unique point mutation , G322A , in the gene encoding the proteasome beta5 subunit ( P28074 ) , likely resulting in conformational changes to the bortezomib-binding pocket of this subunit . KMS-11 parental cells transfected to express mutated P28074 also showed reduced bortezomib-induced apoptosis compared with those expressing wild-type P28074 or the parental cells . Expression of mutated P28074 was associated with the prevention of the accumulation of unfolded proteins . Thus , a fraction of MM cells may acquire bortezomib resistance by suppressing apoptotic signals through the inhibition of unfolded protein accumulation and subsequent excessive ER stress by a mutation of the P28074 gene . P37268 inhibitors : An update on the search for new antihyperlipidemic and antiatherosclerotic agents . Atherosclerosis and related heart disease is strongly associated with elevated blood levels of total ( and LDL ) cholesterol . Due to the widespread incidence as well as severity of this pathological condition , major efforts have been made for the discovery and development of hypocholesteroleamic agents . In the past few decades , P04035 inhibitors ( statins ) are being extensively used as lipid lowering drugs . These agents act predominantly by inhibiting the enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase ( HMGR ) that is the rate limiting step of cholesterol biosynthesis . Both the success as well as drawbacks of HMGRIs , have led to the investigation and design of inhibitors of other ( downstream ) enzymes involved in the multistep cholesterol biosynthetic pathway . One such class of agents consists of the squalene sythase inhibitors which act at the first and solely committed step towards the biosynthesis of the cholesterol nucleus . This target is considered not to interfere with the biosynthesis of other biologically important molecules and thus a better side-effect profile is expected for these inhibitors . Several classes of squalene synthase inhibitors ( SQSIs ) , such as substrate or transition-state analogues , zaragozic acids or 2,8- dioxabicyclo[3.2.1]octane derivatives , dicarboxylic acid and quinuclidine derivatives , 4,1-benzoxazepine as well as substituted morpholine derivatives , have been studied as potent inhibitors of squalene synthase . So far only one benzoxazepine derivative ( DB05317 ) has been evaluated in advanced clinical trials . In this article we review the up to date research and literature on the therapeutic potential of this relatively new class of compounds , the drug discovery efforts towards the development of active squalene synthase inhibitors , their activity profile and effectiveness , as well as their structure-activity relationships . Opposite effect of Hsp90α and Hsp90β on P29474 ability to produce nitric oxide or superoxide anion in human embryonic kidney cells . Heat shock protein 90 subfamily is composed by two cytosolic isoforms known as Hsp90α and Hsp90β . Endothelial nitric oxide synthase ( P29474 ) is regulated by Hsp90 , however the specific role of each Hsp90 isoform on NO production has not been established . This study was designed to evaluate the effect of Hsp90α and Hsp90β over-expression on P29474 /NO pathway . Rat Hsp90α and Hsp90β were cloned into pcDNA3.1(+) and transfected in human embryonic kidney cells ( P29320 -293 ) . Hsp90α and Hsp90β transfection was corroborated by Western blot analysis and their effect on NO production ( NO(2)/NO(3) ) , P29474 protein and its phosphorylation at Ser1177 and Thr495 , as well as Akt/ P31749 Ser473 phosphorylation was determined . The interaction of Hsp90α and Hsp90β with P29474 and the dimer/monomer ratio of Hsp90 , as well as O(2)(-) generation were also assessed . After transfection , Hsp90α and Hsp90β levels were significantly increased in P29320 -293 cells . The Hsp90α over-expression induced a significant increase in NO(2)/NO(3) levels , an effect that was associated with increased phosphorylation of P29474 DB00133 1177 and Akt/ P31749 Ser473 , as well as with a greater Hsp90α dimerization . Noteworthy , pcHsp90β transfection reduced significantly NO(2)/NO(3) and increased O(2)(-) generation . These effects were associated with a reduction of P29474 dimeric conformation , increased P29474 Thr495 phosphorylation , reduced Akt/ P31749 phosphorylation , and by a greater amount of monomeric Hsp90β conformation . These data show for first time that Hsp90α and Hsp90β differentially modulate NO and O(2)(-) generation by P29474 through promoting changes in P29474 conformation and phosphorylation state . P08246 inhibitors as treatment for P48444 . Chronic obstructive pulmonary disease , characterised by a slowly progressive , irreversible airways limitation , is a major worldwide cause of chronic morbidity and mortality . The imbalance between human neutrophil elastase and endogenous antiproteases may cause excess human neutrophil elastase in pulmonary tissues , which may be considered a major pathogenic factor in chronic obstructive pulmonary disease . Great effort has been devoted to finding a method to restore the balance , resulting in the discovery of potent two-typed small-molecular-weight human neutrophil elastase inhibitors . In the application of chronic obstructive pulmonary disease therapy , the human neutrophil elastase inhibitors mainly focused upon include ONO-5046 , MR-889 , L-694,458 , CE-1037 , GW-311616 and TEI-8362 as the acyl-enzyme inhibitors ; and DB03925 , AE-3763 , FK-706 , ICI-200,880 , ZD-0892 and ZD-8321 as the transition-state inhibitors . In this review , various problems that remain to be solved in the clinical use of human neutrophil elastase inhibitors are discussed . The time course of Akt and P29323 activation on P98170 expression in P29320 293 cell line . The cell growth is controlled by the interaction of survival and cell growth arrest pathways as well as apoptosis mechanisms which determine the outcome of cell faith as proliferation or apoptosis . In this study , we have studied the activity of survival pathways , i.e. , Akt and P27361 /2 with regard to P98170 ( inhibitor of apoptosis ) in serum starved and stimulated conditions . The P29320 -293 cells were cultured in RPMI + 10 % FBS . The cells were serum starved by switching to medium with 1 % FBS for 24 h and serum stimulated by changing the medium to 10 % FBS following serum starvation . The expression of p-Akt , p- P29323 , Akt , P29323 and P98170 was studied in various time points using western blot . The apoptosis was evaluated by DNA condensation using Hoechst 33258 and P42574 assay . In serum starved condition expression of p-Akt and P98170 is very low . Serum stimulation increases p-Akt and p- P29323 within 5 min and sustains a high level for 30 min . The expression of total Akt and P27361 /2 has not changed significantly for 24 h . P98170 expression starts at 6 h after serum stimulation , reaches to maximum level at 12 h and decreases to baseline within 24 h . Furthermore , serum starvation for 24 h does not induced apoptosis and DNA condensation . Taken together , the results indicate that serum activates Akt and P29323 pathways earlier than P98170 expression . Furthermore , P98170 expression is low in serum starvation unlike p- P29323 which suggests a survival role for P29323 in serums starvation . The expression pattern of P98170 indicates induction by Akt and/or P29323 activation which requires further studies . Neuroprotective gene expression profiles in ischemic cortical cultures preconditioned with DB01277 or P09038 . The mechanisms underlying growth factor preconditioning of neurons are only partially elucidated , and no studies have been conducted in this area using a gene profiling approach . We used cDNA microarrays to compare the transcriptional profiles of cells preconditioned either with insulin-like growth factor I ( DB01277 ) or basic fibroblast growth factor ( P09038 ) , to identify differentially regulated genes that may function in growth factor signaling , response to oxygen-glucose deprivation ( OGD ) , and most importantly , cell survival . Primary rat cortical cultures were treated with P09038 or DB01277 for 2 , 24 , or 24 h followed by OGD for 90 min , and compared with cells that were subject to OGD without growth factor pretreatment . Although the majority of surveyed genes were unchanged in all experimental treatments , 175 genes ( 10 % of the cDNAs on the chip ) were found to be differentially regulated in at least one of the treatment conditions . Hierarchical clustering of these 175 genes was used to identify four expression clusters : DB01277 regulated , P09038 regulated , OGD regulated , and putative neuroprotective genes . Further analysis using realtime RT-PCR confirmed that we had identified genes that are regulated by single growth factors , as well as several more that are co-regulated by both DB01277 and P09038 . These genes can influence neuronal survival by affecting diverse pathways such as growth factor signal transduction ( P16070 , Q99075 , Q16828 , Q12929 , P17936 ) , DNA repair and transcription ( Q92949 ) , metabolic homeostasis ( P20936 , P34897 ) , cytoskeletal stability ( P26038 , P10636 ) and cholesterol biosynthesis ( P37268 , P14324 ) .

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P10275 inducing bladder cancer progression by promoting an epithelial-mesenchymal transition . The study investigated the role of androgen receptor ( AR ) as a potential target for the treatment of bladder cancer in regulating epithelial-mesenchymal transition or transformation ( EMT ) . Cell proliferation , and migration capacity were determined in bladder cancer T24 cells treated with small interfering RNA directed against AR , and expression levels of P12830 , β-catenin and N- cadherin were assessed using quantitative reverse transcription PCR ( qRT-PCR ) . Tumour cell growth was evaluated in vivo in T24 tumour-bearing nude mice receiving electroporation-assisted administration of anti-AR small interfering RNA . It was found that low AR expression decreased proliferation and migration of bladder cancer cells . In vivo experiments showed that silencing AR expression significantly suppressed AR-positive bladder tumour growth with decreased cell proliferation . Low AR level of T24 bladder cancer cells treated with DB01541 ( DB02901 ) decreased expression of P12830 , β-catenin and P19022 expression , indicating a strong sensitivity to the EMT and In cells with low AR content , TGF-β induced down-regulation of P12830 and β-catenin . It is concluded that suppression of AR expression decreased the production of TGF-β , inhibiting EMT and bladder cancer cell growth in vitro and in vivo , implying that its use might be a potential therapeutic target for the treatment of bladder cancer . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients . Increased plasma thyroid hormone concentrations in P01130 deficient mice may be explained by inhibition of aryl hydrocarbon receptor-dependent expression of hepatic UDP-glucuronosyltransferases . BACKGROUND : Overexpression of P36956 causes a repression of hepatic genes involved in phase II metabolism . In P01130 deficient ( P01130 (-/-) ) mice , active levels of P36956 in the liver are increased . We investigated the hypothesis that P01130 (-/-) mice have increased concentrations of thyroid hormones in plasma due to a reduced hepatic glucuronidation . METHODS : Female P01130 (-/-) and wild-type mice were used to study the effect of the P01130 (-/-) genotype on thyroid hormone metabolism . RESULTS : P01130 (-/-) mice had a higher concentration of nuclear P36956 , higher concentrations of thyroxine and triiodothyronine in plasma , a lower expression of relevant P22309 isoforms , reduced activities of pNP- P78381 , T(3)- P78381 and T(4)- P78381 and a lower mRNA and protein concentration of P35869 in the liver than wild-type mice ( P < 0.05 ) . Plasma concentration of DB00024 , mRNA concentrations of various genes involved in thyroid hormone synthesis in the thyroid , activity of deiodinase and mRNA concentrations of two thyroid hormone responsive genes , P22680 and Na(+)/K(+)-ATPase , in the liver did not differ between both genotypes . CONCLUSIONS : This study shows that P01130 (-/-) mice have increased concentrations of thyroid hormones in plasma . This effect is probably due to an inhibition of thyroid hormone glucuronidation , which might be caused by down-regulation of P78381 genes due to a reduced expression of P35869 . However , with respect to plasma DB00024 concentration and expression of thyroid hormone responsive genes no overt hyperthyroidism was detected . GENERAL SIGNIFICANCE : P01130 deficiency leads to a reduced glucuronidation of thyroid hormones in the liver which causes a moderate increase of plasma thyroid hormone concentrations . Selection of suitable reference genes for quantitative real-time polymerase chain reaction in human meningiomas and arachnoidea . FINDINGS : At first 32 housekeeping genes were analyzed in six randomly chosen meningiomas , brain and dura mater using geNorm , NormFinder , Bestkeeper-1 software and the comparative ΔCt method . Reference genes were ranked according to an integration tool for analyzing reference genes expression based on those four algorithms . Eight highest ranked reference genes ( O15234 , Q14232 , O15397 , P49406 , P00558 , POP4 , P62937 , and P61513 ) plus P04406 and P60709 were then analyzed in 35 meningiomas , arachnoidea , dura mater and normal brain . NormFinder and Bestkeeper-1 identified P61513 as the most stable expressed gene in meningiomas and their normal control tissue . NormFinder also determined the best combination of genes : P61513 and Q14232 . Commonly used reference genes P04406 and P60709 were considered least stable genes . The critical influence of reference genes on qPCR data analysis is shown for P15692 transcription patterns . BACKGROUND : In meningiomas quantitative real-time reverse transcription-polymerase chain reaction ( qPCR ) is most frequently used for accurate determination of gene expression using various reference genes . Although meningiomas are a heterogeneous group of tissue , no data have been reported to validate reference genes for meningiomas and their control tissues . CONCLUSIONS : P61513 is the optimal single reference gene for normalization of gene expression in meningiomas and their control tissues , although the use of the combination of P61513 and Q14232 would provide more stable results . The O60656 enzyme is a peroxisome proliferator-activated receptor alpha and gamma target gene . Peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma are ligand-activated transcription factors belonging to the nuclear receptor family . Q07869 alpha mediates the hypolipidemic action of the fibrates , whereas Q07869 gamma is a receptor for the antidiabetic glitazones . In the present study , the UDP-glucuronosyltransferase ( P78381 ) 1A9 enzyme is identified as a Q07869 alpha and Q07869 gamma target gene . UGTs catalyze the glucuronidation reaction , which is a major pathway in the catabolism and elimination of numerous endo- and xenobiotics . Among the P22309 family enzymes , O60656 metabolizes endogenous compounds , including catecholestrogens , and xenobiotics , such as fibrates and to a lesser extent troglitazone . Treatment of human hepatocytes and macrophages and murine adipocytes with activators of Q07869 alpha or Q07869 gamma resulted in an enhanced O60656 expression and activity . In addition , disruption of the Q07869 alpha gene in mice completely abolished the Q07869 alpha agonist-induced O60656 mRNA and activity levels . A Q07869 response element was identified in the promoter of O60656 at positions -719 to -706 bp by transient transfection and electromobility shift assays . Considering the role of O60656 in catecholestrogen metabolism , Q07869 alpha and Q07869 gamma activation may contribute to the protection against genotoxic catecholestrogens by stimulating their inactivation in glucuronide derivatives . Furthermore , since O60656 is involved in the catabolism of fibrates , these results suggest that Q07869 alpha and Q07869 gamma may control the intracellular level of active fibrates . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 , CYP2B1/2 , P05181 , CYP3A , catechol-O-methyltransferase ( P21964 ) and microsomal glutathione transferase 1 ( P10620 ) . All but actin and soluble P21964 were positively detected at ≥1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 . The binding correlated well with inhibition of CYP activities , except for P05181 whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 antibodies showed a significant reduction in binding at ≥1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults . DB04873 ( SB 207266 ) , a selective Q13639 receptor antagonist , reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle . The aim of this study is to evaluate the potency of piboserod ( SB 207266 ) , a selective 5-HT(4) receptor antagonist , at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin ( 5-HT ) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations ( O43281 ) . Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and O43281 ( 20 Hz , 1 ms duration at 300 mA for 5 s ) was applied continuously at 1 min intervals . After stabilization of the O43281 -induced contractions , concentration-response curves to 5-HT ( 0.1 nM-100 microM ) were constructed in the absence or presence of 1 or 100 nM of piboserod . The experiments were performed in the presence of methysergide ( 1 microM ) and ondansetron ( 3 microM ) to block 5HT(1)/5HT(2) and 5-HT(3) receptors , respectively . 5-HT potentiated the contractile responses to O43281 of human bladder strips in a concentration-dependent manner , with a maximum mean of 60.0+/-19.9 % of the basal O43281 -evoked contractions . DB04873 did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to O43281 . In presence of 1 and 100 nM of piboserod , the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7 % , respectively . A mean apparent antagonist dissociation constant value ( K(B) ) of 0.56+/-0.09 nM was determined . These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally-mediated contractions of isolated human bladder strips . Therefore , the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder . P00734 kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation . We have shown that prothrombin kringle-2 ( pKr-2 ) , a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro . However , little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic ( DA ) neurons in vivo . To address this question , pKr-2 was injected into the rat substantia nigra ( SN ) . Tyrosine hydroxylase ( TH ) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2 . In parallel , pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry . Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-2 ( P35354 ) and several proinflammatory cytokines . The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride , and the P35354 inhibitor DuP-697 . P27361 /2 , c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection , and localized within microglia . Inhibition of these kinases led to attenuation of mRNA expression of P35228 , P35354 and several proinflammatory cytokines , and rescue of DA neurons in the SN . Intriguingly , following treatment with pKr-2 in vitro , neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia , but not microglia-free neuron-enriched mesencephalic cultures , indicating that microglia are required for pKr-2 neurotoxicity . Our results strongly suggest that microglia activated by endogenous compound(s) , such as pKr-2 , are implicated in the DA neuronal cell death in the SN . Genetic variation modifies risk for neurodegeneration based on biomarker status . BACKGROUND : While a great deal of work has gone into understanding the relationship between Cerebrospinal fluid ( P04141 ) biomarkers , brain atrophy , and disease progression , less work has attempted to investigate how genetic variation modifies these relationships . The goal of this study was two-fold . First , we sought to identify high-risk vs. low-risk individuals based on their P04141 tau and Aβ load and characterize these individuals with regard to brain atrophy in an AD-relevant region of interest . Next , we sought to identify genetic variants that modified the relationship between biomarker classification and neurodegeneration . METHODS : Participants were categorized based on established cut-points for biomarker positivity . Mixed model regression was used to quantify longitudinal change in the left inferior lateral ventricle . Interaction analyses between single nucleotide polymorphisms ( SNPs ) and biomarker group status were performed using a genome wide association study ( GWAS ) approach . Correction for multiple comparisons was performed using the Bonferroni procedure . RESULTS : One intergenic SNP ( rs4866650 ) and one SNP within the O15269 gene ( rs7849530 ) modified the association between amyloid positivity and neurodegeneration . A transcript variant of WDR11-AS1 gene ( rs12261764 ) modified the association between tau positivity and neurodegeneration . These effects were consistent across the two sub-datasets and explained approximately 3 % of variance in ventricular dilation . One additional SNP ( rs6887649 ) modified the association between amyloid positivity and baseline ventricular volume , but was not observed consistently across the sub-datasets . CONCLUSIONS : Genetic variation modifies the association between AD biomarkers and neurodegeneration . Genes that regulate the molecular response in the brain to oxidative stress may be particularly relevant to neural vulnerability to the damaging effects of amyloid-β . Activation of intestinal peroxisome proliferator-activated receptor-α increases high-density lipoprotein production . AIMS : Peroxisome proliferator-activated receptor ( Q07869 ) -α is a transcription factor controlling lipid metabolism in liver , heart , muscle , and macrophages . Peroxisome proliferator-activated receptor-α activation increases plasma HDL cholesterol and exerts hypotriglyceridaemic actions via the liver . However , the intestine expresses Q07869 -α , produces HDL and chylomicrons , and is exposed to diet-derived Q07869 -α ligands . Therefore , we examined the effects of Q07869 -α activation on intestinal lipid and lipoprotein metabolism . METHODS AND RESULTS : The impact of Q07869 -α activation was evaluated in term of HDL-related gene expression in mice , ex vivo in human jejunal biopsies and in Caco-2/TC7 cells . Apolipoprotein-AI/HDL secretion , cholesterol esterification , and trafficking were also studied in vitro . In parallel to improving plasma lipid profiles and increasing liver and intestinal expression of fatty acid oxidation genes , treatment with the dual Q07869 -α/δ ligand DB05187 resulted in a more pronounced increase in plasma HDL compared with fenofibrate in mice . DB05187 , but not fenofibrate , increased the expression of HDL production genes such as apolipoprotein-AI and DB00171 -binding cassette A1 transporter in murine intestines . A similar increase was observed upon Q07869 -α activation of human biopsies and Caco-2/TC7 cells . Additionally , HDL secretion by Caco-2/TC7 cells increased . Moreover , Q07869 -α activation decreased the cholesterol esterification capacity of Caco-2/TC7 cells , modified cholesterol trafficking , and reduced apolipoprotein-B secretion . CONCLUSION : Peroxisome proliferator-activated receptor-α activation reduces cholesterol esterification , suppresses chylomicron , and increases HDL secretion by enterocytes . These results identify the intestine as a target organ of Q07869 -α ligands with entero-hepatic tropism to reduce atherogenic dyslipidaemia . Effects on thrombin generation of single injections of DB02351 in patients with calf vein thrombosis . STUDY OBJECTIVE : To determine whether single injections of DB02351 , a direct thrombin inhibitor , can inhibit thrombin generation in patients with calf vein thrombosis and , if so , if the inhibition is sustained . DESIGN : Phase II open label cohort study . SETTING : Tertiary-care referral centres , university affiliated hospitals . PATIENTS : 10 patients with venographically-demonstrated calf vein thrombosis . INTERVENTION : Patients received a single injection of DB02351 , either 1.0 mg/kg subcutaneously or 0.6 mg/kg as a 15 min intravenous infusion . P00734 fragment ( F1++2 ) levels , as an index of thrombin generation , were measured before as well as 6 h post- and 24 h post- DB02351 administration . Patients were followed with non-invasive tests to detect thrombus extension into the proximal veins . RESULTS : There was a significant reduction in the levels of F1+2 with both regimens , 6 h after DB02351 . The F1+2 levels 24 h post- DB02351 showed a significant increase relative to the 6 h post- DB02351 results . One patient developed thrombus extension into the popliteal vein and was treated with conventional anticoagulants . CONCLUSION : The single injections of DB02351 used in the study produced incomplete and temporary suppression of F1+2 . Complete and permanent inhibition of thrombin generation with DB02351 in patients with calf vein thrombosis may require higher doses , multiple subcutaneous injections and/or prolonged intravenous infusion . Acute pharmacodynamic and antivascular effects of the vascular endothelial growth factor signaling inhibitor DB04849 in Calu-6 human lung tumor xenografts . The vascular endothelial growth factor-A ( P15692 ) signaling pathway , a key stimulant of solid tumor vascularization , is primarily dependent on the activation of the endothelial cell surface receptor P15692 receptor-2 ( P35968 ) . DB04849 is an oral , highly potent small-molecule inhibitor of VEGFR tyrosine kinase activity that inhibits angiogenesis and the growth of human tumor xenografts in vivo . Here , we show pharmacodynamic changes in P35968 phosphorylation induced by DB04849 . In mouse lung tissue , a single dose of DB04849 at 6 mg/kg inhibited P15692 -stimulated P35968 phosphorylation by 87 % at 2 h with significant inhibition ( > or=60 % ) maintained to 24 h . To examine inhibition of P35968 phosphorylation in tumor vasculature by immunohistochemistry , a comprehensive assessment of antibodies to various phosphorylation sites on the receptor was undertaken . Antibodies to the phosphotyrosine epitopes pY1175/1173 and pY1214/1212 were found suitable for this application . Calu-6 human lung tumor xenografts , from mice receiving DB04849 or vehicle treatment ( p.o. , once daily ) , were examined by immunohistochemistry . A significant reduction in tumor vessel staining of phosphorylated P35968 ( pVEGFR-2 ) was evident within 28 h of DB04849 treatment ( 6 mg/kg ) . This effect preceded a significant reduction in tumor microvessel density , which was detectable following 52 h of DB04849 treatment . These data show that DB04849 is a potent inhibitor of P35968 activation in vivo and suggest that DB04849 delivers therapeutic benefit in Calu-6 tumors by targeting vessels dependent on P35968 signaling for survival . In addition , this work highlights the utility of measuring either pY1175/1173 or pY1214/1212 on P35968 as a pharmacodynamic marker of P35968 activation . Detection of anti-isoniazid and anti-cytochrome P450 antibodies in patients with isoniazid-induced liver failure . Isoniazid ( DB00951 ) -induced hepatotoxicity remains one of the most common causes of drug-induced idiosyncratic liver injury and liver failure . This form of liver injury is not believed to be immune-mediated because it is not usually associated with fever or rash , does not recur more rapidly on rechallenge , and previous studies have failed to identify anti- DB00951 antibodies ( Abs ) . In this study , we found Abs present in sera of 15 of 19 cases of DB00951 -induced liver failure . Anti- DB00951 Abs were present in 8 sera ; 11 had anti-cytochrome P450 (CYP)2E1 Abs , 14 had Abs against P05181 modified by DB00951 , 14 had anti- P08684 antibodies , and 10 had anti- P11712 Abs . DB00951 was found to form covalent adducts with P05181 , P08684 , and P11712 . None of these Abs were detected in sera from DB00951 -treated controls without significant liver injury . The presence of a range of antidrug and autoAbs has been observed in other drug-induced liver injury that is presumed to be immune mediated . CONCLUSION : These data provide strong evidence that DB00951 induces an immune response that causes DB00951 -induced liver injury . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . DB01045 Does not Significantly Affect the Expression of Small Heterodimer Partner in Primary Human Hepatocytes . The small/short heterodimer partner ( Q15466 , Q15466 ) is a nuclear receptor corepressor lacking a DNA binding domain . Q15466 is induced by bile acid-activated farnesoid X receptor ( Q96RI1 ) resulting in P22680 gene suppression . In contrast , O75469 ( O75469 ) activation by its ligands was recently suggested to inhibit Q15466 gene transactivation to maximize the induction of O75469 target genes . However , there are also conflicting reports in literature whether O75469 or rodent Pxr activation down-regulates Q15466 /Shp expression . Moreover , the O75469 -mediated regulation of the Q15466 gene has been studied only at the Q15466 mRNA and transactivation ( gene reporter assay ) levels . In this study , we studied the effect of rifampicin , a prototype O75469 ligand , on Q15466 mRNA , and protein expression in three primary human hepatocyte cultures . We found that Q15466 mRNA is not systematically down-regulated in hepatocyte in culture after 24 h treatment with rifampicin . Consistently , we did not observe down-regulation of Q15466 protein in primary human hepatocytes after 24 and 48 h of incubation with rifampicin . We can conclude that although we observed slight down-regulation of Q15466 mRNA and protein in several hepatocyte preparations , the phenomenon is unlikely critical for O75469 -mediated induction of its target genes . A water-soluble homodimeric serine palmitoyltransferase from Sphingomonas paucimobilis EY2395T strain . Purification , characterization , cloning , and overproduction . DB00133 palmitoyltransferase ( P21549 , EC ) is a key enzyme in sphingolipid biosynthesis and catalyzes the decarboxylative condensation of l-serine and palmitoyl-coenzyme A to 3-ketodihydrosphingosine . We found that the Gram-negative obligatory aerobic bacteria Sphingomonas paucimobilis EY2395(T) have significant P21549 activity and purified P21549 to homogeneity . This enzyme is a water-soluble homodimeric protein unlike eukaryotic enzymes , known as heterodimers composed of tightly membrane-bound subunits , named O15269 and O15270 . The purified P21549 shows an absorption spectrum characteristic of a pyridoxal 5'-phosphate-dependent enzyme . The substrate specificity of the Sphingomonas P21549 is less strict than the P21549 complex from Chinese hamster ovary cells . We isolated the P21549 gene encoding 420 amino acid residues ( M(r) 45,041 ) and succeeded in overproducing the P21549 protein in Escherichia coli , in which the product amounted to about 10-20 % of the total protein of the cell extract . Sphingomonas P21549 shows about 30 % homology with the enzymes of the alpha-oxamine synthase family , and amino acid residues supposed to be involved in catalysis are conserved . The recombinant P21549 was catalytically and spectrophotometrically indistinguishable from the native enzyme . This is the first successful overproduction of an active enzyme in the sphingolipid biosynthetic pathway . Sphingomonas P21549 is a prototype of the eukaryotic enzyme and would be a useful model to elucidate the reaction mechanism of P21549 . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Critical role of P21453 and integrin β4 in P14210 /c- DB00134 -mediated increases in vascular integrity . Vascular endothelial cell ( EC ) barrier integrity is critical to vessel homeostasis whereas barrier dysfunction is a key feature of inflammatory disorders and tumor angiogenesis . We previously reported that hepatocyte growth factor ( P14210 ) -mediated increases in EC barrier integrity are signaled through a dynamic complex present in lipid rafts involving its receptor , c- DB00134 . We extended these observations to confirm that P21453 ( sphingosine 1-phosphate receptor 1 ) and integrin β4 ( P16144 ) are essential participants in P14210 -induced EC barrier enhancement . Immunoprecipitation experiments demonstrated P14210 -mediated recruitment of c- DB00134 , P16144 and P21453 to caveolin-enriched lipid rafts in human lung EC with direct interactions of c- DB00134 with both P21453 and P16144 accompanied by c- DB00134 -dependent P21453 and P16144 transactivation . Reduced P21453 expression ( siRNA ) attenuated both P16144 and Rac1 activation as well as c- DB00134 / P16144 interaction and resulted in decreased transendothelial electrical resistance . Furthermore , reduced P16144 expression attenuated P14210 -induced c- DB00134 activation , c- DB00134 / P21453 interaction , and effected decreases in Q14703 - and P14210 -induced EC barrier enhancement . Finally , the c- DB00134 inhibitor , DB05030 , suppressed P14210 -induced c- DB00134 activation as well as P21453 and P16144 transactivation . These results support a critical role for P21453 and P16144 transactivation as rate-limiting events in the transduction of P14210 signals via a dynamic c- DB00134 complex resulting in enhanced EC barrier integrity . Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . Induction of P22309 and P20813 by an antimitogenic factor in HepG2 cells is mediated through suppression of cyclin-dependent kinase 2 activity : cell cycle-dependent expression . P14210 ( P14210 ) , an antimitogenic factor for HepG2 cells , increased mRNA and protein levels of P22309 and P20813 , as well as the endogenous cyclin-dependent kinase ( CDK ) inhibitors p16 , P38936 , and p27 in HepG2 cells but not in HuH6 , Caco2 , or MCF7 cells . Treatment with 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene ( U0126 ) ( an extracellular signal-regulated kinase inhibitor ) suppressed the P14210 -induced expression of P22309 and P20813 , as well as p16 , P38936 , and p27 in HepG2 cells . The CDK inhibitor roscovitine also enhanced the expression of P22309 , P20813 , and P08684 . Transfection of anti- P24941 siRNA led to elevated levels of P22309 , P20813 , and P08684 in HepG2 and SW480 cells , whereas anti- P11802 small interfering RNA ( siRNA ) did not significantly enhance the expression of these enzymes . In fact , P24941 activity was decreased in P14210 -treated HepG2 cells . In cells arrested in S phase by a thymidine block and then released into a synchronous cell cycle , there was a clear dissociation among the activation of P24941 and the expression of P22309 , P20813 , and P08684 . Furthermore , the induction of P08684 but not P22309 or P20813 mRNA expression by roscovitine was repressed in pregnane X receptor ( O75469 ) siRNA-transfected HepG2 cells . Transfection with constitutive androstane receptor siRNA or O75469 siRNA in HepG2 cells did not repress the P14210 -stimulated expression of P22309 mRNA . Taken together , our results show that the expression of P22309 and P20813 is negatively regulated through a P24941 signaling pathway linked to cell cycle progression in HepG2 and SW480 cells , the mechanism of which may differ from that of P08684 expression through O75469 phosphorylated by P24941 . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors . CpG island methylation of Q12983 predicts resistance against S-1/CPT-11 combined therapy in colorectal cancer patients . Aberrant gene methylation is frequently observed in various cancers and plays an important role in carcinogenesis , cancer progression and drug responsiveness . The aim of this study is to identify colorectal cancer specific gene methylation determining chemosensitivity to S-1/CPT-11 therapy . The gene methylation of Q96EP1 , p16 , Q13761 , P12830 , P16455 , hMLH1 , Q9UNQ0 , P22309 and Q12983 genes were analyzed in 27 colorectal cancer tissues by quantitative methylation-specific PCR ( q-MSP ) . All 27 patients were postoperatively treated by S-1/CPT-11 therapy targeting the metastatic lesion and the recurrent tumor . Thereafter , the patients were divided into a responder group ( RG ) or a non-responder group ( DB04223 ) according to the effect of the chemotherapy . There were 13 cases of RG ( 48.1 % ) and 14 cases of DB04223 ( 51.9 % ) . The methylation level in Q96EP1 , Q13761 and Q12983 was significantly higher in cancer lesions in comparison to the non-cancerous lesion . Only methylation of the Q12983 gene was significantly higher in primary cancer tissue of the DB04223 than the RG . The correlation between the Q12983 methylation status and time to progression ( TTP ) suggested that the low methylation group ( n=16 ) resulted in a significantly longer TTP , in comparison to the high methylation group ( n=11 ; P=0.004 ) . The methylation level of Q12983 showed a significant inverse correlation with the mRNA expression suggesting the DNA methylation suppressed Q12983 expression ( r=-0.466 , P=0.021 ) . In conclusion , Q12983 gene methylation is a possible marker predicting a poor response to the S-1/CPT-11 combined therapy in colorectal cancer .

MH_train_1269

MH_train_1269

MH_train_1269

interacts_with DB00834?

multiple_choice

PARP and CHK inhibitors interact to cause DNA damage and cell death in mammary carcinoma cells . The present studies examined viability and DNA damage levels in mammary carcinoma cells following P09874 and O14757 inhibitor drug combination exposure . P09874 inhibitors [ AZD2281 ; ABT888 ; DB02690 ; AG014699 ] interacted with O14757 inhibitors [ P55089 -01 ; AZD7762 ; LY2603618 ] to kill mammary carcinoma cells . P09874 and O14757 inhibitors interacted to increase both single strand and double strand DNA breaks that correlated with increased γ P16104 phosphorylation . Treatment of cells with O14757 inhibitors increased the phosphorylation of O14757 and P27361 /2 . Knock down of Q13315 suppressed the drug-induced increases in O14757 and P27361 /2 phosphorylation and enhanced tumor cell killing by P09874 and O14757 inhibitors . Expression of dominant negative Q02750 enhanced drug-induced DNA damage whereas expression of activated Q02750 suppressed both the DNA damage response and tumor cell killing . Collectively our data demonstrate that P09874 and O14757 inhibitors interact to kill mammary carcinoma cells and that increased DNA damage is a surrogate marker for the response of cells to this drug combination . Structural determinants of phosphoinositide 3-kinase inhibition by wortmannin , LY294002 , quercetin , myricetin , and staurosporine . The specific phosphoinositide 3-kinase ( PI3K ) inhibitors wortmannin and LY294002 have been invaluable tools for elucidating the roles of these enzymes in signal transduction pathways . The X-ray crystallographic structures of P48736 bound to these lipid kinase inhibitors and to the broad-spectrum protein kinase inhibitors quercetin , myricetin , and staurosporine reveal how these compounds fit into the DB00171 binding pocket . With a nanomolar IC50 , wortmannin most closely fits and fills the active site and induces a conformational change in the catalytic domain . Surprisingly , LY294002 and the lead compound on which it was designed , quercetin , as well as the closely related flavonoid myricetin bind PI3K in remarkably different orientations that are related to each other by 180 degrees rotations . DB02010 /PI3K interactions are reminiscent of low-affinity protein kinase/staurosporine complexes . These results provide a rich basis for development of isoform-specific PI3K inhibitors with therapeutic potential . Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . P06401 activation of extranuclear signaling pathways in regulating p53 expression in vascular endothelial cells . We previously showed that progesterone ( P4 ) inhibited the proliferation of human umbilical vein endothelial cells ( HUVECs ) through a p53-dependent pathway . Now we investigated further the molecular mechanism underlying the hormone activity . In cultured HUVECs , P4 increased the protein levels of phosphorylated Src ( p-Src ) , P04049 , and P29323 . The levels of p-Src and p-Src-progesterone receptor complex in HUVECs were increased by P4 treatment . These effects were blocked by pretreatment with a progesterone receptor antagonist , DB00834 . The P4-induced increase in p53 transactivity was abolished by pretreatment with Src kinase inhibitors . Moreover , administration with cSrc antisense oligonucleotide prevented the P4-induced increases of the levels of p53 mRNA and protein . These data suggest that P4-induced up-regulation of p53 might be mediated through activation of cSrc . Pretreatment with Src kinase inhibitors also prevented P4-induced membrane translocation of Kras and increases of the protein levels of phosphorylated Raf and phosphorylated P29323 . Transfection with dominant-negative P28482 prevented the P4-induced increases of protein level and promoter activity of p53 and a decrease of thymidine incorporation . P4 also increased nuclear factor-κB ( NF-κB ) nuclear translocation and NF-κB binding onto the p53 promoter . These effects were abolished by pretreatment with P29323 inhibitors . The P4-induced up-regulation of the p53 promoter activity was prevented by preadministration with dominant-negative P28482 or NF-κB inhibitors . Taken together , our data suggest that the cSrc/Kras/ P04049 / P28482 /NF-κB signaling pathway contributes to the P4-induced up-regulation of p53 in HUVECs . These findings highlight progesterone receptor activation of extranuclear signaling pathways in regulating p53 and cell cycle progression in HUVECs . A transcription-independent function of Q12778 in inhibition of androgen-independent activation of the androgen receptor in prostate cancer cells . Increasing evidence suggests that aberrant activation of the androgen receptor ( AR ) plays a pivotal role in the development and progression of androgen depletion-independent prostate cancer ( PCa ) after androgen deprivation therapy . Here , we show that loss of the P60484 tumor suppressor gene is associated with hyperactivation of the AR in human PCa cell lines . This effect is mediated primarily by its downstream effector Q12778 . In addition to the inhibition of androgenic activation of the AR , forced expression of Q12778 in P60484 -negative PCa cells also inhibits androgen-independent activation of the AR in a manner independent of Q12778 transcriptional function . In contrast , silencing of Q12778 in P60484 -positive cells not only increases the basal activity of the AR in the absence of androgens , it also markedly sensitizes the AR activation by low levels of androgens or nonandrogenic factors such as interleukin-6 . Q12778 -mediated inhibition of the AR is partially attenuated by the histone deacetylase ( HDAC ) inhibitor trichostatin A . Accordingly , Q12778 interacts with O15379 as shown by coimmunoprecipitation assays , and cotransfection of cells with Q12778 and O15379 , but not Q13547 and Q92769 , results in a greater inhibition of AR activity than in cells transfected with Q12778 or O15379 individually . Together , our findings define a novel corepressor function of Q12778 in inhibition of androgen-independent activation of the AR . DB08901 may overcome resistance of P36888 -ITD harbouring additional point mutations , notably the previously refractory F691I mutation . Fms-like tyrosine kinase ( P36888 ) mutations are the most frequent mutations in patients with acute myeloid leukaemia ( AML ) that confer a poor prognosis . Constitutively active P36888 -ITD ( internal tandem duplications ) mutations define a promising target for therapeutic approaches using small molecule inhibitors . However , several point mutations of the P36888 tyrosine kinase domain ( P36888 -TKD ) have been identified to mediate resistance towards P36888 tyrosine kinase inhibitors ( P36888 -TKI ) , including secondary mutations of P36888 . We investigated the cellular effects of the recently characterised P36888 -TKI ponatinib ( DB08901 ) on murine myeloid cells transfected with P36888 -ITD with or without additional point mutations of the P36888 -TKD including the ( so far ) multi-resistant F691I mutation . DB08901 effectively induced apoptosis not only in the parental P36888 -ITD cell line but also in all stably transfected subclones harbouring additional P36888 -TKD point mutations ( N676D , F691I or G697R ) . These observations correlated with a strong inhibition of P36888 -ITD and its downstream targets P42229 , AKT and P27361 /2 upon ponatinib incubation , as determined by Western blotting . We conclude that ponatinib represents a promising P36888 -TKI that should be further investigated in clinical trials . The targeted therapy of P36888 -ITD-positive AML with ponatinib might be associated with a lower frequency of secondary resistance caused by acquired P36888 -TKD mutations . Cbl-b is a critical regulator of macrophage activation associated with obesity-induced insulin resistance in mice . We previously reported the potential involvement of casitas B-cell lymphoma-b ( Cbl-b ) in aging-related murine insulin resistance . Because obesity also induces macrophage recruitment into adipose tissue , we elucidated here the role of Cbl-b in obesity-related insulin resistance . Cbl-b(+/+) and Cbl-b(-/-) mice were fed a high-fat diet ( HFD ) and then examined for obesity-related changes in insulin signaling . The HFD caused recruitment of macrophages into adipose tissue and increased inflammatory reaction in Cbl-b(-/-) compared with Cbl-b(+/+) mice . Peritoneal macrophages from Cbl-b(-/-) mice and Cbl-b-overexpressing RAW264.7 macrophages were used to examine the direct effect of saturated fatty acids ( FAs ) on macrophage activation . In macrophages , Cbl-b suppressed saturated FA-induced O00206 ( O00206 ) signaling by ubiquitination and degradation of O00206 . The physiological role of Cbl-b in vivo was also examined by bone marrow transplantation and DB04933 , a O00206 antagonist . Hematopoietic cell-specific depletion of the Cbl-b gene induced disturbed responses on insulin and glucose tolerance tests . Blockade of O00206 signaling by DB04933 reduced fasting blood glucose and serum interleukin-6 levels in obese Cbl-b(-/-) mice . These results suggest that Cbl-b deficiency could exaggerate HFD-induced insulin resistance through saturated FA-mediated macrophage activation . Therefore , inhibition of O00206 signaling is an attractive therapeutic strategy for treatment of obesity-related insulin resistance . Dexamethasone inhibits interleukin-1β-induced matrix metalloproteinase-9 expression in cochlear cells . OBJECTIVES : To investigate the effect of interleukin ( IL ) -1β on matrix metalloproteinase ( MMP ) -9 expression in cochlea and regulation of IL-1β-mediated P14780 expression by dexamethasone and the molecular and signaling mechanisms involved . METHODS : House ear institute-organ of Corti 1 ( HEI-OC1 ) cells were used and exposed to IL-1β with/without dexamethasone . P04150 antagonist , DB00834 , was used to see the role of dexamethasone . PD98059 ( an extracellular signal-regulated kinases [ ERKs ] inhibitor ) , SB203580 ( a p38 mitogen-activated protein kinases [ MAPK ] inhibitor ) , SP600125 ( a c-Jun N-terminal kinase [ JNK ] inhibitor ) were also used to see the role of MAPKs signaling pathway(s) in IL-1β-induced P14780 expression in HEI-OC1 cells . Reverse transcription-polymerase chain reaction and gelatin zymography were used to measure mRNA expression level of P14780 and activity of P14780 , respectively . RESULTS : Treatment with IL-1β-induced the expression of P14780 in a dose- and time-dependent manner . IL-1β ( 1 ng/mL ) -induced P14780 expression was inhibited by dexamethasone . Interestingly , p38 MAPK inhibitor , SB203580 , significantly inhibited IL-1β-induced P14780 mRNA and P14780 activity . However , inhibition of JNKs and ERKs had no effect on the IL-1β-induced P14780 expression . CONCLUSION : These results suggest that the pro-inflammatory cytokine IL-1β strongly induces P14780 expression via activation of p38 MAPK signaling pathway in HEI-OC1 cells and the induction was inhibited by dexamethasone . Oleuropein , an anti-oxidant polyphenol constituent of olive promotes α-secretase cleavage of the amyloid precursor protein ( AβPP ) . Over the past decade , intense focus has been dedicated on investigating processes involved in the proteolysis of amyloid precursor protein ( AβPP ) and β-amyloid ( Aβ ) peptide metabolism , as possible targets for Alzheimer 's disease ( AD ) therapy . To this goal , considerable research has been targeted on potential therapeutic use of compounds promoting non-amyloidogenic processing of AβPP . One of these compounds , oleuropein , a polyphenol constituent of extra virgin olive oil exhibiting a wide range of pharmacological properties , was shown to interact non-covalently with Aβ , an interaction that might be related to a potential protective role of oleuropein against Aβ aggregation . In the present study , it was demonstrated that oleuropein treatment of HEK293 cells stably transfected with the isoform 695 of human AβPP ( APP695 ) leads to markedly elevated levels of sAPPα and to significant reduction of Aβ oligomers . These effects were associated with increased activity of matrix metalloproteinase 9 ( P14780 ) , whereas no significant alterations in the expression of secretases P78536 , ADAM-10 or P56817 -1 were observed . Similar results were obtained using the human neuroblastoma cell line SK-N-SH . The experimental data reveal an anti-amyloidogenic effect of oleuropein and suggest a possible protective role for oleuropein against AD , extending the spectrum of beneficial properties of this naturally occurring polyphenol . Relationship between calcium-activated chloride channel 1 and P98088 in goblet cell hyperplasia induced by interleukin-13 in human bronchial epithelial cells . BACKGROUND : Interleukin ( IL ) -13 has recently been reported as the major T-helper 2 cytokine involved in mucus overproduction and oversecretion in allergic airways . However , the relationship between human calcium-activated chloride channel-1 ( A8K7I4 ) and P98088 induced by P35225 in vitro has not been fully investigated . OBJECTIVES : The present study examines whether P35225 induces the expression of A8K7I4 in normal human bronchial epithelial ( NHBE ) cells . We also investigated the relationship between A8K7I4 and P98088 expression and the development of goblet cell hyperplasia ( P30793 ) . METHODS : NHBE cells were isolated from human bronchi , and cultured with an air-liquid interface . A8K7I4 and P98088 gene and protein expression , as well as P30793 were examined in the cells after exposure to P35225 . RESULTS : Incubation with P35225 for 14 and 21 days increased the total number of epithelial cells , the number of periodic acid-Schiff ( DB00233 ) -stained epithelial cells , the number of goblet cells , as well as expression of mRNA and protein of A8K7I4 and P98088 . The number of goblet cells with secretory granules also increased after 21 days of incubation with P35225 . Niflumic acid , a chloride channel inhibitor , reduced mRNA expression of A8K7I4 and P98088 , and reduced the number of DB00233 -positive cells after incubation with P35225 . NHBE cells exposed or not to P35225 expressed P35225 receptor alpha(1) ( IL-13Ralpha(1) ) , and an antibody to P35225 Ralpha(1) also reduced the number of DB00233 -positive cells after exposure to P35225 . CONCLUSIONS : P35225 might induce the expression of P98088 and A8K7I4 gene and protein in well-differentiated NHBE cells . These cells might also differentiate into goblet cells and become hyperplastic . P30047 -dependent and -independent inhibitors of P30793 . P30047 ( P30047 ) mediates the feedback inhibition of P30793 activity by ( 6R ) -L-erythro- DB00360 ( BH4 ) through protein complex formation . Since guanine and BH4 have a common pyrimidine ring structure , we examined the inhibitory effect of guanine and its analogs on the enzyme activity . DB02377 , 8-hydroxyguanine , 8-methylguanine , and 8-bromoguanine inhibited the enzyme activity in a P30047 -dependent and pH-dependent manner and induced complex formation between P30793 and P30047 . The type of inhibition by this group is a mixed type . All these properties were shared with BH4 . In striking contrast , inhibition by DB01667 and 8-mercaptoguanine was P30047 -independent and pH-independent . The type of inhibition by DB01667 and 8-mercaptoguanine was a competitive type . The two compounds did not induce complex formation between the enzyme and P30047 . These results demonstrate that guanine compounds of the first group bind to the BH4-binding site of the P30793 / P30047 complex , whereas DB01667 and 8-mercaptoguanine bind to the active site of the enzyme . Finally , the possible implications in Lesch-Nyhan syndrome and Parkinson diseases of the inhibition of P30793 by guanine and 8-hydroxyguanine are discussed . Short and long access to cocaine self-administration activates tyrosine phosphatase P54829 and attenuates GluN expression but differentially regulates GluA expression in the prefrontal cortex . RATIONALE : Dephosphorylation of extracellular signal-regulated kinase ( P29323 ) and cyclic AMP response element binding protein ( CREB ) in the dorsomedial prefrontal cortex ( dmPFC ) at the end of short access ( ShA ) cocaine self-administration is implicated in cocaine seeking . However , what receptors and phosphatases mediate this effect and whether P29323 /CREB and related phospho-proteins in the dmPFC react similarly during early withdrawal from long access ( LgA ) cocaine self-administration are unknown . OBJECTIVES : The effects of ShA vs. LgA cocaine self-administration on the phosphorylation of protein phosphatase 2A ( PP2A ) and striatal-enriched protein tyrosine phosphatase ( P54829 ) , as well as GluN and GluA receptor subtype expression in the dmPFC during early withdrawal , were compared . METHODS : Rats self-administered cocaine or received saline during 2- or 6-h daily sessions for 10-11 days . Two hours after the final session , the dmPFC was dissected out and processed for immunoblotting . RESULTS : Similar to previous findings after ShA cocaine , phospho- P29323 and phospho-CREB in the dmPFC were decreased after LgA cocaine . Cocaine elevated phospho-PP2A ( deactivation ) and decreased phospho- P54829 ( activation ) in both ShA and LgA cocaine rats . Q05586 , Q13224 , and phospho- Q13224 Tyr1472 in the dmPFC were decreased after ShA and LgA cocaine . Further , a significant reduction of P42262 , P42261 , and phospho- P42261 Ser845 was found only in LgA rats . CONCLUSIONS : Activation of phospho- P54829 may underlie P29323 and CREB dephosphorylation in the dmPFC as well as internalization and degradation of GluN complexes during early withdrawal from both ShA and LgA cocaine self-administration , whereas differential alteration of AMPA receptor subunits after ShA and LgA cocaine self-administration depends on cocaine intake . P04150 antagonism disrupts the reconsolidation of social reward-related memories in rats . Reconsolidation is the process whereby consolidated memories are destabilized upon retrieval and restabilized to persist for later use . Although the neurobiology of the reconsolidation of both appetitive and aversive memories has been intensively investigated , reconsolidation of memories of physiologically relevant social rewards has received little attention . Social play , the most characteristic social behaviour displayed by young mammals , is highly rewarding , illustrated by the fact that it can induce conditioned place preference ( CPP ) . Here , we investigated the role of signalling mechanisms implicated in memory processes , including reconsolidation , namely glucocorticoid , mineralocorticoid , DB01221 glutamatergic and P21554 cannabinoid receptors , in the reconsolidation of social play-induced CPP in rats . Systemic treatment with the glucocorticoid receptor antagonist mifepristone before , but not immediately after , retrieval disrupted the reconsolidation of social play-induced CPP . DB00834 did not affect social play-induced CPP in the absence of memory retrieval . Treatment with the DB01221 receptor antagonist MK-801 modestly affected the reconsolidation of social play-induced CPP . However , the reconsolidation of social play-induced CPP was not affected by treatment with the mineralocorticoid and P21554 cannabinoid receptor antagonists spironolactone and rimonabant , respectively . We conclude that glucocorticoid neurotransmission mediates the reconsolidation of social reward-related memories in rats . These data indicate that the neural mechanisms of the reconsolidation of social reward-related memories only partially overlap with those underlying the reconsolidation of other reward-related memories . Environment-related adaptive changes of gut commensal microbiota do not alter colonic toll-like receptors but modulate the local expression of sensory-related systems in rats . Pathogenic and protective roles have been attributed to gut commensal microbiota ( GCM ) in gastrointestinal inflammatory and functional disorders . We have shown that the adaptation to a new environment implies specific changes in the composition of GCM . Here we assessed if environment-related adaptive changes of GCM modulate the expression of colonic Toll-like receptors ( TLRs ) and sensory-related systems in rats . Adult male SD rats were maintained under different environmental conditions : barrier-breed-and-maintained , barrier-breed adapted to conventional conditions or conventional-breed-and-maintained . Fluorescent in situ hybridization and real-time quantitative PCR ( qPCR ) were used to characterize luminal ceco-colonic microbiota . Colonic expression of O60603 , O00206 , O60602 , and Q9NYK1 , cannabinoid receptors ( P21554 /CB2 ) , μ-opioid receptor ( MOR ) , transient receptor potential vanilloid ( Q8NER1 , Q8NET8 , and Q9HBA0 ) , protease-activated receptor 2 ( P55085 ) , and calcitonin gene-related peptide were quantified by RT-qPCR . P21554 , CB2 and MOR expression , was evaluated also by immunohistochemistry . In rats , housing-related environmental conditions induce specific changes of GCM , without impact on the expression of TLR-dependent bacterial recognition systems . Expression of sensory-related markers ( MOR , Q8NET8 , P55085 , and CB2 ) decreased with the adaptation to a conventional environment , correlating with changes in Bacteroides spp. , Lactobacillus spp. , and Bifidobacterium spp. counts . This suggests an interaction between GCM and visceral sensory mechanisms , which might be part of the mechanisms underlying the beneficial effects of some bacterial groups on functional and inflammatory gastrointestinal disorders . DB09210 defines a new binding site for allosteric modulators of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid ( AMPA ) receptors . Glutamate receptors are the most prevalent excitatory neurotransmitter receptors in the vertebrate central nervous system and are important potential drug targets for cognitive enhancement and the treatment of schizophrenia . Allosteric modulators of AMPA receptors promote dimerization by binding to a dimer interface and reducing desensitization and deactivation . The pyrrolidine allosteric modulators , piracetam and aniracetam , were among the first of this class of drugs to be discovered . We have determined the structure of the ligand binding domain of the AMPA receptor subtypes P42262 and P42263 with piracetam and a corresponding structure of P42263 with aniracetam . Both drugs bind to P42262 and P42263 in a very similar manner , suggesting little subunit specificity . However , the binding sites for piracetam and aniracetam differ considerably . DB04599 binds to a symmetrical site at the center of the dimer interface . DB09210 binds to multiple sites along the dimer interface with low occupation , one of which is a unique binding site for potential allosteric modulators . This new site may be of importance in the design of new allosteric regulators . The expression level of P21554 and CB2 receptors determines their efficacy at inducing apoptosis in astrocytomas . BACKGROUND : Cannabinoids represent unique compounds for treating tumors , including astrocytomas . Whether CB(1) and CB(2) receptors mediate this therapeutic effect is unclear . PRINCIPAL FINDINGS : We generated astrocytoma subclones that express set levels of CB(1) and CB(2) , and found that cannabinoids induce apoptosis only in cells expressing low levels of receptors that couple to P27361 /2 . In contrast , cannabinoids do not induce apoptosis in cells expressing high levels of receptors because these now also couple to the prosurvival signal AKT . Remarkably , cannabinoids applied at high concentration induce apoptosis in all subclones independently of CB(1) , CB(2) and AKT , but still through a mechanism involving P27361 /2 . SIGNIFICANCE : The high expression level of CB(1) and CB(2) receptors commonly found in malignant astrocytomas precludes the use of cannabinoids as therapeutics , unless AKT is concomitantly inhibited , or cannabinoids are applied at concentrations that bypass CB(1) and CB(2) receptors , yet still activate P27361 /2 . Hyperecho-turbo spin-echo sequences at 3T : clinical application in neuroradiology . BACKGROUND AND PURPOSE : Hyperecho-turbo spin-echo ( hyperTSE ) sequences were developed to reduce the specific absorption rate ( SAR ) , especially at high fields such as 3T and above . The purpose of this study was to quantitatively and qualitatively assess the detection of neuroradiologic pathologies by hyperTSE in comparison with standard turbo spin-echo ( TSE180 degrees ) sequences . MATERIALS AND METHODS : TSE180 degrees and hyperTSE images with parameters adapted for equal P24752 contrast were acquired on a 3T whole-body system in 51 patients with 54 cerebral pathologies . Region-of-interest analysis was performed of signal intensities of pathologies , normal white and gray matter , P04141 , and the SD of noise . Signal intensity-to-noise ratios ( SNRs ) and contrast-to-noise ratios ( CNRs ) for healthy tissues and pathologies were determined . A qualitative rating concerning artifacts , lesion conspicuity , and image quality was performed by 2 experienced neuroradiologists . RESULTS : HyperTSE sequences were equivalent to standard TSE180 degrees sequences for the P21554 of pathologies and of the contrast between gray and white matter . The SNR of gray and white matter and P04141 were also the same . The CNRs of the pathologies in hyperTSE and TSE180 degrees images were strongly correlated with each other ( r = 0.93 , P = .001 ) . The visual rating of images revealed no significant differences between hyperTSE and TSE180 degrees . CONCLUSION : HyperTSE sequences proved to be qualitatively and quantitatively equivalent to TSE180 degrees sequences in the detection of high- and low-signal-intensity lesions . They provide equal P21554 of pathologies and of gray minus white matter and reduce the imaging restrictions of conventional TSE180 degrees imposed by SAR limitations at 3T . Inhibition of O00329 restores glucocorticoid function in smoking-induced airway inflammation in mice . RATIONALE : There is an increasing prevalence of reduced responsiveness to glucocorticoid therapy in severe asthma and chronic obstructive pulmonary disease ( P48444 ) . The molecular mechanism of this remains unknown . Recent studies have shown that histone deacetylase activity , which is critical to glucocorticoid function , is altered by oxidant stress and may be involved in the development of glucocorticoid insensitivity . OBJECTIVES : To determine the role of phosphoinositol-3-kinase ( PI3K ) in the development of cigarette smoke-induced glucocorticoid insensitivity . METHODS : Wild-type , P48736 knock-out and O00329 kinase dead knock-in transgenic mice were used in a model of cigarette smoke-induced glucocorticoid insensitivity . Peripheral lung tissue was obtained from six healthy nonsmokers , nine smokers with normal lung function , and eight patients with P48444 . MEASUREMENTS AND MAIN RESULTS : In vitro oxidative stress activates PI3K and induced a relative glucocorticoid resistance , which is restored by PI3K inhibition . In vivo , cigarette smoke exposure in mice increased tyrosine nitration of histone deacetylase 2 in the lung , correlating with reduced histone deacetylase 2 activity and reduced glucocorticoid function . Q92769 activity and the antiinflammatory effects of glucocorticoids were restored in O00329 kinase dead knock-in but not P48736 knock-out smoke-exposed mice compared with wild type mice , correlating with reduced histone deacetylase 2 tyrosine nitration . P04150 expression was significantly reduced in smoke-exposed mice , in smokers with normal lung function , and in patients with P48444 . CONCLUSIONS : These data show that therapeutic inhibition of O00329 may restore glucocorticoid function in oxidative stress-induced glucocorticoid insensitivity . The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . P08183 , Q9UNQ0 , and P60484 determine the response of glioblastoma to temozolomide and ABT-888 therapy . PURPOSE : Little is known about the optimal clinical use of ABT-888 ( veliparib ) for treatment of glioblastoma . ABT-888 is a PARP inhibitor undergoing extensive clinical evaluation in glioblastoma , because it may synergize with the standard-of-care temozolomide ( DB00853 ) . We have elucidated important factors controlling ABT-888 efficacy in glioblastoma . EXPERIMENTAL DESIGN : We used genetically engineered spontaneous glioblastoma mouse models and allograft models that were orthotopically transplanted into wild-type ( WT ) and Abcb1/Abcg2-deficient ( KO ) recipients . RESULTS : ABT-888/ DB00853 is not efficacious against p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR allografts in wild-type recipients , indicating inherent resistance . Abcb1/Abcg2 mediated efflux of ABT-888 at the blood-brain barrier ( BBB ) causes a 5-fold reduction of ABT-888 brain penetration ( P < 0.0001 ) that was fully reversible by elacridar . Efficacy studies in WT and KO recipients and/or concomitant elacridar demonstrate that Abcb1/Abcg2 at the BBB and in tumor cells impair DB00853 /ABT-888 combination treatment efficacy . DB04881 also markedly improved DB00853 /ABT-888 combination treatment in the spontaneous p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR glioblastoma model . Importantly , ABT-888 does enhance DB00853 efficacy in Pten deficient glioblastoma allografts and spontaneous tumors , even in Abcb1/Abcg2 proficient wild-type mice . Loss of P60484 occurs frequently in glioblastoma ( 36 % ) and in silico analysis on patient with glioblastoma samples revealed that it is associated with a worse overall survival ( 310 days vs. 620 days , n = 117 ) . CONCLUSIONS : The potential of ABT-888 in glioblastoma can best be demonstrated in patients with P60484 null tumors . Therefore , clinical trials with ABT-888 should evaluate these patients as a separate group . Importantly , inhibition of P08183 and Q9UNQ0 ( by elacridar ) may improve the efficacy of DB00853 /ABT-888 therapy in all glioblastoma patients . P04150 and histone deacetylase-2 mediate dexamethasone-induced repression of P98088 gene expression . Airway occlusion in obstructive airway diseases is caused in part by the overproduction of secretory mucin glycoproteins through the up-regulation of mucin ( MUC ) genes by inflammatory mediators . Some pharmacological agents , including the glucocorticoid dexamethasone ( DB00514 ) , repress mucin concentrations in lung epithelial cancer cells . Here , we show that DB00514 reduces the expression of P98088 , a major airway mucin gene , in primary differentiated normal human bronchial epithelial ( NHBE ) cells in a dose-dependent and time-dependent manner , and that the DB00514 -induced repression is mediated by the glucocorticoid receptor ( GR ) and two glucocorticoid response elements ( GREs ) in the P98088 promoter . The pre-exposure of cells to DB00834 , a GR antagonist , and mutations in either the GRE3 or GRE5 cis-sites abolished the DB00514 -induced repression . Chromatin immunoprecipitation ( ChIP ) assays showed a rapid temporal recruitment of GR to the GRE3 and GRE5 cis-elements in the P98088 promoter in NHBE and in A549 cells . Immunofluorescence showed nuclear colocalization of GR and histone deacetylase-2 ( Q92769 ) in P98088 -expressing NHBE cells . ChIP also showed a rapid temporal recruitment of Q92769 to the GRE3 and GRE5 cis-elements in the P98088 promoter in both cell types . The knockdown of Q92769 by Q92769 -specific short interfering RNA prevented the DB00514 -induced repression of P98088 in NHBE and A549 cells . These data demonstrate that GR and Q92769 are recruited to the GRE3 and GRE5 cis-sites in the P98088 promoter and mediate the DB00514 -induced cis repression of P98088 gene expression . A better understanding of the mechanisms whereby glucocorticoids repress P98088 gene expression may be useful in formulating therapeutic interventions in chronic lung diseases .

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P01375 -α-mediated cardiorenal injury after rhabdomyolysis in rats . The P01375 -α serum level increases after rhabdomyolysis and is involved in the subsequent cardiorenal injury . In the present study , we investigated the P01375 -α-dependent cell signaling pathways implicated in cellular injury in these organs . Rhabdomyolysis was induced by intramuscular glycerol injection in rats . Renal function , cardiac and renal pathology , and activation of caspases were evaluated during the first 24 h after glycerol injection . P01375 -α blockade with infliximab reduced tubular necrosis and cardiorenal apoptosis . Cellular Fas-associated protein with death domain-like IL-1β-converting enzyme inhibitory protein ( cFLIP ) , an inhibitor of caspase-8 , was overexpressed in the kidney but not in the heart . The inhibitory effect of cFLIP blunted caspase-8 activation in the kidney . In this condition , the cellular response to the P01375 -α stimulus was driven to receptor-interacting protein-1 ( Q13546 ) -mediated necroptosis . Treatment with Q13546 inhibitor ( necrostatin-1 ) isolated or in combination with infliximab showed a similar reduction in tubular necrosis , underscoring the importance of P01375 -α-mediated tubular necroptosis in this model . P01375 -α played a positive regulatory role in the transcription of proapoptotic Bax and p53-upregulated modulator of apoptosis ( PUMA ) proteins . DB00065 treatment reduced caspase-9-mediated apoptosis in both organs . Treatment with a caspase-8 inhibitor showed that caspase-8 participated in the process of apoptosis only in the heart , upstream of caspase-9 activation . P01375 -α-mediated necroptosis is the predominant form of tubular injury observed in the glycerol model . P01375 -α up regulates Bax and PUMA proapoptotic proteins , resulting in activation of the intrinsic pathway of apoptosis in the kidney and heart . Activation of farnesoid X receptor attenuates liver injury in systemic lupus erythematosus . To investigate the expression and effect of farnesoid X receptor ( Q96RI1 ) on systemic lupus erythematosus ( SLE ) liver dysfunction and indicate its hepatoprotective role and the immunomodulatory property . mRNA and protein levels of Q96RI1 were determined on the liver specimens of SLE patients with liver injury as well as MRL/lpr rodent models . The Q96RI1 agonist chenodeoxycholic acid ( DB06777 ) was administrated to MRL/lpr mice and the control BALB/C with concanavalin A ( ConA ) -induced liver injury . Blood samples were taken 0 , 4 , 8 , 12 , 16 , and 24 h after ConA injection for the detection of serum ALT , Q9NRA2 , IFN-γ , P01375 -α , and P05231 . Q96RI1 was down-regulated at both mRNA and protein levels in the liver specimens of SLE patients with liver injury as well as MRL/lpr mice . MRL/lpr was more susceptible to ConA than BALB/C indicated by significantly higher levels of aminotransferase and inflammatory cytokines . Activation of Q96RI1 by DB06777 significantly reduced aminotransferase and inflammatory cytokines IFN-γ , P01375 -α , and P05231 caused by ConA injection in MRL/lpr mice . Q96RI1 was down-regulated in SLE patients as well as MRL/lpr lupus models with liver dysfunction . Q96RI1 activation ameliorated liver injury and suppressed inflammatory cytokines , thereby showing its protective function in SLE . Our findings raised the promising potential target for the treatment of SLE liver injury . Heterogeneous expression of cyclooxygenase-2 and inducible nitric oxide synthase within colorectal tumors : correlation with tumor angiogenesis . BACKGROUND : Recent studies have shown that the cyclooxygenase ( P36551 ) and the inducible nitric oxide synthase ( P35228 ) pathways are involved in the development of tumor angiogenesis in human cancers . AIMS : To investigate whether a different pattern of P35354 and P35228 expression/activity exists within different areas of colorectal tumors and to analyze the relationship between these two enzymes and tumor angiogenesis . METHODS : Microvessel density ( P53602 ) and P35354 , P35228 , vascular endothelial growth factor ( P15692 ) and P15692 receptor-2 ( P35968 ) protein expression were evaluated at both the invasive front ( IF ) and the tumor center ( TC ) in 46 human colorectal cancer specimens . We also investigated the concentration of DB00917 and NO at the same sites . RESULTS : P35354 and P35228 protein expression and activity were significantly higher within the IF than the TC of the tumor specimens . Similarly , P53602 and P15692 / P35968 expression significantly increased from the TC to the IF . Only P35354 expression was significantly correlated with P53602 and P15692 / P35968 expression at both the TC and the IF . CONCLUSION : Our study shows a heterogeneous expression of P35354 and P35228 in colorectal cancer . The up-regulation of P35354 at the IF parallels an increase in vessel density and P15692 / P35968 expression , thus supporting the hypothesis that the tumor periphery is the most aggressive portion of a colorectal tumor . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Diet induced regulation of genes involved in cholesterol metabolism in rat liver parenchymal and Kupffer cells . BACKGROUND/AIMS : Feeding rodents atherogenic diets enriched in cholesterol or cholic acid changes hepatic cholesterol metabolism . In the present study , the effect of an atherogenic diet enriched in cholesterol and cholic acid on cellular hepatic cholesterol metabolism was studied . METHODS : Gene and protein expression analysis was performed on parenchymal , endothelial , and Kupffer cells isolated from rats fed a chow or atherogenic diet using quantitative real-time PCR and immunoblotting , respectively . RESULTS : The atherogenic diet raised the serum cholesterol concentration 11-fold , mostly in the VLDL fraction , and led to heavy lipid loading of rat liver parenchymal and Kupffer cells . Only moderate changes in the expression of genes involved in cholesterol metabolism were observed in parenchymal cells on the diet , while Q07869 delta expression was 6.8-fold decreased . Kupffer cells , however , showed a highly adaptive response with a 2- to 9-fold induction of Q8WTV0 , O95477 , and Q9H222 / Q9UBA6 , and an 82-fold induction in P22680 mRNA expression , respectively . CONCLUSIONS : Heavy lipid loading of parenchymal cells leads to moderate gene expression changes , while Kupffer cells respond in a highly adaptive fashion by stimulating the expression of genes involved in cholesterol metabolism and transport . P03372 alpha augments changes in hemostatic gene expression in HepG2 cells treated with estradiol and phytoestrogens . Phytoestrogens are popular alternatives to estrogen therapy however their effects on hemostasis in post-menopausal women are unknown . The aim of this study was to determine the effect of the phytoestrogens , genistein , daidzein and equol on the expression of key genes from the hemostatic system in human hepatocyte cell models and to determine the role of estrogen receptors in mediating any response seen . HepG2 cells and Hep89 cells ( expressing estrogen receptor alpha ( ERα ) ) were incubated for 24 h with 50 nM 17β-estradiol , genistein , daidzein or equol . Tissue plasminogen activator ( tPA ) , plasminogen activator inhibitor-1 ( P05121 ) , Factor VII , fibrinogen γ , protein C and protein S mRNA expression were determined using TaqMan PCR . DB01645 and equol increased tPA and P05121 expression in Hep89 cells with fold changes greater than those observed for estradiol . In HepG2 cells ( which do not express ERα ) , P05121 and tPA expression were unchanged . Increased expression of Factor VII was observed in phytoestrogen treated Hep89 cells but not in similarly treated HepG2s . P00734 gene expression was increased in equol and daidzein treated HepG2 cells in the absence of the classical estrogen receptors . These data suggest that phytoestrogens can regulate the expression of coagulation and fibrinolytic genes in a human hepatocyte cell line ; an effect which is augmented by ERα . A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 and P35354 ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg . Phytoestrogens induce apoptosis through a mitochondria/caspase pathway in human breast cancer cells . OBJECTIVE : To explore the effect and pathway of phytoestrogens on the growth of breast cancer cell line MCF-7 . METHODS : MCF-7 cells ( human estrogen receptor-positive and progesterone receptor-positive breast cancer cells ) were cultured in serum-free medium for 24 h and then treated with genistein , resveratrol , and quercetin ( 10(-10)-10(-4) mol/l ) . After further incubation for 24 , 48 , 72 , and 92 h , the cells were harvested and extracted for 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ( MTT ) assay . According to the above results , the proteins involving proliferative and apoptotic pathways were evaluated by Western blot analysis . RESULTS : DB01645 , resveratrol , and quercetin significantly inhibited cellular proliferation in a dose- and time-dependent manner . Significantly elevated caspase-3 activity was noted after treatment with genistein ( 10(-9)-10(-7) mol/l ) , as well as with resveratrol and quercetin ( 10(-9)-10(-5) mol/l ) . Significant reduction of PI3K and AKT protein and significant increase of P48023 , Fas-associated protein with death domain , cytochrome C , truncated Bid , caspase-9 , and caspase-3 were all noted after genistein , resveratrol , and quercetin treatment . 17β-Estradiol induced completely opposite results . P03372 ( ER ) α expression was significantly increased with 17β-estradiol , whereas ERβ expression was significantly elevated in the cultures with genistein , resveratrol , and quercetin . CONCLUSIONS : These data demonstrate that genistein , resveratrol , and quercetin have antiproliferative effects on breast cancer cells . Their induction of apoptosis involves the activation of both the intrinsic and extrinsic apoptotic pathways , which may be related to the differential affinity to ERα and ERβ . Whether phytoestrogens have similar effects on normal breast cells remains to be examined . Apigenin inhibits release of inflammatory mediators by blocking the NF-κB activation pathways in the HMC-1 cells . Apigenin is a plant flavonoid and a pharmacologically active agent that has been isolated from several plant species . However , the molecular mechanism of apigenin-mediated immune modulation has not been fully understood . One of the possible mechanisms of its protective effects is the down-regulation of inflammatory responses . In this study , we used cells from the human mast cell line ( HMC-1 ) to investigate this effect . Apigenin significantly inhibits the inductive effect of phorbol 12-myristate 13-acetate ( PMA ) plus A23187 on the production of inflammatory cytokines such as tumor necrosis factor ( P01375 ) -α , interleukin ( IL ) -8 , P05231 , and granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) . Moreover , apigenin attenuated the cyclooxygenase ( P36551 ) -2 expression and intracellular Ca(2+) level . In activated HMC-1 cells , apigenin inhibited the PMA plus A23187-induced activation of nuclear factor ( NF ) -κB , IκB degradation , and luciferase activity . Furthermore , apigenin suppressed the expression of P01375 -α , P10145 , P05231 , GM- P04141 , and P35354 by decreasing the intracellular Ca(2+) level and inhibiting NF-κB activation . These results indicate that apigenin has a potential regulatory effect on inflammatory reactions that are mediated by mast cells . Autocrine stimulation of P35968 activates human leukemic cell growth and migration . Emerging data suggest that P15692 receptors are expressed by endothelial cells as well as hematopoietic stem cells . Therefore , we hypothesized that functional P15692 receptors may also be expressed in malignant counterparts of hematopoietic stem cells such as leukemias . We demonstrate that certain leukemias not only produce P15692 but also express functional P35968 in vivo and in vitro , resulting in the generation of an autocrine loop that may support leukemic cell survival and proliferation . Approximately 50 % of freshly isolated leukemias expressed mRNA and protein for P35968 . P15692 (165) induced phosphorylation of P35968 and increased proliferation of leukemic cells , demonstrating these receptors were functional . P15692 (165) also induced the expression of P14780 by leukemic cells and promoted their migration through reconstituted basement membrane . The neutralizing mAb DB06101 , specific to human P35968 , inhibited leukemic cell survival in vitro and blocked P15692 (165)-mediated proliferation of leukemic cells and P15692 -induced leukemic cell migration . Xenotransplantation of primary leukemias and leukemic cell lines into immunocompromised nonobese diabetic mice resulted in significant elevation of human , but not murine , P15692 in plasma and death of inoculated mice within 3 weeks . Injection of DB06101 inhibited proliferation of xenotransplanted human leukemias and significantly increased the survival of inoculated mice . Interruption of signaling by VEGFRs , particularly P35968 , may provide a novel strategy for inhibiting leukemic cell proliferation . Oleanane triterpenoid CDDO-Me inhibits growth and induces apoptosis in prostate cancer cells by independently targeting pro-survival Akt and P42345 . BACKGROUND : Synthetic triterpenoids are potent anticancer agents , but their therapeutic efficacy or mechanism of action for prostate cancer has not been investigated . The goal of this study was to determine the antitumor activity and the mechanism of action of methyl-2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oate ( CDDO-Me ) , a oleanane-derived synthetic triterpenoid for human prostate cancer cells . METHODS : The antitumor activity of CDDO-Me for hormone-refractory PC-3 ( AR(-) ) and C4-2 ( AR(+) ) prostate cancer cell lines was determined by effects on cell growth and induction of apoptosis , identification of molecular targets , and therapeutic efficacy in vivo in PC-3 xenograft model . RESULTS : CDDO-Me inhibited the growth and induced apoptosis in PC-3 and C4-2 cells at extremely low concentrations . The antitumor activity of CDDO-Me was associated with the inhibition of p-Akt , mammalian target of rapamycin ( P42345 ) , and nuclear factor kappa B ( NF-kappaB ) signaling proteins and their downstream targets such as p-Bad and p-Foxo3a ( Akt ) ; p- P23443 , p- P06730 and p- Q13541 ( P42345 ) ; and P35354 , P15692 and cyclin D1(NF-kappaB) . Silencing of Akt sensitized the PC-3 cells to CDDO-Me , whereas overexpression of Akt induced resistance to CDDO-Me . Targeted silencing of Akt showed that Akt does not regulate P42345 activation in PC-3 cells , but targeted silencing of P42345 sensitized PC-3 cells to CDDO-Me mediated growth inhibition . Further , treatment with CDDO-Me inhibited the growth of PC-3 xenografts in nude mice . CONCLUSIONS : This study demonstrated potent antitumor activity of CDDO-Me against prostate cancer cells both in vitro and in vivo . Data also identified Akt and P42345 as molecular targets of CDDO-Me in prostate cancer cells . Fluid secretion caused by aerolysin-like hemolysin of Aeromonas sobria in the intestines is due to stimulation of production of prostaglandin E2 via cyclooxygenase 2 by intestinal cells . To clarify the mechanisms of diarrheal disease induced by Aeromonas sobria , we examined whether prostaglandin E2 ( DB00917 ) was involved in the intestinal secretory action of A. sobria hemolysin by use of a mouse intestinal loop model . The amount of DB00917 in jejunal fluid and the fluid accumulation ratio were directly related to the dose of hemolysin . The increase over time in the level of DB00917 was similar to that of the accumulated fluid . In addition , hemolysin-induced fluid secretion and DB00917 synthesis were inhibited by the selective cyclooxygenase 2 ( P35354 ) inhibitor NS-398 but not the P23219 inhibitor SC-560 . Western blot analysis revealed that hemolysin increased the P35354 protein levels but reduced the P23219 protein levels in mouse intestinal mucosa in vivo . These results suggest that DB00917 functions as an important mediator of diarrhea caused by hemolysin and that DB00917 is produced primarily through a P35354 -dependent mechanism . Subsequently , we examined the relationship between DB00917 , cyclic AMP ( DB02527 ) , and cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channels in mouse intestinal mucosa exposed to hemolysin . Hemolysin increased the levels of DB02527 in the intestinal mucosa . NS-398 inhibited the increase in DB02527 production , but SC-560 did not . In addition , H-89 , a DB02527 -dependent protein kinase A ( PKA ) inhibitor , and glibenclamide , a P13569 inhibitor , inhibited fluid accumulation . Taken together , these results indicate that hemolysin activates DB00917 production via P35354 and that DB00917 stimulates DB02527 production . DB02527 then activates PKA , which in turn stimulates P13569 Cl- channels and finally leads to fluid accumulation in the intestines . Defined inflammatory states in astrocyte cultures : correlation with susceptibility towards CD95-driven apoptosis . A complete cytokine mix ( CCM ) or its individual components tumour necrosis factor-alpha ( P01375 ) , interleukin-1beta ( IL-1beta ) and interferon-gamma ( P01579 ) were used to switch resting murine astrocytes to reactive states . The transformation process was characterized by differential up-regulation of interleukin-6 ( P05231 ) , cyclooxygenase-2 ( P35354 ) and inducible nitric oxide synthetase ( P35228 ) mRNA and protein and a subsequent release of prostaglandin E2 , nitric oxide ( NO ) and P05231 . Both P48023 and anti-CD95 antibodies triggered caspase activation followed by apoptotic death in fully pro-inflammatory astrocytes , whereas resting cells were totally resistant . Two other death-inducing ligands , P01375 and P50591 ( P50591 ) did not induce apoptosis in reactive astrocytes . The switch in astrocyte sensitivity was accompanied by up-regulation of caspase-8 and CD95 as well as the capacity to recruit Fas-associated death domain ( Q13158 ) to the activated death receptor complex . Neither CD95-mediated death , nor other inflammatory parameters were affected by inhibition of P35228 or P36551 , respectively . Accordingly , P01579 was absolutely essential for up-regulation of P35228 , but not for the switch in apoptosis sensitivity . In contrast , p38 kinase activity was identified as an important controller of both the inflammatory reaction and apoptosis both in astrocytes stimulated with CCM and in glia exposed to P01375 and IL-1 only . Establishment and characterization of a new granulocyte-macrophage colony-stimulating factor-dependent and interleukin-3-dependent human acute myeloid leukemia cell line ( GF- P41226 ) . A novel factor-dependent human myeloid leukemia cell line ( GF- P41226 ) was established from the peripheral blood of an 82-year-old man suffering from acute myeloblastic leukemia ( AML ) . By morphology , cytochemical staining , and analysis of surface antigens , GF- P41226 cells are myeloblasts of immature progenitor origin . The consensus karyotype is 45 , XY , -5 , 7q- , inv(7) ( q31.2q36 ) , 8q+ , +8q+ , 11q+ , 12p- , -15 , -17 , + marker . The long-term survival and proliferation of GF- P41226 cells is dependent on the presence of either granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) or interleukin-3 ( P08700 ) . Weak colony growth was observed after exposure of GF- P41226 cells to stem cell factor ( P21583 ) but not after exposure to granulocyte- P04141 ( DB00099 ) , macrophage- P04141 ( P09603 ) , P01584 , P60568 , P05113 , or tumor necrosis factor-alpha ( P01375 ) . GM- P04141 - and P08700 -induced proliferation is dose dependent , with significant growth observed at concentrations as low as 0.1 ng/mL , but the combination of both factors has no synergistic effect . A significant proliferation is induced by GM- P04141 and P08700 even in serum-deprived cultures , although with a slightly decreased efficiency . GF- P41226 cells were shown to express specific messenger RNAs for the alpha chains of the GM- P04141 and P08700 receptors as well as for the beta chain , common to both receptors . Interestingly , despite the absence of biologic response to DB00099 , specific transcripts for the Q99062 gene were similarly identified by reverse polymerase chain reaction analysis . GF- P41226 cells represent a useful tool for studying chromosome abnormalities of human AML as well as the regulation of myeloid proliferation and differentiation in vitro . DB05767 ( Andrographis paniculata extract ) prevents development of murine colitis by inhibiting T-cell proliferation and Q8IXH7 /TH17 responses . BACKGROUND : Extracts of the plant Andrographis paniculata have been used to treat inflammatory diseases in Asian countries . A recent double-blind , placebo-controlled trial of DB05767 ( A. paniculata extract ) has demonstrated its safety and effectiveness for induction of clinical response , remission , and mucosal healing in patients with mild to moderate ulcerative colitis ( UC ) . We aimed to determine if DB05767 could prevent the development of T-cell-dependent murine colitis and to define its in vivo mechanism(s) of action . METHODS : CD(+)4CD45RB(high) T cells were transferred into Rag1(-/-) mice and gavaged daily with DB05767 or methyl cellulose ( MC ) . Severity of colitis was evaluated by weight loss , histology , and cytokine expression . RESULTS : Mice treated with MC developed colitis within 4-7 weeks , as evaluated by weight loss , and severe intestinal inflammation . DB05767 -treated mice did not lose weight and displayed only very mild intestinal inflammation . P01375 alpha ( P01375 -α ) , interleukin ( IL ) -1β , interferon-gamma ( IFN-γ ) , and Q9GZX6 expression were significantly decreased in DB05767 -treated mice . We observed higher percentages of naïve P01730 (+) T cells in the lamina propria of DB05767 -treated mice . At early timepoints DB05767 -treated mice have significantly reduced splenic cell counts , reduced P01730 (+) , and Q16552 (+) , and IFN-γ T(+) cells . Furthermore , DB05767 inhibited the proliferation of P01730 T cells and differentiation into Q8IXH7 /TH17 cells in vitro . CONCLUSIONS : DB05767 inhibits the development of chronic colitis by affecting early T-cell proliferation , differentiation , and TH(1)/TH(17) responses in a T-cell-driven model of colitis , presenting a unique mechanism of action . Our data suggest that DB05767 could be an attractive herbal therapeutic for inflammatory bowel disease . Microarray analysis reveals characteristic changes of host cell gene expression in response to attenuated modified vaccinia virus Ankara infection of human HeLa cells . The potential use of the modified vaccinia virus Ankara ( MVA ) strain as a live recombinant vector to deliver antigens and elicit protective immune responses against infectious diseases demands a comprehensive understanding of the effect of MVA infection on human host gene expression . We used microarrays containing more than 15,000 human cDNAs to identify gene expression changes in human HeLa cell cultures at 2 , 6 , and 16 h postinfection . Clustering of the 410 differentially regulated genes identified 11 discrete gene clusters with altered expression patterns after MVA infection . Clusters 1 and 2 ( accounting for 16.59 % [ 68 of 410 ] of the genes ) contained 68 transcripts showing a robust induction pattern that was maintained during the course of infection . Changes in cellular gene transcription detected by microarrays after MVA infection were confirmed for selected genes by Northern blot analysis and by real-time reverse transcription-PCR . Upregulated transcripts in clusters 1 and 2 included 20 genes implicated in immune responses , including interleukin 1A ( IL-1A ) , P05231 , P13232 , P10145 , and P40933 genes . MVA infection also stimulated the expression of NF-kappaB and components of the NF-kappaB signal transduction pathway , including p50 and TRAF-interacting protein . A marked increase in the expression of histone family members was also induced during MVA infection . Expression of the Wiskott-Aldrich syndrome family members P42768 , Q92558 , and the small GTP-binding protein P31749 -1 , which are involved in actin cytoskeleton reorganization , was enhanced after MVA infection . This study demonstrates that MVA infection triggered the induction of groups of genes , some of which may be involved in host resistance and immune modulation during virus infection . Farnesoid X receptor responds to bile acids and represses cholesterol 7alpha-hydroxylase gene ( P22680 ) transcription . DB04540 7alpha-hydroxylase gene ( P22680 ) transcription is repressed by bile acids . The goal of this study is to elucidate the mechanism of P22680 transcription by bile acid-activated farnesoid X receptor ( Q96RI1 ) in its native promoter and cellular context and to identify Q96RI1 response elements in the gene . In Chinese hamster ovary cells transfected with retinoid X receptor alpha ( RXRalpha ) / Q96RI1 , only chenodeoxycholic acid ( DB06777 ) and deoxycholic acid ( DB08809 ) were able to stimulate a heterologous promoter/reporter containing an ecdysone response element . In HepG2 cells , all bile acids ( 25 microM ) were able to repress P22680 /luciferase reporter activity , and only DB06777 and DB08809 further repressed reporter activity when cotransfected with RXRalpha/ Q96RI1 . The concentration of DB06777 required to inhibit 50 % of reporter activity ( IC(50) ) was determined to be approximately 25 microM without Q96RI1 and 10 microM with Q96RI1 . Deletion analysis revealed that the bile acid response element located between nucleotides -148 and -128 was the Q96RI1 response element , but RXRalpha/ Q96RI1 did not bind to this sequence . These results suggest that bile acid-activated Q96RI1 exerts its inhibitory effect on P22680 transcription by an indirect mechanism , in contrast to the stimulation and binding of Q96RI1 to intestinal bile acid-binding protein gene promoter . Results also reveal that bile acid receptors other than Q96RI1 are present in HepG2 cells . Involvement of protein kinase Cepsilon in the negative regulation of Akt activation stimulated by granulocyte colony-stimulating factor . Stimulation of cells with G- P04141 activates multiple signaling cascades , including the serine/threonine kinase Akt pathway . We show in this study that G- P04141 -induced activation of Akt in myeloid 32D was specifically inhibited by treatment with PMA , a protein kinase C ( PKC ) activator . PMA treatment also rapidly attenuated sustained Akt activation mediated by a carboxy truncated Q99062 , expressed in patients with acute myeloid leukemia evolving from severe congenital neutropenia . The inhibitory effect of PMA was abolished by pretreatment of cells with specific PKC inhibitor GF109203X , suggesting that the PKC pathway negatively regulates Akt activation . Ro31-8820 , a PKCepsilon inhibitor , also abrogated PMA-mediated inhibition of Akt activation , whereas rottlerin and Go6976 , inhibitors of PKCdelta and PKCalphabetaI , respectively , exhibited no significant effects . Furthermore , overexpression of the wild-type and a constitutively active , but not a kinase-dead , forms of PKCepsilon markedly attenuated Akt activation , and inhibited the proliferation and survival of cells in response to DB00099 . The expression of PKCepsilon was down-regulated with G- P04141 -induced terminal granulocytic differentiation . Together , these results implicate PKCepsilon as a negative regulator of Akt activation stimulated by G- P04141 and indicate that PKCepsilon plays a negative role in cell proliferation and survival in response to DB00099 . Randomised clinical trial : herbal extract DB05767 in active ulcerative colitis - a double-blind comparison with sustained release mesalazine . BACKGROUND : Andrographis paniculata is an herbal mixture used to treat inflammatory diseases . An extract of the herb , DB05767 , inhibits P01375 -α and IL-1β , and prevents colitis in animal models . AIM : To determine the efficacy and safety of DB05767 in patients with mild-to-moderate ulcerative colitis . METHODS : A randomised , double-blind , multicentre , 8-week parallel group study was conducted using DB05767 1200 mg/day compared with 4500 mg/day of slow release mesalazine ( mesalamine ) granules in patients with mild-to-moderately active ulcerative colitis . Disease activity was assessed at baseline and every 2 weeks for clinical response , and at baseline and 8 weeks by colonoscopy . RESULTS : One hundred and twenty patients at five centres in China were randomised and dosed . Clinical remission and response were seen in 21 % and 76 % of DB05767 -treated patients , and 16 % and 82 % of mesalazine-treated patients . By colonoscopy , remission and response were seen in 28 % and 74 % of DB05767 -treated patients and 24 % and 71 % of mesalazine-treated patients , respectively . There was no significant difference between the two treatment groups . CONCLUSION : DB05767 may be an efficacious alternative to mesalazine in ulcerative colitis . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . P11926 as a target for chemoprevention . l- P11926 ( ODC ) is essential for polyamine synthesis and growth in mammalian cells ; it provides putrescine that is usually converted into the higher polyamines , spermidine and spermine . Many highly specific and potent inhibitors of ODC are based on the lead compound alpha-difluoromethylornithine ( DB06243 ) , which is an enzyme-activated irreversible inhibitor . DB06243 is accepted as a substrate by ODC and is decarboxylated , leading to the formation of a highly reactive species that forms a covalent adduct with either cysteine-360 ( 90 % ) or lysine-69 ( 10 % ) . Both modifications inactivate the enzyme . ODC activity is normally very highly regulated at both transcriptional and post-transcriptional levels according to the growth state of the cell and the intracellular polyamine content . Experimental over-production of ODC can be caused by either transfection with plasmids containing the ODC cDNA with part of the 5'-untranslated region ( 5'UTR ) deleted under the control of a very strong viral promoter , or transfection of plasmids that cause the overproduction of P06730 , reported to be a limiting factor in the translation of mRNAs with extensive secondary structures in the 5'UTR . In both cases , unregulated overexpression of ODC transforms NIH 3T3 cells to a neoplastic state . Along with studies showing that many tumor promoters increase ODC activity and that a number of preneoplastic conditions and tumor samples show high levels of ODC , these results suggest that ODC may act as an oncogene in an appropriate background . This provides a rationale for the possible use of ODC inhibitors as chemopreventive agents. ( ABSTRACT TRUNCATED AT 250 WORDS ) A validated high-performance liquid chromatographic assay for determination of lumiracoxib in human plasma . DB01283 { 2-[(2-fluoro-6-chlorophenyl)amino]-5-methyl- DB09269 } is a highly selective and potent cyclooxygenase-2 ( P35354 ) inhibitor , which is chemically distinct from other coxibs in that it contains a carboxylic group and is weakly acidic . In the present study , a liquid-liquid extraction-based reversed-phase HPLC method with UV detection was validated and applied for the analysis of lumiracoxib in human plasma . The analyte was separated on a reversed-phase column with acetonitrile and 0.05 % trichloracetic acid in water ( 35:65 , v/v ) as mobile phase at a flow rate of 1 mL/min , and UV detection at 270 nm . The retention times for lumiracoxib and niflumic acid ( internal standard ) were 16.9 and 10.4 min , respectively . The validated quantitation range for lumiracoxib was 10-10,000 ng/mL . The developed procedure was applied to assess the pharmacokinetics of lumiracoxib following administration of a single oral 200 mg dose to a healthy male volunteer . P01375 and its inhibitors in cancer . P01375 ( P01375 ) -alpha is implicated in the same time in apoptosis and in cell proliferation . P01375 not only acts as pro-inflammatory cytokine conducing to wide spectrum of human diseases including inflammatory diseases , but can also induce tumor development . The molecular mechanisms of P01375 functions have been intensively investigated . In this review we covered P01375 , the molecule , its signaling pathway , and its therapeutic functions . We provide a particular insight in its paradoxical role in tumor promotion and in its use as anti-tumor agent . This review considers also the recent findings regarding P01375 inhibitors , their pharmacokinetics , and their pharmacodynamics . Six P01375 inhibitors have been considered here : DB00065 , DB00051 , Golimumab , DB08904 , CDP571 , DB00005 , and Thalidomide . We discussed the clinical relevance of their functions in treatment of several diseases such as advanced inflammatory rheumatic and bowel disease , with a focus in cancer treatment . Targeting P01375 by these drugs has many side effects like malignancies development , and the long-term sequels are not very well explored . Their efficacy and their safety were discussed , underscoring the necessity of close patients monitoring and of their caution use . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . Targeting autocrine and paracrine P15692 receptor pathways inhibits human lymphoma xenografts in vivo . The role of angiogenesis in lymphoproliferative diseases is not well established . We demonstrate here that human lymphoma cells secrete vascular endothelial growth factor ( P15692 ) and express P15692 receptor 1 ( P17948 ) and P35968 . Proliferation of non-Hodgkin lymphoma ( Q9NZ71 ) cells under serum-free conditions was enhanced by the addition of P15692 and was blocked by P17948 - and P35968 -specific antibodies . To differentiate between P15692 -mediated autocrine and paracrine effects on lymphoma growth , NOD/SCID mice engrafted with human diffuse large B-cell lymphoma ( DLBCL ) were treated with species-specific antibodies against human P17948 ( 6.12 ) , human P35968 ( DB06101 ) , murine P17948 ( MF-1 ) , or murine P35968 ( DC101 ) . Treatment with 6.12 or DC101 ( targeting tumor P17948 and host P35968 ) reduced established DLBCL xenograft growth , whereas treatment with DB06101 or MF-1 ( targeting tumor P17948 and host P17948 ) had no effect . Decreased tumor volumes after 6.12 and DC101 treatment correlated with increased tumor apoptosis and reduced vascularization , respectively , supporting the presence of autocrine P17948 - and paracrine P35968 -mediated pathways in lymphomagenesis . Inhibition of paracrine P15692 interactions ( DC101 ) in these models was equivalent to their inhibition with rituximab . Combining DC101 with therapeutic agents ( rituximab , 6.12 , methotrexate ) consistently improved tumor responses over those of single-agent therapy . These data support the further clinical development of VEGFR-targeted approaches for the therapy of aggressive DLBCL . DB01645 stimulates duodenal HCO(3)(-) secretion through PI3K pathway in mice . DB01645 has been proposed as a promising pharmacotherapeutic for cystic fibrosis . We recently found that genistein stimulates murine duodenal HCO(3)(-) secretion through cystic fibrosis transmembrane conductance regulator ( P13569 ) . The aim of the present study was to determine the intracellular signal pathways involved in genistein-stimulated duodenal HCO(3)(-) secretion . Murine duodenal mucosal HCO(3)(-) secretion was examined in vitro in Ussing chambers by the pH-stat technique . The results showed that neither DB02527 -dependent signal pathway inhibitors MDL-12330A and KT-5720 , nor cGMP signal pathway inhibitors NS2028 and KT5823 , nor calcium signal pathway inhibitors verapamil and W-13 , altered genistein-stimulated duodenal HCO(3)(-) secretion . In calcium-free solution , genistein-stimulated duodenal HCO(3)(-) secretion was not altered either . Vanadate , an inhibitor of protein tyrosine phosphatase , only partially inhibited genistein-stimulated duodenal HCO(3)(-) secretion . However , both wortmannin and LY294002 , two structurally and mechanistically distinct phosphatidylinositol 3-kinase ( PI3K ) inhibitors , markedly inhibited genistein-stimulated duodenal HCO(3)(-) secretion . DB01645 increased duodenal mucosal PI3K activity and induced the phosphorylation of Akt , a signaling molecule downstream of PI3K , which was again inhibited by wortmannin . P03372 antagonist , ICI182,780 , also markedly inhibited genistein-stimulated duodenal HCO(3)(-) secretion and genistein-induced PI3K activity increase in duodenal mucosa . These results demonstrate that genistein stimulates duodenal HCO(3)(-) secretion mainly through estrogen receptor and PI3K-dependent pathway . These findings contribute to the understanding of the molecular mechanism of genistein-induced anion secretion and further pharmacotherapeutic development and use of genistein or related substances in the treatment of diseases of epithelial tissues . Regulation of P01160 secretion from isolated atria by prostaglandins and cyclooxygenase-2 . Cyclooxygenase ( P36551 ) is a key enzyme regulating the production of various prostaglandins ( PGs ) from arachidonic acid . Angiotensin II has been reported to be an important inflammatory mediator , which increases P35354 . The aim of this study was to determine the role of various PGs and P35354 in the regulation of atrial natriuretic peptide ( P01160 ) secretion . PGF2alpha and PGD2 caused dose-dependent increases in P01160 release and intra-atrial pressure . The potency for the stimulation of P01160 secretion by PGF2alpha was higher than that by PGD2 . In contrast , DB00917 , DB01240 , PGJ2 , and thromboxane A2 did not show any significant effects . The increases in intra-atrial pressure and P01160 secretion induced by PGF2alpha and PGD2 were significantly attenuated by the pretreatment with an inhibitor of PGF2alpha receptor . By the pretreatment with an inhibitor for phospholipase C ( P98160 ) , inositol 3-phosphate ( IP3 ) receptor , protein kinase C ( PKC ) , or myosin light chain kinase ( MLCK ) , PGF2alpha-mediated increase in P01160 secretion and positive inotropy were attenuated . Inhibitor for P23219 or P35354 did not cause any significant effects on atrial parameters . In hypertrophied rat atria , PGF2alpha-induced positive inotropy and P01160 secretion were markedly attenuated whereas P35354 inhibitor stimulated P01160 secretion . The expression of P35354 increased and the expression of PGF2alpha receptor mRNA decreased in hypertrophied rat atria . These results suggest that PGF2alpha increased the P01160 secretion and positive inotropy through P98160 -IP3-PKC-MLCK pathway , and the modulation of P01160 secretion by P35354 inhibitor and PGF2alpha may partly relate to the development of renal hypertension . P11926 activity in human endometrium and endometrial cancer cells . P11926 ( ODC ) activities were significantly higher in proliferative endometrium during the estrogen-dominated follicular phase of the menstrual cycle than in secretory endometrium after the formation of the progesterone-secreting corpus luteum . The enzymatic activity was increased about fivefold by renewal of the medium during incubations of endometrial fragments or isolated endometrial glands . Endometrial adenocarcinoma cells ( O14777 -1 , O14777 -50 ) , both in monolayers and suspension , also responded to medium renewal by increasing ODC activity about 10-fold after 4 h , with subsequent reduction to control levels after 7 h . These effects were blocked by actinomycin D and cycloheximide . Endometrial stromal cells exhibited highly variable ODC activities at different passages . Difluoromethylornithine ( DB06243 ) and sodium molybdate had marked antiproliferative effects in O14777 -50 cultures , reducing cell numbers to 10 to 20 % of control values 11 d after plating and inhibiting ODC activity by approximately 80 % on Day 7 . The antiproliferative effect of DB06243 , but not that of molybdate , was reversed by 10 microM putrescine , the product of ODC activity . In contrast to DB06243 , molybdate had no effect on ODC activity of cell homogenates . Molybdate did not elicit antizyme formation in O14777 -50 cells under conditions in which putrescine did . These results indicate that ODC activity , present in both epithelial and stromal cells , as shown analytically and also by autoradiography after labeling with [3H] DB06243 , may be related to cell proliferation in vivo and that proliferation of human endometrial cancer cells in culture can be arrested by DB06243 and by molybdate . DB01283 ( Novartis ) . DB01283 , an inhibitor of cyclooxygenase 2 ( P35354 ) , is under development by Novartis for the potential treatment of osteoarthritis , rheumatoid arthritis and pain . By late December 2000 , phase III trials had been initiated and were ongoing in December 2001 . Regulation and therapeutic targeting of peptide-activated receptor guanylyl cyclases . Cyclic GMP is a ubiquitous second messenger that regulates a wide array of physiologic processes such as blood pressure , long bone growth , intestinal fluid secretion , phototransduction and lipolysis . Soluble and single-membrane-spanning enzymes called guanylyl cyclases ( GC ) synthesize cGMP . In humans , the latter group consists of P16066 , P20594 , P25092 , GC-E and P51841 , which are also known as P16066 , P20594 , StaR , Ret1-GC and Ret2-GC , respectively . Membrane GCs are activated by peptide ligands such as atrial natriuretic peptide ( P01160 ) , B-type natriuretic peptide ( DB04899 ) , P23582 ( P09543 ) , guanylin , uroguanylin , heat stable enterotoxin and GC-activating proteins . DB04899 and carperitide are clinically approved peptide-based drugs that activate P16066 . CD-NP is an experimental heart failure drug that primarily activates P20594 but also activates P16066 at high concentrations and is resistant to degradation . Inactivating mutations in P20594 cause acromesomelic dysplasia type Maroteaux dwarfism and chromosomal mutations that increase P09543 concentrations are associated with Marfanoid-like skeletal overgrowth . Pump-based P09543 infusions increase skeletal growth in a mouse model of the most common type of human dwarfism , which supports P09543 / P20594 -based therapies for short stature diseases . DB08890 is a peptide activator of P25092 that stimulates intestinal motility and is in late-stage clinical trials for the treatment of chronic constipation . This review discusses the discovery of cGMP , guanylyl cyclases , the general characteristics and therapeutic applications of P16066 , P20594 and P25092 , and emphasizes the regulation of transmembrane guanylyl cyclases by phosphorylation and DB00171 . Anti-tumor necrosis factor monoclonal antibody therapy for gastrointestinal Behçet 's disease : a case report . Behçet 's disease ( BD ) is a multisystem immune-mediated inflammatory disorder that involves the intestine in 3 % -26 % of cases . Corticosteroids , DB00244 derivatives , immunomodulators , and more recently thalidomide and pentoxifylline have been used to treat BD with varying degrees of success . P01375 ( P01375 ) -alpha is believed to play a pivotal role in this T helper cell type 1 ( Th1 ) -mediated disease . DB00065 , a chimeric monoclonal antibody to P01375 , has been demonstrated to be an effective therapy for Crohn 's disease and rheumatoid arthritis , 2 other Th1-mediated disorders . We describe a patient with chronically active , steroid-dependent BD involving the gastrointestinal tract who received 4 doses of infliximab during a 6-month period . Because most of her symptoms were gastrointestinal , the Crohn 's Disease Activity Index ( CDAI ) was used to assess response . A rapid and dramatic improvement in both gastrointestinal and extraintestinal symptoms was observed . The CDAI score decreased from 270 points ( preinfusion ) to 13 points by week 2 , and remission was sustained despite complete withdrawal of steroids . Colonoscopy performed 10 weeks after the first infusion showed marked endoscopic and histologic improvement . This report suggests that infliximab may be an effective new therapy for gastrointestinal BD , and perhaps other manifestations of BD as well . DB08890 - a secretagogue and antihyperalgesic agent - what next ? Ongoing clinical trials suggest that linaclotide , a first-in-class , 14-amino acid peptide guanylate cyclase-C ( P25092 ) receptor agonist and intestinal secretagogue is an effective treatment for chronic constipation . A study in this issue of the Journal suggests that linaclotide also has antihyperalgesic effects in three common rat models of inflammation- and stress-induced hypersensitivity ( i.e. , acute trinitrobenzene sulfonic acid colitis , water avoidance stress [ P42768 ] , and restraint-induced stress ) but not in naïve animals . In mice , linaclotide at least partly reduces hyperalgesia via P25092 receptors . Dose-effect relationships of linaclotide were complicated and non-linear . This viewpoint discusses human clinical trials with linaclotide and the results of this study . Potential mechanisms and clinical significance of these findings are explored . Collectively , these data suggest that P25092 receptors exert other , as yet poorly understood , effects on gastrointestinal sensitivity in conditions associated with inflammation and/or stress-induced increased intestinal permeability . However , the data need to be confirmed in humans and in long-term animal models . Further studies are also necessary to elucidate the mechanisms as these effects can not be explained by linaclotide 's known effects on epithelial P25092 receptors . Guanylate cyclase C-mediated antinociceptive effects of linaclotide in rodent models of visceral pain . BACKGROUND DB08890 is a novel , orally administered investigational drug currently in clinical development for the treatment of constipation-predominant irritable bowel syndrome ( IBS-C ) and chronic idiopathic constipation . Visceral hyperalgesia is a major pathophysiological mechanism in IBS-C . Therefore , we investigated the anti-nociceptive properties of linaclotide in rodent models of inflammatory and non-inflammatory visceral pain and determined whether these pharmacological effects are linked to the activation of guanylate cyclase C ( P25092 ) . METHODS Orally administered linaclotide was evaluated in non-inflammatory acute partial restraint stress ( PRS ) and acute water avoidance stress ( P42768 ) models in Wistar rats , and in a trinitrobenzene sulfonic acid ( TNBS ) -induced inflammatory model in Wistar rats and P25092 null mice . KEY RESULTS In TNBS-induced colonic allodynia , linaclotide significantly decreased the number of abdominal contractions in response to colorectal distension without affecting the colonic wall elasticity change in response to distending pressures after TNBS . However , linaclotide had no effect on visceral sensitivity under basal conditions . In addition , linaclotide significantly decreased colonic hypersensitivity in the PRS and P42768 models . In wild type ( wt ) and P25092 null mice , the instillation of TNBS induced similar hyperalgesia and allodynia . However , in post-inflammatory conditions linaclotide significantly reduced hypersensitivity only in wt mice , but not in P25092 null mice . CONCLUSIONS & INFERENCES These findings indicate that linaclotide has potent anti-nociceptive effects in several mechanistically different rodent models of visceral hypersensitivity and that these pharmacological properties of linaclotide are exerted through the activation of the P25092 receptor . Therefore , linaclotide may be capable of decreasing abdominal pain in patients suffering from IBS-C . Studies on induction of lamotrigine metabolism in transgenic UGT1 mice . A transgenic ' knock-in ' mouse model expressing a human UGT1 locus ( P01266 -UGT1 ) was recently developed and validated . Although these animals express mouse P22309 proteins , P22310 is a pseudo-gene in mice . Therefore , P01266 -UGT1 mice serve as a ' humanized ' P22310 animal model . Lamotrigine ( LTG ) is primarily metabolized to its N-glucuronide ( LTGG ) by hUGT1A4 . This investigation aimed at examining the impact of pregnane X receptor ( O75469 ) , constitutive androstane receptor ( CAR ) and peroxisome proliferator-activated receptor ( Q07869 ) activators on LTG glucuronidation in vivo and in vitro . P01266 -UGT1 mice were administered the inducers phenobarbital ( CAR ) , pregnenolone-16alpha-carbonitrile ( O75469 ) , WY-14643 ( Q07869 ) , ciglitazone ( P37231 ) , or L-165041 ( Q03181 ) , once daily for 3 or 4 days . Thereafter , LTG was administered orally and blood samples were collected over 24 h . LTG was measured in blood and formation of LTGG was measured in pooled microsomes made from the livers of treated animals . A three-fold increase in in vivo LTG clearance was seen after phenobarbital administration . In microsomes prepared from phenobarbital-treated P01266 -UGT1 animals , 13-fold higher CL(int) ( Vmax/K(m) ) value was observed as compared with the untreated transgenic mice . A trend toward induction of catalytic activity in vitro and in vivo was also observed following pregnenolone-16alpha-carbonitrile and WY-14643 treatment . This study demonstrates the successful application of P01266 -UGT1 mice as a novel tool to study the impact of induction and regulation on metabolism of P22310 substrates . P11926 is a mediator of c-Myc-induced apoptosis . c-Myc plays a central role in the regulation of cell cycle progression , differentiation , and apoptosis . However , the proteins which mediate c-Myc function(s) remain to be determined . Enforced c-myc expression rapidly induces apoptosis in interleukin-3 ( P08700 ) -dependent 32D.3 murine myeloid cells following P08700 withdrawal , and this is associated with the constitutive , growth factor-independent expression of ornithine decarboxylase ( ODC ) , a rate-limiting enzyme of polyamine biosynthesis . Here we have examined the role of ODC in c-Myc-induced apoptosis . Enforced expression of ODC , like c-myc , is sufficient to induce accelerated death following P08700 withdrawal . ODC induced cell death in a dose-dependent fashion , and alpha-difluoromethylornithine ( DB06243 ) , an irreversible inhibitor of ODC enzyme activity , effectively blocked ODC-induced cell death . ODC-induced cell death was due to the induction of apoptosis . We also demonstrate that ODC is a mediator of c-Myc-induced apoptosis . 32D.3-derived c-myc clones have augmented levels of ODC enzyme activity , and their rates of death were also a function of their ODC enzyme levels . Importantly , the rates of death of c-myc clones were inhibited by treatment with DB06243 . These findings demonstrate that ODC is an important mediator of c-Myc-induced apoptosis and suggest that ODC mediates other c-Myc functions . Regulation of cyclooxygenase-2 and endogenous cytokine expression by bacterial lipopolysaccharide that acts in synergy with c-kit ligand and Fc epsilon receptor I crosslinking in cultured mast cells . Emerging evidence has suggested the pivotal role of mast cells in a host defense against bacterial infection . In this paper , we report that bacterial lipopolysaccharide ( LPS ) is a potent enhancer of the cytokine- and IgE-dependent delayed responses of P08700 -dependent mouse bone marrow-derived cultured mast cells ( BMMC ) . LPS , although showing minimal effects , significantly augmented the c-kit ligand ( KL ) - or IgE-dependent expression of cyclooxygenase ( P36551 ) -2 and the attendant delayed PGD2 generation , with P22301 and P05112 acting as potentiating and inhibitory cytokines , respectively . The P35354 -inducing activity of LPS was mimicked by exogenous P01584 . Assessment of endogenous cytokine induction revealed that P01584 expression was stimulated by either LPS or exogenous P01584 . P05231 expression occurred in parallel with P35354 expression . P22301 expression , which lagged behind P35354 expression , depended on exogenous P22301 , but not on LPS and P01584 . Thus , LPS and P01584 exhibited similar biological activities in terms of P35354 and endogenous cytokine expression . However , adding an antibody against the type I IL-1 receptor to BMMC , which abrogated the effects of P01584 , failed to neutralize the effects of LPS . These results suggest that LPS activates BMMC through the signal transduction pathway shared with exogenous P01584 , rather than exerting its action indirectly via the production of endogenous P01584 . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . Andrographis paniculata extract ( DB05767 ) for active ulcerative colitis . OBJECTIVES : Andrographis paniculata has in vitro inhibitory activity against P01375 -α , IL-1β and NF-κB . A pilot study of A. paniculata extract ( DB05767 ) suggested similar efficacy to mesalamine for ulcerative colitis . METHODS : A randomized , double-blind , placebo-controlled trial evaluated the efficacy of A. paniculata extract ( DB05767 ) in 224 adults with mild-to-moderate ulcerative colitis . Patients were randomized to A. paniculata extract ( DB05767 ) 1,200 mg or 1,800 mg daily or placebo for 8 weeks . RESULTS : In total , 45 and 60 % of patients receiving A. paniculata 1,200 mg and 1,800 mg daily , respectively , were in clinical response at week 8 , compared with 40 % of those who received placebo ( P=0.5924 for 1,200 mg vs. placebo and P=0.0183 for 1,800 mg vs. placebo ) . In all , 34 and 38 % of patients receiving A. paniculata 1,200 mg and 1,800 mg daily , respectively , were in clinical remission at week 8 , compared with 25 % of those who received placebo ( P=0.2582 for 1,200 mg vs. placebo and P=0.1011 for 1,800 mg vs. placebo ) . Adverse events developed in 60 and 53 % of patients in the A. paniculata 1,200 mg and 1,800 mg daily groups , respectively , and 60 % in the placebo group . CONCLUSIONS : Patients with mildly to moderately active ulcerative colitis treated with A. paniculata extract ( DB05767 ) at a dose of 1,800 mg daily were more likely to achieve clinical response than those receiving placebo . Metronomic chemotherapy dosing-schedules with estramustine and temozolomide act synergistically with anti- P35968 antibody to cause inhibition of human umbilical venous endothelial cell growth . The effects of ' metronomic ' or extended chemotherapy dosing schedules ( ECS ) are mediated through poorly understood anti-angiogenic mechanisms . ECS combined with biological anti-angiogenic agents have produced promising pre-clinical results . MATERIALS AND METHODS : We have expanded the list of agents with an in vitro ECS profile to include the methylating agent temozolomide ( DB00853 ) and the anti-mitotic agent estramustine ( Estracyt ) . These agents were also combined with a specific anti-angiogenic inhibitor DB06101 and a non-specific agent with anti-angiogenic properties , Compound 5h . The in vitro HUVEC ECS model system was optimised and cell proliferation assays undertaken . RESULTS : As a single agent , estramustine inhibited endothelial cell proliferation with an IC50 of 4.5 microM and was active at 10-33 % of the maximum tolerated dose ( MTD ) from clinical schedules , whilst temozolomide had IC50 of 6.6 microM and was active at 1-6 % of MTD . In combination , significant synergy was seen with DB06101 in combination with either drug , whilst modest additive effects were observed with Compound 5h . None of the combinations resulted in significant cytotoxicity or apoptosis . DISCUSSION : The results show that ECS of temozolomide and estramustine can be significantly enhanced when combined with specific anti-angiogenic inhibitors in an in vitro HUVEC system . Clinical pharmacology of novel selective P35354 inhibitors . Novel coxibs ( i.e. etoricoxib , valdecoxib , parecoxib and lumiracoxib ) with enhanced biochemical cyclooxygenase ( P36551 ) -2 selectivity over that of rofecoxib and celecoxib have been recently developed . They have the potential advantage to spare P23219 activity , thus reducing gastrointestinal toxicity , even when administered at high doses to improve efficacy . They are characterized by different pharmacodynamic and pharmacokinetics features . The higher biochemical selectivity of valdecoxib than celecoxib , evidenced in vitro , may be clinically relevant leading to an improved gastrointestinal safety . Interestingly , parecoxib , a pro-drug of valdecoxib , is the only injectable coxib . DB01628 shows only a slightly improved P35354 selectivity than rofecoxib , a highly selective P35354 inhibitor that has been reported to halve the incidence of serious gastrointestinal toxicity compared to nonselective nonsteroidal antiinflammatory drugs ( NSAIDs ) . DB01283 , the most selective P35354 inhibitor in vitro , is the only acidic coxib . The hypothesis that this chemical property may lead to an increased and persistent drug accumulation in inflammatory sites and consequently to an improved clinical efficacy , however , remains to be verified . Several randomized clinical studies suggest that the novel coxibs have comparable efficacy to nonselective NSAIDs in the treatment of osteoarthritis , rheumatoid arthritis and acute pain , but they share similar renal side-effects . The apparent dose-dependence of renal toxicity may limit the use of higher doses of the novel coxibs for improved efficacy . Large-size randomized clinical trials are ongoing to define the gastrointestinal and cardiovascular safety of the novel coxibs . G- P04141 -mediated inhibition of JNK is a key mechanism for Lactobacillus rhamnosus-induced suppression of P01375 production in macrophages . Lactobacillus rhamnosus is a human commensal with known immunomodulatory properties . To date the mechanism of these immunomodulatory effects is not well understood . To unravel the immunomodulatory signalling mechanism , we investigated the effects of two strains of L. rhamnosus , L. rhamnosus GG and GR-1 , in modulating production of tumour necrosis factor-alpha ( P01375 ) in human monocytic cell line THP-1 and mouse macrophages . Live L. rhamnosus GG and GR-1 or their spent culture supernatant induced minuscule amounts of P01375 production but large quantities of granulocyte-colony stimulating factor ( DB00099 ) in macrophages compared with those induced by pathogenic Escherichia coli GR-12 and Enterococcus faecalis . By using neutralizing antibodies and Q99062 knockout mice , we demonstrated that G- P04141 secreted from L. rhamnosus GG- and GR-1-exposed macrophages suppressed P01375 production induced by E. coli- or lipopolysaccharide-activated macrophages through a paracrine route . The suppression of P01375 production by G- P04141 was mediated through activation of P40763 and subsequent inhibition of c-Jun-N-terminal kinases ( JNKs ) . The inhibition of JNK activation required STAT3alpha-mediated de novo protein synthesis . This demonstrates a novel role of G- P04141 in L. rhamnosus-triggered anti-inflammatory effects and its mechanism in the suppression of P01375 production in macrophages . Regulation of the human bile acid P35503 by the farnesoid X receptor and bile acids . BACKGROUND & AIMS : Cholestasis is a serious complication of many liver diseases leading to increased serum bile acids ( BA ) and their conjugates . Chenodeoxycholic ( DB06777 ) acid is a substrate of the human hepatic UDP-glucuronosyltransferase ( P78381 ) 1A3 . P35503 may , therefore , be a BA-inducible gene relevant to BA regulation . METHODS : BA and human bile were used to induce P35503 in HepG2 cells . Genomic DNA was analyzed by PCR amplification and sequencing . Transcriptional regulation was studied by DNA mutagenesis , RT-PCR , luciferase reporter gene constructs and electrophoretic mobility shift assays ( EMSA ) . RESULTS : DB06777 differentially induced P35503 but not P22310 expression . Bile from ursodeoxycholic acid ( DB01586 ) -treated and untreated patients differentially induced P35503 . A farnesoid X receptor ( Q96RI1 ) half-site DNA motif was identified in the P35503 5' upstream region . The Q96RI1 inducer GW4064 activated P35503 transcription , and electrophoretic mobility shift assays identified P35503 as a Q96RI1 target gene . CONCLUSIONS : Transcriptional regulation of the human bile acid and xenobiotic P35503 by its substrate DB06777 and Q96RI1 is shown . DB06777 glucuronidation can be controlled by feed back inhibition proceeding via the glucuronidation of DB06777 . DB01586 does not induce P35503 transcription . Since P35503 is significantly induced by xenobiotics this physiologically links xenobiotic and bile acid metabolism to cholestasis . DB00107 increases invasive properties of endometrial cancer cells through phosphatidylinositol 3-kinase/AKT-dependent up-regulation of cyclooxygenase-1 , -2 , and P98170 . Traditionally , oxytocin ( OT ) is well known to play a crucial role in the regulation of cyclic changes in the uterus , implantation of the embryo , and parturition . Recently , an additional role for OT has been identified in several types of cancer cells in which OT acts as a growth regulator . In endometrial cancer cells , OT is known to efficiently inhibit cellular proliferation . In the present study , we show that OT increases invasiveness of human endometrial carcinoma ( O14777 ) cells , which are otherwise resistant to the growth-inhibiting effects of OT . Using pharmacological inhibitors , invasion assay , RNA interference , and immunofluorescence , we found that OT enhances the invasive properties of O14777 cells through up-regulation of P98170 ( P98170 ) , matrix-metalloproteinase 2 ( P08253 ) , and matrix-metalloproteinase 14 ( P50281 ) . In addition , we show that OT-mediated invasion is both cyclooxygenase 1 ( P23219 ) and cyclooxygenase-2 ( P35354 ) dependent via the phosphatidylinositol 3-kinase/AKT ( PIK3/AKT ) pathway . P35354 knockdown by shRNA resulted in P98170 down-regulation . We also show that OT receptor is overexpressed in grade I to III endometrial cancer . Taken together , our results describe for the first time a novel role for OT in endometrial cancer cell invasion .

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interacts_with DB04868?

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MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens . We propose the Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout ( MAGeCK ) method for prioritizing single-guide RNAs , genes and pathways in genome-scale CRISPR/Cas9 knockout screens . MAGeCK demonstrates better performance compared with existing methods , identifies both positively and negatively selected genes simultaneously , and reports robust results across different experimental conditions . Using public datasets , MAGeCK identified novel essential genes and pathways , including P00533 in vemurafenib-treated A375 cells harboring a P15056 mutation . MAGeCK also detected cell type-specific essential genes , including P11274 and P00519 , in KBM7 cells bearing a P11274 - P00519 fusion , and P08069 in HL-60 cells , which depends on the insulin signaling pathway for proliferation . Blockade of interleukin-6 signalling with siltuximab enhances melphalan cytotoxicity in preclinical models of multiple myeloma . Signalling through the interleukin ( IL ) -6 pathway induces proliferation and drug resistance of multiple myeloma cells . We therefore sought to determine whether the P05231 -neutralizing monoclonal antibody siltuximab , formerly CNTO 328 , could enhance the activity of melphalan , and to examine some of the mechanisms underlying this interaction . DB09036 increased the cytotoxicity of melphalan in KAS-6/1 , Q16352 -6 , ANBL-6 , and RPMI 8226 human myeloma cell lines ( HMCLs ) in an additive-to-synergistic manner , and sensitized resistant RPMI 8226.LR5 cells to melphalan . These anti-proliferative effects were accompanied by enhanced activation of drug-specific apoptosis in HMCLs grown in suspension , and in HMCLs co-cultured with a human-derived stromal cell line . DB09036 with melphalan enhanced activation of caspase-8 , caspase-9 , and the downstream effector caspase-3 compared with either of the single agents . This increased induction of cell death occurred in association with enhanced Bak activation . Neutralization of P05231 also suppressed signalling through the phosphoinositide 3-kinase/Akt pathway , as evidenced by decreased phosphorylation of Akt , P08133 S6 kinase and Q13541 . Importantly , the siltuximab/melphalan regimen demonstrated enhanced anti-proliferative effects against primary plasma cells derived from patients with myeloma , monoclonal gammopathy of undetermined significance , and amyloidosis . These studies provide a rationale for translation of siltuximab into the clinic in combination with melphalan-based therapies . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . Genetic determinants influencing human serum metabolome among African Americans . Phenotypes proximal to gene action generally reflect larger genetic effect sizes than those that are distant . The human metabolome , a result of multiple cellular and biological processes , are functional intermediate phenotypes proximal to gene action . Here , we present a genome-wide association study of 308 untargeted metabolite levels among African Americans from the Atherosclerosis Risk in Communities ( ARIC ) Study . Nineteen significant common variant-metabolite associations were identified , including 13 novel loci ( p < 1.6 × 10(-10) ) . These loci were associated with 7-50 % of the difference in metabolite levels per allele , and the variance explained ranged from 4 % to 20 % . Fourteen genes were identified within the nineteen loci , and four of them contained non-synonymous substitutions in four enzyme-encoding genes ( P03952 , Q9HAT2 , P31327 , and Q9UHE5 ) ; the other significant loci consist of eight other enzyme-encoding genes ( P12821 , P50440 , Q96HD9 , Q68CK6 , Q5T1C6 , P08319 , P22309 , O43280 ) , a transporter gene ( Q9NSD5 ) and a polycystin protein gene ( Q9P0L9 ) . In addition , four potential disease-associated paths were identified , including two direct longitudinal predictive relationships : Q9UHE5 with N-acetylornithine , N-acetyl-1-methylhistidine and incident chronic kidney disease , and O43280 with trehalose and incident diabetes . These results highlight the value of using endophenotypes proximal to gene function to discover new insights into biology and disease pathology . Characterization of gonadotropin-releasing hormone analogs based on a sensitive cellular luciferase reporter gene assay . A novel cellular assay for the functional characterization of agonistic and antagonistic analogs of gonadotropin-releasing hormone ( DB00644 ) was developed . This assay is based on a fusion of the c-fos immediate-early gene promoter to Photinus pyralis luciferase ( Luc ) as a reporter gene , stably transfected in a recombinant cell line expressing the human P30968 . Transcription of endogenous c-fos and fos-Luc fusion gene are transiently induced quite similar by fetal calf serum or the superagonistic analog [ D-Trp6 ] DB00644 in a selected cell line . The reporter gene was therefore used to monitor agonist-induced signaling via the human P30968 . Whereas Luc activity was induced in a dose-dependent manner by DB00644 or [ D-Trp6 ] DB00644 , different antagonistic peptides completely inhibited this stimulation . The antagonistic potency ( IC50 ) of various peptides with DB00050 and Antarelix as lead compounds in general correlated well with the binding affinity ( KD ) as determined from ligand binding experiments . The specificity of an inhibitory effect was confirmed by P30968 -independent stimulation with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate or basic fibroblast growth factor . Since this new reporter gene assay is sensitive and simple and can be performed in a microtiter plate , it will significantly facilitate screening and functional characterization of DB00644 analogs . Differential effects of the vascular endothelial growth factor receptor inhibitor PTK787/ZK222584 on tumor angiogenesis and tumor lymphangiogenesis . Halting tumor growth by interfering with tumor-induced angiogenesis is an attractive therapeutic approach . Such treatments include humanized antibodies blocking the activity of vascular endothelial growth factor ( P15692 ) -A ( bevacizumab ) , soluble P15692 receptor ( VEGFR ) constructs ( P15692 - Q8NHU6 ) , or small-molecule inhibitors of VEGFR signaling , including PTK787/ZK222584 ( DB04879 ) , sorafenib , and sunitinib . DB04879 has been shown previously to specifically block P15692 -induced phosphorylation of P17948 , -2 and -3 and thereby to inhibit endothelial cell proliferation , differentiation , and tumor angiogenesis . We have investigated the effect of DB04879 on tumor angiogenesis and tumor lymphangiogenesis using the Rip1Tag2 transgenic mouse model of pancreatic beta cell carcinogenesis . In Rip1Tag2 mice , tumor angiogenesis is predominantly mediated by P15692 , and as expected , DB04879 efficiently impaired tumor blood vessel angiogenesis and tumor growth . Double-transgenic Rip1Tag2;Rip1VEGF-C and Rip1Tag2;Rip1VEGF-D mice not only exhibit P15692 -dependent blood vessel angiogenesis but also tumor lymphangiogenesis induced by the transgenic expression of P49767 or -D . In these mouse models , DB04879 also repressed tumor blood vessel angiogenesis and tumor growth yet failed to affect tumor lymphangiogenesis and lymphogenic metastasis . Adenoviral delivery of soluble P35916 also did not prevent tumor lymphangiogenesis in these mice . In contrast , spontaneous tumor lymphangiogenesis , as observed by the stochastic expression of P49767 and -D in tumors of neural cell adhesion molecule-deficient Rip1Tag2 mice , was repressed by DB04879 and soluble P35916 . The results indicate that the time of onset and the levels of P49767 /D expression may be critical variables in efficiently repressing tumor lymphangiogenesis and that pathways other than VEGFR signaling may be involved in tumor lymphangiogenesis . P10275 signals regulate UDP-glucuronosyltransferases in the urinary bladder : a potential mechanism of androgen-induced bladder carcinogenesis . UDP-glucuronosyltransferases ( UGTs ) , major phase II drug metabolism enzymes , play an important role in urinary bladder cancer initiation by detoxifying carcinogens . We aimed to determine if androgens regulate P78381 expression via the androgen receptor ( AR ) pathway in the bladder . Real-time reverse transcription-polymerase chain reaction and Western blot analyses were used to assess P22309 levels in the normal urothelium SVHUC cell line stably expressed with AR and in bladder tissues from AR knockout ( ARKO ) and castrated male mice . Immunohistochemistry was also performed in radical cystectomy specimens . DB02901 ( DB02901 ) treatment in SVHUC-AR reduced mRNA expression of all the P22309 subtypes ( 19-75 % decrease ) , and hydroxyflutamide antagonized the DB02901 effects . In contrast , DB02901 showed only marginal effects on P22309 expression in SVHUC-Vector . Of note were higher expression levels of UGT1As in SVHUC-Vector than in SVHUC-AR . In ARKO mice , all the Ugt1a subtypes were up-regulated , compared to wild-type littermates . In wild-type male mice , castration increased the expression of Ugt1a8 , Ugt1a9 , and Ugt1a10 . Additionally , wild-type female mice had higher levels of Ugt1a than wild-type males . Immunohistochemical studies showed strong ( 3+ ) P22309 staining in 11/24 ( 46 % ) cancer tissues , which was significantly lower than in corresponding benign tissues [ 17/18 ( 94 % ) cases ( P = 0.0009 ) ] . These results suggest that androgen-mediated AR signals promote bladder carcinogenesis by down-regulating the expression of UGTs in the bladder . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . Inactivation of caspase-1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin-1 ( IL-1 ) -beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL-1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL-1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE/caspase-1 ) or through caspase-1 gene deletion . METHODS : P29466 was selectively blocked by using pralnacasan or DB05507 . IL-1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL-1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 inhibition reduced the release of IL-1beta in organotypic slices exposed to LPS+ DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 ( intraperitoneal , 25-200 mg/kg ) to rats blocked seizure-induced production of IL-1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase-1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase-1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL-1beta . DB00644 induction of extracellular-signal regulated kinase is blocked by inhibition of calmodulin . Our previous studies demonstrate that DB00644 -induced P29323 activation required influx of extracellular Ca2+ in alphaT3-1 and rat pituitary cells . In the present studies , we examined the hypothesis that calmodulin ( Cam ) plays a fundamental role in mediating the effects of Ca2+ on P29323 activation . Cam inhibition using W7 was sufficient to block DB00644 -induced reporter gene activity for the c-Fos , murine glycoprotein hormone alpha-subunit , and MAPK phosphatase ( MKP ) -2 promoters , all shown to require P29323 activation . Inhibition of Cam ( using a dominant negative ) was sufficient to block DB00644 -induced P29323 but not c-Jun N-terminal kinase activity activation . The Cam-dependent protein kinase ( CamK ) II inhibitor KN62 did not recapitulate these findings . DB00644 -induced phosphorylation of Q02750 and c-Raf kinase was blocked by Cam inhibition , whereas activity of phospholipase C was unaffected , suggesting that Ca2+/Cam modulation of the P29323 cascade potentially at the level of c-Raf kinase . Enrichment of Cam-interacting proteins using a Cam agarose column revealed that c-Raf kinase forms a complex with Cam . Reconstitution studies reveal that recombinant c-Raf kinase can associate directly with Cam in a Ca2+-dependent manner and this interaction is reduced in vitro by addition of W7 . Cam was localized in lipid rafts consistent with the formation of a Ca2+-sensitive signaling platform including the P30968 and c-Raf kinase . These data support the conclusion that Cam may have a critical role as a Ca2+ sensor in specifically linking Ca2+ flux with P29323 activation within the DB00644 signaling pathway . Therapeutic targeting of CPT-11 induced diarrhea : a case for prophylaxis . CPT-11 ( irinotecan ) , a P11387 inhibitor is one of the main treatments for colorectal cancer . The main dose limiting toxicities are neutropenia and late onset diarrhea . Though neutropenia is manageable , CPT-11 induced diarrhea is frequently severe , resulting in hospitalizations , dose reductions or omissions leading to ineffective treatment administration . Many potential agents have been tested in preclinical and clinical studies to prevent or ameliorate CPT-11 induced late onset diarrhea . It is predicted that prophylaxis of CPT-11 induced diarrhea will reduce sub-therapeutic dosing as well as hospitalizations and will eventually lead to dose escalations resulting in better response rates . This article reviews various experimental agents and strategies employed to prevent this debilitating toxicity . Covered topics include schedule/dose modification , intestinal alkalization , structural/chemical modification , genetic testing , anti-diarrheal therapies , transporter ( P08183 , Q92887 , Q96JK2 ) inhibitors , enzyme ( β-glucuronidase , P22309 , P08684 , carboxylesterase , P35354 ) inducers and inhibitors , probiotics , antibiotics , adsorbing agents , cytokine and growth factor activators and inhibitors and other miscellaneous agents . Effect of oncostatin M on uridine diphosphate-5'-glucuronosyltransferase 1A1 through cross talk with constitutive androstane receptor . Hyperbilirubinemia remains a common condition in neonates . The constitutive androstane receptor ( CAR ) is an orphan nuclear receptor that has been shown to participate in the activation of the uridine diphosphate-5'-glucuronosyltransferase 1A1 ( P22309 ) gene , which plays an important role in bilirubin clearance . Oncostatin M ( P13725 ) , a member of the P05231 family , is involved in the maturation of fetal hepatocytes . We have demonstrated that low P13725 levels are a potential indicator of neonatal jaundice and the need for phototherapy . In this study we examined the effects of P13725 on CAR-mediated signaling to investigate its potential role in neonatal jaundice via the CAR- P22309 pathway . We observed that P13725 positively augmented the CAR and P22309 expressions and CAR-mediated signaling in vivo and in vitro , through cross talk between the nuclear CAR receptor and the plasma membrane P13725 receptor , via the MAPK cascade . These data suggest that P13725 might play a role in bilirubin metabolism via the CAR- P22309 pathway . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . m.8993T > G-Associated Leigh Syndrome with Hypocitrullinemia on Newborn Screening . DB00155 is among the metabolites measured by expanded newborn screening ( O60934 ) . While hypocitrullinemia can be a marker for deficiency of proximal urea cycle enzymes such as ornithine transcarbamylase ( P00480 ) , only a handful of state newborn screening programs in the United States officially report a low citrulline value for further work-up due to low positive predictive value . We report a case of a male infant who was found to have hypocitrullinemia on O60934 . After excluding proximal urea cycle disorders by DNA sequencing , his O60934 result was felt to be a false positive . At 4 months of age , he developed poor feeding , failure to thrive , apnea and infantile spasms with a progression to intractable seizures , as well as persistent hypocitrullinemia . He was diagnosed with Leigh syndrome due to a maternally inherited homoplasmic m.8993T > G mutation in the ATPase 6 gene . DB00117 mother , who had previously been diagnosed with cerebral palsy , was concurrently diagnosed with neuropathy , ataxia , and retinitis pigmentosa ( NARP ) due to heteroplasmy of the same mutation . She had progressive muscle weakness , ataxia , and speech dyspraxia . The m.8993T > G mutation causes mitochondrial DB00171 synthase deficiency and it is hypothesized to undermine the synthesis of citrulline by P31327 . In addition to proximal urea cycle disorders , the evaluation of an infant with persistent hypocitrullinemia should include testing for the m.8993T > G mutation and other disorders that cause mitochondrial dysfunction . Diversity and versatility of GTPase activating proteins for the p21rho subfamily of ras G proteins detected by a novel overlay assay . The P01112 superfamily , involved in diverse processes including cell growth and intracellular trafficking , possesses intrinsic GTPase activity and cycles between GTP-bound active and GDP-bound quiescent states . This intrinsic activity , which results in down-regulation , is accelerated by GTPase activating proteins ( GAPs ) . Other proteins regulating the GDP/GTP cycle include exchange proteins and dissociation inhibitors . The p21s rho , rac , and cdc42Hs constitute a subfamily implicated in cytoskeletal organization . P11274 and n-chimaerin are prototypes of a new P20936 family for these p21s . To investigate proteins modulating GTP hydrolysis of the three p21s , we developed a novel overlay assay applicable to tissue extracts . Diverse GAPs with different specificities were identified in all rat tissues . Brain contained rac1 GAPs of 45 , 50 , 85 , 100 , and 150 kDa . The p50 and p150 GAPs also act on rhoA and cdc42Hs and are ubiquitous , while the P29466 - P20936 , n-chimaerin , is brain- and testis-specific and acts preferentially on rac1 ; the Q9ULW0 P20936 acts on both rac1 and cdc42Hs and is brain-specific . A new class of P38936 -interacting proteins was also identified . This diversity , versatility , and tissue specificity of GAPs may be required for fine control of the down-regulation of GTP-bound p21s and the suggested specific downstream effects of individual GAPs , which could involve " cross-talk " between GAPs and p21s . Beneficial effect of the non-psychotropic plant cannabinoid cannabigerol on experimental inflammatory bowel disease . Inflammatory bowel disease ( Q9UKU7 ) is an incurable disease which affects millions of people in industrialized countries . Anecdotal and scientific evidence suggests that Cannabis use may have a positive impact in Q9UKU7 patients . Here , we investigated the effect of cannabigerol ( CBG ) , a non-psychotropic Cannabis-derived cannabinoid , in a murine model of colitis . Colitis was induced in mice by intracolonic administration of dinitrobenzene sulphonic acid ( DNBS ) . Inflammation was assessed by evaluating inflammatory markers/parameters ( colon weight/colon length ratio and myeloperoxidase activity ) , by histological analysis and immunohistochemistry ; interleukin-1β , interleukin-10 and interferon-γ levels by ELISA , inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) by western blot and RT-PCR ; CuZn-superoxide dismutase ( SOD ) activity by a colorimetric assay . Murine macrophages and intestinal epithelial cells were used to evaluate the effect of CBG on nitric oxide production and oxidative stress , respectively . CBG reduced colon weight/colon length ratio , myeloperoxidase activity , and P35228 expression , increased SOD activity and normalized interleukin-1β , interleukin-10 and interferon-γ changes associated to DNBS administration . In macrophages , CBG reduced nitric oxide production and P35228 protein ( but not mRNA ) expression . DB06155 ( a P21554 receptor antagonist ) did not change the effect of CBG on nitric oxide production , while SR144528 ( a CB2 receptor antagonist ) further increased the inhibitory effect of CBG on nitric oxide production . In conclusion , CBG attenuated murine colitis , reduced nitric oxide production in macrophages ( effect being modulated by the CB2 receptor ) and reduced ROS formation in intestinal epithelial cells . CBG could be considered for clinical experimentation in Q9UKU7 patients . P10275 repression of gonadotropin-releasing hormone gene transcription via enhancer 1 . DB00644 ( DB00644 ) plays a major role in the hypothalamic-pituitary-gonadal ( HPG ) axis , and synthesis and secretion of DB00644 are regulated by gonadal steroid hormones . Disruptions in androgen levels are involved in a number of reproductive defects , including hypogonadotropic hypogonadism and polycystic ovarian syndrome . Androgens down-regulate DB00644 mRNA synthesis in vivo and in vitro via an androgen receptor ( AR ) -dependent mechanism . DB02998 ( R1881 ) , a synthetic AR agonist , represses DB00644 expression through multiple sites in the proximal promoter . In this study , we show AR also represses DB00644 transcription via the major enhancer ( DB00644 -E1 ) . A multimer of the -1800/-1766 region was repressed by R1881 treatment . Mutation of two bases , -1792 and -1791 , resulted in decreased basal activity and a loss of AR-mediated repression . AR bound to the -1796/-1791 sequence in electrophoretic mobility shift assays , indicating a direct interaction with DNA or other transcription factors in this region . We conclude that AR repression of DB00644 -E1 acts via multiple AR-responsive regions , including the site at -1792/-1791 . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment .

MH_train_1272

MH_train_1272

MH_train_1272

interacts_with DB01024?

multiple_choice

Heme oxygenase-1 attenuates glucose-mediated cell growth arrest and apoptosis in human microvessel endothelial cells . Heme oxygenase-1 ( P09601 ) is a stress protein that has been suggested to participate in defense mechanisms against agents that may induce oxidative injury , such as heme and inflammatory molecules . Incubation of endothelial cells in a high-glucose ( 33 mmol/L ) medium for 7 days resulted in a decrease of HO activity by 34 % and a decrease in P09601 and P30519 proteins compared with cells exposed to low glucose ( 5 mmol/L ) ( P < 0.05 ) or cells exposed to mannitol ( 33 mmol/L ) . Overexpression of P09601 was coupled with an increase in HO activity and carbon monoxide synthesis , decreased cellular heme , and acceleration in all phases of the cell cycle ( P < 0.001 ) . The rate of cell cycle or cell birth rate was increased by 29 % ( P < 0.05 ) in cells overexpressing P09601 but decreased by 23 % ( P < 0.05 ) in cells underexpressing P09601 compared with control cells . Exposure to high glucose significantly decreased cell-cycle progression in control cells and in cells underexpressing P09601 but did not decrease cell-cycle progression in cells overexpressing P09601 . High glucose induced P38936 and p27 in control cells but not in cells overexpressing P09601 . The addition of DB04912 ( DB04912 ) , an inhibitor of HO activity , reversed the P09601 -mediated decrease of P38936 and p27 in cells overexpressing P09601 . These findings identify a novel effect of P09601 on endothelial cell growth and indicate that heme metabolism and P09601 expression regulate signaling systems in cells exposed to high glucose , which controls cell-cycle progression . P62937 interacts with domain II of hepatitis C virus NS5A and stimulates RNA binding in an isomerase-dependent manner . NS5A plays a critical , yet poorly defined , role in hepatitis C virus genome replication . The protein consists of three domains , each of which is able to bind independently to the 3' untranslated region ( UTR ) of the viral positive strand genomic RNA . The peptidyl-prolyl isomerase cyclophilin A ( CypA ) binds to domain II , catalyzing cis-trans isomerization . CypA inhibitors such as cyclosporine ( DB00091 ) have been shown to inhibit hepatitis C virus ( HCV ) replication . We show here that CypA stimulated domain II RNA binding activity , and this stimulation was abrogated by DB00091 . An isomerase mutant of CypA ( H126Q ) failed to bind to domain II and did not stimulate RNA binding . Finally , we demonstrate that the RNA binding of two domain II mutants , the D316E and D316E/Y317N mutants , previously shown to exhibit CypA independence for RNA replication , was unaffected by CypA . This study provides an insight into the molecular mechanism of CypA activity during HCV replication and further validates the use of CypA inhibitors in HCV therapy . Further insights on the inhibition of HIV type 1 infection in vitro by P01730 -modified synthetic peptides containing phenylalanine . DB00120 -containing peptides from P01730 were synthesized on the basis of chemical similarity with active P01730 (81-92)-benzylated peptides . Systematic replacement of amino acids of these peptides bearing the benzyl group by phenylalanine , afforded several peptides that were able to block the binding of gp120 to P01730 and to inhibit HIV-induced syncytium formation . These experiments showed that substitution of residues 81 and 85 by phenylalanine was the most important for activity . Following optimization of the length of phenylalanine-substituted peptides it was found that FYICFVED and FYICFVEDE were the most active . Their IC50 for the inhibition of syncytium formation was around 1. P35326 .6 microM . This activity is at least 30 times higher than that of the parent peptide FYIFFVEDQKEEDD previously reported ( Lasarte et al. , J Acquir Immune Defic Syndr 1994;7:129-134 ) . Binding competition experiments with two different anti-peptide antisera recognizing the V3 region of gp120 and FYICFVEDE , show that the active peptides bind to V3 or to a sterically near region of V3 . None of the active peptides was toxic to cells in vitro . The enhanced activity and simplicity of these new phenylalanine-substituted P01730 peptides might be a good starting point for the development of mimotopes of potential use for the treatment of AIDS . Comprehensive and temporal analysis of secreted proteins in the medium from P05231 exposed human hepatocyte . We have previously identified intracellular secretory acute phase response ( sAPR ) proteins in human hepatocytes following interleukin-6 ( P05231 ) induction by fluorogenic derivatization ( FD ) -liquid chromatography ( LC ) -tandem mass spectrometry ( MS/MS ) . In this report , we utilized this method , which uses 7-chloro-N-[2-(dimethylamino)ethyl]-2,1,3-benzoxadiazole-4-sulfonamide ( DAABD-Cl ) as the FD reagent , to comprehensively and time-dependently analyze secreted proteins in the medium , including sAPR proteins . Since DAABD-Cl selectively reacts with thiol moieties of cysteinyl residues , direct derivatization , high-resolution LC separation and identification of the secreted proteins in the culture medium were successfully achieved without a pretreatment step . As a result , 14 sAPR proteins were identified simultaneously during a 72 h induction by P05231 . The secretion levels of 11 proteins increased , whereas the secretion levels of three important transport proteins decreased ( albumin , retinol-binding protein 4 and transthyretin ) . In addition , the secretion level of a haptoglobin was found to increase significantly between 0 and 6 h by 1.88-fold compared with the control sample . The secretion levels of four cytoplasmic proteins increased : LDH , a known marker for cell damage , and P08263 , P07148 and P00325 , which are marker proteins for hepatocellular damage . The secretion levels of the other two newly identified cytoplasmic proteins , profilin-1 and P04179 , were also found to increase , suggesting that these two proteins represent novel markers for cell damage . These results suggest that the FD-LC-MS/MS proteomics method can be used to analyze comprehensively and time-dependently the secreted proteins and thereby can offer information that aids our understanding of the dynamics of protein secretion affected by the exposure of cytokines such as P05231 . Genetic polymorphism and activities of human lung alcohol and aldehyde dehydrogenases : implications for ethanol metabolism and cytotoxicity . DB00898 dehydrogenase ( DB00067 ) and aldehyde dehydrogenase ( ALDH ) exhibit genetic polymorphism and tissue specificity . DB00067 and ALDH isozyme phenotypes from 39 surgical Chinese lung specimens were identified by agarose isoelectric focusing . The identity of the lung beta-ADHs was further demonstrated by their characteristic pH-activity profiles for ethanol oxidation , Km values for NAD and ethanol , and inhibition by DB01213 or 1,10-phenanthroline . The beta 2 allele , coding for beta 2 polypeptide , was found to be predominant in the lung specimens studied . The DB00067 activities in the lungs with the homozygous phenotype P00325 2-2 ( exhibiting beta 2 beta 2 ) and P00325 1-1 ( exhibiting beta 1 beta 1 ) and the heterozygous phenotype P00325 P35326 ( exhibiting beta 2 beta 2 , beta 2 beta 1 , and beta 1 beta 1 ) were determined to be 999 +/- 77 , 48 +/- 17 , and 494 +/- 61 nmol/min/g tissue , respectively . Fifty-one percent of the specimens studied lacked the P05091 activity band on the isoelectric focusing gels . The activities in the lung tissues with the P05091 -active phenotype and the inactive phenotype were determined to be 30 +/- 3 and 17 +/- 1 nmol/min/g tissue , respectively . These findings indicate that human pulmonary ethanol-metabolizing activities differ significantly with respect to genetic polymorphism at both the P00325 and the P05091 loci . The results suggest that individuals with high Vmax beta 2- DB00067 and deficient in low-Km mitochondrial P05091 , accounting for approximately 45 % of the Chinese population , may end up with acetaldehyde accumulation during alcohol consumption , rendering them vulnerable to tissue injury caused by this highly reactive and toxic metabolite . DB03128 and vasoactive intestinal peptide in cerebral blood vessels : effect of extirpation of the sphenopalatine ganglion . The innervation of cerebral blood vessels by nerve fibers containing acetylcholinesterase ( P22303 ) and vasoactive intestinal peptide ( P01282 ) and the vasomotor effects of the two neurotransmitters have been analyzed in the rat following the uni- or bilateral removal of the sphenopalatine ganglion ( Q9HBG6 ) , which is thought to be the major origin of this innervation . Histochemistry of P22303 -positive nerve fibers and the immunoreactivity toward P01282 revealed only a 30 % reduction in the innervation pattern of the rostral part of the cerebral circulation following the operation . At approximately 4 weeks postoperatively , the original nerve network was restored . Quantitative measurements of cholineacetyltransferase activity and P01282 revealed similar reductions in the levels of collected large cerebral arteries at the base of the brain and in small pial vessels overlying the cerebral cortex at the various postoperative times following uni- or bilateral removal of the Q9HBG6 . The two techniques thus complemented each other . Vasomotor reactivity to acetylcholine ( ACh ) and P01282 was examined in proximal segments of the middle cerebral artery at the various postoperative times . Generally , the removal of the Q9HBG6 had no effect on the responses to ACh or P01282 . The evidence indicates that only approximately one-third of the cholinergic/ P01282 innervation of the rostral part of the cerebral circulation originates in the Q9HBG6 . Genomic imbalance of O75330 / O75330 regulates the sensitivity and response of malignant peripheral nerve sheath tumour cells to aurora kinase inhibition . Malignant peripheral nerve sheath tumours ( MPNST ) are rare , hereditary cancers associated with neurofibromatosis type I . MPNSTs lack effective treatment options as they often resist chemotherapies and have high rates of disease recurrence . O14965 ( O14965 ) is an emerging target in cancer and an aurora kinase inhibitor ( AKI ) , termed DB05220 , shows promise against MPNST cell lines in vitro and in vivo . Here , we test DB05220 against two primary human MPNST grown in vivo as xenotransplants and find that treatment results in tumour cells exiting the cell cycle and undergoing endoreduplication , which cumulates in stabilized disease . Targeted therapies can often fail in the clinic due to insufficient knowledge about factors that determine tumour susceptibilities , so we turned to three MPNST cell-lines to further study and modulate the cellular responses to AKI . We find that the sensitivity of cell-lines with amplification of O14965 depends upon the activity of the kinase , which correlates with the expression of the regulatory gene products Q9ULW0 and O75330 / O75330 . Silencing of O75330 / O75330 , but not Q9ULW0 , augments O14965 activity and sensitizes MPNST cells to AKI . Furthermore , we find that O14965 activity is critical to the propagation and self-renewal of sphere-enriched MPNST cancer stem-like cells . AKI treatment significantly reduces the formation of spheroids , attenuates the self-renewal of spheroid forming cells , and promotes their differentiation . Moreover , silencing of O75330 / O75330 is sufficient to endow MPNST cells with an ability to form and maintain sphere culture . Collectively , our data indicate that O14965 is a rationale therapeutic target for MPNST and tumour cell responses to AKI , which include differentiation , are modulated by the abundance of O75330 / O75330 . Expression of cyclooxygenase-2 ( P35354 ) in tumour and stroma compartments in cervical cancer : clinical implications . This study aims at investigating the relationship between cyclooxygenase-2 expression in tumour vs stroma inflammatory compartment and its possible clinical role . The study included 99 stage IB-IV cervical cancer patients : immunostaining of tumour tissue sections was performed with rabbit antiserum against cyclooxygenase-2 . CD3 , P01730 , CD8 , CD25 , Mast Cell Tryptase monoclonal antibodies were used to characterise stroma inflammatory cells in nine cervical tumours . An inverse relation was found between cyclooxygenase-2 levels ( cyclooxygenase-2 IDV ) of tumour vs stroma compartment ( r=-0.44 , P < 0.0001 ) . The percentage of cases showing high tumour/stromal cyclooxygenase-2 IDV ratio was significantly higher in patients who did not respond to treatment ( 93.3 % ) with respect to patients with partial ( 60.5 % ) , and complete ( 43.7 % ) response ( P= 0.009 ) . Cases with a high tumour/stroma cyclooxygenase-2 IDV ratio had a shorter overall survival rate than cases with a low tumour/stroma cyclooxygenase-2 IDV ( P < 0.0001 ) . In the multivariate analysis advanced stage and the status of tumour/stroma cyclooxygenase-2 IDV ratio retained an independent negative prognostic role . The proportion of CD3(+) , P01730 (+) , and CD25(+) cells was significantly lower in tumours with high tumour/stroma cyclooxygenase-2 IDV ratio , while a higher percentage of mast cells was detected in tumours showing high tumour/stroma cyclooxygenase-2 IDV ratio . Our study showed the usefulness of assessing cyclooxygenase-2 status both in tumour and stroma compartment in order to identify cervical cancer patients endowed with a very poor chance of response to neoadjuvant therapy and unfavourable prognosis . P62937 regulates HIV-1 infectivity , as demonstrated by gene targeting in human T cells . The human immunodeficiency virus type 1 ( HIV-1 ) Gag polyprotein binds most members of the cyclophilin family of peptidyl-prolyl isomerases . Of 15 known human cyclophilins , cyclophilin A ( CypA ) has been the focus of investigation because it was detected in HIV-1 virions . To determine whether CypA promotes HIV-1 replication , we deleted the gene encoding CypA ( P62937 ) in human P01730 (+) T cells by homologous recombination . HIV-1 replication in P62937 (-/-) cells was decreased and not inhibited further by cyclosporin or gag mutations that disrupt Gag 's interaction with cyclophilins , indicating that no other cyclophilin family members promote HIV-1 replication . The defective replication phenotype was specific for wild-type HIV-1 since HIV-2/SIV isolates , as well as HIV-1 bearing a gag mutation that confers cyclosporin resistance , replicated the same in P62937 (+/+) and P62937 (-/-) cells . Stable re-expression of CypA in P62937 (-/-) cells restored HIV-1 replication to an extent that correlated with steady-state levels of CypA . Finally , virions from P62937 (-/-) cells possessed no obvious biochemical abnormalities but were less infectious than virions from wild-type cells . These data formally demonstrate that CypA regulates the infectivity of HIV-1 virions . DB00439 potentiates nitric oxide release and enos expression through inhibition of isoprenoids synthesis . Endothelium dysfunction , which is often defined as a decrease in NO bioavailability , is one of the earliest manifestations of endothelium-impaired function disorders , including atherosclerosis . Although improvement in NO bioavailability has been attributed to the lowering of serum cholesterol levels , recent studies suggest that P04035 inhibitors , statins , may have direct effects on NO bioavailability by little known mechanisms that are independent of serum cholesterol levels . The long-term effect of cerivastatin on NO release from endothelial cells was determined by using highly sensitive electrochemical microsensors and was correlated with endothelial NO synthase ( P29474 ) levels . To explore whether changes in isoprenoid synthesis affect NO bioavailability and P29474 expression , human endothelial cells were treated with cerivastatin , L-mevalonate ( MVA ; 1.5 mmol/L ) , geranylgeranylpyrophosphate ( GGPP ; 1 mg/mL ) and farnesylpyrophosphate ( FPP ; 1 mg/mL ) . DB00439 increased spontaneous ( by 53 % +/- 6 ) and an P29474 -stimulated NO release ( by 41 +/- 6 % for calcium ionophore and by 47 +/- 5 % acetylcholine ) as well as P29474 expression ( by 118 +/- 6 % ) in the same concentration-range . DB00439 -dependent increase in both NO release and P29474 expression was revealed after approximately 4 h of exposure reaching the maximum after approximately 10 h . Co-treatment with MVA or GGPP , but not FPP or LDL , reversed the effects of cerivastatin . These findings indicate that the long-term effect of cerivastatin resulting in enhanced NO bioavailabilty in endothelial cell is , at least in part , due to up-regulation of P29474 by blocking isoprenoids synthesis . Increased cellular senescence and vascular rarefaction exacerbate the progression of kidney fibrosis in aged mice following transient ischemic injury . Recent findings indicate that elderly patients with acute kidney injury ( AKI ) have an increased incidence of progression to chronic kidney disease ( CKD ) due to incomplete recovery from an acute insult . In the current study , a co-morbid model of AKI was developed to better mimic the patient population and to investigate whether age exacerbates the fibrosis and inflammation that develop in the sequelae of progressive kidney disease following acute injury . Young ( 8-10 weeks ) and aged ( 46-49 weeks ) C57BL/6 mice were subjected to 30 min bilateral renal ischemia-reperfusion ( I/R ) to induce AKI . The aged animals have greater mortality and prolonged elevation of plasma creatinine correlating with less tubular epithelial cell proliferation compared to the young . Six weeks post-reperfusion , interstitial fibrosis is greater in aged kidneys based on picrosirius red staining and immunolocalization of cellular fibronectin , collagen III and collagen IV . Aged kidneys 6 weeks post-reperfusion also express higher levels of p53 and P38936 compared to the young , correlating with greater increases in senescence associated ( SA ) β-galactosidase , a known marker of cellular senescence . A higher influx of F4/80(+) macrophages and P01730 (+) T lymphocytes is measured and is accompanied by increases in mRNA of monocyte chemoattractant protein-1 ( P13500 ) and tumor necrosis factor-α ( P01375 -α ) . Importantly , microvascular density is significantly less , correlating with an increase in nitro-tyrosine , a marker of oxidative stress . Collectively , these data demonstrate that prolonged acute injury in the aged animals results in an accelerated progression of kidney disease in a chronic state . High-fat diet exacerbates renal dysfunction in SHR : reversal by induction of P09601 -adiponectin axis . High-dietary fat intake is a major risk factor for development of metabolic and cardiovascular-renal dysfunction including obesity , coronary artery disease , hypertension , and chronic renal failure . We examined the effect of a high-fat diet on renal function and morphology in spontaneously hypertensive rats ( SHR ) , a phenotype designed to mimic metabolic syndrome . High-fat diet induced increase ( P < 0.05 ) in blood pressure , body weight , and renal lipid deposition in these rats . This increase in body weight was accompanied by elevations ( P < 0.05 ) of blood glucose and low-density lipoprotein ( LDL ) levels , a decrease ( P < 0.05 ) in adiponectin and increases ( P < 0.05 ) in plasma monocyte chemotactic protein-1 ( P13500 ) along with renal macrophage infiltration . These pathophysiological perturbations were attenuated ( P < 0.05 ) by heme oxygenase-1 ( P09601 ) induction by treatment with cobalt protoporphyrin ( CoPP ) . Further effects of CoPP included increased ( P < 0.05 ) renal expression of adiponectin along with enhancement ( P < 0.05 ) of pAKT , pAMPK , and p- P29474 in SHRs fed a high-fat diet . Prevention of such beneficial effects of CoPP by the concurrent administration of the heme-HO inhibitor stannous mesoporphyrin ( DB04912 ) corroborates the role of HO system in mediating such effects . Taken together , our results demonstrate that high-fat diet induces a metabolic syndrome-like phenotype in hypertensive rats , which is amenable to rescue by increases in P09601 - and adiponectin-dependent pathways . Sirtuin activators and inhibitors . Sirtuins 1-7 ( Q96EB6 -7 ) belong to the third class of deacetylase enzymes , which are dependent on NAD(+) for activity . Sirtuins activity is linked to gene repression , metabolic control , apoptosis and cell survival , DNA repair , development , inflammation , neuroprotection , and healthy aging . Because sirtuins modulation could have beneficial effects on human diseases there is a growing interest in the discovery of small molecules modifying their activities . We review here those compounds known to activate or inhibit sirtuins , discussing the data that support the use of sirtuin-based therapies . Almost all sirtuin activators have been described only for Q96EB6 . DB02709 is a natural compound which activates Q96EB6 , and may help in the treatment or prevention of obesity , and in preventing tumorigenesis and the aging-related decline in heart function and neuronal loss . Due to its poor bioavailability , reformulated versions of resveratrol with improved bioavailability have been developed ( resVida , Longevinex(®) , DB05073 ) . Molecules that are structurally unrelated to resveratrol ( SRT1720 , SRT2104 , SRT2379 , among others ) have been also developed to stimulate sirtuin activities more potently than resveratrol . Sirtuin inhibitors with a wide range of core structures have been identified for Q96EB6 , Q8IXJ6 , Q9NTG7 and Q9NXA8 ( splitomicin , sirtinol , AGK2 , cambinol , suramin , tenovin , salermide , among others ) . Q96EB6 inhibition has been proposed in the treatment of cancer , immunodeficiency virus infections , Fragile X mental retardation syndrome and for preventing or treating parasitic diseases , whereas Q8IXJ6 inhibitors might be useful for the treatment of cancer and neurodegenerative diseases . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Characterization of human P01730 helper T cell responses against O14965 . O14965 ( Aurora-A ) is a cell cycle-associated serine-threonine kinase that is overexpressed by various types of cancer and is highly associated with poor prognosis . Since the expression of Aurora-A in normal tissues has been shown to be significantly lower as compared to tumor cells , this protein is being considered as a potential tumor-associated antigen for developing immunotherapies . The goal in the present study was to identify P01730 helper T lymphocyte ( HTL ) epitopes for Aurora-A for the design of T cell-based immunotherapies against Aurora-A-expressing tumors . Synthetic peptides corresponding to potential HTL epitopes were identified from Aurora-A and used to stimulate P01730 T lymphocytes in vitro to generate antigen-specific HTL clones that were evaluated for antigen specificity , MHC restriction and for their ability to interact with Aurora-A-expressing tumor cells . The results show that two peptides ( Aurora-A(161-175) and Aurora-A(233-247) ) were effective in generating HTL responses that were restricted by more than one MHC class II allele ( i.e. , promiscuous responses ) . The P01730 HTL clones were able to directly recognize Aurora-A-expressing tumor cells in an antigen-specific and MHC class II-restricted manner and some of the clones displayed cytolytic activity toward Aurora-A + tumor cells . Both of these peptides were capable of stimulating in vitro T cell responses in patients with bladder cancer . P01730 receptor binding peptides that block HIV infectivity cause human monocyte chemotaxis . Relationship to vasoactive intestinal polypeptide . The octapeptide Ala- DB00133 - DB00156 - DB00156 - DB00156 - DB00174 - DB00135 - DB00156 ( peptide T ) and two structural analogs are potent agonists of human monocyte chemotaxis , evincing identical rank potency orders as was previously shown for their inhibition of human immunodeficiency virus ( HIV ) envelope binding and T cell infectivity . Chemotactic activity could be inhibited by anti- P01730 monoclonal antibodies ( Mabs ) , but not other mononuclear cell Mabs . The core peptide required for chemotactic activity is a pentapeptide related to the sequence DB00156 - DB00156 - DB00174 - DB00135 - DB00156 . Homologous pentapeptides , identified by computer search , were detected in several other non-HIV-related viruses as well as the neuropeptide vasoactive intestinal polypeptide ( P01282 ) . The P01730 molecule , therefore , appears to be a recognition molecule for a small signal peptide ligand whose active sequence is a homolog of peptide T and which may be the neuropeptide P01282 . T cell-mediated vascular dysfunction of human allografts results from P01579 dysregulation of NO synthase . Allograft vascular dysfunction predisposes to arteriosclerosis and graft loss . We examined how dysfunction develops in transplanted human arteries in response to circulating allogeneic T cells in vivo using immunodeficient murine hosts . Within 7-9 days , transplanted arteries developed endothelial cell ( EC ) dysfunction but remained sensitive to exogenous NO . By 2 weeks , the grafts developed impaired contractility and desensitization to NO , both signs of VSMC dysfunction . These T cell-dependent changes correlated with loss of P29474 and expression of P35228 -- the latter predominantly within infiltrating T cells . Neutralizing P01579 completely prevented both vascular dysfunction and changes in NOS expression ; neutralizing P01375 reduced P01579 production and partially prevented dysfunction . Inhibiting P35228 partially preserved responses to NO at 2 weeks and reduced graft intimal expansion after 4 weeks in vivo . In vitro , memory P01730 + T cells acted on allogeneic cultured ECs to reduce P29474 activity and expression of protein and mRNA . These effects required T cell activation by class II MHC antigens and costimulators ( principally lymphocyte function-associated antigen-3 , or LFA-3 ) on the ECs and were mediated by production of soluble mediators including P01579 and P01375 . We conclude that P01579 is a central mediator of vascular dysfunction and , through dysregulation of NOS expression , links early dysfunction with late arteriosclerosis . In vitro treatment of human monocytes/macrophages with myristoylated recombinant Nef of human immunodeficiency virus type 1 leads to the activation of mitogen-activated protein kinases , IkappaB kinases , and interferon regulatory factor 3 and to the release of beta interferon . The viral protein Nef is a virulence factor that plays multiple roles during the early and late phases of human immunodeficiency virus ( HIV ) replication . Nef regulates the cell surface expression of critical proteins ( including down-regulation of P01730 and major histocompatibility complex class I ) , T-cell receptor signaling , and apoptosis , inducing proapoptotic effects in uninfected bystander cells and antiapoptotic effects in infected cells . It has been proposed that Nef intersects the P29965 signaling pathway in macrophages , leading to modification in the pattern of secreted factors that appear able to recruit and activate T lymphocytes , rendering them susceptible to HIV infection . There is also increasing evidence that in vitro cell treatment with Nef induces signaling effects . Exogenous Nef treatment is able to induce apoptosis in uninfected T cells , maturation in dendritic cells , and suppression of P25942 -dependent immunoglobulin class switching in B cells . Previously , we reported that Nef treatment of primary human monocyte-derived macrophages ( MDMs ) induces a cycloheximide-independent activation of NF-kappaB and the synthesis and secretion of a set of chemokines/cytokines that activate P42224 and P40763 . Here , we show that Nef treatment is capable of hijacking cellular signaling pathways , inducing a very rapid regulatory response in MDMs that is characterized by the rapid and transient phosphorylation of the alpha and beta subunits of the O15111 complex and of JNK , P27361 /2 , and p38 mitogen-activated protein kinase family members . In addition , we have observed the activation of interferon regulatory factor 3 , leading to the synthesis of beta interferon mRNA and protein , which in turn induces P52630 phosphorylation . All of these effects require Nef myristoylation . Q8NFT8 bromodomain inhibition as a novel strategy for reactivation of HIV-1 . The persistence of latent HIV-1 remains a major challenge in therapeutic efforts to eradicate infection . We report the capacity for HIV reactivation by a selective small molecule inhibitor of Q8NFT8 family bromodomains , JQ1 , a promising therapeutic agent with antioncogenic properties . JQ1 reactivated HIV transcription in models of latent T cell infection and latent monocyte infection . We also tested the effect of exposure to JQ1 to allow recovery of replication-competent HIV from pools of resting P01730 (+) T cells isolated from HIV-infected , O00253 -treated patients . In one of three patients , JQ1 allowed recovery of virus at a frequency above unstimulated conditions . JQ1 potently suppressed T cell proliferation with minimal cytotoxic effect . Transcriptional profiling of T cells with JQ1 showed potent down-regulation of T cell activation genes , including CD3 , P10747 , and P61073 , similar to HDAC inhibitors , but JQ1 also showed potent up-regulation of chromatin modification genes , including Q96EB6 , Q9UBN7 , and multiple lysine demethylases ( KDMs ) . Thus , JQ1 reactivates HIV-1 while suppressing T cell activation genes and up-regulating histone modification genes predicted to favor increased Tat activity . Thus , JQ1 may be useful in studies of potentially novel mechanisms for transcriptional control as well as in translational efforts to identify therapeutic molecules to achieve viral eradication . P04035 inhibitor cerivastatin inhibits interleukin-6 expression and secretion in human adipocytes . Human adipose tissue is a main contributor to plasma levels of pro-inflammatory cytokine P05231 . How P05231 expression is regulated in adipocytes remains unclear . In the current study , we investigated the effect of the P04035 inhibitor , cerivastatin , on the production of P05231 from cultured human adipocytes . DB00439 reduced both P05231 mRNA and secretion in a dose- and time-dependent manner . The inhibitory effect on P05231 mRNA was prevented by the intermediates of the cholesterol synthesis pathway , mevalonate and geranyl-geranyl-phyrophosphate ( GGPP ) but not by farnesyl-pyrophosphate . This suggests the involvement of geranylgeranyl-modified intermediates in the effect of cerivastatin on P05231 . Moreover , cerivastatin induced an inactivation of the phosphorylation of the p65 subunit of NFkappaB which was prevented by GGPP . Our data suggest that cerivastatin exerts an anti-inflammatory effect by down-regulating P05231 levels in adipocytes , which seems to be mediated by reduced production of GGPP and interference with the NFkappaB pathway . Sulfasalazine inhibits the growth of primary brain tumors independent of nuclear factor-kappaB . Nuclear factor-kappaB ( NF-kappaB ) is a pleiotropic transcription factor that generally enhances cellular resistance to apoptotic cell death . It has been shown to be constitutively active in some cancers and is being pursued as potential anticancer target . Sulfasalazine which is used clinically to treat Crohn 's disease has emerged as a potential inhibitor of NF-kappaB and has shown promising results in two pre-clinical studies to target primary brain tumors , gliomas . Once digested , sulfasalazine is cleaved into sulfapyridine and DB00244 ( DB00244 ; mesalamine ) by colonic bacteria , and the latter , too , is reported to suppress NF-kappaB activity . We now show that glioma cells obtained from patient biopsies or glioma cell lines do not show significant constitutive NF-kappaB activation , unless exposed to inflammatory cytokines . This does not change when gliomas are implanted into the cerebrum of severe combined immun-deficient mice . Nevertheless , sulfasalazine but not its cleaved form DB00244 caused a dose-dependent inhibition of glioma growth . This effect was entirely attributable to the inhibition of cystine uptake via the system x(c)(-) cystine-glutamate transporter . It could be mimicked by S-4-carboxy-phenylglycine ( S-4-CPG ) a more specific system x(c)(-) inhibitor , and lentiviral expression of a constitutively active form of O15111 b was unable to overcome the growth retarding effects of sulfasalazine or S-4-CPG . Both drugs inhibited cystine uptake causing a chronic depletion of intracellular DB00143 and consequently compromised cellular redox defense which stymied tumor growth . This data suggests that system x(c)(-) is a promising therapeutic target in gliomas and possibly other cancers and that it can be pharmacologically inhibited by Sulfasalazine , an FDA-approved drug . DB02709 protects against peripheral deficits in a mouse model of Huntington 's disease . Sirtuins are NAD-dependent deacetylases that regulate important biologic processes including transcription , cell survival and metabolism . Activation of Q96EB6 , a mammalian sirtuin , extends longevity and increases neuronal survival . An important substrate of Q96EB6 is peroxisome proliferator-activated receptor gamma co-activator-1alpha ( P20142 -1alpha ) , a principal regulator of energy metabolism , whose function is significantly impaired in Huntington 's disease ( HD ) . We studied the effects of a pharmacological preparation of the Q96EB6 activator resveratrol ( DB05073 -M ) , in the N171-82Q transgenic mouse model of HD . We analyzed motor performance , survival , central and peripheral pathology and levels of P20142 -1alpha expression . Administration of DB05073 -M increased expression of P20142 -1alpha , as well as its downstream targets , nuclear respiratory factor-1 ( Q16656 ) and uncoupling protein-1 ( P25874 -1 ) in brown adipose tissue ( Q14032 ) , but there was no effect on P20142 -1alpha , Q16656 or the mitochondrial transcription factor ( Tfam ) in the striatum . DB05073 -M administration also reduced Q14032 vacuolation and decreased elevated blood glucose levels . However , there was no significant improvement in weight loss , motor performance , survival and striatal atrophy . Activation of the P20142 -1alpha signaling pathway via resveratrol-induced activation of Q96EB6 , therefore , is an effective therapy in Q14032 , but not in the central nervous system of HD transgenic mice . 5- DB00233 Inhibits Acute Clostridium difficile Toxin A-Induced Colitis in Rats . We tested the hypothesis that DB00244 ( DB00244 ) inhibits toxin A-induced generation of colonic leukotriene B4 ( LTB4 ) and toxin A colitis in rats . Isolated colonic segments in anesthetized rats were treated intraluminally with toxin A for 3 hours with or without 30 minutes of pretreatment with either DB00244 or sulfapyridine and then colonic tissue levels of LTB4 were measured and inflammation was assessed . Separately , sulfasalazine was administered to rats in their drinking water for 5 days , isolated colonic segments were then prepared , toxin A was administered , and inflammation was assessed as before . Pretreatment with DB00244 inhibited toxin A-induced increased tissue LTB4 concentration in the colon . Sulfasalazine and DB00244 but not sulfapyridine significantly inhibited toxin A colitis . However , pretreatment with DB00244 did not protect against direct Q8NER1 -mediated colitis caused by capsaicin . Toxin A stimulated the release of DB05875 ( SP ) , and this effect was also inhibited by sulfasalazine and DB00244 but not by sulfapyridine . Thus , toxin A stimulates colonic LTB4 resulting in activation of Q8NER1 , release of SP , and colitis . Inhibition of P09917 by DB00244 disrupts this pathway and supports the concept that LTB4 activation of Q8NER1 plays a role in toxin A colitis . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . P09917 is a key determinant of acute myocardial inflammation and mortality during Trypanosoma cruzi infection . This study provides evidence supporting the idea that although inflammatory cells migration to the cardiac tissue is necessary to control the growth of Trypanosoma cruzi , the excessive influx of such cells during acute myocarditis may be deleterious to the host . Production of lipid mediators of inflammation like leukotrienes ( LTs ) along with cytokines and chemokines largely influences the severity of inflammatory injury in response to tissue parasitism . T. cruzi infection in mice deficient in P09917 ( P09917 ) , the enzyme responsible for the synthesis of LTs and other lipid inflammatory mediators , resulted in transiently increased parasitemia , and improved survival rate compared with WT mice . Myocardia from P09917 (-/-) mice exhibited reduced inflammation , collagen deposition , and migration of P01730 (+) , CD8(+) , and P01579 -producer cells compared with WT littermates . Moreover , decreased amounts of P01375 , P01579 , and nitric oxide synthase were found in the hearts of P09917 (-/-) mice . Interestingly , despite of early higher parasitic load , P09917 (-/-) mice survived , and controlled T. cruzi infection . These results show that efficient parasite clearance is possible in a context of moderate inflammatory response , as occurred in P09917 (-/-) mice , in which reduced myocarditis protects the animals during T. cruzi infection . [ P11511 inhibitors -- theoretical concept and present experiences in the treatment of endometriosis ] . The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies of endometriosis-associated pain or recurrent disease is primarily aimed at downregulating the ovarian function or at antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is delivering powerful tools for the possible development of new , specific treatment modalities . Recently , aromatase overexpression has been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for conversion of C19 androgens to estrogen in several human tissues . P11511 activity gives rise to local estrogen biosynthesis , which , in turn , stimulates prostaglandin E(2) production by upregulation of cyclooxygenase-2 ( P35354 ) , thus establishing a positive feedback cycle . Another abnormality in endometriosis , i. e. the deficiency in 17 beta-hydroxysteroiddehydrogenase type-II ( 17 beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations collectively favour accumulation of increasing amounts of local estradiol and prostaglandin E(2) in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance , and even invasiveness . Consequently , aromatase and P35354 are promising new therapeutic targets . In summary , specific aromatase inhibitors ( such as Letrozole , DB01217 or Exemestan ) or selective P35354 inhibitors ( e.g. Celecoxib , DB00533 ) are of great interest to be studied in clinical trials in premenopausal woman with endometriosis to extend the spectrum of currently available treatment options . Neuromuscular therapeutics by RNA-targeted suppression of P22303 gene expression . RNA-targeted therapeutics offers inherent advantages over small molecule drugs wherever one out of several splice variant enzymes should be inhibited . Here , we report the use of Monarsen , a 20-mer acetylcholinesterase-targeted antisense agent with three 3'-2'o-methyl-protected nucleotides , for selectively attenuating the stress-induced accumulation of the normally rare , soluble " readthrough " acetylcholinesterase variant P22303 -R . DB03128 hydrolysis by P22303 -R may cause muscle fatigue and moreover , limit the cholinergic anti-inflammatory blockade , yielding inflammation-associated pathology . Specific P22303 -R targeting by Monarsen was achieved in cultured cells , experimental animals , and patient volunteers . In rats with experimental autoimmune myasthenia gravis , oral delivery of Monarsen improved muscle action potential in a lower dose regimen ( nanomolar versus micromolar ) , rapid and prolonged manner ( up to 72 h versus 2-4 h ) as compared with the currently used small molecule anticholinesterases . In central nervous system neurons of both rats and cynomolgus monkeys , systematic Monarsen treatment further suppressed the levels of the proinflammatory cytokines interleukin-1 ( IL-1 ) and P05231 . Toxicology testing and ongoing clinical trials support the notion that Monarsen treatment would offer considerable advantages over conventional cholinesterase inhibitors with respect to dosing , specificity , side effects profile , and duration of efficacy , while raising some open questions regarding its detailed mechanism of action . O75330 , a receptor for hyaluronan-mediated motility , on normal human lymphocytes , thymocytes and malignant B cells : a mediator in B cell malignancy ? O75330 ( Receptor for HA Mediated Motility ) is a novel HA receptor that has been linked to regulating cell locomotion and density dependent contact inhibition of fibroblasts , smooth muscle cells , macrophages , lymphocytes , astrocytes and sperm . The ubiquitous expression of O75330 suggests the existence of multiple isoforms , and indeed , O75330 is found in various cellular compartments , namely nuclear , cytosolic , membrane-bound and extracellular . In this review , we emphasize the evolving role of O75330 in B cell malignancies , and examine the function of O75330 in T cell development in the thymic microenvironment . Both the motile behaviour of progenitor thymocytes ( CD3- P01730 -CD8- ) and malignant B cells from multiple myeloma ( MM ) , plasma cell leukemia , and hairy cell leukemia was blocked by monoclonal antibodies to O75330 , suggesting that motility may correlate with increased expression of O75330 at the cell surface . Interestingly , the soluble form of O75330 is able to inhibit fibroblast locomotion , and it is likely that a balance between expression of both forms determines , in part the motility of cells . O75330 appears to play a fundamental role in the immune system and the ability of O75330 to function as a motility receptor is likely to be due to complex variables including the extent to which soluble O75330 is secreted . O75330 expression characterizes circulating monoclonal B cells as abnormal. potentially invasive and/or metastatic components of myeloma and may underlie the malignant behavior of these cells . Oxidation of alcohols and reduction of aldehydes derived from methyl- and dimethylpyrenes by cDNA-expressed human alcohol dehydrogenases . Some methylated pyrenes can form DNA adducts in rat tissues after benzylic hydroxylation and sulpho conjugation . However , oxidation of the intermediate alcohols to carboxylic acids is an important competing pathway leading to detoxification . We previously showed that co-administration of ethanol or DB01213 strongly enhances DNA adduct formation by 1-hydroxymethylpyrene , indicating an involvement of alcohol dehydrogenases ( ADHs ) in the detoxification . This mechanism may be involved in the observed synergism of smoking and alcohol consumption in certain human cancers . In a preceding study , cDNA-expressed human P00325 efficiently oxidised 1- , 2- and 4-hydroxymethylpyrene ; these reactions were inhibited in the presence of ethanol or DB01213 . Here we report that P00326 , P00326 and P08319 also show substantial activity towards these substrates and two further congeners , 1-hydroxymethyl-6-methylpyrene and 1-hydroxymethyl-8-methylpyrene . All four DB00067 forms also catalysed the reverse reaction , implying that the aldehydes have to be sequestered by other enzymes , such as aldehyde dehydrogenases , for final detoxification . P00326 and P08319 activities towards hydroxymethylpyrenes were more strongly inhibited in the presence of ethanol and DB01213 than those of P00325 . P00326 was only inhibited at very high concentrations of the modulators . In conclusions , several human ADHs are capable of detoxifying benzylic alcohols of alkylated polycyclic aromatic hydrocarbons . However , some competing substrates and inhibitors may affect all these redundant detoxification systems , although to various extents . The investigational O14965 inhibitor DB05220 induces defects in cell viability and cell-cycle progression in malignant bladder cancer cells in vitro and in vivo . PURPOSE : Despite more than 70,000 new cases of bladder cancer in the United States annually , patients with advanced disease have a poor prognosis due to limited treatment modalities . We evaluated O14965 , identified as an upregulated candidate molecule in bladder cancer , as a potential therapeutic target . EXPERIMENTAL DESIGN : Gene expression in human bladder cancer samples was evaluated using RNA microarray and quantitative reverse transcriptase PCR . Effects of the O14965 inhibitor DB05220 ( Millennium ) on cell dynamics in malignant T24 and UM-UC-3 and papilloma-derived RT4 bladder cells were evaluated in vitro and in vivo in a mouse xenograft model . RESULTS : A set of 13 genes involved in the mitotic spindle checkpoint , including Aurora kinases A and B , were upregulated in human urothelial carcinoma compared with normal urothelium . The O14965 inhibitor DB05220 induced cell-cycle arrest , aneuploidy , mitotic spindle failure , and apoptosis in the human bladder cancer cell lines T24 and UM-UC-3 . DB05220 also arrested tumor growth when administered orally over 4 weeks in a mouse bladder cancer xenograft model . Finally , in vitro sequential administration of DB05220 with either paclitaxel or gemcitabine resulted in synergistic cytotoxic effects in T24 cells . CONCLUSIONS : Mitotic spindle checkpoint dysfunction is a common characteristic of human urothelial carcinoma and can be exploited with pharmacologic Aurora A inhibition . Given our demonstration of the ability of the Aurora A inhibitor DB05220 to inhibit growth of bladder cancer in vitro and in vivo , we conclude that Aurora kinase inhibitors warrant further therapeutic investigation in bladder cancer . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . HIV protease substrate conformation : modulation by cyclophilin A . P62937 ( CyPA ) , a cytosolic peptidyl-prolyl trans-cis isomerase can accelerate the trans-cis isomerization of Xxx-Pro peptide bonds . One- and two-dimensional 1H-NMR spectroscopy were used to determine that the heptapeptide DB00133 -Gln- DB00174 - DB00135 -Pro- DB00167 - DB00161 , a model peptide of an HIV-1 protease cleavage site in the gag polyprotein of HIV-1 , is a substrate for CyPA . Experiments revealed a slow exchange about the DB00135 -Pro peptide bond with 30 +/- 5 % in the cis conformation ( pH 1-9 ) . While the interconversion rate is too slow to measure by kinetic NMR methods in the absence of CyPA , these methods , saturation transfer and NOE experiments , established that CyPA enhanced the rate of trans-cis interconversion , a process inhibited by cyclosporin A ( DB00091 ) . With a substrate:CyPA ratio of 40:1 , an interconversion rate of 2.5 s(-1) at 25 degrees C was observed .

MH_train_1273

MH_train_1273

MH_train_1273

interacts_with DB00603?

multiple_choice

Arterial reocclusion and persistent distal occlusion after thrombus aspiration . BACKGROUND AND PURPOSE : Early reocclusion of intracranial arteries can lead to poor clinical outcome . We report reocclusion detection after endovascular clot aspiration , followed by administration of P08514 -IIIa antagonist under continuous ultrasound monitoring . CASE DESCRIPTION : A 73-year-old man developed the right middle cerebral artery ( MCA ) occlusion with NIHSS 17 points , 6 days after aortic valve replacement . Recanalization was achieved with Penumbra™ system and reocclusion was detected with transcranial Doppler ( P24386 ) 30 minutes postcompletion of intra-arterial procedure . Proximal recanalization was achieved with the second thrombus aspiration while M2 MCA occlusion persisted beyond the reach of the device . Intravenous abciximab was administered under continuous P24386 monitoring . Recanalization with Thrombolysis in Brain Ischemia ( TIBI ) flow grade 4 was observed at 60 minutes postintervention accompanied with clinical recovery to NIHSS 3 points . DB00054 was given for 12 hours with no hemorrhagic transformation on repeat CT scan . Patient was discharged home with mild left pronator drift and facial droop , and his modified ranking score was 1 at 6-week follow-up visit . CONCLUSIONS : Early arterial reocclusion can occur after successful thrombus aspiration while P08514 -IIIa antagonist administration may lead to subsequent recanalization of persisting distal occlusions not amenable to mechanical removal . Steroid hormone receptors and coregulators in endocrine-resistant and estrogen-independent breast cancer cells . Resistance to hormonal therapy is often a problem in the treatment of breast cancer patients . It has been suggested that resistance could be explained by altered nuclear hormone receptor or coregulator levels or inappropriately increased agonist activity of selective estrogen receptor modulator ( SERM ) . To test these hypotheses , we have established novel MCF-7 cell line-derived in vitro models of anti-estrogen- and progestin-resistant and estrogen-independent breast cancer by long-term culture in the presence of toremifene and medroxyprogesterone acetate ( DB00603 ) and in the absence of estradiol , respectively . Using cell growth and multiprobe ribonuclease protection assays , the expression of 5 nuclear hormone receptors and 9 coregulators as well as the alterations in the cell proliferation and target gene transcription in response to hormonal treatments were studied . P06401 ( PR ) expression was decreased and silencing mediator for retinoid acid and thyroid hormone receptors ( Q9Y618 ) and amplified in breast cancer-1 ( Q9Y6Q9 ) expression increased in anti-estrogen-resistant cells . Estrogen caused PR and ERbeta upregulation in all cell lines , but we did not observe increased agonist activity of anti-estrogen measured by regulation of these estrogen target genes . Basal ERalpha levels and estrogenic growth response were decreased and p300/CBP-associated factor ( pCAF ) and Q9Y6Q9 upregulated by estrogen in progestin-resistant cells , but coregulator levels were unchanged . Estrogen-independent cells were still estrogen-responsive and PR , nuclear receptor corepressor ( O75376 ) and Q9Y618 expression was increased whereas steroid receptor coactivator-1 ( P12931 -1a ) and CBP-related protein p300 ( p300 ) expression decreased . Their growth was inhibited by toremifene , but estradiol was able to abrogate this effect , which might have interesting clinical implications concerning the use of postmenopausal hormone replacement therapy . Increased miR-21 expression during human monocyte differentiation into DCs . Differentiation of monocytes into dendritic cells ( DCs ) is characterised by marked changes in gene expression . The role of microRNAs ( miRNAs ) , a new class of small endogenous non-coding regulatory RNAs , in this process is still unclear . We identified miR-223 , miR-16 , miR-191 , miR-24 , let-7b , and miR-21 as differentially expressed between monocytes and monocyte derived DCs . We evaluated the expression levels of computationally predicted target genes of miR-21 in human monocytes following stimulation with GM- P04141 and P05112 . Moreover , transfection of monocytes with synthetic miR-21 inhibited the expression of a set of genes that were also repressed during monocyte differentiation to DCs in response to GM- P04141 and P05112 . Among these , we identified genes that are involved in cell cycle , apoptosis and differentiation such as Q07343 , Q53EL6 , P04083 , Q9NXR8 , Q8N3U4 , Q15582 , P80511 , LAT2 and P48552 . Collectively , the present study highlights the involvement of miRNAs , particularly miR-21 in monocyte differentiation to DCs and identifies potential miR-21 target genes . DB06186 : first global approval . DB06186 ( Yervoy® ) is an anti-cytotoxic T-lymphocyte antigen ( CTLA ) -4 monoclonal antibody that has been approved in the US for the first- or second-line treatment of patients with malignant melanoma . In the EU , it is awaiting approval as second-line therapy for melanoma . DB06186 blocks the effects of the negative T-cell regulator P16410 , which may in turn augment T-cell responses to tumour cells . Preclinical studies have indicated that antibody blocking of P16410 can lead to potent immune responses . DB06186 is also in development as first- and second-line therapy for prostate cancer where it has progressed to phase III clinical trials worldwide , and it is in phase II development for non-small cell lung cancer . DB06186 was originated by the University of California , Berkeley , in the US and subsequently licensed to Medarex , which was later acquired by Bristol-Myers Squibb . This article summarizes the milestones in the development of intravenous ipilimumab leading to this first approval . This profile has been extracted from Wolters Kluwer 's R & D Insight drug pipeline database . R & D Insight tracks drug development worldwide through the entire development process , from discovery , through pre-clinical and clinical studies to market launch . Stimulation of growth hormone secretion by central administration of atrial natriuretic polypeptide in the rat . Intracerebroventricular ( icv ) injection of alpha-human atrial natriuretic polypeptide ( alpha hANP ) or alpha-rat P01160 ( 0.6 and 3 nmol/rat ) elicited an increase in plasma GH levels both in conscious freely moving rats and in urethane-anesthetized rats when given at the trough of spontaneous GH secretion . Antiserum specific for rat P01286 did not affect the plasma GH increase induced by icv injection of alpha hANP . Intracerebroventricular injection of alpha P01160 ( 3 nmol/rat ) failed to stimulate GH secretion in conscious rats pretreated with cysteamine ( 30 mg/100 g BW , sc ) , a depletor of somatostatin ( SRIF ) , and in conscious rats during constant iv infusion of SRIF ( 55 ng/ml ) . GH release induced by iv injection of synthetic rat P01286 ( 200 ng/100 g BW ) was exaggerated by alpha hANP ( 3 nmol/rat , icv ) in conscious rats . These results suggest that central P01160 stimulates pituitary GH secretion possibly by inhibiting SRIF release from the hypothalamus in the rat . Hypothalamic-pituitary-adrenal axis hyperactivity accounts for anxiety- and depression-like behaviors in rats perinatally exposed to bisphenol A . Accumulating studies have proved that perinatal exposure to environmental dose causes long-term potentiation in anxiety/depression-related behaviors in rats . Hyperactivity of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis is one of the most consistent biological findings in anxiety- and depression-related disorders . The Q9Y251 axis is reported to be susceptible to developmental reprogramming . The present study focused on Q9Y251 reactivity in postnatal day ( P01160 ) 80 male rats exposed perinatally to environmental-dose Q03001 . When female breeders were orally administered 2 μg/(kg.day) Q03001 from gestation day 10 to lactation day 7 , their offspring ( P01160 80 Q03001 -exposed rats ) showed obvious anxiety/depression-like behaviors . Notably , significant increase in serum corticosterone and adrenocorticotropin , and corticotropin-releasing hormone mRNA were detected in Q03001 -exposed rats before or after the mild stressor . Additionally , the level of glucocorticoid receptor mRNA in the hippocampus , but not the hypothalamus , was decreased in Q03001 -exposed rats . The levels of hippocampal mineralocorticoid receptor mRNA , neuronal nitric oxide synthase and phosphorylated DB02527 response element binding protein were increased in Q03001 -exposed rats . In addition , the testosterone level was in Q03001 -exposed rats . The results indicate that reprogramming-induced hyperactivity of the Q9Y251 axis is an important link between perinatal Q03001 exposure and persistent potentiation in anxiety and depression . P03372 -immunoreactive neurons contain calcitonin gene-related peptide , methionine-enkephalin or tyrosine hydroxylase in the female rat preoptic area . We have shown in our previous studies that estrogen treatment selectively influences calcitonin gene-related peptide ( P80511 ) - , methionine-enkephalin ( DB00134 -Enk ) - and tyrosine hydroxylase ( TH ) -immunoreactive ( IR ) intensities in the neurons of the periventricular preoptic nucleus ( Q9H237 ) and the medial preoptic area ( DB00603 ) of the female rat . In the present study , we examined whether estrogen receptor ( ER ) -IR neurons in the Q9H237 and DB00603 contain P80511 , DB00134 -Enk , or TH using a double-labeling immunohistochemical method and investigated changes in the number of double-labeling cells upon treatment with estrogen . Brain sections of ovariectomized rats and ovariectomized and estrogen-treated rat were stained using the avidin-biotin-peroxidase complex method followed by the peroxidase-anti-peroxidase method . The sections were first incubated with an anti-ER antibody in conjunction with nickel diaminobenzidine which produces a dark blue reaction product in the nucleus . Subsequently , P80511 , DB00134 -Enk or TH antisera were applied to these sections and the resulting brown diaminobenzidine reaction product in the cytoplasm was examined . Neurons that were double-labeled for ER and P80511 , DB00134 -Enk or TH were investigated in the Q9H237 and DB00603 . The number of doubly labeled ER/ P80511 - and ER/TH-IR neurons was large , whereas the number of ER/ DB00134 -Enk-IR neurons was small . These results suggest that ER in the Q9H237 and DB00603 may be more closely related to the mechanism of changes in P80511 - and TH-IR intensities upon estrogen treatment than that in DB00134 -Enk-IR intensity . Inhibition of aldehyde dehydrogenase 2 activity enhances antimycin-induced rat cardiomyocytes apoptosis through activation of MAPK signaling pathway . Aldehyde dehydrogenase 2 ( P05091 ) , a mitochondrial-specific enzyme , has been proved to be involved in oxidative stress-induced cell apoptosis , while little is known in cardiomyocytes . This study was aimed at investigating the role of P05091 in antimycin A-induced cardiomyocytes apoptosis by suppressing P05091 activity with a specific P05091 inhibitor DB02115 . Antimycin A ( 40μg/ml ) was used to induce neonatal cardiomyocytes apoptosis . DB02115 ( 60μM ) effectively inhibited P05091 activity by 50 % without own effect on cell apoptosis , and significantly enhanced antimycin A-induced cardiomyocytes apoptosis from 33.5±4.4 to 56.5±6.4 % ( Hochest method , p < 0.05 ) , and from 57.9±1.9 to 74.0±11.9 % ( FACS , p < 0.05 ) . Phosphorylation of activated MAPK signaling pathway , including extracellular signal-regulated kinase ( P27361 /2 ) , c-Jun NH2-terminal kinase ( JNK ) and p38 was also increased in antimycin A and daidzin treated cardiomyocytes compared to the cells treated with antimycin A alone . These findings indicated that modifying mitochondrial P05091 activity/expression might be a potential therapeutic option on reducing oxidative insults induced cardiomyocytes apoptosis . The role of tetrahydrobiopterin in superoxide generation from P29474 : enzymology and physiological implications . DB00360 ( BH4 ) is a ubiquitous pteridine metabolite that serves as a NOS cofactor . Recently , we showed that BH4 efficiently inhibits superoxide generation from the heme group at the oxygenase domain of P29474 . This role indicates that BH4 acts as a redox switch in the catalytic mechanism of the enzyme , which may have important consequences in the physiology of the endothelium . Here the mechanism by which BH4 inhibits superoxide release from P29474 and the " uncoupling " effects of oxidized BH4 metabolites are presented . The implications of the disparate actions of fully reduced and oxidized BH4 metabolites in the control of P29474 biochemistry are discussed in the light of clinical data indicating that BH4 levels are important in the regulation of superoxide levels and of endothelial reactivity . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Exploration and optimization of substituted triazolothiadiazines and triazolopyridazines as DB05876 inhibitors . An expansion of structure-activity studies on a series of substituted 7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine DB05876 inhibitors and the introduction of a related [1,2,4]triazolo[4,3-b]pyridazine based inhibitor of DB05876 is presented . The development of SAR included strategic incorporation of known substituents on the critical catachol diether moiety of the 6-phenyl appendage on each heterocyclic core . From these studies , ( R ) -3-(2,5-dimethoxyphenyl)-6-(4-methoxy-3-(tetrahydrofuran-3-yloxy)phenyl)-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazine ( 10 ) and ( R ) -3-(2,5-dimethoxyphenyl)-6-(4-methoxy-3-(tetrahydrofuran-3-yloxy)phenyl)-[1,2,4]triazolo[4,3-b]pyridazine ( 18 ) were identified as highly potent P27815 inhibitors . Each of these analogues was submitted across a panel of 21 PDE family members and was shown to be highly selective for DB05876 isoforms ( P27815 , Q07343 , Q08493 , Q08499 ) . Both 10 and 18 were then evaluated in divergent cell-based assays to assess their relevant use as probes of DB05876 activity . Finally , docking studies with selective ligands ( including 10 and 18 ) were undertaken to better understand this chemotypes ability to bind and inhibit DB05876 selectively . Autoradiographic localization of specific atrial natriuretic peptide binding sites on immunocytochemically identified cells in cultures from rat and guinea-pig hearts . Dissociated cell culture preparations from rat and guinea-pig atria and interatrial septum , and from rat ventricles were studied using a combined autoradiographic and immunocytochemical approach . Alpha-atrial natriuretic peptide ( 125I-ANP1-128 ) bindings sites were confined to subpopulations of identified non-neuronal cells in each type of culture preparation , and had distinct patterns of labelling . The density of ANP1-28 binding sites was substantially greater in guinea-pig cultures than in rat cultures and was least in rat ventricular cultures . ANP1-28-labelled subpopulations of S-100-like immunoreactive glial cells were only seen in guinea-pig cultures . Von Willebrand factor ( P04275 ) -like immunoreactive endothelial cells and P04275 -negative endothelioid cells expressed ANP1-28 binding sites in both the guinea-pig and rat atrial cultures , but were unlabelled in rat ventricular cultures . In contrast , labelled subpopulations of fibronectin-like immunoreactive fibroblasts were present in all of the three types of culture preparation studied . P01160 -like immunoreactive myocytes were present in both atrial and ventricular cultures . These cells did not , however , express ANP1-28 binding sites . Dermatological toxicity associated with targeted therapies in cancer : optimal management . Targeted therapies have developed rapidly over the last few years in the field of oncology thanks to a better understanding of carcinogenesis . They target pathways involved in signal transduction ( P00533 , P04626 , P21860 , Q15303 , P36888 , DB01367 , RAF , MEK , P10721 , P07949 , P42345 , P12931 , P21709 , P21583 ) , tumor angiogenesis ( VEGFR , Q02763 ) , and tumor microenvironment ( P09619 , FGFR ) . They rarely cause the systemic adverse reactions generally associated with chemotherapy , but frequently cause disabling and specific skin toxicity . The impact on patient quality of life can be important both in terms of symptoms caused and of potentially aesthetic consequences . Inappropriate management can increase the risk of dose reduction or discontinuation of the cancer treatment . In this review , we will discuss skin toxicity associated with the main drug classes- P00533 , P15056 , MEK , P42345 , c- P10721 , P16410 , and SMO inhibitors , and anti-angiogenic agents . Targeted therapy-induced skin toxicities will be detailed in terms of symptoms , frequency , evolution , complications , and topical and oral treatments in order to improve their diagnosis and management . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . Effect of progesterone on intracellular Ca2+ homeostasis in human myometrial smooth muscle cells . Although it is well known that progesterone alters uterine contractility and plays an important role in maintenance of pregnancy , the biochemical mechanisms by which progesterone alters uterine contractility in human gestation are less clear . In this investigation we sought to identify progesterone-induced adaptations in human myometrial smooth muscle cells that may alter Ca2+ signaling in response to contractile agents . Cells were treated with vehicle or the progesterone analog medroxyprogesterone acetate ( DB00603 ) for 5 days , and intracellular free Ca2+ concentration ( [Ca2+]i ) was quantified after treatment with oxytocin ( OX ) or endothelin ( ET ) -1 . OX- and ET-1-induced increases in [Ca2+]i were significantly attenuated in cells pretreated with DB00603 in a dose-dependent manner . P06401 antagonists prevented the attenuated Ca2+ transients induced by DB00603 . P25101 and ETB receptor subtypes were expressed in myometrial cells , and treatment with DB00603 resulted in significant downregulation of P25101 and ETB receptor binding . DB00603 did not alter ionomycin-stimulated increases in [Ca2+]i and had no effect on inositol trisphosphate-dependent or -independent release of Ca2+ from internal Ca2+ stores . We conclude that adaptations of Ca2+ homeostasis in myometrial cells during pregnancy may include progesterone-induced modification of receptor-mediated increases in [Ca2+]i . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Minimal influence of tocilizumab on P01579 synthesis by tuberculosis antigens . P01579 ( P01579 ) production is a critical step of antituberculosis ( anti-TB ) immune response . The purpose of this study was to determine the influences of biologics , including the interleukin ( IL ) -6 receptor-inhibitor tocilizumab ( TCZ ) , and tumor necrosis factor ( P01375 ) antagonists infliximab ( P27352 ) and etanercept ( P25101 ) , on Mycobacterium tuberculosis ( MTB ) antigen-induced P01579 production . MTB antigen ( ESAT-6 and P27918 -10 ) -induced P01579 -releasing assay was performed with or without addition of biologics ( TCZ , P25101 , and P27352 ) using whole blood from patients with active TB . P25101 and P27352 inhibited P01579 production in a dose-dependent manner . In whole blood from TB patients , ESAT-6 stimulated significant production of P01579 ( 1.30 +/- 1.95 IU/ml ) , and TCZ did not inhibit P01579 production ( 1.56 +/- 1.88 IU/ml ) . P01579 production by ESAT-6 was inhibited by P25101 and P27352 ( 0.98 +/- 1.74 , 0.75 +/- 1.66 IU/ml , respectively ) . P27918 -10 stimulated significant production of P01579 ( 1.46 +/- 1.60 IU/ml ) , and TCZ did not inhibit P01579 production ( 1.51 +/- 1.77 IU/ml ) . P01579 production by P27918 -10 was inhibited by P25101 and P27352 ( 0.91 +/- 0.99 , 0.72 +/- 0.88 IU/ml , respectively ) . P24386 did not inhibit MTB antigen-induced P01579 production . As P01579 production is important in antimycobacterial host defenses , the minimal influence of TCZ on P01579 -releasing assay suggests a low risk of latent TB infection reactivation during tocilizumab therapy . DB06268 ( Q9Y6W8 -Texas Biotechnology ) . Q9Y6W8 -Texas Biotechnology is developing the endothelin A ( P25101 ) receptor antagonist , sitaxsentan , for the potential treatment of pulmonary hypertension , congestive heart failure ( CHF ) , chronic obstructive pulmonary disease and subarachnoid hemorrhage [ 205713 ] , [ 302200 ] . The compound is in phase IIa trials as an iv formulation for CHF and has completed phase I safety trials as an oral formulation [ 272071 ] . Phase II/III trials for pulmonary hypertension are planned for the first quarter of 2001 [ 3945711 ] . In June 2000 , Q9Y6W8 and Texas Biotechnology established a joint venture to develop and commercialize endothelin antagonists [ 370007 ] . US-05591761 was issued to Texas in January 1997 , covering TBC-11251 and several related isomers [ 2309301 . Pharmacokinetic , pharmacodynamic and clinical profile of novel antiplatelet drugs targeting vascular diseases . Platelet inhibitors are the mainstay treatment for patients with vascular diseases . The current ' gold standard ' antiplatelet agent clopidogrel has several pharmacological and clinical limitations that have prompted the search for more effective platelet antagonists . The candidates include various blockers of the purinergic Q9H244 receptor such as prasugrel , an oral irreversible thienopyridine ; two adenosine triphosphate analogues that bind reversibly to the Q9H244 receptor : ticagrelor ( oral ) and cangrelor ( intravenous ) ; elinogrel , a direct-acting reversible Q9H244 receptor inhibitor ( the only antiplatelet compound that can be administered both intravenously and orally ) ; BX 667 , an orally active and reversible small-molecule Q9H244 receptor antagonist ; P35240 530348 , P35240 205831 , P35240 602539 and E5555 , highly selective and orally active antagonists on the protease-activated receptor 1 . A number of drugs also hit new targets : terutroban , an oral , selective and specific inhibitor of the thromboxane receptor ; DB05202 , a second-generation , nuclease resistant aptamer which inhibits P04275 -dependent platelet aggregation ; Q96JZ2 -0081 , a bivalent humanized nanobody targeting the GPIb binding site of P04275 and AJW200 , an IgG4 monoclonal antibody of P04275 . The pharmacology and clinical profiles of new platelet antagonists indicate that they provide more consistent , more rapid and more potent platelet inhibition than agents currently used . Whether these potential advantages will translate into clinical advantages will require additional comparisons in properly powered , randomized , controlled trials . Psychosocial stress affects the involvement of prostaglandins and nitric oxide in the lipopolysaccharide-induced hypothalamic-pituitary-adrenal response . The role of prostaglandins and nitric oxide ( NO ) , generated after peripheral lipopolysaccharide ( LPS ) administration , in the adaptation of hypothalamic-pituitary-adrenal ( Q9Y251 ) axis under stressful circumstances remains to be elucidated . The aim of the present study was to assess the effect of chronic repetitive restraint or social crowding stress on the involvement of nitric oxide and prostaglandins in the LPS-induced pituitary-adrenocortical response . Male Wistar rats were restrained in metal tubes 2 x 10 min/day or crowded in cages for 7 days prior to treatment . All compounds were injected i.p. , cyclooxygenase ( P36551 ) and nitric oxide synthase ( NOS ) inhibitors 15 min before LPS . Two hrs after injection LPS induced a significant increase in DB01285 and corticosterone secretion . Repeated restraint impaired more potently than crowding stress the LPS-induced Q9Y251 -response . Indomethacin , a non-selective P36551 inhibitor , considerably reduced the LPS-induced Q9Y251 response in non-stressed rats and to a lesser extent diminished this response in repeatedly restrained or crowded rats . P29475 inhibitor , Nomega-nitro-L-arginine decreased the LPS-induced Q9Y251 response , more potently in control than crowded rats . DB02533 , an P35228 inhibitor , diminished the LPS-elicited DB01285 response in crowded rats . These results indicate that prostaglandins and NO generated by neuronal and inducible NOS are involved in the LPS-induced Q9Y251 axis response under basal conditions and during its adaptation to chronic social stress circumstances . Atrial natriuretic peptide : a possible mediator involved in dexamethasone 's inhibition of cell proliferation in multiple myeloma . Atrial natriuretic peptide ( P01160 ) has been recognized for several decades for its role of regulating blood pressure . Recently , cumulating evidences show that P01160 plays an anticancer role in various solid tumors via blocking the kinase cascade of Ras- Q02750 /2- P27361 /2 with the result of inhibition of DNA synthesis . P01160 , as well as its receptors ( P16066 and P17342 ) has been identified present in the embryonic stem cell and a wide range of cancer cells . Various lymphoid organs , such as lymph nodes , have been detected the presence of P01160 . Multiple myeloma ( MM ) , though the therapies have evolved significantly , is still an incurable disease as B lymphocyte cell neoplasm . Dexamethasone is the cornerstone in treatment of MM via inactivation of Ras- Q02750 /2- P27361 /2 cascade reaction . Coincidently , dexamethasone can increase the expression of P01160 markedly . Nevertheless , the role of P01160 in MM is unclear . Based on these results above , we raise the hypothesis that P01160 is involved in mediating dexamethasone 's inhibition of proliferation in MM cells , which suggests that P01160 may be a potential agent to treat MM .

MH_train_1274

MH_train_1274

MH_train_1274

interacts_with DB09280?

multiple_choice

Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . Structure-function studies of linear and cyclized peptide antagonists of the P30968 . Structurally new analogs of the peptidic P30968 antagonist DB00050 as well as conformationally constrained cyclized deca- or pentapeptides were synthesized and selected peptides evaluated comprehensively . To understand how structural variations of the antagonistic peptide effect pharmacodynamic properties , binding affinities and antagonistic potencies toward the human and rat P30968 were determined . Whereas large substituents in position 6 of linear peptides are compatible with high binding affinity ( K(D) < 0.5 nM ) , all cyclized peptides except the cyclo[3-10] analog D-52391 depicted low binding affinity ( K(D) > 10 nM ) . Binding affinity and antagonistic potency in vitro correlated for all peptides and surprisingly no discrimination between human and rat receptor proteins was observed . Since receptor residues W(101) and N(102) are involved in agonist and antagonist binding , equally potent but structurally different antagonists were tested for binding to the respective W(101)A and N(102)A mutants . In contrast to linear decapeptides , residues N(102) and W(101) are not involved in binding of D-23938 and W(101) is the critical residue for D-52391 binding . We conclude that although equally potent , peptidic P30968 antagonists do have distinct interactions within the ligand binding pocket . Finally , selected antagonists were tested for testosterone suppression in male rats . The duration of testosterone suppression below castration levels differed largely from 1 day for DB06785 to 27 days for D-23487 . Systemic availability became evident as the most important parameter for in vivo efficacy . Improvement of chloride transport defect by gonadotropin-releasing hormone ( DB00644 ) in cystic fibrosis epithelial cells . Cystic fibrosis ( CF ) , the most common autosomal recessive disease in Caucasians , is due to mutations in the P13569 gene . F508del , the most frequent mutation in patients , impairs P13569 protein folding and biosynthesis . The F508del- P13569 protein is retained in the endoplasmic reticulum ( ER ) and its traffic to the plasma membrane is altered . Nevertheless , if it reaches the cell surface , it exhibits a Cl(-) channel function despite a short half-life . Pharmacological treatments may target the F508del- P13569 defect directly by binding to the mutant protein or indirectly by altering cellular proteostasis , and promote its plasma membrane targeting and stability . We previously showed that annexine A5 ( AnxA5 ) directly binds to F508del- P13569 and , when overexpressed , promotes its membrane stability , leading to the restoration of some Cl(-) channel function in cells . Because Gonadotropin-Releasing Hormone ( DB00644 ) increases AnxA5 expression in some cells , we tested it in CF cells . We showed that human epithelial cells express DB00644 -receptors ( P30968 ) and that DB00644 induces an AnxA5 overexpression and an increased Cl(-) channel function in F508del- P13569 cells , due to an increased stability of the protein in the membranes . Beside the numerous physiological implications of the P30968 expression in epithelial cells , we propose that a topical use of DB00644 is a potential treatment in CF . P01375 polymorphisms as a potential modifier gene in the cystic fibrosis . Modifier genes , as the P01375 -α gene , can modulate the cystic fibrosis ( CF ) severity . Thus , -238G > A and -308G > A polymorphisms of P01375 -α gene were analyzed as modifiers of CF . In this context , the present study enrolled 49 CF patients ( diagnosis performed by sweat test and complete P13569 screening ) . The -238G > A polymorphism analysis was performed by Q9ULH0 -PCR , and -308G > A , by PCR-RFLP . In our data , the -238G > A polymorphism was not associated with clinical variability . The AA genotype for -308G > A polymorphism was a risk factor for early gastrointestinal symptoms ( OR=5.98 , 95 % CI=1.06-49.68 ) and protection for the first Pseudomonas aeruginosa ( OR=0.05 , 95 % CI=0.0003-0.007 ) . For the first P. aeruginosa , GA genotype was a risk factor ( OR=10.2 , 95 % CI=1.86-84.09 ) ; for the same genotype , the diagnosis was made in minor time than the AA genotype ( p=0.031 ) . Considering the -308G > A polymorphism alleles , the G allele was a risk factor for early pulmonary symptoms ( OR=3.81 , 95 % CI=1.13-12.97 ) and P. aeruginosa ( OR=66.77 , 95 % CI=15.18-482.7 ) ; however , the same allele showed better transcutaneous oxygen saturation ( OR=9.24 , 95 % CI=1.53-206.1 ) . The A allele was a protective factor for early pulmonary symptoms ( OR=12.26 , 95 % CI=0.08-0.89 ) and P. aeruginosa ( OR=12.15 , 95 % CI=0002-0007 ) , however , the same allele was a risk factor for worst transcutaneous oxygen saturation ( OR=7.01 , 95 % CI=1.14-157.4 ) . As conclusion , the -308G > A polymorphism of the P01375 -α gene was associated with the CF severity . Effect of endogenous kinins , prostanoids , and NO on kinin B1 and B2 receptor expression in the rabbit . To determine whether kinin receptor expression is regulated by kinins , prostaglandins , and/or nitric oxide ( NO ) , rabbits were treated with a B(1) receptor ( B(1)R ) antagonist , a B2 receptor ( P30411 ) antagonist , a prostacyclin mimetic , or inhibitors of NO synthase , cyclooxygenase , or angiotensin-converting enzyme . The mRNA concentrations for P46663 and P30411 ( multiplex RT-PCR ) were measured in several organs . The P30411 mRNA expression was not significantly upregulated by any of the treatments ; it was notably downregulated by angiotensin-converting enzyme or cyclooxygenase blockade or P30411 antagonism in the heart and duodenum . A treatment with bacterial lipopolysaccharide ( LPS ) , known to induce P46663 expression , has also been applied and was the most consistent in upregulating the expression of P46663 mRNA ( kidney , duodenum , and striated muscle ) . The contractile responses mediated by kinin receptors in blood vessels isolated from the treated rabbits also indicated that LPS was the only P46663 inducer ( aorta ) . DB06196 , a nonequilibrium antagonist of the rabbit P30411 , was the sole tested drug to alter the contractions mediated by the P30411 in the jugular vein or the intensity of the immunohistochemical P30411 staining in several organs ( inhibition in both cases ) . P30411 mRNA expression was downregulated in some organs by several of the applied treatments , but the data did not support generally applicable feedback for the regulation of P30411 expression involving endogenous kinins , prostanoids , or NO . There was no indication of compensatory or reciprocal regulation of B1Rs , relative to B2Rs , inasmuch as P46663 expression was restricted to LPS-treated animals . Characterization of disease-associated mutations affecting an exonic splicing enhancer and two cryptic splice sites in exon 13 of the cystic fibrosis transmembrane conductance regulator gene . Sequences in exons can play an important role in constitutive and regulated pre-mRNA splicing . Since exonic splicing regulatory sequences are generally poorly conserved and their mechanism of action is not well understood , the consequence of exonic mutations on splicing can only be determined empirically . In this study , we have investigated the consequence of two cystic fibrosis ( CF ) disease-causing mutations , E656X and 2108delA , on the function of a putative exonic splicing enhancer ( ESE ) in exon 13 of the P13569 gene . We have also determined whether five other CF mutations D648V , D651N , G654S , E664X and T665S located near this putative ESE could lead to aberrant splicing of exon 13 . Using minigene constructs , we have demonstrated that the E656X and 2108delA mutations could indeed cause aberrant splicing in a predicted manner , supporting a role for the putative ESE sequence in pre-mRNA splicing . In addition , we have shown that D648V , E664X and T665S mutations could cause aberrant splicing of exon 13 by improving the polypyrimidine tracts of two cryptic 3' splice sites . We also provide evidence that the relative levels of two splicing factors , hTra2alpha and Q07955 /ASF , could alter the effect on splicing of some of the exon 13 disease mutations . Taken together , our results suggest that the severity of CF disease could be modulated by changes in the fidelity of P13569 pre-mRNA splicing . Drosophila Answers to Q13148 Proteinopathies . Initially implicated in the pathogenesis of P13569 and HIV-1 transcription , nuclear factor Q13148 was subsequently found to be involved in the origin and development of several neurodegenerative diseases . In 2006 , in fact , it was reported for the first time the cytoplasmic accumulation of Q13148 in ubiquitin-positive inclusions of P35858 and FTLD patients , suggesting the presence of a shared underlying mechanism for these diseases . Today , different animal models of Q13148 proteinopathies are available in rodents , nematodes , fishes , and flies . Although these models recapitulate several of the pathological features found in patients , the mechanisms underpinning the progressive neuronal loss observed in Q13148 proteinopathies remain to be characterized . Compared to other models , Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies . At present , the development of Q13148 -related Drosophila models has further strengthened the hypothesis that both Q13148 " loss-of-function " and " gain-of-function " mechanisms can contribute to disease . The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies , along with the information they have generated , towards a better understanding of the mechanisms underlying Q13148 proteinopathies . Inflammatory signaling compromises cell responses to interferon alpha . DB05258 ( IFNα ) is widely used for treatment of melanoma and certain other malignancies . This cytokine as well as the related IFNβ exerts potent anti-tumorigenic effects ; however , their efficacy in patients is often suboptimal . Here , we report that inflammatory signaling impedes the effects of IFNα/β . Melanoma cells can secrete pro-inflammatory cytokines that inhibit cellular responses to IFNα/β via activating the ligand-independent pathway for the phosphorylation and subsequent ubiquitination and accelerated degradation of the P17181 chain of type I IFN receptor . Catalytic activity of the p38 protein kinase was required for P17181 downregulation and inhibition of IFNα/β signaling induced by proinflammatory cytokines such as interleukin 1 ( IL-1 ) . Activation of p38 kinase inversely correlated with protein levels of P17181 in clinical melanoma specimens . Inhibition of p38 kinase augmented the inhibitory effects of IFNα/β on cell viability and growth in vitro and in vivo . The roles of inflammation and p38 protein kinase in regulating cellular responses to IFNα/β in normal and tumor cells are discussed . Specific interaction between P23443 and Q13057 : a potential link between the P42345 /S6K pathway , DB01992 biosynthesis and energy metabolism . Ribosomal protein S6 kinase ( S6K ) is a key regulator of cell size and growth . It is regulated via phosphoinositide 3-kinases ( PI3K ) and the mammalian target of rapamycin ( P42345 ) signaling pathways . We demonstrate for the first time that Q13057 associates specifically with P23443 . The association was observed between native and transiently overexpressed proteins in vivo , as well as by BIAcore analysis in vitro . The sites of interaction were mapped to the C-terminal regions of both Q13057 and P23443 . In vitro studies indicated that the interaction does not affect their enzymatic activities and that Q13057 is not a substrate for S6 kinase . This study uncovers a potential link between mTor/S6K signaling pathway and energy metabolism through DB01992 and its thioester derivatives , but its physiological relevance should be further elucidated . 3-Hydroxy-3-methylglutaryl coenzyme A synthase-1 of Blattella germanica has structural and functional features of an active retrogene . Blattella germanica has two cytosolic 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) synthase genes , HMG- Q13057 -1 and -2 . HMG- Q13057 -1 gene shows several features of processed genes ( retroposons ) : it contains no introns but has a short direct-repeat sequence ( ATTATTATT ) at both ends . An atypical feature is the presence at both ends of the gene of short inverse repeats flanked by direct repeats . There is neither a TATA box nor a CAAT box in the 5' region . Comparative analysis with other species suggests that the HMG- Q13057 -1 gene derives from HMG- Q13057 -2 . Cultured embryonic B. germanica UM-BGE-1 cells express HMG- Q13057 -1 but not HMG- Q13057 -2 , suggesting that the intron-less gene is functional . In addition , it can complement MEV-1 cell line , which is auxotrophic for mevalonate . We show that compactin and mevalonate do not significantly affect the mRNA levels of HMG- Q13057 -1 in UM-BGE-1 cells . DB06693 induces a 6.7-fold increase in P04035 activity , which is restored to normal levels by mevalonate . HMG- Q13057 activity is not modified by either of these effectors , suggesting that the mevalonate pathway in this insect cell line is regulated by post-transcriptional mechanisms affecting P04035 but not HMG- Q13057 . P25025 -dependent mucosal neutrophil influx protects against colitis-associated diarrhea caused by an attaching/effacing lesion-forming bacterial pathogen . Enteropathogenic Escherichia coli ( EPEC ) is a major cause of diarrheal disease in young children , yet symptoms and duration are highly variable for unknown reasons . Citrobacter rodentium , a murine model pathogen that shares important functional features with EPEC , colonizes mice in colon and cecum and causes inflammation , but typically little or no diarrhea . We conducted genome-wide microarray studies to define mechanisms of host defense and disease in C. rodentium infection . A significant fraction of the genes most highly induced in the colon by infection encoded CXC chemokines , particularly P09341 /2/5 and Q07325 /10 , which are ligands for the chemokine receptors P25025 and P49682 , respectively . CD11b(+) dendritic cells were the major producers of P09341 , P42830 , and Q07325 , while P19875 was mainly induced in macrophages . Infection of gene-targeted mice revealed that P49682 had a significant but modest role in defense against C. rodentium , whereas P25025 had a major and indispensable function . P25025 was required for normal mucosal influx of neutrophils , which act as direct antibacterial effectors . Moreover , P25025 loss led to severe diarrhea and failure to express critical components of normal ion and fluid transport , including ATPase beta(2)-subunit , P13569 , and P40879 . The antidiarrheal functions were unique to P25025 , since other immune defects leading to increased bacterial load and inflammation did not cause diarrhea . Thus , P25025 -dependent processes , particularly mucosal neutrophil influx , not only contribute to host defense against C. rodentium , but provide protection against infection-associated diarrhea . DB01819 cycling via mitochondrial phosphoenolpyruvate carboxykinase links anaplerosis and mitochondrial GTP with insulin secretion . Pancreatic beta-cells couple the oxidation of glucose to the secretion of insulin . Apart from the canonical K( DB00171 )-dependent glucose-stimulated insulin secretion ( GSIS ) , there are important K( DB00171 )-independent mechanisms involving both anaplerosis and mitochondrial GTP ( mtGTP ) . How mtGTP that is trapped within the mitochondrial matrix regulates the cytosolic calcium increases that drive GSIS remains a mystery . Here we have investigated whether the mitochondrial isoform of phosphoenolpyruvate carboxykinase ( Q16822 ) is the GTPase linking hydrolysis of mtGTP made by succinyl- DB01992 synthetase ( SCS-GTP ) to an anaplerotic pathway producing phosphoenolpyruvate ( PEP ) . Although cytosolic PEPCK ( P35558 ) is absent , Q16822 message and protein were detected in P01308 -1 832/13 cells , rat islets , and mouse islets . PEPCK enzymatic activity is half that of primary hepatocytes and is localized exclusively to the mitochondria . Novel (13)C-labeling strategies in P01308 -1 832/13 cells and islets measured substantial contribution of Q16822 to the synthesis of PEP . As high as 30 % of PEP in P01308 -1 832/13 cells and 41 % of PEP in rat islets came from Q16822 . The contribution of Q16822 to overall PEP synthesis more than tripled with glucose stimulation . Silencing the Q16822 gene completely inhibited GSIS underscoring its central role in mitochondrial metabolism-mediated insulin secretion . Given that mtGTP synthesized by SCS-GTP is an indicator of TCA flux that is crucial for GSIS , Q16822 is a strong candidate to link mtGTP synthesis with insulin release through anaplerotic PEP cycling . Identification of flanking markers for the familial amyotrophic lateral sclerosis gene ALS1 on chromosome 21 . Amyotrophic lateral sclerosis ( P35858 ) is a progressive , adult-onset , neurodegenerative disorder characterized by the death of large motor neurons from the cerebral cortex , brainstem , and spinal cord . The etiology of P35858 remains unknown ; however , approximately 10 % of the cases are familial in nature . In the majority of these families , the mode of transmission is autosomal dominant . Recently , linkage of an autosomal dominant familial P35858 ( FALS ) gene to the locus ALS1 on chromosome 21q was established . In addition , evidence was provided for genetic heterogeneity , with approximately 55 % of families most likely linked to chromosome 21 . The development of a number of highly informative simple sequence repeat polymorphisms in the region of linkage-21q21 through 21q22.1-has permitted us to confirm both the assignment of ALS1 to 21q and the genetic heterogeneity of FALS . In addition , we have been able to refine the mapping of ALS1 , based on recombination events in two of the linked families . Flanking markers for the FALS gene are D21S213 on the centromeric side and D21S219 on the telomeric side . The candidate region is approximately 4 Mb and contains the genes copper/zinc superoxide dismutase ( CuZnSOD ) ; the fourth member of the class II cytokine receptor family ( Q08334 ) ; and the interferon-alpha receptor ( P17181 ) . P01308 is involved in transcriptional regulation of NKCC and the P13569 Cl(-) channel through PI3K activation and P29323 inactivation in renal epithelial cells . It is is well known that insulin stimulates glucose transport and epithelial Na(+) channel ( ENaC ) -mediated Na(+) reabsorption ; however , the action of insulin on Cl(-) secretion is not fully understood . In this study , we investigated the action of insulin on Na(+)-K(+)-2Cl(-) cotransporter ( NKCC ) -mediated Cl(-) secretion in epithelial A6 cells . Interestingly , insulin treatment remarkably enhanced the forskolin-stimulated Cl(-) secretion associated with an increase in apical Cl(-) conductance by upregulating mRNA expression of both P13569 and NKCC , although insulin treatment alone had no effect on the basal Cl(-) secretion or apical Cl(-) conductance without forskolin application . We next elucidated a role of phosphoinositide 3-kinase ( PI3K ) in the insulin-induced enhancement of the Cl(-) secretion , since insulin actually activated PI3K , resulting in activation of Akt , a downstream molecule of PI3K . LY294002 ( a PI3K inhibitor ) reduced the Cl(-) secretion by suppressing mRNA expression of NKCC , whereas insulin still had a stimulatory action on mRNA expression of P13569 even in the presence of LY294002 . On the other hand , we found that a MEK inhibitor ( PD98059 ) further enhanced the insulin-stimulated P13569 mRNA expression and the Cl(-) secretion in forskolin-stimulated A6 cells and that insulin induced slight , transient activation of P29323 followed by significant inactivation of P29323 . These observations suggest that : ( 1 ) insulin respectively upregulates mRNA expression of NKCC and P13569 through activation of PI3K and inactivation of P29323 ; ( 2 ) insulin signals on mRNA expression of NKCC and P13569 are not enough to stimulate transepithelial Cl(-) secretion , but enhance the stimulatory action of DB02527 on transepithelial Cl(-) secretion . DB00107 -stimulated NFAT transcriptional activation in human myometrial cells . DB00107 ( P01178 ) is a peptide hormone that binds the P01178 receptor on myometrial cells , initiating an intracellular signaling cascade , resulting in accumulation of intracellular calcium and smooth muscle contraction . In other systems , an elevation of intracellular Ca(2+) stimulates nuclear translocation of the transcription factor , nuclear factor of activated T cells ( NFAT ) , which is transcriptionally active in arterial and ileal smooth muscle . Here we have investigated the role of NFAT in the mechanism of action of P01178 . Human myometrial cells expressed all five NFAT isoforms ( O95644 -C4 and -5 ) . Myometrial cells were transduced with a recombinant adenovirus expressing a O95644 - Q14258 reporter , and a semi-automated imaging system was used to monitor effects of P01178 on reporter localization in live cells . P01178 induced a concentration-dependent nuclear translocation of O95644 - Q14258 in a reversible manner , which was inhibited by P01178 antagonists and calcineurin inhibitors . Pulsatile stimulation with P01178 caused intermittent , pulse-frequency-dependent , nuclear translocation of O95644 - Q14258 , which was more efficient than sustained stimulation . P01178 induced nuclear translocation of endogenous NFAT that was transcriptionally active , because P01178 stimulated activity of a NFAT-response element-luciferase reporter and induced calcineurin-NFAT dependent expression of P41220 , P53805 , and P35354 ( P35354 ) mRNA . Furthermore , P01178 -dependent transcription was dependent on protein neosynthesis ; cycloheximide abolished P41220 transcription but augmented P53805 and P35354 transcriptional readouts . This study identifies a novel signaling mechanism within the myometrium , whereby calcineurin-NFAT signaling mediates P01178 -induced transcriptional activity . Furthermore , we show O95644 - Q14258 is responsive to pulses of P01178 , a mechanism by which myometrial cells could decode P01178 pulse frequency . DB04998 inhibits activation of nuclear factor-kappaB ( NF-kappaB ) by forming a complex with NF-kappaB essential modulator ( Q9Y6K9 ) and nucleolin . DB04998 , also known as DB04998 , is an experimental anticancer drug that recently entered human clinical trials . It is a member of a novel class of antiproliferative agents known as G-rich oligonucleotides ( P09341 ) , which are non-antisense , guanosine-rich phosphodiester oligodeoxynucleotides that form stable G-quadruplex structures . The biological activity of GROs results from their binding to specific cellular proteins as aptamers . One important target protein of GROs has been previously identified as nucleolin , a multifunctional protein expressed at high levels by cancer cells . Here , we report that DB04998 also associates with nuclear factor-kappaB ( NF-kappaB ) essential modulator ( Q9Y6K9 ) , which is a regulatory subunit of the inhibitor of kappaB ( IkappaB ) kinase ( IKK ) complex , and also called IKKgamma . In the classic NF-kappaB pathway , the IKK complex is required for phosphorylation of P25963 and subsequent activation of the transcription factor NF-kappaB . We found that treatment of cancer cells with DB04998 inhibits IKK activity and reduces phosphorylation of P25963 in response to tumor necrosis factor-alpha stimulation . Using a reporter gene assay , we showed that DB04998 blocks both tumor necrosis factor-alpha-induced and constitutive NF-kappaB activity in human cancer cell lines derived from cervical , prostate , breast , and lung carcinomas . In addition , we showed that , in DB04998 -treated cancer cells , Q9Y6K9 is coprecipitated by nucleolin , indicating that both proteins are present in the same complex . Our studies suggest that abrogation of NF-kappaB activity may contribute to the anticancer effects of DB04998 and that nucleolin may play a previously unknown role in regulating the NF-kappaB pathway . Three adjacent genes of African swine fever virus with similarity to essential poxvirus genes . Nucleotide sequencing of the right end of the SalIj fragment of the highly virulent Malawi Lil20/1 strain of African swine fever virus ( ASFV ) has revealed three adjacent genes with similarity to : serine-threonine protein kinases ; members of the putative helicase superfamily Q07955 ; and the vaccinia virus 56 kDa abortive late protein . All three genes are transcribed to the left with respect to the orientation of the ASFV genome . Gene L19IL predicts a protein similar to serine-threonine protein kinases including vaccinia virus gene P46663 . Gene L19KL predicts a protein that is likely to be a nucleic acid-dependent ATPase , as it has similarity to both the poxvirus 70 kDa early transcription factor subunit and the poxvirus nucleoside triphosphatase I gene . Gene L19LL has extensive similarity to the vaccinia virus 56 kDa abortive late protein . Meloxicam . Meloxicam ( DB00814 trade mark , Boehringer Ingelheim ) is a relatively new oral non-steroidal anti-inflammatory drug ( NSAID ) approved for the treatment of osteoarthritis in the US . It has also been evaluated for the treatment of rheumatoid arthritis , ankylosing spondylitis and acute ' rheumatic ' pain . Meloxicam has been shown to be P35354 preferential , particularly at its lowest therapeutic dose , and is anti-inflammatory by inhibiting prostanoid synthesis in inflammatory cells . Since it is P35354 preferential , it would be expected to have less gastrointestinal toxicity than non-selective NSAIDs . In clinical trials of meloxicam in osteoarthritis , it was found to be as effective as piroxicam , diclofenac and naproxen with less clinical gastrointestinal symptoms and less perforations , obstructions and bleeds by meta-analysis . Adverse events , including peripheral oedema and hypertension , occurred at a similar rate as with traditional NSAIDs . Activating AMP-activated protein kinase ( AMPK ) slows renal cystogenesis . Renal cyst development and expansion in autosomal dominant polycystic kidney disease ( ADPKD ) involves both fluid secretion and abnormal proliferation of cyst-lining epithelial cells . The chloride channel of the cystic fibrosis transmembrane conductance regulator ( P13569 ) participates in secretion of cyst fluid , and the mammalian target of rapamycin ( P42345 ) pathway may drive proliferation of cyst epithelial cells . P13569 and P42345 are both negatively regulated by AMP-activated protein kinase ( AMPK ) . Metformin , a drug in wide clinical use , is a pharmacological activator of AMPK . We find that metformin stimulates AMPK , resulting in inhibition of both P13569 and the P42345 pathways . Metformin induces significant arrest of cystic growth in both in vitro and ex vivo models of renal cystogenesis . In addition , metformin administration produces a significant decrease in the cystic index in two mouse models of ADPKD . Our results suggest a possible role for AMPK activation in slowing renal cystogenesis as well as the potential for therapeutic application of metformin in the context of ADPKD . ICE/ P29466 inhibitors as novel anti-inflammatory drugs . In recent years , several strategies that selectively inhibit pro-inflammatory cytokines , have yielded effective protein-based therapies for inflammatory disorders , validating the therapeutic hypothesis that intervention in cytokine signalling can provide clinical benefit . However , these protein-based products must be administered by injection , a constraint associated with inconvenience , adverse effects and expense for patients , caregivers and insurers . Besides interfering with the effects of cytokines such as P01375 or IL-1beta that have already been produced , inhibition of pro-inflammatory cytokine production or signalling with low-molecular weight orally-active drugs would combine the convenience of conventional pharmaceuticals with the focused efficacy of the protein therapies . Reducing IL-1beta and Q14116 production by inhibition of IL-1beta converting enzyme ( ICE , caspase-1 ) is one promising strategy because of the key roles of these cytokines in many inflammatory diseases . DB04875 , the first orally available , potent and selective ICE inhibitor to enter clinical trials , is currently under investigation in rheumatoid arthritis . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability .

MH_train_1275

MH_train_1275

MH_train_1275

interacts_with DB08899?

multiple_choice

P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . P10275 -induced tumor suppressor , B2CW77 , inhibits breast cancer growth and transcriptionally activates p53/p73-mediated apoptosis in breast carcinomas . P10275 ( AR ) expression by immunohistochemistry correlates with better prognosis and survival among breast cancer patients . We and others have shown that AR inhibits proliferation and induces apoptosis in breast cancer cells . However , the mechanism of AR 's anti-tumor effect in breast cancer is still not fully understood . Our recent study indicates that AR upregulates expression of tumor suppressor gene P60484 by promoter activation in breast cancer . B2CW77 , encoding B2CW77 protein , is a newly identified gene , which shares a bidirectional promoter with P60484 and is transcribed in the opposite direction . So far , the function of B2CW77 has never been studied in tumorigenesis . Here , we define B2CW77 as a tumor suppressor in breast carcinomas , which inhibits tumor growth and invasiveness . After analyzing 188 normal breast and 1247 malignant breast cancer tissues , we observed the loss of B2CW77 in multiple breast cancer subtypes and this decreased B2CW77 expression associates with tumor progression and increasing histological grade in invasive carcinomas . We characterize B2CW77 , for the first time , as a transcription factor , directly promoting the expression of P04637 and O15350 , with consequent elevated apoptosis and cell cycle arrest in breast cancer cells . We demonstrate , in vitro and in murine xenograph models , that both B2CW77 and P60484 are AR-target genes , mediating androgen-induced growth inhibition and apoptosis in breast cancer cells . Our observations suggest that B2CW77 might be used as a potential prognostic marker and novel therapy target for breast carcinomas . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . Hyperhemolysis syndrome in anemia of chronic disease . BACKGROUND : Occasional cases of delayed hemolytic transfusion reaction ( P10275 ) demonstrate severe and persistent hemolysis and are referred to as hyperhemolysis syndrome . This syndrome usually occurs in patients with sickle cell disease and possibly thalassemia who receive multiple transfusions . There are few such clinical reports in patients without hemoglobinopathies . CASE REPORT : A 67-year-old woman with anemia and a history of four previous transfusions was admitted with shortness of breath and a hematocrit ( Hct ) level of 27 percent . The patient was group O with a negative antibody screen . She received 1 unit of electronically cross-matched red blood cells ( RBCs ) and was discharged . Thirteen days later she returned to hospital with weakness and a Hct level of 23 percent . The antibody screen now demonstrated anti-K alloantibody . The direct antigloblulin test ( Q01959 ) was positive with both anti-immunoglobulin G and anti-complement ( P01024 ) . Anti-K was recovered in the eluate . The previously transfused RBC unit was positive for presence of the K antigen . The patient 's RBCs were negative for the presence of K antigen . Other laboratory data confirmed ongoing hemolysis , and a diagnosis of P10275 was made . She continued to display findings of active hemolysis for 9 more weeks requiring 19 units of RBCs . Thirty-four days after the original transfusion , her Q01959 remained positive and both the plasma sample and a RBC eluate demonstrated anti-K . CONCLUSION : The delayed hemolytic transfusion reaction with hyperhemolysis can occur among patients without hemoglobinopathies . Activation of peroxisome proliferator-activated receptor-{delta} enhances regenerative capacity of human endothelial progenitor cells by stimulating biosynthesis of tetrahydrobiopterin . The mechanisms underlying the regenerative capacity of endothelial progenitor cells ( EPCs ) are not fully understood . We hypothesized that biosynthesis of tetrahydrobiopterin is an important mechanism responsible for the stimulatory effects of peroxisome proliferator-activated receptor-δ ( PPARδ ) activation on regenerative function of human EPCs . Treatment of human EPCs with a selective PPARδ agonist GW501516 for 24 hours increased the levels of mRNA , protein , and enzymatic activity of P30793 ( GTPCH I ) , as well as the production of tetrahydrobiopterin . The effects of GW501516 were mediated by suppression of P60484 expression , thereby increasing phosphorylation of AKT . The AKT signaling also mediated GW501516-induced phosphorylation of endothelial NO synthase . In addition , activation of PPARδ significantly enhanced proliferation of EPCs . This effect was abolished by the GTPCH I inhibitor , 2,4-diamino-6-hydroxypyrimidine , or genetic inactivation of GTPCH I with small interfering RNA but not by inhibition of endothelial NO synthase with N(G)-nitro-l-arginine methyl ester . Supplementation of NO did not reverse 2,4-diamino-6-hydroxypyrimidine-inhibited 5-bromodeoxyuridine incorporation . Furthermore , transplantation of human EPCs stimulated re-endothelialization in a mouse model of carotid artery injury . Pretreatment of EPCs with GW501516 significantly enhanced the ability of transplanted EPCs to repair denuded endothelium . GTPCH I-small interfering RNA transfection significantly inhibited in vivo regenerative capacity of EPCs stimulated with GW501516 . Thus , in human EPCs , activation of PPARδ stimulates expression and activity of GTPCH I and biosynthesis of tetrahydrobiopterin via P60484 -AKT signaling pathway . This effect enhances the regenerative function of EPCs . DB02709 inhibits PDGF receptor mitogenic signaling in mesangial cells : role of P18031 . Mesangioproliferative glomerulonephritis is associated with overactive PDGF receptor signal transduction . We show that the phytoalexin resveratrol dose dependently inhibits PDGF-induced DNA synthesis in mesangial cells with an IC(50) of 10 microM without inducing apoptosis . Remarkably , the increased Q96EB6 deacetylase activity induced by resveratrol was not necessary for this inhibitory effect . DB02709 significantly blocked PDGF-stimulated c-Src and Akt kinase activation , resulting in reduced cyclin D1 expression and attenuated P06400 phosphorylation and cyclin-dependent kinase-2 ( P24941 ) activity . Furthermore , resveratrol inhibited P09619 phosphorylation at the PI 3 kinase and Grb-2 binding sites tyrosine-751 and tyrosine-716 , respectively . This deficiency in P09619 phosphorylation resulted in significant inhibition of PI 3 kinase and Erk1/2 MAPK activity . Interestingly , resveratrol increased the activity of protein tyrosine phosphatase P18031 , which dephosphorylates PDGF-stimulated phosphorylation at tyrosine-751 and tyrosine-716 on P09619 with concomitant reduction in Akt and Erk1/2 kinase activity . P18031 significantly inhibited PDGF-induced DNA synthesis without inducing apoptosis . These results for the first time provide evidence that the stilbene resveratrol targets P18031 to inhibit P09619 mitogenic signaling . Genetically rescued tetrahydrobiopterin-depleted mice survive with hyperphenylalaninemia and region-specific monoaminergic abnormalities . One of the possibly mutated genes in DOPA-responsive dystonia ( DRD , Segawa 's disease ) is the gene encoding P30793 , which is the rate-limiting enzyme for tetrahydrobiopterin ( BH4 ) biosynthesis . Based on our findings on 6-pyruvoyltetrahydropterin synthase ( Q03393 ) gene-disrupted ( Pts(-/-) ) mice , we suggested that the amount of tyrosine hydroxylase ( TH ) protein in dopaminergic nerve terminals is regulated by the intracellular concentration of BH4 . In this present work , we rescued Pts(-/-) mice by transgenic introduction of human Q03393 cDNA under the control of the dopamine beta-hydroxylase promoter to examine regional differences in the sensitivity of dopaminergic neurons to BH4-insufficiency . The DPS-rescued ( Pts(-/-) , DPS ) mice showed severe hyperphenylalaninemia . Human Q03393 was efficiently expressed in noradrenergic regions but only in a small number of dopaminergic neurons . DB03886 and dopamine contents , and TH activity in the striatum were poorly restored compared with those in the midbrain . TH-immunoreactivity in the lateral region of the striatum was far weaker than that in the medial region or in the nucleus accumbens . We concluded that dopaminergic nerve terminals projecting to the lateral region of the striatum are the most sensitive to BH4-insufficiency . Biochemical and pathological changes in DPS-rescued mice were similar to those in human malignant hyperphenylalaninemia and DRD . Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking . Binge drinking of alcoholic beverages among adolescents is a common practice that can have serious health consequences . DB00898 is a potent immunomodulator that alters a wide range of immune responses . However , it is unclear whether there is a differential immune response to alcoholic beverages with a high versus low concentration of ethanol . In this study , we used a PCR array containing 46 primer pairs of selected genes to compare mRNA expression in the spleen , an immune system organ , of adolescent rats following binge drinking of alcohol solutions containing either 20 % or 52 % ethanol ( v/v , 4.8 g/kg daily dosage ) , or water ( control ) for 3 d . We found that , expression of IL-1β , P05231 , P13500 , and GABA(A) receptor α2 subunit in the spleen were decreased , and P41594 and P46098 receptor expression were increased after administration of an ethanol solution containing 52 % ethanol , but not one with 20 % ethanol . Our data suggest that alcohol-mediated immunomodulatory effects are , in part , dependent on the ethanol by volume concentration . This is the first study to show that exposure to a high ethanol percentage beverage can have more profound effects on immune responses than one with a low percentage of ethanol . Telomere shortening is associated with reduced duodenal HCOFormula secretory but normal gastric acid secretory capacity in aging mice . The incidence of duodenal ulcer , especially Helicobacter pylori-negative duodenal ulcer , strongly increases with age . In humans , telomere length shortening is considered to be one critical factor in cellular senescence and organ survival . In this study , we compared basal and stimulated gastric acid and duodenal HCO(3)(-) secretory rates in aged late-generation ( G(3) ) telomerase-deficient ( mTERC(-/-) ) mice , which are characterized by severe telomere dysfunction due to the inability to elongate telomeres during cell division . We found that basal and forskolin-stimulated HCO(3)(-) secretion and short-circuit current ( I(sc) ) in isolated duodenal mucosa of G(3) mTERC(-/-) mice were markedly reduced compared with age-matched wild-type mice . In contrast , basal and forskolin-stimulated acid secretory rates in isolated G(3) mTERC(-/-) gastric mucosa were not significantly altered . Correspondingly , duodenal mucosa of G(3) mTERC(-/-) mice showed slimming and shortening of villi , whereas gastric mucosal histology was not significantly altered . However , the ratios of cystic fibrosis transmembrane conductance regulator ( P13569 ) and solute-linked carrier 26 gene family ( Slc26a6 ) mRNA expression in relation to cytokeratin-18 were not altered in duodenal mucosa . The further knockout of P38936 , which is a downstream effector of telomere shortening-induced senescence , rescued villus atrophy of duodenal mucosa , and basal and forskolin-stimulated duodenal HCO(3)(-) secretion and I(sc) in mTERC(-/-) P38936 (-/-) double-knockout mice were not different from wild-type controls . In conclusion , genetic ablation of telomerase resulted in P38936 -dependent duodenal mucosal atrophy and reduced duodenal HCO(3)(-) secretory capacity , whereas gastric morphology and acid secretory function were preserved . This suggests that telomere shortening during aging may result in an imbalance between aggressive and protective secretions against duodenal mucosa and thus predispose to ulcer formation . Human bronchial smooth muscle cells express adenylyl cyclase isoforms 2 , 4 , and 6 in distinct membrane microdomains . Adenylyl cyclases ( AC ) are important regulators of airway smooth muscle function , because β-adrenergic receptor ( AR ) agonists stimulate AC activity and increase airway diameter . We assessed expression of AC isoforms in human bronchial smooth muscle cells ( hBSMC ) . Reverse transcriptase-polymerase chain reaction and immunoblot analyses detected expression of AC2 , AC4 , and AC6 . DB02587 -stimulated AC activity in membranes from hBSMC displayed Ca(2+)-inhibited and G(βγ)-stimulated AC activity , consistent with expression of AC6 , AC2 , and AC4 . DB01064 -stimulated AC activity was inhibited by Ca(2+) but unaltered by G(βγ) , whereas butaprost-stimulated AC activity was stimulated by G(βγ) but unaffected by Ca(2+) addition . Using sucrose density centrifugation to isolate lipid raft fractions , we found that only AC6 localized in lipid raft fractions , whereas AC2 and AC4 localized in nonraft fractions . Immunoisolation of caveolae using caveolin-1 antibodies yielded Ca(2+)-inhibited AC activity ( consistent with AC6 expression ) , whereas the nonprecipitated material displayed G(βγ)-stimulated AC activity ( consistent with expression of AC2 and/or AC4 ) . Overexpression of AC6 enhanced DB02527 production in response to isoproterenol and beraprost but did not increase responses to prostaglandin E(2) or butaprost . β(2)AR , but not prostanoid EP(2) or EP(4) receptors , colocalized with O95622 /6 in lipid raft fractions . Thus , particular G protein-coupled receptors couple to discreet AC isoforms based , in part , on their colocalization in membrane microdomains . These different DB02527 signaling compartments in airway smooth muscle cells are responsive to different hormones and neurotransmitters and can be regulated by different coincident signals such as Ca(2+) and G(βγ) . Changing paradigms in management of metastatic Castration Resistant Prostate Cancer ( mCRPC ) . Recently , the standard of care for metastatic Castration Resistant Prostate Cancer ( mCRPC ) has changed considerably . Persistent androgen receptor ( AR ) signaling has been identified as a target for novel therapies and reengages the fact that AR continues to be the primary target responsible for metastatic prostate cancer . P10275 gene amplification and over expression have been found to result in a higher concentration of androgen receptors on tumor cells , making them extremely sensitive to low levels of circulating androgens . Additionally , prostate cancer cells are able to maintain dihydrotestosterone ( DB02901 ) concentration in excess of serum concentrations to support tumor growth . For many years ketoconazole was the only P05093 inhibitor that was used to treat mCRPC . However , significant toxicities limit its use . Newly approved chemotherapeutic agents such as DB05812 ( an oral selective inhibitor of CYP17A ) , which blocks androgen biosynthesis both within and outside the prostate cancer cells ) , and enzalutamide ( blocks AR signaling ) have improved overall survival . There are also ongoing phase III trials for Orteronel ( P50750 - 700 ) , ARN- 509 and Galeterone ( TOK-001 ) , which targets androgen signaling . In this review , we will present the rationale for the newly approved hormonal treatments , their indications and complications , and we will discuss ongoing trials that are being done to improve the efficacy of the approved agents . Finally , we will talk about the potential upcoming hormonal treatments for mCRPC . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Structure and function of eritadenine and its 3-deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d- DB03769 ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 ) and has hypocholesterolemic activity . We have hypothesized that 3-deaza-DEA ( P01024 -DEA ) and its analogues retain high level of P23526 inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 -DEA analogues would have much higher hypocholesterolemic activity . P01024 -DEA , and its methyl ester ( P01024 -OMeDEA ) and its methyl amido ( P01024 -NMeDEA ) were synthesized to examine their P23526 inhibitory and hypocholesterolemic activities . A crystal structure of P23526 containing P01024 -DEA was determined and confirmed that DEA and P01024 -DEA bound to the same site of P23526 with the same binding mode . The P23526 inhibitory activities of P01024 -DEA ( K(I)=1.5 microM ) and P01024 -OMeDEA ( K(I)=1.5 microM ) are significantly lower than that of DEA ( K(I)=30 nM ) , while rats fed by P01024 -DEA and P01024 -OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 -NMeDEA lost most of the P23526 inhibitory activity ( K(I)=30 microM ) and dietary P01024 -NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 -DEA analogues are neither substrates nor inhibitors of adenosine deaminase . Novel treatments of GERD : focus on the lower esophageal sphincter . Up to 50 % of patients with gastroesophageal reflux disease ( GERD ) still suffer from GERD symptoms despite proton pump inhibitor ( PPI ) therapy , indicating a need for new treatments . The lower esophageal sphincter ( LES ) plays a crucial role in maintaining the mechanical barrier necessary for prevention of gastric reflux . Transient LES relaxation ( TLESR ) is an important factor behind the occurrence of reflux , and preclinical studies have identified a number of targets for pharmacologic modification of TLESR . However , only gamma-aminobutyric acid ( GABA ) type B receptor ( GABA(B) ) agonists and metabotropic glutamate receptor 5 ( P41594 ) modulators have shown positive proof of concept in the clinical setting . The P41594 negative allosteric modulator DB05070 improved symptoms in GERD patients , but was associated with central side effects such as dizziness . DB00181 , a GABA(B) receptor agonist , reduces the incidence of TLESR and improves GERD symptoms in both adult and pediatric GERD patients . However , the utility of baclofen is similarly limited by poor tolerability and recent research has focused on the development of GABA(B) receptor agonists with improved tolerability . DB05031 , a prodrug of R-baclofen , reduced the number of reflux episodes in a dose-ranging study and was similarly tolerated to placebo . AZD3355 and AZD9343 are GABA(B) receptor agonists with limited central nervous system activity that have been shown in preclinical studies to reduce the incidence of TLESR and decrease esophageal acid exposure ; data from clinical studies of these agents in GERD patients are awaited with interest . Agents that target TLESR activity may therefore offer a promising new add-on treatment for patients who suffer from GERD symptoms despite PPI therapy . Analog of somatostatin vapreotide exhibits biological effects in vitro via interaction with neurokinin-1 receptor . OBJECTIVES : DB04894 , a synthetic analog of somatostatin , has analgesic activity most likely mediated through the blockade of neurokinin-1 receptor ( P25103 ) , the DB05875 ( SP ) -preferring receptor . The ability of vapreotide to interfere with other biological effects of SP has yet to be investigated . METHODS : We studied the ability of vapreotide to antagonize P25103 in three different cell types : immortalized U373MG human astrocytoma cells , human monocyte-derived macrophages ( MDM ) and a human embryonic kidney cell line , HEK293 . Both U373MG and MDM express endogenous P25103 while HEK293 cells , which normally do not express P25103 , are stably transformed to express human P25103 ( HEK293- P25103 ) . RESULTS : DB04894 attenuates SP-triggered intracellular calcium increases and nuclear factor-κB activation in a dose-dependent manner . DB04894 also inhibits SP-induced interleukin-8 and monocyte chemotactic protein-1 production in HEK293- P25103 and U373MG cell lines . DB04894 inhibits HIV-1 infection of human MDM in vitro , an effect that is reversible by SP pretreatment . CONCLUSIONS : Our findings indicate that vapreotide has P25103 antagonist activity and may have a potential application as a therapeutic intervention in HIV-1 infection . Technique appraisement of comparative proteomics and screening of differentiation-related protein in gastric carcinoma . BACKGROUND/AIMS : Different differentiations of cancer have resulted in its unique biological characteristics . We screen and appraise differentially expressed proteins in different differentiated gastric adenocarcinoma with comparative proteomics technology in order to find regulatory factors of tumor differentiation related and finally reach the purpose of tumor differentiation reversal . METHODOLOGY : With two-dimensional fluorescence difference gel electrophoresis ( 2-D DIGE ) and liquid chromatography in conjunction with tandem mass spectrometry ( LC–MS/MS ) , the differentially expressed proteins from 8 patients with different differentiated gastric adenocarcinoma were identified and some factors identified were verified with application of QPCR and Western blot techniques . RESULTS : Significant differences in 35 protein spots were found and 48 kinds of proteins were identified . Other than structural proteins and non-specific protein , six possible proteins associated with tumor differentiation were determined - the serine protease inhibitor B1 ( serine protease inhibitor , clade B , member 1 , P30740 ) , calcium-phospholipid binding protein III ( annexin A3 ) , transcription factor Nm23-H1 , adenine phosphoribosyl-transferase enzyme P07741 ( DB00173 Phosphoribosyltransferase in APO and AMP ) , glutathione S-transferase P1-1 ( Q86UG4 -π-1 ) , antimicrobial peptides P81605 -lL . The identified P30740 , annexin A3 , Nm23-H1 and P07741 were verified , confirming the expression of these factors was in line with proteomics identification . CONCLUSIONS : Protein expression in different differentiated gastric adenocarcinoma was varied . Neurokinin-1 receptor blockade and murine lung tumorigenesis . RATIONALE : Analogous to the adenoma-carcinoma sequence in the colon , it has been proposed that adenocarcinoma ( AC ) in the lung arises from adenomatous hyperplasia that progresses through atypical adenomatous hyperplasia to AC . However , the data supporting this sequence are largely circumstantial and the almost impossible task of identifying these lesions before resection rules out any longitudinal study in humans . OBJECTIVES , METHODS , AND RESULTS : We show in mice that the loss of function of the neurokinin-1 receptor ( P25103 ) -due to either a pharmacologic or genetic manipulation-results in a sequence of morphologic changes in response to bleomycin treatment that precede the development of AC . We also demonstrate that a series of alterations in gene expression of proliferation markers ( i.e. , P12004 and Ki-67 ) and cell cycle regulators ( i.e. , P49789 , p53 , and P38936 ) characterizes the sequence of the precursor lesions . The loss of function of the P25103 results in changes of the apoptotic rate and in a delay of DNA break recovery of alveolar epithelial cells following bleomycin treatment . The P25103 blockade interferes with a caspase-independent pathway of apoptosis by affecting both the translocation of P22736 into the cytoplasm and the expression of some important Bcl2 family members such as Bcl2 and Bak . CONCLUSIONS : To our knowledge , this is the first model to demonstrate a role for P25103 in lung epithelial cell death and tumorigenesis . This animal model may provide new information on the biology of AC and will facilitate designing and testing of new therapeutic interventions . A novel extract SB-300 from the stem bark latex of Croton lechleri inhibits P13569 -mediated chloride secretion in human colonic epithelial cells . An oligomeric proanthocyanidin ( DB04941 ) extracted from the bark latex of the tree Croton lechleri ( family Euphorbiaceae ) is a potent inhibitor of cholera toxin-induced fluid accumulation and chloride secretion . The manufacturing process for DB04941 was optimized and simplified to produce an increased yield of the herbal extract . The novel extract ( named SB-300 ) contained on average 70.6+/-7.2 % DB04941 by weight ( mean +/- S.D. ; n=56 lots ) . Here , we describe the effectiveness of SB-300 on DB02527 -regulated chloride secretion , which is mediated by the cystic fibrosis transmembrane conductance regulator Cl- channel ( P13569 ) in human colonic T84 cells . Exposure of the apical surface to SB-300 blocked forskolin-stimulated Cl- secretion by 92.2+/-3.0 % with a half-maximal inhibition constant ( KB ) of 4.8+/-0.8 microM . For DB04941 , stimulated Cl- currents were decreased by 98.0+/-7.2 % and KB averaged 4.1+/-1.3 microM . There was no significant difference between the blocking kinetics of DB04941 and SB-300 . DB02587 -stimulated whole cell Cl- currents were effectively blocked by extracellular addition of SB-300 ( 63+/-8.5 % ; n=3 ) and to a similar extent by DB04941 ( 83 +/- 0.6 % ; n=2 ; at 50 microM each ) . Both extracts inhibited a time- and voltage-independent Cl- conductance , which indicated the involvement of P13569 Cl- channels . We conclude that both DB04941 ( used in DB04941 ) and SB-300 ( used in P46459 Normal Stool Formula ) are novel natural products that target the P13569 Cl- channel . SB-300 is a low cost herbal extract and may present a complementary and alternative medicine approach for the treatment of fluid loss in watery diarrhea . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . Adenylyl cyclases 5 and 6 underlie PIP3-dependent regulation . Many different neurotransmitters and hormones control intracellular signaling by regulating the production of the second messenger DB02527 . The function of the broadly expressed adenylyl cyclases ( ACs ) 5 and 6 is regulated by either stimulatory or inhibitory G proteins . By analyzing a well-known rebound stimulation phenomenon after withdrawal of Gi protein in atrial myocytes , we discovered that O95622 and -6 are tightly regulated by the second messenger PIP3 . By monitoring DB02527 levels in real time by means of Förster resonance energy transfer ( FRET ) -based biosensors , we reproduced the rebound stimulation in a heterologous expression system specifically for O95622 or -6 . Strikingly , this DB02527 rebound stimulation was completely blocked by the PI3K inhibitor wortmannin , both in atrial myocytes and in transfected human embryonic kidney cells . Similar effects were observed by heterologous expression of the PIP3 phosphatase and tensin homolog ( P60484 ) . However , general kinase inhibitors or inhibitors of Akt had no effect , suggesting a PIP3-dependent mechanism . These findings demonstrate the existence of a novel general pathway for regulation of O95622 and -6 activity via PIP3 that leads to pronounced alterations of cytosolic DB02527 levels . Synthesis of oleanolic acid derivatives : In vitro , in vivo and in silico studies for P18031 inhibition . Non-insulin dependent diabetes mellitus is a multifactorial disease that links different metabolic routes ; a point of convergence is the enzyme P18031 which turns off insulin and leptin receptors involved in glucose and lipid metabolism , respectively . Pentacyclic acid triterpenes such as oleanolic acid ( OA ) have proved to be excellent P18031 inhibitors , thus , the purpose of current work was to generate a series of derivatives that improve the pharmacological effect of OA . Our findings suggest that the presence of the carboxylic acid and/or its corresponding reduction product carbinol derivative ( H-bond donor ) in C-28 is required to maintain the inhibitory activity ; moreover , this is further enhanced by ester or ether formation on C-3 . The most active derivatives were cinnamoyl ester ( 6 ) and ethyl ether ( 10 ) . DB04088 showed potent in vitro inhibitory activity and significantly decrease of blood glucose levels on in vivo experiments . Meanwhile , 10 showed contrasting outcomes , since it was the compound with higher inhibitory activity and selectivity over P18031 and has improved interaction with site B , according with docking studies , the in vivo antidiabetic effect was similar to oleanolic acid . In conclusion , oleanolic acid derivatives have revealed an enhanced inhibitory effect over P18031 activity by increasing molecular interactions with either catalytic or allosteric sites and producing a hypoglycaemic effect on non insulin dependent diabetes mellitus rat model . P05093 inhibitors -- abiraterone , C17,20-lyase inhibitors and multi-targeting agents . As the first in class steroid 17α-hydroxylase/C17,20-lyase ( P05093 ) inhibitor , abiraterone acetate ( of which the active metabolite is abiraterone ) has been shown to improve overall survival in patients with castration-resistant prostate cancer ( CRPC ) -- in those who are chemotherapy-naive and those previously treated with docetaxel . Furthermore , the clinical success of abiraterone demonstrated that CRPC , which has previously been regarded as an androgen-independent disease , is still driven , at least in part , by androgens . More importantly , abiraterone is a ' promiscuous ' drug that interacts with a number of targets , which dictate its clinical benefits and adverse effects profile . Besides P05093 inhibition , abiraterone acts as an antagonist to the androgen receptor and inhibits 3β-hydroxysteroid dehydrogenase -- two effects that potentially contribute to its antitumour effects . However , the inhibition of the 17α-hydroxylase activity of P05093 , P15538 and a panel of hepatic CYP enzymes leads to adverse effects and toxicities that include secondary mineralocorticoid excess . DB05812 is also associated with increased incidence of cardiac disorders . Under such circumstances , development of new P05093 inhibitors as an additional line of defence is urgently needed . To achieve enhanced clinical benefits , new strategies are being explored that include selective inhibition of the C17,20-lyase activity of P05093 and multi-targeting strategies that affect androgen synthesis and signalling at different points . Some of these strategies-including the drugs orteronel , VT-464 and galeterone -- are supported by preclinical data and are being explored in the clinic . A comparison of inflammatory , cytoprotective and injury gene expression profiles in kidneys from brain death and cardiac death donors . BACKGROUND : The superior long-term survival of kidneys from living donors ( LDs ) compared with kidneys from donation-after-brain-death ( DBD ) and donation-after-cardiac-death ( P81605 ) donors is now well established . However , comparative studies on transcriptional changes that occur at organ retrieval and during and after cold ischemia ( CI ) are sparse . METHODS : Using a rat model , we used qRT-PCR to examine expression levels of inflammatory , cytoprotective , and injury genes at different time points after organ retrieval . Cleaved caspase-3 was used to evaluate early apoptosis in P81605 and DBD kidneys . RESULTS : Immediately after retrieval , we found massive up-regulation of proinflammatory genes interleukin-1β , interleukin-6 , tumor necrosis factor-α , monocyte chemotactic protein-1 , P16109 , and P16581 in DBD compared with LD and P81605 kidneys . A significant increase in the expression of injury markers Kim-1 , P38936 , and the cytoprotective gene heme oxygenase-1 accompanied this . Bax was increased in P81605 kidneys , and Bcl-2 was decreased in DBD kidneys . After 2 hr of CI in the LD group and 18 hr in the DBD and P81605 groups , gene expression levels were similar to those found after retrieval . During 18 hr of cold storage , expression levels of these genes did not change . In P81605 and DBD kidneys , early apoptosis increased after CI . DISCUSSION/CONCLUSION : The gene expression profile in DBD kidneys represents an inflammatory and injury response to brain death . In contrast , P81605 kidneys show only mild up-regulation of inflammatory and injury genes . These results may imply why delayed graft function in P81605 kidneys does not have the deleterious effect it has on DBD kidneys .

MH_train_1276

MH_train_1276

MH_train_1276

interacts_with DB00741?

multiple_choice

Modulation of iron metabolism in monocyte cell line U937 by inflammatory cytokines : changes in transferrin uptake , iron handling and ferritin mRNA . We have investigated the effects of the pro-inflammatory cytokines interleukin 1 beta ( P01584 ) , tumour necrosis factor alpha ( P01375 alpha ) and interferon gamma ( IFN gamma ) on the iron metabolism of the human monocytic cell line U937 . Cells were treated with each cytokine for up to 24 h , and then iron uptake from diferric transferrin was determined . The intracellular distribution of this iron , the expression of the transferrin receptor and levels of mRNA for the two ferritin subunits were also studied . P01584 , P01375 alpha and IFN gamma all decreased transferrin-iron uptake into cells , and all three cytokines had effects on the proportion of iron associated with ferritin . With P01375 alpha there was a marked enhancement of the fraction incorporated into ferritin . P02787 -receptor expression was diminished by P01375 alpha and P01584 , but not IFN gamma , suggesting different effector mechanisms . Both P01375 alpha and IFN gamma increased the amount of cellular mRNA for ferritin H-chain , but not the L-chain ; P01584 affected mRNA for neither ferritin . These data demonstrate that cytokines , which can be present at high concentrations in inflammation , have the capacity to affect macrophage iron uptake , transferrin receptor expression , intracellular iron handling and the relative abundance of ferritin-subunit mRNA , and may therefore be important mediators in the observed perturbations of iron metabolism in inflammatory diseases . [ Concentration of acute phase proteins in serum of children with rheumatoid arthritis ] . An attempts was made at evaluation of the changes of acute phase proteins-seromucoid ( BRS ) , alpha 1-acid glycoprotein ( alpha 1-AGP ) , alpha 1-antitrypsin ( alpha 1-AT ) , haptoglobin ( Hp ) , P01024 protein of complement system ( P01024 ) , P05155 ( DB05341 ) , and transferrin ( Tf ) in the serum of children with rheumatoid arthritis . The patients were divided into clinical and age groups . The studies carried out have shown in ill children increase of BRS , alpha 1-AGP , alpha 1-AT , Hp , and DB05341 levels , During remission increased levels of alpha 1-AGP and Hp persisted . In the studied clinical groups a positive correlation was found between the intensity of changes of the studied indices and the degree of disease activity . In the age groups greater increase of the levels of the studied acute phase proteins was found in the group of preschool children and the group of puberty spurt . The obtained results suggest that the determination of acute phase proteins may be useful in laboratory investigation of children with rheumatoid arthritis . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . Toxicogenetics of antiretroviral therapy : genetic factors that contribute to metabolic complications . Metabolic complications of antiretroviral therapy ( O00253 ) have emerged as a major concern for long-term , successful management of HIV infection . Variability in the response to O00253 between individuals has been increasingly linked to the genetic background of patients , as regards efficacy and susceptibility to adverse reactions ( toxicogenetics ) . This review summarizes the biological and methodological background for the genetic prediction of metabolic toxicity of O00253 . Recent studies are discussed which suggest that single-nucleotide polymorphisms ( SNPs ) in several genes involved in lipid metabolism and lipid transport in the general population ( O95477 , Q6Q788 , P02656 , P02649 , P11597 ) might modulate plasma triglyceride and high-density lipoprotein cholesterol levels in HIV-infected patients . At present , genetic prediction of lipodystrophy is not possible . Lipodystrophy has been linked to an accumulation of mtDNA mutations , a finding causally associated with ageing phenotypes in animal models . No mutations in P02545 , a gene linked to rare , inherited forms of lipodystrophy , have been identified in small studies of patients with lipodystrophy , and a possible link to a P01375 promoter SNP remains to be confirmed . With the rapidly decreasing cost of genetic testing , the main issues that need to be addressed prior to introduction of toxicogenetic prediction in HIV clinical practice include reproducibly high predictive values of SNP associations with clinically relevant and well defined metabolic outcomes , studies that evaluate the contribution of SNPs in the context of multi-SNP and haplotype analysis , and the validation of genetic markers in independent , large patient cohorts . Comprehensive , whole genome approaches are increasingly being used . P02787 enhances the antiproliferative effect of aluminum on osteoblast-like cells . DB01370 ( Al ) retention in the body can cause metabolic bone disease . This disorder is characterized by reductions in the number of osteoblasts , a feature that suggests a disturbance in bone cell proliferation or differentiation . Because Al as well as iron ( Fe ) can bind to transferrin ( TF ) in plasma , the role of TF as a modifier of osteoblast proliferation was examined in UMR-106-01 osteoblast-like cells by measuring the incorporation of tritiated thymidine ( [ 3H ] -TdR ) into DNA ( counts.min-1.microgram cell protein-1 , means +/- SE ) during 48-h incubations in serum-free medium ( SFM ) . In the absence of TF , DNA synthesis decreased when media levels of Al exceeded 6-10 microM . The mitogenic response to physiological levels of unsaturated TF ( apo-TF ) was attenuated however during incubations with TF that was partially saturated with Al ( Al-TF ) . A similar inhibitory response was seen in cells incubated with the antiproliferative agent gallium ( Ga ) when added to SFM as partially saturated Ga-TF . TF produced a shift to the left in the inhibitory dose-response curve to Al in osteoblast-like cells ; thus , DNA synthesis decreased at substantially lower media concentrations of Al in cells grown in SFM containing partially saturated Al-TF . The results indicate that TF is an important determinant of the inhibitory effect of Al on DNA synthesis by osteoblast-like cells at the micromolar levels of Al that can occur in plasma in vivo . P01024 gene expression and regulation in human glomerular epithelial cells . Extra-hepatic synthesis of complement is thought to mediate local tissue inflammatory injury . To investigate this phenomenon in the glomerular epithelial cell ( GEC ) , we examined the biosynthesis and regulation of gene expression of the third component of complement in isolated human GEC derived from normal tissue . Metabolic labelling and immunoprecipitation studies demonstrated that P01024 protein was synthesized , processed and secreted by GEC under basal conditions . The secreted P01024 alpha and beta polypeptide chains had identical electrophoretic mobilities with those of hepatic P01024 . Examination of cellular RNA using semi-quantitative polymerase chain reaction ( PCR ) showed that P01024 gene expression was present in unstimulated GEC and was increased by stimulation with interferon-gamma ( P01579 ) in a time- and dose-dependent manner . Tumour necrosis factor-alpha ( P01375 ) , while mediating an increase in monocyte U937 P01024 expression , revealed no evidence of regulation of GEC P01024 gene expression . These results indicate that human GEC spontaneously express the P01024 gene and that increased gene expression is regulated by P01579 . These observations may reflect part of a wider mechanism of protection against or mediation of local , immune-mediated tissue injury . Effects of pituitary hormones on the prostate . BACKGROUND : Although essential , androgens alone are not sufficient to induce normal growth and functionality of the prostate . Nonandrogenic hormones must also be involved in the proliferation of the prostate cancer cells which do not respond to antiandrogenic therapy and which thus become androgen-independent . P01236 , but also growth hormone and luteinizing hormone , are potentially able to act on both normal and abnormal prostatic cells . METHODS : In this review we summarize data from the literature concerning the physiological and pathological implications of prolactin , growth hormone , and luteinizing hormone on the prostate . RESULTS : In rodent prostates , prolactin and growth hormone can induce a variety of effects independently of androgens ( e.g. , transactivation of certain genes , or synthesis of the major secretion products ) . Moreover , hyperprolactinemia is responsible for inflammation and dysplasia of the gland , while growth hormone promotes the development of prostate tumors in vivo in the mouse and rat . Growth hormone acts on the gland directly , through prostatic growth hormone receptors , and/or indirectly via the stimulation of insulin-like growth factor-I ( P05019 ) synthesis in the liver . P22888 is expressed in rat and human prostates . DB00044 increases the amount of various transcripts in the rat prostate through an androgen-independent pathway . CONCLUSIONS : P01236 , growth hormone , and luteinizing hormone , alone or synergistically with androgens , play physiologically significant roles in the normal prostate . The involvement of these hormones in the development of benign prostatic hyperplasia and prostatic carcinoma is an issue that needs to be addressed . Endocrine modulation of physiological responses to catabolic disease . Disease or endotoxemia alters the plasma concentrations of anabolic hormones , particularly growth hormone ( GH ) and insulin-like growth-factor I ( P05019 ) . In general , these hormones are inhibited during the catabolic disease state . A hypothesis has evolved that anabolic hormones might be useful in patients ' recovery under these and other catabolic circumstances . The treatment of cattle with GH has provided significant improvement in the physiological response of the animals to the subsequent injection of bacterial lypopolysaccharide ( LPS ) , perhaps via inhibition of tumor necrisis factor ( P01375 ) release . However , this improved response to disease was not observed with animals treated with GH and infected with one of two parasitic organisms , Sarcocystis cruzi or Eimeria bovis . Recent attempts with other anabolic hormones , estradiol and progesterone , have proven remarkably effective in improving the adaptive physiological responses of calves to either E. bovis infection or to the injection of LPS . All animals displayed signs of infection , but the intensity and duration of symptoms were reduced . Although a mechanism is not yet known , there were no effects on P01375 ; cortisol ; the percentages of lymphocytes expressing P06729 , 4 , or 8 antigens ; or the production of antibodies . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . DB01599 alleviates atherosclerosis and improves high density lipoprotein function . BACKGROUND : DB01599 is a unique hypolipidemic agent that decreases high density lipoprotein cholesterol ( HDL-C ) . However , it is not definite that whether probucol hinders the progression of atherosclerosis by improving HDL function . METHODS : Eighteen New Zealand White rabbits were randomly divided into the control , atherosclerosis and probucol groups . Control group were fed a regular diet ; the atherosclerosis group received a high fat diet , and the probucol group received the high fat diet plus probucol . Hepatocytes and peritoneal macrophages were isolated for [ (3)H ] labeled cholesterol efflux rates and expression of O95477 and SR-B1 at gene and protein levels ; venous blood was collected for serum paraoxonase 1 , myeloperoxidase activity and lipid analysis . Aorta were prepared for morphologic and immunohistochemical analysis after 12 weeks . RESULTS : Compared to the atherosclerosis group , the paraoxonase 1 activity , cholesterol efflux rates , expression of O95477 and Q8WTV0 in hepatocytes and peritoneal macrophages , and the level of O95477 and Q8WTV0 in aortic lesions were remarkably improved in the probucol group , But the serum HDL cholesterol concentration , myeloperoxidase activity , the IMT and the percentage plaque area of aorta were significantly decreased . CONCLUSION : DB01599 alleviated atherosclerosis by improving HDL function . The mechanisms include accelerating the process of reverse cholesterol transport , improving the anti-inflammatory and anti-oxidant functions . Inhibition of P51587 and DB01643 Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 mediates homologous recombination repair , and P51587 polymorphisms increase cancer risk . However , tumors with P51587 mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 synergistically potentiated drugs with mechanisms of action related to P51587 function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality. " TS knockdown induced complementary lethality to TS-targeting drugs ( 5-FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 -targeting antisense oligdeoxynucleotide ( ASO ) " BR-1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi:10.1038/mtna.2013.7 published online 12 March 2013 . Adipose tissue tumor necrosis factor and interleukin-6 expression in human obesity and insulin resistance . Adipose tissue expresses tumor necrosis factor ( P01375 ) and interleukin ( IL ) -6 , which may cause obesity-related insulin resistance . We measured P01375 and P05231 expression in the adipose tissue of 50 lean and obese subjects without diabetes . P01308 sensitivity ( S(I) ) was determined by an intravenous glucose tolerance test with minimal-model analysis . When lean [ body mass index ( BMI ) < 25 kg/m(2) ] and obese ( BMI 30-40 kg/m(2) ) subjects were compared , there was a 7.5-fold increase in P01375 secretion ( P < 0.05 ) from adipose tissue , and the P01375 secretion was inversely related to S(I) ( r = -0.42 , P < 0.02 ) . P05231 was abundantly expressed by adipose tissue . In contrast to P01375 , plasma ( rather than adipose ) P05231 demonstrated the strongest relationship with obesity and insulin resistance . Plasma P05231 was significantly higher in obese subjects and demonstrated a highly significant inverse relationship with S(I) ( r = -0.71 , P < 0.001 ) . To separate the effects of BMI from S(I) , subjects who were discordant for S(I) were matched for BMI , age , and gender . By use of this approach , subjects with low S(I) demonstrated a 3.0-fold increased level of P01375 secretion from adipose tissue and a 2.3-fold higher plasma P05231 level ( P < 0.05 ) compared with matched subjects with a high S(I) . Plasma P05231 was significantly associated with plasma nonesterified fatty acid levels ( r = 0.49 , P < 0.002 ) . Thus the local expression of P01375 and plasma P05231 are higher in subjects with obesity-related insulin resistance . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS ) [ Cellular adhesion signal transduction network of tumor necrosis factor-alpha induced hepatocellular carcinoma cells ] . OBJECTIVE : To systemically explore the cellular adhesion signal transduction network of tumor necrosis factor-alpha ( P01375 -α ) -induced hepatocellular carcinoma cells with bioinformatics tools . METHODS : Published microarray dataset of P01375 -α-induced HepG2 , human transcription factor database HTRI and human protein-protein interaction database HPRD were used to construct and analyze the signal transduction network . RESULTS : In the signal transduction network , MYC and SP1 were the key nodes of signaling transduction . Several genes from the network were closely related with cellular adhesion. P00533 ( P00533 ) is a possible key gene of effectively regulating cellular adhesion during the induction of P01375 -α . CONCLUSION : P00533 is a possible key gene for P01375 -α-induced metastasis of hepatocellular carcinoma . [ Obesity studies in candidate genes ] . There are more than 430 chromosomic regions with gene variants involved in body weight regulation and obesity development . Polymorphisms in genes related to energy expenditure -- uncoupling proteins ( UCPs ) , related to adipogenesis and insulin resistance -- hormone-sensitive lipase ( Q96M11 ) , peroxisome proliferator-activated receptor gamma ( Q07869 gamma ) , beta adrenergic receptors ( P07550 ,3 ) , and alfa tumor necrosis factor ( P01375 ) , and related to food intake -- ghrelin ( Q9UBU3 ) -- appear to be associated with obesity phenotypes . Obesity risk depends on two factors : a ) genetic variants in candidate genes , and b ) biographical exposure to environmental risk factors . It is necessary to perform new studies , with appropriate control groups and designs , in order to reach relevant conclusions with regard to gene/environmental ( diet , lifestyle ) interactions . The relationship of P04141 and plasma cytokine levels to cerebral white matter injury in the premature newborn . Ischemia and systemic infection are implicated in the etiology of periventricular white matter injury , a major cause of adverse motor and cognitive outcome in preterm infants . Cytokines are signaling proteins that can be produced as part of the inflammatory response to both ischemia and infection . The aim of this study was to relate cerebrospinal fluid ( P04141 ) concentrations of P05231 , P10145 , P22301 , tumor necrosis factor alpha ( P01375 ) , and interferon gamma ( P01579 ) to magnetic resonance-defined white matter injury in preterm infants . Relationships between P04141 and plasma cytokine concentrations were also examined . Preterm infants ( < or=32 wk ) and more mature infants from The Royal Women 's Hospital , Melbourne , Australia , and Christchurch Women 's Hospital , Christchurch , New Zealand , were eligible for study if they required a clinically indicated lumbar puncture . Plasma samples were obtained in a subgroup of Christchurch infants . Preterm infants underwent advanced quantitative volumetric magnetic resonance imaging using a 1.5-Tesla scanner at term equivalent . One hundred forty-six infants were enrolled and 190 P04141 and 42 plasma samples obtained . There was no significant correlation between paired P04141 and plasma concentrations for any cytokine . In comparing plasma and P04141 concentrations , levels of P10145 were significantly higher in P04141 than plasma . Preterm infants with Q9BWK5 -defined cerebral white matter injury had higher levels of P05231 , P22301 , and P01375 in the P04141 than infants without such injury . Plasma cytokine concentrations may not reflect P04141 cytokine levels or inflammatory events within the brain . Elevated P04141 levels of cytokines in infants with white matter injury suggest an altered inflammatory balance . Breast cancer susceptibility variants alter risk in familial ovarian cancer . Recent candidate gene and genome wide association studies have revealed novel loci associated with an increased risk of breast cancer . We evaluated the effect of these breast cancer associated variants on ovarian cancer risk in individuals with familial ovarian cancer both with and without P38398 or P51587 mutations . A total of 158 unrelated white British women ( 54 P38398 /2 mutation positive and 104 P38398 /2 mutation negative ) with familial ovarian cancer were genotyped for P21802 , O15405 / O15405 and Q14790 variants . The p.Asp302His Q14790 variant was associated with reduced ovarian cancer risk in the familial P38398 /2 mutation negative ovarian cancer cases ( P = 0.016 ) . The synonymous O15405 / O15405 ( Ser51 ) variant was present at a significantly lower frequency than in patients with familial P38398 /2 positive breast cancer ( P = 0.0002 ) . Our results indicate that variants in Q14790 and O15405 / O15405 alter the risk of disease in individuals affected with familial ovarian cancer . Second-generation epidermal growth factor receptor tyrosine kinase inhibitors in lung cancers . P00533 mutations identify patients who are more likely to respond to treatment with epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors ( TKIs ) than cytotoxic chemotherapy . The distinct success of the first-generation P00533 TKIs erlotinib and gefitinib has been accompanied by the observation that acquired resistance to these treatments develops after a median of 1 year of treatment . Newer , second-generation P00533 TKIs have been developed with the intent to delay or overcome acquired resistance by the broader inhibition of kinases ( eg , P04626 and vascular endothelial growth factor receptor ) and/or altering the interactions with P00533 through irreversibly binding to the kinase domain . This article discusses many of these agents ( including afatinib , dacomitinib , DB05007 , AP26113 , and CO-1686 ) which have the potential for greater efficacy compared with first-generation P00533 TKIs , and may also have clinical activity against other oncogenic mutations within the P00533 family , including P04626 . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells .

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interacts_with DB00472?

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Pharmacological profile of astemizole-derived compounds at the histamine H1 and H4 receptor -- H1/H4 receptor selectivity . DB00637 , a P35367 antagonist shows high affinity to the histamine H1 receptor but only a moderate affinity to the histamine H4 receptor . This study aims to modify the astemizole to keep high affinity to the histamine H1 receptor and to increase affinity to the histamine H4 receptor . Therefore , 13 astemizole-derived compounds and astemizole-JNJ7777120-derived hybrid compounds were synthesized and pharmacologically characterized at the histamine H1 and H4 receptors . The new compounds show affinity to the histamine H1 receptor in the pK i range from 5.3 to 8.8 , whereas the affinity of these compounds to the histamine H4 receptor was surprisingly rather low ( pK i from 4.4 to 5.6 ) . Three representative compounds were docked into the histamine H1 receptor and molecular dynamic studies were performed to explain the binding mode and the experimental results on a molecular level . Furthermore , taking into account the binding mode of compounds with high affinity to the histamine H4 receptor , a H1/H4-pharmacophore hypothesis was developed . Effects of mitiglinide ( S 21403 ) on Kir6.2/ Q09428 , Kir6.2/SUR2A and Kir6.2/SUR2B types of DB00171 -sensitive potassium channel . 1. We have investigated the mechanism of action of the novel anti-diabetic agent mitiglinide ( S 21403 ) on Kir6.2/ Q09428 , Kir6.2/SUR2A and Kir6.2/SUR2B types of DB00171 -sensitive potassium ( K( DB00171 ) ) channel . These possess a common pore-forming subunit , Kir6.2 , and different regulatory sulphonylurea receptor ( Q09428 ) subunits . It is believed that they correspond to native K( DB00171 ) channels in pancreatic beta-cells , heart and non-vascular smooth muscle , respectively . 2 . Kir6.2 was coexpressed with Q09428 , SUR2A or SUR2B in Xenopus oocytes and macroscopic currents were recorded in giant inside-out membrane patches . DB01252 was added to the intracellular membrane surface . 3 . DB01252 inhibited Kir6.2/ Q09428 currents at two sites : a low-affinity site on Kir6.2 and a high-affinity site on Q09428 . Low-affinity inhibition was similar for all three types of K( DB00171 ) channel but high-affinity inhibition was greater for Kir6.2/ Q09428 currents ( IC(50) , 4 nM ) than for Kir6.2/SUR2A or Kir6.2/SUR2B currents ( IC(50) , 3 and 5 microM , respectively ) . 4 . Inhibition of Kir6.2/ Q09428 currents was only slowly reversible on the time scale of electrophysiological experiments . 5 . Kir6.2/ Q09428 -S1237Y currents , which previously have been shown to lack high affinity tolbutamide inhibition , resembled Kir6.2/SUR2 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 . Comparative effects of the anti-platelet drugs , clopidogrel , ticlopidine , and cilostazol on aspirin-induced gastric bleeding and damage in rats . AIMS : The present study compared the effects of frequently used anti-platelet drugs , such as clopidogrel , ticlopidine , and cilostazol , on the gastric bleeding and ulcerogenic responses induced by intraluminal perfusion with 25 mM aspirin acidified with 25 mM HCl ( acidified ASA ) in rats . MAIN METHODS : The stomach was perfused with acidified ASA at a rate of 0.4 ml/min for 60 min under urethane anesthesia , and gastric bleeding was measured as the concentration of hemoglobin in the luminal perfusate , which was collected every 15 min . DB00758 ( 10-100mg/kg ) , ticlopidine ( 10-300 mg/kg ) , or cilostazol ( 3-30 mg/kg ) was given p.o . 24h or 90 min before the perfusion of acidified ASA , respectively . KEY FINDINGS : Perfusion of the stomach with acidified ASA alone led to slight bleeding and lesions in the stomach . The pretreatment with clopidogrel , even though it did not cause bleeding or damage by itself , dose-dependently increased the gastric bleeding and ulcerogenic responses induced by acidified ASA . DB00208 also aggravated the severity of damage by increasing gastric bleeding , and the effects of ticlopidine at 300 mg/kg were equivalent to those of clopidogrel at 100mg/kg . In contrast , cilostazol dose-dependently decreased gastric bleeding and damage in response to acidified ASA . SIGNIFICANCE : These results demonstrated that clopidogrel and ticlopidine , Q9H244 receptor inhibitors , increased gastric bleeding and ulcerogenic responses to acidified ASA , to the same extent , while cilostazol , a phosphodiesterase III inhibitor , suppressed these responses . Therefore , cilostazol may be safely used in dual anti-platelet therapy combined with ASA , without increasing the risk of gastric bleeding . Lipoic acid inhibits the DNA repair protein O 6-methylguanine-DNA methyltransferase ( P16455 ) and triggers its depletion in colorectal cancer cells with concomitant autophagy induction . Alkylating agents are present in food and tobacco smoke , but are also used in cancer chemotherapy , inducing the DNA lesion O (6)-methylguanine . This critical adduct is repaired by O (6)-methylguanine-DNA methyltransferase ( P16455 ) , resulting in P16455 inactivation and degradation . In the present study , we analyzed the effects of the natural disulfide compound lipoic acid ( LA ) on P16455 in vitro and in colorectal cancer cells . We show that LA , but not its reduced form dihydrolipoic acid , potently inhibits the activity of recombinant P16455 by interfering with its catalytic DB00151 -145 residue , which was partially reversible by N-acetyl cysteine . Incubation of HCT116 colorectal cancer cells with LA altered their glutathione pool and caused a decline in P16455 activity . This was mirrored by LA-induced depletion of P16455 protein , which was not attributable to changes in P16455 messenger RNA levels . Loss of P16455 protein coincided with LA-induced autophagy , a process resulting in lysosomal degradation of proteins , including presumably P16455 . LA-stimulated autophagy in a p53-independent manner as revealed by the response of isogenic HCT116 cell lines . Knockdown of the crucial autophagy component beclin-1 and chemical inhibitors blocked LA-induced autophagy , but did not abrogate LA-triggered P16455 degradation . Concomitant with P16455 depletion , LA pretreatment resulted in enhanced O (6)-methylguanine levels in DNA . It also increased the cytotoxicity of the alkylating anticancer drug temozolomide in temozolomide-resistant colorectal cancer cells . Taken together , our study showed that the natural compound LA inhibits P16455 and induces autophagy . Furthermore , LA enhanced the cytotoxic effects of temozolomide , which makes it a candidate for a supplement in cancer therapy . Tumor marker score for prognostication of early-stage squamous cell cervical cancer . BACKGROUND/AIM : Histopathological and clinical scores to predict prognosis in cervical cancer have been of limited value . In the present study a tumor marker expression score was evaluated for prognostication in early-stage cervical cancer . MATERIALS AND METHODS : The entire study population included 128 women with invasive squamous cell cervical cancer followed-up for at least 10 years . RESULTS : Expression of 12 tumor markers ( epidermal growth factor receptor ( P00533 ) , Ki-67 , c-MYC , p53 , p27 , P12830 , P16070 , vascular endothelial growth factor receptor ( P15692 ) , cyclooxygenase-2 ( P35354 ) , P01730 , and leucine-rich immunoglobulin-like repeats-1 ( Q96JA1 ) and O94898 , considered relevant for cervical cancer prognostication was evaluated by immunohistochemistry . Expression of five markers , Q96JA1 , O94898 , p53 , P35354 and c-MYC were useful to make a prognostication score , ranging from 0 to 5 . Score 0-1 correlated to less than 5 % 10-year mortality , while the mortality rate of those with score 4-5 approached 70 % ; those with score 2 formed an intermediate group . Using different models , a high sensitivity , specificity , positive predictive value and negative predictive value was attained . CONCLUSION : Tumor marker scoring could be an adjunct to histopathological and clinical parameters in prognostication of early-stage cervical cancer . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . DB08818 Synthesis , Catabolism , and Signaling in Neurodegenerative Diseases . The glycosaminoglycan hyaluronan ( HA ) , a component of the extracellular matrix , has been implicated in regulating neural differentiation , survival , proliferation , migration , and cell signaling in the mammalian central nervous system ( CNS ) . HA is found throughout the CNS as a constituent of proteoglycans , especially within perineuronal nets that have been implicated in regulating neuronal activity . HA is also found in the white matter where it is diffusely distributed around astrocytes and oligodendrocytes . Insults to the CNS lead to long-term elevation of HA within damaged tissues , which is linked at least in part to increased transcription of HA synthases . HA accumulation is often accompanied by elevated expression of at least some transmembrane HA receptors including P16070 . Hyaluronidases that digest high molecular weight HA into smaller fragments are also elevated following CNS insults and can generate HA digestion products that have unique biological activities . A number of studies , for example , suggest that both the removal of high molecular weight HA and the accumulation of hyaluronidase-generated HA digestion products can impact CNS injuries through mechanisms that include the regulation of progenitor cell differentiation and proliferation . These studies , reviewed here , suggest that targeting HA synthesis , catabolism , and signaling are all potential strategies to promote CNS repair . P40933 affects serotonin system and exerts antidepressive effects through IL15Rα receptor . Contrary to the reduction of depressive-like behavior observed in several strains of cytokine receptor knockout mice , mice lacking the specific receptor for interleukin ( IL ) -15 showed increased immobility in tail suspension and modified forced swimming tests . There was also a reduction in social interactions . The hippocampus of the IL15Rα knockout mice had decreased mRNA for 5-HT(1A) , increased mRNA for 5-HT(2C) , and region-specific changes of serotonin reuptake transporter ( P31645 ) immunoreactivity . DB00472 ( the classic antidepressant DB00472 , which inhibits 5-HT(2C) and P31645 ) reduced the immobility of the IL15Rα knockout mice in comparison with their pretreatment baseline . Together with the unchanged performance of the IL15Rα knockout mice on the rotarod , this response to fluoxetine indicates that the immobility reflects depression . Wildtype mice responded to P40933 treatment with improvement of immobility induced by forced swimming , whereas the knockout mice failed to respond . Thus , the cognate P40933 receptor is necessary for the antidepressive activity of P40933 . In ex vivo studies , P40933 decreased synaptosomal uptake of 5-HT , and modulated the expression of 5-HT(2C) and P31645 in cultured neurons in a dose- and time-dependent manner . Thus , the effect of P40933 on serotonin transmission may underlie the depressive-like behavior of IL15Rα knockout mice . We speculate that P40933 is essential to maintain neurochemical homeostasis and thereby plays a role in preventing neuropsychiatric symptoms . Genetics of antipsychotic-induced weight gain : update and current perspectives . Antipsychotic medications are used to effectively treat various symptoms for different psychiatric conditions . Unfortunately , antipsychotic-induced weight gain ( AIWG ) is a common side effect that frequently results in obesity and secondary medical conditions . Twin and sibling studies have indicated that genetic factors are likely to be highly involved in AIWG . Over recent years , there has been considerable progress in this area , with several consistently replicated findings , as well as the identification of new genes and implicated pathways . Here , we will review the most recent genetic studies related to AIWG using the Medline database ( PubMed ) and Google Scholar . Among the steadiest findings associated with AIWG are serotonin 2C receptors ( P28335 ) and leptin promoter gene variants , with more recent studies implicating P42898 and , in particular , P32245 genes . Additional support was reported for the P35367 , P23560 , P01303 , P21554 , Q9UBU3 , Q9C0B1 and AMPK genes . Notably , some of the reported variants appear to have relatively large effect sizes . These findings have provided insights into the mechanisms involved in AIWG and will help to develop predictive genetic tests in the near future . Q07869 agonists for the treatment of cardiovascular disease in patients with diabetes . Diabetes is a complex disease defined by hyperglycaemia ; however , strong associations with abdominal obesity , hypertension and dyslipidaemia contribute to the high risk of cardiovascular disease . Although aggressive glycaemic control reduces microvascular complications , the evidence for macrovascular complications is less certain . The theoretical benefits of the mode of action of peroxisome proliferator-activated receptor ( Q07869 ) agonists are clear . In clinical practice , Q07869 -α agonists such as fibrates improve dyslipidaemia , while Q07869 -γ agonists such as thiazolidinediones improve insulin resistance and diabetes control . However , although these agents are traditionally classed according to their target , they have different and sometimes conflicting clinical benefit and adverse event profiles . It is speculated that this is because of differing properties and specificities for the Q07869 receptors ( each of which targets specific genes ) . This is most obvious in the impact on cardiovascular outcomes -- in clinical trials pioglitazone appeared to reduce cardiovascular events , whereas rosiglitazone potentially increased the risk of myocardial infarction . The development of a dual Q07869 -α/γ agonist may prove beneficial in effectively managing glycaemic control and improving dyslipidaemia in patients with type 2 diabetes . Yet , development of agents such as muraglitazar and tesaglitazar has been hindered by various serious adverse events . DB08915 , a balanced dual Q07869 -α/γ agonist , is currently the most advanced in clinical development and has shown promising results in phase II clinical trials with beneficial effects on glucose and lipid variables . A phase III study , ALECARDIO , is ongoing and will establish whether improvements in laboratory test profiles translate into an improvement in cardiovascular outcomes . Phase I evaluation of DB05243 , an oral , potent , and selective O60674 inhibitor . This phase I study evaluated selective O60674 inhibitor DB05243 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 was approximately 21 h , with steady state reached by Day 8 . International Working Group defined responses were seen in three ( 10 % ) patients . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Specific Biomarkers Are Associated with Docetaxeland Gemcitabine-Resistant NSCLC Cell Lines . Five-year survival rate for lung cancer is limited to 10 % to 15 % . Therefore , the identification of novel therapeutic prognostic factors is an urgent requirement . The aim of this study is thus to highlight specific biomarkers in chemoresistant non-small cell lung cancer cell lines . Therefore , we checked-in the control condition as well as after short-term pharmacological treatment with either docetaxel or gemcitabine-the expression of genes such as tumor suppressor genes ( CDKN2A , P53355 , P49789 , P09211 , P16455 , RARβ2 , RASSF1A , and P35625 ) , genes associated with drug resistance ( P38398 , P35354 , P07992 , P17936 , P23921 , and Q13509 ) , and stemness-related genes ( CD133 , Q01860 , and O43623 ) in two cellular models of squamous carcinoma ( CAEP ) and adenocarcinoma ( RAL ) of the lung originally established . Their promoter methylation profile was also evaluated . Drug-related genes were upregulated . DB00515 resistance matched with high levels of P38398 and P07992 in both cell lines ; docetaxel sensitivity of CAEP cells was associated to levels of Q13509 lower than RAL cells . Although CAEP cells were more sensitive to gemcitabine , both cell lines showed high levels of P23921 . Stemness-related genes were downregulated in the control condition but became upregulated in docetaxel-resistant cells , indicating the selection of a population with stemness features . We did not find an unequivocal correspondence between gene expression and respective DNA promoter methylation status , suggesting the involvement of additional mechanisms of gene expression regulation . These results highlight specific biomarkers consistent with the different responses of the two cell lines to standard pharmacological treatments and indicate specific molecular traits for their chemoresistance . Q13261 drives antagonistic mechanisms of cancer development and immune control in lymphocyte-enriched triple-negative breast cancers . Despite its aggressive nature , triple-negative breast cancer ( TNBC ) often exhibits leucocyte infiltrations that correlate with favorable prognosis . In this study , we offer an explanation for this apparent conundrum by defining TNBC cell subsets that overexpress the P40933 immune receptor Q13261 . This receptor usually forms a heterotrimer with the P60568 receptors P14784 and P31785 , which regulates the proliferation and differentiation of cytotoxic T cells and NK cells . However , unlike Q13261 , the P14784 and P31785 receptors are not upregulated in basal-like TNBC breast cancer cells that express Q13261 . Mechanistic investigations indicated that Q13261 signaling activated P23458 , P42224 , P52630 , AKT , Q96B36 , and P27361 /2 in the absence of P14784 and P31785 , whereas neither P42229 nor O60674 were activated . RNAi-mediated attenuation of Q13261 established its role in cell growth , apoptosis , and migration , whereas expression of the P40933 cytokine in Q13261 -expressing cells stimulated an autocrine signaling cascade that promoted cell proliferation and migration and blocked apoptosis . Notably , coexpression of Q13261 and P40933 was also sufficient to activate peripheral blood mononuclear cells upon coculture in a paracrine signaling manner . Overall , our findings offer a mechanistic explanation for the paradoxical association of some high-grade breast tumors with better survival outcomes , due to engagement of the immune stroma . DB01373 -induced P29323 activation in human T lymphocytes occurs via p56(Lck) and P62158 -kinase . We previously demonstrated that stimulation of human T-lymphocytes with calcium ionophores induced the phosphorylation and enzymatic activation of P28482 . We now report on the mechanism by which calcium-ionophore-induced activation of P27361 and 2 occurs in these cells . The activation of P27361 and 2 by increases in intracellular calcium was inhibited by calmidazolium suggesting the involvement of calmodulin in this response . To further elucidate the mechanism by which calcium-induced P29323 activation occurs , we used the P62158 -kinase inhibitor KN-93 and an inactive analog of KN-93 ( KN-92 ) . KN-93 , but not KN-92 , blocked ionomycin-induced activation of P27361 and 2 in human T lymphocytes . We previously demonstrated that stimulation of T lymphocytes with ionomycin or A23187 resulted in a P62158 -kinase-dependent shift in the mobility of p56(Lck) . To determine if p56(Lck) was involved in calcium-induced P29323 activation , we stimulated the p56(Lck) negative Jurkat cell derivatives , J.CaM1.6 and J.CaM1/Rep3 , with ionomycin . In these p56(Lck) negative cell lines , activation of P27361 and 2 in response to ionomycin was only minimally detected . When J.CaM1 cells were reconstituted with p56(Lck) , ionomycin induced P27361 and 2 activation . Treatment of Jurkat cells with Q99463 , an inhibitor of p56(Lck) , inhibited calcium-induced , but not PMA-induced , P27361 and 2 activation . Treatment of Jurkat cells with the MEK inhibitor PD98059 blocked ionomycin-induced P29323 activation , but not the shift in the mobility of p56(Lck) . Our data suggests that increases in intracellular calcium induce the activation of P27361 and 2 in human T lymphocytes via sequential activation of P62158 -kinase and phosphorylation of p56(Lck) . DB00472 -induced proliferation and differentiation of neural progenitor cells isolated from rat postnatal cerebellum . Previous studies have shown that the serotonin-reuptake inhibitor ( SSRI ) fluoxetine affects neural progenitors derived from postnatal cerebellum or adult hippocampus and stimulates their proliferation . In the human cerebellum , the proliferation of cerebellar granule cells ( CGC ) continues until the 11th postnatal month and could be influenced in infants by breastfeeding-delivered SSRIs . Current information about fluoxetine effects on postnatal cerebellar neural progenitors is limited . Here we report the characterization of fluoxetine actions on rat postnatal cerebellar neural progenitors . RT-PCR and immunostaining revealed the expression of serotonin transporter ( P31645 ) , 5HT(1A) receptors , tryptophan hydroxylase ( P17752 ) , and serotonin ( 5HT ) . Protracted in vitro fluoxetine treatment increased cell proliferation and differentiation . The proliferative effects of fluoxetine , 5HT , and the selective agonist of 5HT(1A) receptors trans-8-hydroxy-2-(N-n-propyl-N-3'-iodo-2'-propenyl)aminotetralin ( 8-OH-PIPAT ) were abolished by the selective antagonist of 5HT(1A) receptors , N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohexanecarboxamide trihydrochloride ( WAY-100635 ) . Furthermore , fluoxetine-induced activation of both the DB02527 -response element-binding ( CREB ) protein and extracellular signal-regulated protein kinases ( P27361 /2 ) , which was abolished by the selective inhibitor of Q96HU1 kinase kinase ( MEK ) 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene ( U0126 ) , and increased cyclin D1 expression . All these effects were prevented by WAY-100635 . Collectively , our results demonstrate that rat postnatal cerebellum contains neural progenitors capable of proliferating and differentiating in response to fluoxetine exposure , possibly through the activation of 5HT(1A) receptors . The relevance of these findings for possible SSRI effects on the developing postnatal/infant human cerebellum should be explored . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . DB01892 is a dual inhibitor of cyclooxygenase-1 and P09917 . The acylphloroglucinol derivative hyperforin is the major lipophilic constituent in the herb Hypericum perforatum ( St. John 's wort ) . The aim of the present study was to investigate if hyperforin as well as extracts of H. perforatum can suppresses the activities of P09917 ( P09917 ) and cyclooxygenases ( P36551 ) , key enzymes in the formation of proinflammatory eicosanoids from arachidonic acid ( AA ) . In freshly isolated human polymorphonuclear leukocytes stimulated with Ca(2+) ionophore A23187 , hyperforin inhibited P09917 product formation with IC(50) values of about 1-2 microM , in the absence or presence of exogenous AA ( 20 microM ) , respectively , being almost equipotent to the well-documented P09917 inhibitor zileuton ( IC(50) = 0.5-1 microM ) . Experiments with purified human P09917 demonstrate that hyperforin is a direct P09917 inhibitor ( IC(50) approximately 90 nM ) , acting in an uncompetitive fashion . In thrombin- or ionophore-stimulated human platelets , hyperforin suppressed P23219 product ( 12(S)-hydroxyheptadecatrienoic acid ) formation with an IC(50) of 0.3 and 3 microM , respectively , being about 3- to 18-fold more potent than aspirin . At similar concentrations , hyperforin suppressed P23219 activity in platelets in presence of exogenous AA ( 20 microM ) as well as in cell-free systems . DB01892 could not interfere with P35354 product formation and did not significantly inhibit 12- or 15-LO in platelets or leukocytes , respectively . We conclude that hyperforin acts as a dual inhibitor of P09917 and P23219 in intact cells as well as on the catalytic activity of the crude enzymes , suggesting therapeutic potential in inflammatory and allergic diseases connected to eicosanoids .

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A vitamin A deficient diet enhances proinflammatory cytokine , P35372 , and HIV-1 expression in the HIV-1 transgenic rat . The HIV-1 ( HIV ) transgenic ( Tg ) rat develops several immune abnormalities in association with clinical impairments that are similar to what are seen with HIV infection in humans . In HIV infection , retinoids and opioids can have separate and potentially combined effects on the clinical course of HIV disease . In these studies , the effects of a vitamin A deficient diet on T cell proinflammatory cytokine and mu opioid receptor ( MOR ) expression were examined in the Tg and in wild-type ( WT ) rats . The effects of the diet on HIV gene expression were also analyzed in the Tg rats . Phytohemagglutinin-stimulated T cells from WT rats on the vitamin A diet and from Tg rats on either diet were more likely to either produce increased percentages of T cells expressing intracytoplasmic P01579 , secrete higher levels of P01375 , and express higher levels of MOR mRNA and surface MOR . Mitogen stimulation also increased Tg rat HIV env , tat , and nef mRNA expression with even higher env and nef mRNA produced in association with the vitamin A deficient diet . All together , these data suggest that a vitamin A deficient diet can result in cellular effects that increase T cell proinflammatory responses and HIV expression , which may alter the course of disease in the HIV Tg rat model . Development of drugs to target interactions between leukocytes and endothelial cells and treatment algorithms for inflammatory bowel diseases . Increased understanding of the pathogenesis of inflammatory bowel diseases ( IBDs ) has led to new therapeutic strategies . One of these is to target the molecules that regulate interactions between leukocytes and endothelial cells at sites of inflammation ( mainly leukocyte integrins and endothelial cell adhesion molecules of the immunoglobulin superfamily ) . These molecules have been validated as therapeutic targets for Q9UKU7 ; several have shown efficacy , and 2 have been approved by the Food and Drug Administration for treatment of Q9UKU7 . DB00108 , the first anti-integrin antibody tested for treatment of Q9UKU7 , blocks the α4 subunit . Although it is effective , its clinical use has been limited by its association with risk of progressive multifocal leukoencephalopathy . Other , allegedly more selective drugs that affect leukocyte recruitment in the gastrointestinal tract have been developed or are under investigation and could increase safety . These include vedolizumab and Q99217 181 ( antibodies against α4β7 ) , etrolizumab ( anti-β7 ) , and PF-00547659 ( anti-mucosal vascular addressin cell adhesion molecule 1 ) . Other agents have been developed to block α4 ( the small molecule AJM300 ) , P51686 ( the small molecule DB05005 -B ) , and P02778 ( the antibody eldelumab ) . We review the scientific rationale for inhibiting interactions between leukocytes and endothelial cells to reduce intestinal inflammation and analyze the clinical studies that have been performed to test these new molecules , with particular attention to safety . We propose an evidence-based clinical positioning of this class of drugs . Genes involved in carnitine synthesis and carnitine uptake are up-regulated in the liver of sows during lactation . BACKGROUND : Convincing evidence exist that carnitine synthesis and uptake of carnitine into cells is regulated by peroxisome proliferator-activated receptor α ( Q07869 ) , a transcription factor which is physiologically activated during fasting or energy deprivation . Sows are typically in a negative energy balance during peak lactation . We investigated the hypothesis that genes involved in carnitine synthesis and uptake in the liver of sows are up-regulated during peak lactation . FINDINGS : Transcript levels of several PPARα target genes involved in fatty acid uptake ( P15090 , O43772 ) , fatty acid oxidation ( Q15067 , CYP4A24 ) and ketogenesis ( P54868 , Q9NSA1 ) were elevated in the liver of lactating compared to non-lactating sows ( P < 0.05 ) . In addition , transcript levels of genes involved in carnitine synthesis ( P49189 , Q9NVH6 , O75936 ) and carnitine uptake ( O76082 ) in the liver were greater in lactating than in non-lactating sows ( P < 0.05 ) . DB00583 concentrations in liver and plasma were about 20 % and 50 % , respectively , lower in lactating than in non-lactating sows ( P < 0.05 ) , which is likely due to an increased loss of carnitine via the milk . CONCLUSIONS : The results of the present study show that PPARα is activated in the liver of sows during lactation which leads to an up-regulation of genes involved in carnitine synthesis and carnitine uptake . The PPARα mediated up-regulation of genes involved in carnitine synthesis and uptake in the liver of lactating sows may be regarded as an adaptive mechanism to maintain hepatic carnitine levels at a level sufficient to transport excessive amounts of fatty acids into the mitochondrion . Glial response to lipopolysaccharide : possible role of endothelins . Glial inflammation plays a major role in the development of neurodegenerative diseases . Although endothelins ( ETs ) are known as modulators of inflammation in the periphery , little is known about their possible role in brain inflammation . Previously , we demonstrated that all three endothelins ( ET-1 , P20800 and P14138 ) enhanced unstimulated synthesis of the glial pro-inflammatory mediators , prostaglandin E₂ ( PGE₂ ) and nitric oxide ( NO ) . In the present study , glial cells were stimulated in an in vitro model of inflammation by incubation with the bacterial endotoxin lipopolysaccharide ( LPS ) . Indeed , the present study shows that ETs regulate basal and LPS-induced glial inflammation in an opposite fashion . Here we demonstrate that ETs significantly inhibited the LPS-induced glial synthesis of PGE₂ and NO , and each of the selective antagonists for P25101 and ETB receptors ( BQ123 and BQ788 respectively ) , significantly inhibited the ETs effects in LPS-treated cells . Similar results were observed when expression of key enzymes namely , cyclooxygenase-2 ( P35354 ) and inducible nitric oxide synthase ( P35228 ) in PG and NO synthesis respectively , was measured . ET-1 significantly enhanced the expression of both P35354 and P35228 . Whereas , it inhibited the LPS-induced expression of both enzymes . These observations suggest a novel neuro-immune feedback pathway through which inflammatory mediators ' synthesis is initially enhanced by ETs and are eventually blocked by the same neuropeptide when excessive production of inflammatory mediators occurs following an inflammatory insult . Pomegranate flower extract diminishes cardiac fibrosis in Zucker diabetic fatty rats : modulation of cardiac endothelin-1 and nuclear factor-kappaB pathways . The diabetic heart shows increased fibrosis , which impairs cardiac function . Endothelin ( ET ) -1 and nuclear factor-kappaB ( NF-kappaB ) interactively regulate fibroblast growth . We have recently demonstrated that Punica granatum flower ( P49763 ) , a Unani anti-diabetic medicine , is a dual activator of peroxisome proliferator-activated receptor ( Q07869 ) -alpha and -gamma , and improves hyperglycemia , hyperlipidemia , and fatty heart in Zucker diabetic fatty ( ZDF ) rat , a genetic animal model of type 2 diabetes and obesity . Here , we demonstrated that six-week treatment with P49763 extract ( 500 mg/kg , p.o. ) in Zucker diabetic fatty rats reduced the ratios of van Gieson-stained interstitial collagen deposit area to total left ventricular area and perivascular collagen deposit areas to coronary artery media area in the heart . This was accompanied by suppression of overexpressed cardiac fibronectin and collagen I and III mRNAs . Punica granatum flower extract reduced the up-regulated cardiac mRNA expression of ET-1 , P25101 , inhibitor-kappaBbeta and c-jun , and normalized the down-regulated mRNA expression of inhibitor-kappaBalpha in Zucker diabetic fatty rats . In vitro , Punica granatum flower extract and its components oleanolic acid , ursolic acid , and gallic acid inhibited lipopolysaccharide-induced NF-kappaB activation in macrophages . Our findings indicate that Punica granatum flower extract diminishes cardiac fibrosis in Zucker diabetic fatty rats , at least in part , by modulating cardiac ET-1 and NF-kappaB signaling . DB00559 , an endothelin receptor antagonist , ameliorates collagen-induced arthritis : the role of P01375 -α in the induction of endothelin system genes . OBJECTIVE : Endothelins ( ETs ) are involved in several inflammatory events . The present study investigated the efficacy of DB00559 , a dual P25101 /ETB receptor antagonist , in collagen-induced arthritis ( CIA ) in mice . TREATMENT : CIA was induced in DBA/1J mice . Arthritic mice were treated with DB00559 ( 100 mg/kg ) once a day , starting from the day when arthritis was clinically detectable . METHODS : CIA progression was assessed by measurements of visual clinical score , paw swelling and hypernociception . Histological changes , neutrophil infiltration and pro-inflammatory cytokines were evaluated in the joints . Gene expression in the lymph nodes of arthritic mice was evaluated by microarray technology . PreproET-1 mRNA expression in the lymph nodes of mice and in peripheral blood mononuclear cells ( PBMCs ) was evaluated by real-time PCR . The differences were evaluated by one-way Q9UNW9 or Student 's t test . RESULTS : Oral treatment with DB00559 markedly ameliorated the clinical aspects of CIA ( visual clinical score , paw swelling and hyperalgesia ) . DB00559 treatment also reduced joint damage , leukocyte infiltration and pro-inflammatory cytokine levels ( IL-1β , TNFα and Q16552 ) in the joint tissues . Changes in gene expression in the lymph nodes of arthritic mice returned to the levels of the control mice after DB00559 treatment . PreproET mRNA expression increased in PBMCs from rheumatoid arthritis ( RA ) patients but returned to basal level in PBMCs from patients under anti- P01375 therapy . In-vitro treatment of PBMCs with TNFα upregulated ET system genes . CONCLUSION : These findings indicate that ET receptor antagonists , such as DB00559 , might be useful in controlling RA . Moreover , it seems that ET mediation of arthritis is triggered by TNFα . Essential role of p38 mitogen-activated protein kinase in contact hypersensitivity . The present study was designed to elucidate the role of p38 mitogen-activated protein kinase ( p38 ) in the pathogenesis of inflammation , using a mouse contact hypersensitivity ( Q99698 ) model induced by 2,4-dinitro-1-fluorobenzene ( DNFB ) . Ear swelling was induced by challenge with DNFB , accompanied by infiltration of mononuclear cells , neutrophils , and eosinophils and a marked increase in mRNA levels of cytokines such as interleukin ( IL ) -2 , interferon ( IFN ) -gamma , P05112 , P05113 , IL-1beta , Q14116 , and tumor necrosis factor-alpha in the challenged ear skin . Both ear swelling and the number of infiltrated cells in DNFB-challenged ear skin were significantly inhibited by treatment with SB202190 , a p38 inhibitor . Furthermore , the DNFB-induced expression of all cytokines except P05112 was significantly inhibited by treatment with SB202190 . Ribonuclease protection assay revealed that the mRNA levels of chemokines such as P02778 and P13500 in ear skin were markedly increased at 24 h after challenge with DNFB . The induction of these chemokines was significantly inhibited by treatment with SB202190 . In p38alpha +/- mice , both ear swelling and infiltration of cells induced by DNFB were reduced compared with those in wild-type mice . However , induction of cytokines by DNFB was also observed in p38alpha +/- mice , although the induction of P01579 , P05113 , and Q14116 was typically reduced compared with that in wild-type mice . Challenge with DNFB slightly induced P02778 and P13500 mRNA in p38alpha +/- mice , with weaker signals than those in SB202190-treated wild-type mice . These results suggest that p38 plays a key role in Q99698 and is an important target for the treatment of Q99698 . Dual inhibition of P09917 and cyclooxygenases 1 and 2 by DB04725 reduces joint destruction in adjuvant arthritis . OBJECTIVE : To search for potential new therapies to inhibit the progression of joint destruction in patients with rheumatoid arthritis . METHODS : We evaluated the dual acting antiinflammatory drug DB04725 ( 2,2-dimethyl-6-(4-chlorophenyl)-7-phenyl-2,3-dihydro- H-pyrrolizine-5-yl ) acetic acid , a dual inhibitor of P09917 ( 5- P28300 ) as well as both cyclooxygenases ( P23219 and P35354 ) in the rat model of adjuvant arthritis . On Day 0 , female Lewis rats ( 5 per group ) were injected intradermally with complete Freund 's adjuvant at base of the tail . Treatment began on Day 2 ; the rats received DB04725 ( 20 or 80 mg/kg/day ) twice daily 7 h apart for 28 days and were then sacrificed . To reduce pain , the positive control group and 2 treatment groups received paracetamol ( 3 mg/ml water ) . Joint histology was scored for synovial cell proliferation , fibroproliferative pannus , and cartilage and bone erosions , as well as diffuse leukocyte infiltrates . RESULTS : Daily doses of 20 or 80 mg/kg DB04725 significantly reduced the arthritis associated deficiency of body growth , the edema/erythema score , and splenomegaly . In the ankle joint , DB04725 significantly reduced the overall histological score , synovial cell proliferation , and bone/cartilage erosions , and inhibited the appearance of fibroproliferative pannus . The addition of paracetamol in the drinking water had no influence . No side effects were noted . CONCLUSION : DB04725 is an antiarthritic drug with a high gastrointestinal tolerability , which can reduce synovial cell proliferation and joint erosion and is capable of markedly suppressing prostaglandin synthesis . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . A conserved mechanism for gating in an ionotropic glutamate receptor . Ionotropic glutamate receptor ( iGluR ) channels control synaptic activity . The crystallographic structure of P42262 , the prototypical iGluR , reveals a clamshell-like ligand-binding domain ( LBD ) that closes in the presence of glutamate to open a gate on the pore lining α-helix . How LBD closure leads to gate opening remains unclear . Here , we show that bending the pore helix at a highly conserved alanine residue ( Ala-621 ) below the gate is responsible for channel opening . Substituting Ala-621 with the smaller more flexible glycine resulted in a basally active , nondesensitizing channel with ∼39-fold increase in glutamate potency without affecting surface expression or binding . On P42262 (A621G) , the partial agonist kainate showed efficacy similar to a full agonist , and competitive antagonists CNQX and DB03759 acted as a partial agonists . DB00134 -629 in P42262 sits above the gate and is critical in transmitting LBD closure to the gate . Substituting DB00134 -629 with the flexible glycine resulted in reduced channel activity and glutamate potency . The pore regions in potassium channels are structurally similar to iGluRs . Whereas potassium channels typically use glycines as a hinge for gating , iGluRs use the less flexible alanine as a hinge at a similar position to maintain low basal activity allowing for ligand-mediated gating . A whole genome Bayesian scan for adaptive genetic divergence in West African cattle . BACKGROUND : The recent settlement of cattle in West Africa after several waves of migration from remote centres of domestication has imposed dramatic changes in their environmental conditions , in particular through exposure to new pathogens . West African cattle populations thus represent an appealing model to unravel the genome response to adaptation to tropical conditions . The purpose of this study was to identify footprints of adaptive selection at the whole genome level in a newly collected data set comprising 36,320 SNPs genotyped in 9 West African cattle populations . RESULTS : After a detailed analysis of population structure , we performed a scan for SNP differentiation via a previously proposed Bayesian procedure including extensions to improve the detection of loci under selection . Based on these results we identified 53 genomic regions and 42 strong candidate genes . Their physiological functions were mainly related to immune response ( MHC region which was found under strong balancing selection , P11912 , P61073 , P80370 , P48380 , Q9H3S1 , Q8IUC6 and P19474 ) , nervous system ( Q96NK8 , O95897 , MAGI1 , Q9H3S1 and Q13639 ) and skin and hair properties ( P24530 , TRSP1 and Q8IUC2 ) . CONCLUSION : The main possible underlying selective pressures may be related to climatic conditions but also to the host response to pathogens such as Trypanosoma(sp) . Overall , these results might open the way towards the identification of important variants involved in adaptation to tropical conditions and in particular to resistance to tropical infectious diseases . DB00158 and thiamine transporters mediated by facilitative carriers ( P41440 -3 and Q96NT5 ) and folate receptors . The reduced folate carrier ( P41440 , P41440 ) , thiamine transporter-1 ( O60779 , O60779 ) and thiamine transporter-2 ( Q9BZV2 , Q9BZV2 ) evolved from the same family of solute carriers . P41440 transports folates but not thiamine . O60779 and Q9BZV2 transport thiamine but not folates . P41440 and O60779 deliver their substrates to systemic tissues ; Q9BZV2 mediates intestinal thiamine absorption . The proton-coupled folate transporter ( Q96NT5 , Q96NT5 ) is the mechanism by which folates are absorbed across the apical-brush-border membrane of the proximal small intestine . Two folate receptors ( P15328 and P14207 ) mediate folate transport across epithelia by an endocytic process . DB00158 transporters are routes of delivery of drugs for the treatment of cancer and inflammatory diseases . There are autosomal recessive disorders associated with mutations in genes encoded for Q96NT5 ( hereditary folate malabsorption ) , P15328 ( cerebral folate deficiency ) , O60779 ( thiamine-responsive megaloblastic anemia ) , and Q9BZV2 ( biotin-responsive basal ganglia disease ) . A gene expression signature of invasive potential in metastatic melanoma cells . BACKGROUND : We are investigating the molecular basis of melanoma by defining genomic characteristics that correlate with tumour phenotype in a novel panel of metastatic melanoma cell lines . The aim of this study is to identify new prognostic markers and therapeutic targets that might aid clinical cancer diagnosis and management . PRINCIPAL FINDINGS : Global transcript profiling identified a signature featuring decreased expression of developmental and lineage specification genes including O75030 , P24530 , P40126 , and P14679 , and increased expression of genes involved in interaction with the extracellular environment , such as Q03405 , P13611 , and HIF1a . Migration assays showed that the gene signature correlated with the invasive potential of the cell lines , and external validation by using publicly available data indicated that tumours with the invasive gene signature were less melanocytic and may be more aggressive . The invasion signature could be detected in both primary and metastatic tumours suggesting that gene expression conferring increased invasive potential in melanoma may occur independently of tumour stage . CONCLUSIONS : Our data supports the hypothesis that differential developmental gene expression may drive invasive potential in metastatic melanoma , and that melanoma heterogeneity may be explained by the differing capacity of melanoma cells to both withstand decreased expression of lineage specification genes and to respond to the tumour microenvironment . The invasion signature may provide new possibilities for predicting which primary tumours are more likely to metastasize , and which metastatic tumours might show a more aggressive clinical course . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Inhibition of the invasion capacity of carcinoma cells by DB05476 , a novel synthetic inhibitor of the urokinase-type plasminogen activator system . The overall survival rate of patients suffering from carcinomas has remained poor and nearly unchanged over the last decades . This is mainly due to the so-called minimal residual disease , i.e. , remaining tumor cells that overcome surgery and/or radiotherapy and are the cause of locoregional and distant metastases . To metastasize , tumor cells take advantage of proteases to invade and remodel surrounding tissues . Here , we analyzed the efficiency of DB05476 , a novel 3-amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 expression was characterized in different carcinoma cell lines , including SCCHN , breast and cervical carcinoma . Thereafter , the invasive potential of these cell lines was determined using Matrigel invasion chambers and a spheroid cocultivation model with human fibroblasts . Q03405 expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 . A decrease of tumor cell invasion by up to 50 % was achieved in both models with the SCCHN line FaDu and the cervical carcinoma line HeLa after treatment with DB05476 . Thus , our results demonstrate the potential of DB05476 in vitro as a promising adjuvant antimetastatic therapy of carcinomas . Possible role of cholecystokinin-A receptors in regulation of thyrotropin ( DB00024 ) secretion in male rats . We studied the importance of cholecystokinin ( CCK ) system in the regulation of thyrotropin ( DB00024 ) and prolactin ( PRL ) secretion in male rats . To this end , we tested the effects of both unselective CCK agonists CCK-8 and caerulein , and CCK-B selective agonists Q13308 and pentagastrin as well as the selective CCK antagonists ( devazepide and L-365,260 ) at wide dose-ranges on the cold-stimulated and TRH-induced DB00024 and PRL secretion . DB00403 , given s.c . 15 min before sacrifice , decreased DB00024 levels at 5 micrograms/kg . In time course-studies , the maximum inhibition was seen at 15 min but the effect lasted at least 30 , but less than 60 min . Also CCK-8 decreased DB00024 levels at the doses of 20 and 50 micrograms/kg at 15 min . Devazepide and L-365,260 did not affect DB00024 or PRL levels at any dose . The effect of caerulein ( 5 micrograms/kg ) was antagonized by devazepide , a CCK-A antagonist , at 100 micrograms/kg , but not by a CCK-B antagonist L-365,260 tested at a wide dose range . PRL levels were not affected by any treatment . DB00403 ( 5 micrograms/kg ) , given at the same time as TRH ( 500 ng/kg ) , inhibited the TRH-induced DB00024 levels at 15 min , but not at 30 or 60 min . CCK-8 ( 50 micrograms/kg ) , Q13308 ( 100 micrograms/kg ) and pentagastrin ( 500 micrograms/kg ) did not affect the TRH-induced DB00024 secretion . The results probably indicate that P32238 stimulation inhibits DB00024 secretion at the level of the anterior pituitary gland . PRL levels in male rats are not affected by CCK system . [ Localization of the NotI clones from human chromosome 3 on quail microchromosomes ] . For the purpose of comparative mapping of quail ( Coturnix c. japonica ) and human ( Homo sapiens ) genomes , DNA fragments from human chromosome 3 ( HSA3p14-21 and HSA3q13-23 ) were localized on quail mitotic chromosomes . Using the method of double-color fluorescence DNA-DNA in situ hybridization , these fragments were mapped to two different microchromosomes . Earlier , similar studies were performed using chicken mitotic chromosomes . There it was demonstrated that the clones of interest were distributed among three microchromosomes ( GGA12 , GGA14 , and GGA15 ) . Thus , interspecific difference in the location of human chromosome 3 DNA fragments in the genomes of closely related avian species was discovered . A new confirmation of the hypothesis on the preferable localization of the gene-rich human chromosome regions on avian microchromosomes was obtained . At the same time , a suggestion on the localization of some orthologous genes in the genome of the organism under study was made : P18085 , Q14524 , Q9BWX1 , Q9BV23 , Q9NYG2 , Q16644 , ADSYNA ( homolog of chicken chromosome 12 ) , P14416 , PP2C- P25101 , P51149 , P32238 , and PKD1 ( homolog of chicken chromosome 15 ) . However , localization of the corresponding quail genes needs to be confirmed , as far as the sequences used were only the orthologs of the corresponding chicken genes . P01375 alpha mediates GABA(A) receptor trafficking to the plasma membrane of spinal cord neurons in vivo . The proinflammatory cytokine TNFα contributes to cell death in central nervous system ( CNS ) disorders by altering synaptic neurotransmission . TNFα contributes to excitotoxicity by increasing P42262 -lacking AMPA receptor ( AMPAR ) trafficking to the neuronal plasma membrane . In vitro , increased AMPAR on the neuronal surface after TNFα exposure is associated with a rapid internalization of GABA(A) receptors ( GABA(A)Rs ) , suggesting complex timing and dose dependency of the CNS 's response to TNFα . However , the effect of TNFα on GABA(A)R trafficking in vivo remains unclear . We assessed the effect of TNFα nanoinjection on rapid GABA(A)R changes in rats ( N = 30 ) using subcellular fractionation , quantitative western blotting , and confocal microscopy . GABA(A)R protein levels in membrane fractions of TNFα and vehicle-treated subjects were not significantly different by Western Blot , yet high-resolution quantitative confocal imaging revealed that TNFα induces GABA(A)R trafficking to synapses in a dose-dependent manner by 60 min . TNFα-mediated GABA(A)R trafficking represents a novel target for CNS excitotoxicity . DB00158 uptake by the human syncytiotrophoblast is affected by gestational diabetes , hyperleptinemia , and P01375 -α . BACKGROUND : The mechanisms whereby gestational diabetes mellitus ( GDM ) increases the risk of fetal overgrowth and development of metabolic diseases later in life are likely to involve changes in nutrient supply to the fetus . Hence , in this work , we hypothesize that GDM may affect folic acid ( FA ) supply to the placenta and fetus . METHODS : We compared (3)H-FA uptake by human cytotrophoblasts isolated from normal pregnancies ( normal trophoblasts ; NTB cells ) and GDM pregnancies ( diabetic trophoblasts ; DTB cells ) and investigated the effect of GDM hallmarks on (3)H-FA uptake by BeWo cells . RESULTS : (3)H-FA uptake by NTB and DTB cells was time dependent and acidic pH stimulated . When compared with NTB , (3)H-FA uptake by DTB cells was more sensitive to acidic pH changes and to 5-methyltetrahydrofolate and pemetrexed ( PTX ) inhibition , indicating a proportionally greater involvement of the proton-coupled folate transporter ( Q96NT5 ) . A 4-h exposure of BeWo cells to lipopolysaccharide ( LPS , 1-10 μg/ml ) or to high levels of tumor necrosis factor-α ( P01375 -α , 300 ng/l ) significantly reduced (3)H-FA uptake . Moreover , hyperleptinemic conditions ( 100 ng/ml leptin ) decreased (3)H-FA uptake by BeWo cells in a time-dependent manner when compared with normoleptinemic conditions ( 1 ng/ml leptin ) . CONCLUSION : GDM modulates (3)H-FA uptake by the syncytiotrophoblast , and leptin as well as P01375 -α downregulate it . DB08810 protects against ethanol-induced gastric mucosal injury in rats : role of 5-hydroxytryptamine , prostaglandins and sulfhydryl compounds . This study was designed to determine the gastroprotective properties of cinitapride ( CNT ) , a novel prokinetic benzamide derivative agonist of Q13639 and 5-HT1 receptors and 5-HT2 antagonist , on mucosal injury produced by 50 % ( v/v ) ethanol . Results were compared with those for 5-hydroxytryptamine ( 5-HT : 10 mg kg-1 ) . The possible involvements of gastric mucus secretion , endogenous prostaglandins ( PGs ) and sulfhydryl compounds ( SH ) in the protection mediated by CNT were also examined . Intraperitoneal administration of CNT ( 0.50 and 1 mg kg-1 ) , 30 min before ethanol , significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus . CNT ( 1 mg kg-1 ) also produced a significant increase in gastric mucosal levels of DB00917 , but did not induce any significant changes in SH values . On the contrary , pretreatment with 5-HT worsened ethanol-induced erosions , however , did not affect gastric mucus secretion , glycoprotein content or DB00917 levels , although the non-protein SH fraction was significantly decreased . The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism . We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved . Role of Q14116 in overt pain-like behaviour in mice . There are evidences that targeting Q14116 might be beneficial to inhibit inflammatory symptoms , including hypernociception ( decrease in nociceptive threshold ) . The mechanism of Q14116 mechanical hypernociception depends on endothelin in rats and mice . However , the role of Q14116 in overt pain-like behaviour remains undetermined . Therefore , we addressed the role of Q14116 in writhing response induced by intraperitoneal ( i.p. ) injection of phenyl-p-benzoquinone ( PBQ ) and acetic acid in mice . Firstly , it was detected that PBQ and acetic acid i.p. injection induced a dose-dependent number of writhes in Balb/c mice . Subsequently , it was observed that the PBQ - but not the acetic acid-induced writhes were diminished in Q14116 deficient ( ( -/- ) ) mice . Therefore , considering that P01579 , endothelin and prostanoids mediate Q14116 -induced mechanical hypernociception , we also investigated the role of these mediators in the same model of writhing response in which Q14116 participates . It was noticed that PBQ-induced writhes were diminished in P01579 (-/-) mice and by the treatment with DB00559 ( mixed endothelin P25101 /ETB receptor antagonist ) , BQ 123 ( cyclo[DTrp-DAsp-Pro-DVal- DB00149 ] , selective endothelin P25101 receptor antagonist ) , BQ 788 ( N-cys-2,6 dimethylpiperidinocarbonyl-l-methylleucyl-d-1-methoxycarboyl-d-norleucine , selective endothelin ETB receptor antagonist ) or indomethacin ( cycloxigenase inhibitor ) . Thus , Q14116 , P01579 , endothelin acting on endothelin P25101 and ETB receptors , and prostanoids mediate PBQ-induced writhing response in mice . To conclude , these results further advance the understanding of the physiopathology of overt pain-like behaviour , and suggest for the first time a role for Q14116 in writhing response in mice .

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interacts_with DB01182?

multiple_choice

[ The efficacy of degarelix on LUTS ( Lower urinary tract symptoms ) relief in patients with prostate cancer ] . Hormonal therapy is one of the treatment options for prostate cancer patients . There are many hormonal treatments modality to block the testosterone effect on prostate cancer cell proliferation . DB06699 is an innovative molecule able to antagonize the P30968 with comparable oncological results to DB00644 agonist , but with less side effects , avoiding the flare up phase , and better efficacy in LUTS relief . These characteristics of degarelix can impact on the clinical decision making to choose a therapy instead of another . Atrial natriuretic peptide : a possible mediator involved in dexamethasone 's inhibition of cell proliferation in multiple myeloma . Atrial natriuretic peptide ( P01160 ) has been recognized for several decades for its role of regulating blood pressure . Recently , cumulating evidences show that P01160 plays an anticancer role in various solid tumors via blocking the kinase cascade of Ras- Q02750 /2- P27361 /2 with the result of inhibition of DNA synthesis . P01160 , as well as its receptors ( P16066 and P17342 ) has been identified present in the embryonic stem cell and a wide range of cancer cells . Various lymphoid organs , such as lymph nodes , have been detected the presence of P01160 . Multiple myeloma ( MM ) , though the therapies have evolved significantly , is still an incurable disease as B lymphocyte cell neoplasm . Dexamethasone is the cornerstone in treatment of MM via inactivation of Ras- Q02750 /2- P27361 /2 cascade reaction . Coincidently , dexamethasone can increase the expression of P01160 markedly . Nevertheless , the role of P01160 in MM is unclear . Based on these results above , we raise the hypothesis that P01160 is involved in mediating dexamethasone 's inhibition of proliferation in MM cells , which suggests that P01160 may be a potential agent to treat MM . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . Ontogeny of gene expression in the gonadotroph of the developing female rat . During the infantile period of the female rat ( 8-21 postnatal days [ P01160 ] of age ) , there is a dramatic increase in plasma DB00094 , which is thought to be important in initiating ovarian activity and , perhaps , the onset of puberty . To begin to understand the regulation of this DB00094 surge , we determined the ontogenetic development of LHbeta , FSHbeta , and P30968 ( P30968 ) mRNA levels in the pituitary gland throughout the infantile period of the female rat . Steady-state mRNA levels were determined by an external standard quantitative reverse transcriptase polymerase chain reaction assay . FSHbeta and P30968 mRNA levels increased to peak on P01160 12 ( p < 0.03 ) . LHbeta mRNA levels remained relatively constant until rising on P01160 18 . A DB00644 antagonist ( 10-100 microg/animal ) was administered daily from P01160 8-11 or P01160 11-13 , and animals were killed on P01160 12 or P01160 14 , respectively . FSHbeta , LHbeta , and P30968 mRNAs were not affected by DB00644 antagonist treatment . Plasma DB00094 was selectively reduced in the first group , whereas both plasma LH and DB00094 were suppressed in the second group . These data indicate that gene expression of LHbeta , FSHbeta , and P30968 are differentially regulated in the infantile female rat pituitary . DB00644 is involved in regulating the secretion of DB00094 and LH during the infantile period but not in regulating FSHbeta , LHbeta , or P30968 mRNA gene expression . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . DB01093 -treated HL60 cells : a model of neutrophil-like cells mainly expressing Q07343 subtype . The human promyelocytic HL60 cells acquired a neutrophilic phenotype after a 7- to 10-day DB01093 treatment . Fc gammaRII was up-regulated . Fc gammaRI was also up-regulated by an additional P01579 treatment . These cells are able to produce O2*- by NADPH oxidase activation in the presence of immune complexes or phorbol-12-myristate-13-acetate ( PMA ) . A change of their DB05876 subtype profile was also observed : Q07343 was the predominant isoenzyme , Q08499 was down-regulated and P27815 was no longer detectable . Additionally , the more NADPH oxidase was activated by PMA , the less P27815 was expressed , suggesting that NADPH oxidase activity could be used as a surrogate marker of P27815 down-regulation . DB01954 and DB03849 ( cilomilast ) , two selective DB05876 inhibitors , dose-dependently inhibited receptor-coupled activation of superoxide . These results suggest that Q07343 is the main subtype involved in regulating superoxide induced by Fc gammaRs activation . Furthermore , these cells , expressing almost exclusively Q07343 subtype , could be useful to identify selective Q07343 inhibitors . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . T cells targeting a neuronal paraneoplastic antigen mediate tumor rejection and trigger CNS autoimmunity with humoral activation . Paraneoplastic neurologic diseases ( P01160 ) involving immune responses directed toward intracellular antigens are poorly understood . Here , we examine immunity to the P01160 antigen Nova2 , which is expressed exclusively in central nervous system ( CNS ) neurons . We hypothesized that ectopic expression of neuronal antigen in the periphery could incite P01160 . In our C57BL/6 mouse model , CNS antigen expression limits antigen-specific P01730 + and CD8+ T-cell expansion . Chimera experiments demonstrate that this tolerance is mediated by antigen expression in nonhematopoietic cells . CNS antigen expression does not limit tumor rejection by adoptively transferred transgenic T cells but does limit the generation of a memory population that can be expanded upon secondary challenge in vivo . Despite mediating cancer rejection , adoptively transferred transgenic T cells do not lead to paraneoplastic neuronal targeting . Preliminary experiments suggest an additional requirement for humoral activation to induce CNS autoimmunity . This work provides evidence that the requirements for cancer immunity and neuronal autoimmunity are uncoupled . Since humoral immunity was not required for tumor rejection , B-cell targeting therapy , such as rituximab , may be a rational treatment option for P01160 that does not hamper tumor immunity . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Absolute bioavailability and effect of formulation change , food , or elevated pH with rabeprazole on cobimetinib absorption in healthy subjects . DB05239 is a potent and highly selective inhibitor of Q02750 /2 . Since cobimetinib exhibited absorption variability in cancer patients , a series of single-dose studies in healthy subjects were conducted to determine absolute bioavailability and elucidate potential effects of formulation , food , and elevated gastric pH on cobimetinib bioavailability . Three crossover trials were performed with a 20 mg cobimetinib oral dose : absolute bioavailability using a 2 mg intravenous infusion ( n = 13 ) , relative bioavailability of tablets versus capsules and food effect ( n = 20 ) , and drug interaction with a proton pump inhibitor ( 20 mg of rabeprazole daily for 5 days prior to cobimetinib administration ; n = 20 ) . Absolute bioavailability of cobimetinib was 46.2 % ( 24.2 , CV % ) , likely due to metabolism rather than incomplete absorption . The mean systemic clearance of cobimetinib was low ( 11.7 L/h [ 28.2 , CV % ] ) . Administration of cobimetinib tablets with a high-fat meal delayed drug absorption ( prolonged tmax ) but had no statistically significant effect on cobimetinib exposure ( Cmax and AUC0-∞ ) . Tablet and capsule formulations of cobimetinib showed comparable exposures . DB05239 exhibited delayed absorption ( tmax ) in the presence of rabeprazole , with no statistically significant effects on drug exposure ( Cmax and AUC0-∞ ) in the fasted state . In conclusion , cobimetinib oral absorption was not affected by change in formulation , food , or elevated gastric pH . DB09302 inhibits atherosclerosis , improves the plaque morphology , and enhances the effects of a statin . Q8NBP7 ( Q8NBP7 ) inhibition is a potential novel strategy for treatment of CVD . DB09302 is a fully human Q8NBP7 monoclonal antibody in phase 3 clinical development . We evaluated the antiatherogenic potential of alirocumab in P02649 *3Leiden. P11597 mice . Mice received a Western-type diet and were treated with alirocumab ( 3 or 10 mg/kg , weekly subcutaneous dosing ) alone and in combination with atorvastatin ( 3.6 mg/kg/d ) for 18 weeks . DB09302 alone dose-dependently decreased total cholesterol ( -37 % ; -46 % , P < 0.001 ) and TGs ( -36 % ; -39 % , P < 0.001 ) and further decreased cholesterol in combination with atorvastatin ( -48 % ; -58 % , P < 0.001 ) . DB09302 increased hepatic P01130 protein levels but did not affect hepatic cholesterol and TG content . Fecal output of bile acids and neutral sterols was not changed . DB09302 dose-dependently decreased atherosclerotic lesion size ( -71 % ; -88 % , P < 0.001 ) and severity and enhanced these effects when added to atorvastatin ( -89 % ; -98 % , P < 0.001 ) . DB09302 reduced monocyte recruitment and improved the lesion composition by increasing the smooth muscle cell and collagen content and decreasing the macrophage and necrotic core content . DB09302 dose-dependently decreases plasma lipids and , as a result , atherosclerosis development , and it enhances the beneficial effects of atorvastatin in P02649 *3Leiden. P11597 mice . In addition , alirocumab improves plaque morphology . Biosynthetic labeling of diphthamide in Saccharomyces cerevisiae . DB03223 , the post-translational amino acid derivative in the diphtheria toxin-modification site of protein synthesis elongation factor 2 ( P13639 ) , has the proposed structure 2-[3-carboxyamido-3-(trimethylammonio)propyl]histidine ( Van Ness , B. G. , Howard , J. B. , and Bodley , J. W. ( 1980 ) J. Biol. Chem. 255 , 10710-10716 ) . The identification of the biosynthetic precursors of diphthamide would provide a means of evaluating its proposed structure and determining if the amino acid occurs in proteins other than P13639 . Toward this end , yeast were grown on potential radiolabeled precursors and the resulting radiolabeled protein was hydrolyzed in acid . The acid hydrolysates were subjected to amino acid analysis with a program optimized to resolve diphthine , the acid hydrolysis product of diphthamide , from the common amino acids . Radiolabel from [beta-3H]histidine , [alpha-3H]methionine , and [ Me-3H ] methionine was found to be incorporated into diphthine in a molar ratio of 1:1:3 while that of [35S]methionine was not incorporated . These results are in accord with the proposed structure of diphthamide and suggest that in its biosynthesis the backbone and 3 methyl groups of methionine are added to a histidine residue in the peptide chain of P13639 . These labeling experiments show that diphthine ( diphthamide ) constitutes approximately 6 X 10(-6) mol fraction of the total amino acids in yeast protein hydrolysates . Estimates of the amount of diphthamide present in the diphtheria toxin-modification site of P13639 indicate that it constitutes from 4.5 to 9 X 10(-6) mol fraction of the total amino acids in yeast protein . The present evidence suggests that diphthamide occurs only in P13639 . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . DB04690 inhibits Tat-mediated transactivation of type 1 human immunodeficiency virus . Transcription of type 1 human immunodeficiency virus ( HIV-1 ) is governed by the viral long terminal repeat ( LTR ) . By using HIV-1 LTR-directed reporter gene systems , we found that the P11387 inhibitor camptothecin inhibits Tat-mediated transactivation of HIV-1 LTR . The 293.27.2 cells that carry a stably transfected HIV-1 LTR-directed lacZ gene expression vector ( pNAZ ) were used . Inhibitions of LTR were observed at camptothecin concentrations ( IC50 about 0.03 microM , which was an order of magnitude lower than for Ro 24-7429 ) , which had minor effects on cell survival , expression of the cellular gene gro , or Rous sarcoma virus-directed chloramphenicol acetyltransferase ( CAT ) gene expression . Inhibition was also seen with RPMI 8402 , which is a human P01730 -positive lymphocyte line transiently transfected with a HIV-1 LTR-directed ( CAT ) gene . Experiments with HIV-1 LTR mutants suggest that transactivation response sequence but not NF-kappa B is responsible for the inhibition by camptothecin . The target for camptothecin may be a cellular factor that is important for the activation of HIV-1 LTR by Tat and thus may offer a potential target for therapy of HIV-1 infection . Q9Y5Q5 mutations K317E and S472G from preeclamptic patients alter zymogen activation and cell surface targeting. [ Corrected ] . Q9Y5Q5 is a membrane-bound serine protease that acts as the atrial natriuretic peptide ( P01160 ) convertase in the heart . Recent studies show that corin also activates P01160 in the pregnant uterus to promote spiral artery remodeling and prevent pregnancy-induced hypertension . Two Q9Y5Q5 gene mutations , K317E and S472G , were identified in preeclamptic patients and shown to have reduced activity in vitro . In this study , we carried out molecular modeling and biochemical experiments to understand how these mutations impair corin function . By molecular modeling , the mutation K317E was predicted to alter corin P01130 -2 module conformation . Western blot analysis of K317E mutant in HEK293 cells showed that the mutation did not block corin expression on the cell surface but inhibited corin zymogen activation . In contrast , the mutation S472G was predicted to abolish a β-sheet critical for corin frizzled-2 module structure . In Western blot analysis and flow cytometry , S472G mutant was not detected on the cell surface in transfected HEK293 cells . By immunostaining , the S472G mutant was found in the ER , indicating that the mutation S472G disrupted the β-sheet , causing corin misfolding and ER retention . Thus , these results show that mutations in the Q9Y5Q5 gene may impair corin function by entirely different mechanisms . Together , our data provide important insights into the molecular basis underlying corin mutations that may contribute to preeclampsia in patients . DB01373 -induced folding of a fragment of calmodulin composed of EF-hands 2 and 3 . P62158 ( P62158 ) is an EF-hand protein composed of two calcium ( Ca(2+) ) -binding EF-hand motifs in its N-domain ( EF-1 and P13639 ) and two in its C-domain ( EF-3 and Q8N442 ) . In this study , we examined the structure , dynamics , and Ca(2+)-binding properties of a fragment of P62158 containing only P13639 and EF-3 and the intervening linker sequence ( CaM2/3 ) . Based on NMR spectroscopic analyses , Ca(2+)-free CaM2/3 is predominantly unfolded , but upon binding Ca(2+) , adopts a monomeric structure composed of two EF-hand motifs bridged by a short antiparallel beta-sheet . Despite having an " even-odd " pairing of EF-hands , the tertiary structure of CaM2/3 is similar to both the " odd-even " paired N- and C-domains of Ca(2+)-ligated P62158 , with the conformationally flexible linker sequence adopting the role of an inter-EF-hand loop . However , unlike either P62158 domain , CaM2/3 exhibits stepwise Ca(2+) binding with a K ( d1 ) = 30 +/- 5 microM to EF-3 , and a K ( d2 ) > 1000 microM to P13639 . Binding of the first equivalent of Ca(2+) induces the cooperative folding of CaM2/3 . In the case of native P62158 , stacking interactions between four conserved aromatic residues help to hold the first and fourth helices of each EF-hand domain together , while the loop between EF-hands covalently tethers the second and third helices . In contrast , these aromatic residues lie along the second and third helices of CaM2/3 , and thus are positioned adjacent to the loop between its " even-odd " paired EF-hands . This nonnative hydrophobic core packing may contribute to the weak Ca(2+) affinity exhibited by P13639 in the context of CaM2/3 . DB00162 bound to cellular retinol-binding protein is a substrate for cytosolic retinoic acid synthesis . DB00162 bound to cellular retinol-binding protein ( P09455 ) was found to be oxidized to retinoic acid by a soluble activity from calf liver . Cytosolic retinoic acid synthesis from retinol- P09455 was strictly dependent on the exogenous supply of either NAD or NADP . NAD-supported reactions carried out in the presence or in the absence of dimethyl sulfoxide yielded apparent Km and Vmax values for the retinol- P09455 complex of 3.5 +/- 0.6 microM , 611 +/- 49 pmol h-1 ( mg of protein ) -1 , and 0.84 +/- 0.12 microM , 601 +/- 38 pmol h-1 ( mg of protein ) -1 , respectively . The corresponding values for the oxidation of free retinol , dissolved in dimethyl sulfoxide , were 7.1 +/- 0.3 microM and 948 +/- 47 pmol h-1 ( mg of protein ) -1 . Since the dissociation constant of the bovine retinol- P09455 complex is less than 10(-8) M , whereas the Km for retinol- P09455 is of the same order as the Km for free retinol , synthesis of retinoic acid from retinol- P09455 does not rely on prior dissociation of retinol . ApoCRBP proved to be a specific inhibitor of retinoic acid synthesis from P09455 -bound retinol . Its inhibitory effect was indistinguishable from the dilution of the radioactive retinol- P09455 substrate that was obtained by the addition of unlabeled holoCRBP . In contrast , the oxidation of P09455 -bound retinol was not inhibited by the addition of other retinoid binding proteins nor by the addition of either free retinol or retinol complexed with proteins distinct from P09455 . These results indicate that the protein moiety of holoCRBP is specifically recognized by the cytosolic enzyme system that catalyzes retinoic acid synthesis from P09455 -bound retinol . Atrial natriuretic peptide inhibits tumor necrosis factor-alpha production by interferon-gamma-activated macrophages via suppression of p38 mitogen-activated protein kinase and nuclear factor-kappa B activation . We investigated whether the atrial natriuretic peptide ( P01160 ) might have an inhibitory effect on inflammatory cells . Treatment of RAW264.7 macrophages with interferon-gamma ( IFN- gamma ) caused a significant increase in tumor necrosis factor-alpha ( P01375 ) and nitric oxide ( NO ) production . Activation of p38 mitogen-activated protein ( Q96HU1 ) kinase was observed 30 to 120 min after P01579 , and transcription factor nuclear factor-kappa B ( NF-kappaB ) was activated about 7 to 9 times of the basal activity . Human P01160 (99-126) and a specific p38 Q96HU1 kinase inhibitor SB203580 inhibited the P01579 -induced P01375 production in a dose-dependent manner without affecting NO production . P01160 inhibited the P01579 -induced p38 Q96HU1 kinase activation , and P01160 and SB203580 inhibited NF-kappaB activation . To study the involvement of oxidative stress in this system , the effects of allopurinol and acetovanillone , inhibitors of xanthine oxidase and NADPH oxidase , respectively , were studied . Allopurinol or acetovanillone did not inhibit the P01579 -induced production of P01375 or NO , suggesting little involvement of oxidative stress in this system . This is the first evidence in vitro that P01160 has an anti-inflammatory activity on P01579 -activated macrophages by suppressing signal transduction pathway leading to p38 Q96HU1 kinase and NF-kappaB activation . Traumatic brain injury-induced acute gene expression changes in rat cerebral cortex identified by GeneChip analysis . Proper CNS function depends on concerted expression of thousands of genes in a controlled and timely manner . Traumatic brain injury ( TBI ) in mammals results in neuronal death and neurological dysfunction , which might be mediated by altered expression of several genes . By employing a CNS-specific GeneChip and real-time polymerase chain reaction ( PCR ) , the present study analyzed the gene expression changes in adult rat cerebral cortex in the first 24 hr after a controlled cortical impact injury . Many functional families of genes not previously implicated in TBI-induced brain damage are altered in the injured cortex . These include up-regulated transcription factors ( O14543 , O60674 , P35610 -3 , Q03060 , P10914 , SMN , silencer factor-B , ANIA-3 , ANIA-4 , and DB09106 -1 ) and signal transduction pathways ( cpg21 , Narp , and P09455 ) and down-regulated transmitter release mechanisms ( CITRON , synaptojanin II , ras-related rab3 , neurexin-1beta , and SNAP25A and -B ) , kinases ( IP-3-kinase , Pak1 , Ca(2+)/ P62158 -dependent protein kinases ) , and ion channels ( K(+) channels TWIK , RK5 , X62839 , and Na(+) channel I ) . In addition , several genes previously shown to play a role in TBI pathophysiology , including proinflammatory genes , proapoptotic genes , heat shock proteins , immediate early genes , neuropeptides , and glutamate receptor subunits , were also observed to be altered in the injured cortex . Real-time PCR analysis confirmed the GeneChip data for many of these transcripts . The novel physiologically relevant gene expression changes observed here might explain some of the molecular mechanisms of TBI-induced neuronal damage . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . Examination of ceramic restorative material interfacial debonding using acoustic emission and optical coherence tomography . OBJECTIVE : CAD/ P62158 ceramic restorative material is routinely bonded to tooth substrates using adhesive cement . This study investigates micro-crack growth and damage in the ceramic/dentin adhesive interface under fatigue shear testing monitored using the acoustic emission ( AE ) technique with optical coherence tomography ( O75051 ) . METHODS : Ceramic/dentin adhesive samples were prepared to measure the shear bond strength ( SBS ) under static load . Fatigue shear testing was performed using a modified ISO14801 method . Loads in the fatigue tests were applied at 80 % , 70 % , and 60 % of the SBS to monitor interface debonding . The AE technique was used to detect micro-crack signals in static and fatigue shear bond tests . RESULTS : The results showed that the average SBS value in the static tests was 10.61±2.23MPa ( mean±standard deviation ) . The average number of fatigue cycles in which ceramic/dentin interface damage was detected in 80 % , 70 % and 60 % of the SBS were 152 , 1962 and 9646 , respectively . The acoustic behavior varied according to the applied load level . Events were emitted during 60 % and 70 % fatigue tests . A good correlation was observed between crack location in O75051 images and the number of AE signal hits . SIGNIFICANCE : The AE technique and O75051 images employed in this study could potentially be used as a pre-clinical assessment tool to determine the integrity of cemented load bearing restored ceramic material . Sustainable cyclic load stresses in ceramic/dentin-bonded specimens were substantially lower than the measured SBS . Predicted S-N curve showed that the maximum endured load was 4.18MPa passing 10(6) fatigue cyclic .

MH_train_1280

MH_train_1280

MH_train_1280

interacts_with DB00758?

multiple_choice

The use of the VerifyNow Q9H244 point-of-care device to monitor platelet function across a range of Q9H244 inhibition levels following prasugrel and clopidogrel administration . Variability in response to antiplatelet agents has prompted the development of point-of-care ( POC ) technology . In this study , we compared the VerifyNow Q9H244 ( VN- Q9H244 ) POC device with light transmission aggregometry ( P01374 ) in subjects switched directly from clopidogrel to prasugrel . Healthy subjects on aspirin were administered a clopidogrel 600 mg loading dose ( LD ) followed by a 75 mg/d maintenance dose ( MD ) for 10 days . Subjects were then switched to a prasugrel 60 mg LD and then 10 mg/d MD for 10 days ( n = 16 ) , or to a prasugrel 10 mg/d MD for 11 days ( n = 19 ) . Platelet function was measured by P01374 and VN- Q9H244 at baseline and after dosing . DB00758 600 mg LD/75 mg MD treatment led to a reduction in P2Y(12) reaction units ( PRU ) from baseline . A switch from clopidogrel MD to prasugrel 60 mg LD/10 mg MD produced an immediate decrease in PRU , while a switch to prasugrel 10 mg MD resulted in a more gradual decline . Consistent with the reduction in PRU , device-reported percent inhibition increased during both clopidogrel and prasugrel regimens . Inhibition of platelet aggregation as measured by P01374 showed a very similar pattern to that found with VN- Q9H244 measurement , irrespective of treatment regimens . The dynamic range of VN- Q9H244 appeared to be narrower than that of P01374 . With two different thienopyridines , the VN- Q9H244 device , within a somewhat more limited range , reflected the overall magnitude of change in aggregation response determined by P01374 . The determination of the clinical utility of such POC devices will require their use in clinical outcome studies . Testing the utility of an integrated analysis of copy number and transcriptomics datasets for inferring gene regulatory relationships . Correlation patterns between matched copy number variation and gene expression data in cancer samples enable the inference of causal gene regulatory relationships by exploiting the natural randomization of such systems . The aim of this study was to test and verify experimentally the accuracy of a causal inference approach based on genomic randomization using esophageal cancer samples . Two candidates with strong regulatory effects emerging from our analysis are components of growth factor receptors , and implicated in cancer development , namely P04626 and P21802 . We tested experimentally two P04626 and three P21802 regulated interactions predicted by the statistical analysis , all of which were confirmed . We also applied the method in a meta-analysis of 10 cancer datasets and tested 15 of the predicted regulatory interactions experimentally . Three additional predicted P04626 regulated interactions were confirmed , as well as interactions regulated by Q92747 and O15287 . Overall , two thirds of experimentally tested predictions were confirmed . Novel therapies for chronic lymphocytic leukemia . New therapies for chronic lymphocytic leukemia ( CLL ) are moving away from non-specific cytotoxic to more targeted approaches . The monoclonal antibody alemtuzumab induces responses in 33 % to 43 % of patients with relapsed or refractory disease , with 2-5 % CR . Side effects include infusional reactions as well as immunosuppressive effects . DB00073 has limited activity in relapsed refractory patients , but response rates are comparable to follicular non-Hodgkin 's lymphoma in untreated patients . Other antibodies in early phases of development include anti-CD23 [ IDEC-152 ] , anti- P20273 [ epratuzumab ] , Hu1D10 [ apolizumab ] , and anti- P33681 [ anti- P33681 , DB04901 ] . Other agents that are being studied include denileukin diftitox fusion protein ( Ontak ) , and bcl-2 antisense [ G3139 , Genasense ] . The mechanism of action of the new drugs and their role in CLL , as well as the emergence of new prognostic markers are discussed . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . P01374 1 beta 2 and O43557 induce classical and noncanonical NF-kappa B-dependent proinflammatory gene expression in vascular endothelial cells . Activation of the classical and noncanonical NF-kappaB pathways by ligation of the lymphotoxin ( LT ) -beta receptor ( LTbetaR ) plays a crucial role in lymphoid organogenesis and in the generation of ectopic lymphoid tissue at sites of chronic inflammation . Within these microenvironments , LTbetaR signaling regulates the phenotype of the specialized high endothelial cells . However , the direct effects of LTbetaR ligation on endothelial cells remain unclear . We therefore questioned whether LTbetaR ligation could directly activate endothelial cells and regulate classical and noncanonical NF-kappaB-dependent gene expression . We demonstrate that the LTbetaR ligands O43557 and LTalpha1beta2 activate both NF-kappaB pathways in HUVECs and human dermal microvascular endothelial cells ( HDMEC ) . Classical pathway activation was less robust than P01375 -induced signaling ; however , only O43557 and LTalpha1beta2 and not P01375 activated the noncanonical pathway . O43557 and LTalpha1beta2 induced the expression of classical NF-kappaB-dependent genes in HUVEC , including those encoding the adhesion molecules P16581 , P05362 , and P19320 . Consistent with this stimulation , LTbetaR ligation up-regulated T cell adhesion to HUVEC . Furthermore , the homeostatic chemokine P48061 was up-regulated by O43557 and LTalpha1beta2 but not P01375 in both HUVEC and HDMEC . Using HUVEC retrovirally transduced with dominant negative O15111 alpha , we demonstrate that P48061 expression is regulated by the noncanonical pathway in endothelial cells . Our findings therefore demonstrate that LTbetaR ligation regulates gene expression in endothelial cells via both NF-kappaB pathways and we identify P48061 as a bona fide noncanonical NF-kappaB-regulated gene in these cells . Characterization of haloperidol and trifluperidol as subtype-selective N-methyl-D-aspartate ( DB01221 ) receptor antagonists using [3H] DB01520 and [3H]ifenprodil binding in rat brain membranes . [3H] DB01520 and [3H]ifenprodil binding to N-methyl-D-aspartate ( DB01221 ) receptors in rat forebrain membranes was used to compare the inhibition of haloperidol and trifluperidol with that of ifenprodil and eliprodil . In the [3H] DB01520 binding assay , inhibition curves of ifenprodil , eliprodil , haloperidol and trifluperidol revealed two affinity states in the presence of glutamate , glycine and spermidine . The potency of these agents to inhibit the high-affinity fraction of the binding agreed with the results of other studies investigating their potency to block glutamate-induced current at recombinant NR1a/ Q13224 DB01221 receptors expressed in Xenopus oocytes . These agents also inhibited [3H]ifenprodil binding in a biphasic manner , whether in the absence or the presence of either the sigma site ligand GBR-12909 or spermidine . DB03566 reduced the fraction of high-affinity sites labeled with [3H]ifenprodil . The only alteration in the affinity was a decrease in the IC50 value of haloperidol to inhibit the high-affinity fraction of [3H]ifenprodil binding . GBR-12909 also reduced the fraction of [3H]ifenprodil sites inhibited by these compounds with high affinity , with no change in the affinity for either fraction . These data suggest that spermidine is neither a competitive antagonist at the fraction of the binding inhibited by these agents with high affinity , nor is this fraction of the binding to sigma sites . DB00502 and trifluperidol represent a new class of agent that interacts at a site that is labeled by [3H]ifenprodil as well as [3H] DB01520 in rat brain membranes and that closely reflects ifenprodil 's voltage-independent site on the recombinant NR1a/ Q13224 subtype of the DB01221 receptor . DB08816 as an alternative in clopidogrel-associated neutropenia . DB00945 in combination with platelet Q9H244 receptor blocker has become the mainstay antiplatelet treatment strategy for the prevention of stent thrombosis . DB00208 was the first widely used Q9H244 receptor blockers , but clopidogrel has mostly replaced the use of ticlopidine due to its more favorable adverse event profile on bone marrow . However , when clopidogrel induced bone marrow toxicity occurs , little is known about the efficacy and safety of alternative treatments , and thus , in these cases , medical decisions may be very difficult . We report a case of clopidogrel-induced severe neutropenia in a patient treated with coronary stent and safety of alternative treatment with ticagrelor . Endogenous assays of DNA methyltransferases : Evidence for differential activities of P26358 , O14717 , and DNMT3 in mammalian cells in vivo . While CpG methylation can be readily analyzed at the DNA sequence level in wild-type and mutant cells , the actual DNA ( cytosine-5 ) methyltransferases ( DNMTs ) responsible for in vivo methylation on genomic DNA are less tractable . We used an antibody-based method to identify specific endogenous DNMTs ( P26358 , DNMT1b , O14717 , DNMT3a , and DNMT3b ) that stably and selectively bind to genomic DNA containing DB01262 ( aza-dC ) in vivo . Selective binding to aza-dC-containing DNA suggests that the engaged P26358 is catalytically active in the cell . DNMT1b is a splice variant of the predominant maintenance activity P26358 , while O14717 is a well-conserved protein with homologs in plants , yeast , Drosophila , humans , and mice . Despite the presence of motifs essential for transmethylation activity , catalytic activity of O14717 has never been reported . The data here suggest that O14717 is active in vivo when the endogenous genome is the target , both in human and mouse cell lines . We quantified relative global genomic activity of P26358 , -2 , -3a , and -3b in a mouse teratocarcinoma cell line . P26358 and -3b displayed the greatest in vivo binding avidity for aza-dC-containing genomic DNA in these cells . This study demonstrates that individual DNMTs can be tracked and that their binding to genomic DNA can be quantified in mammalian cells in vivo . The different DNMTs display a wide spectrum of genomic DNA-directed activity . The use of an antibody-based tracking method will allow specific DNMTs and their DNA targets to be recovered and analyzed in a physiological setting in chromatin . DB05332 administration shows reduced megakaryocyte response-capacity and increased myelofibrosis in a mouse model of P35579 -RD . Macrothrombocytopenia in P35579 -related disease ( P35579 -RD ) results from defects in nonmuscular myosin-IIA function . P40238 agonists ( eltrombopag ; romiplostim ) seem to improve hemostasis , but little is known about their biologic effects in P35579 -RD . We administered romiplostim to Myh9(-/-) mice ( 100 μg/kg , every 3 days , during 1 month ) . MKs increased to similar numbers in Myh9(-/-) and wild-type ( WT ) mice ( with an increase in immature MKs ) , but Myh9(-/-) platelet count response was much less ( 2.5-fold vs 8-fold increase ) . A strong increase in MK nuclei emboli in the lung , in WT and Myh9(-/-) mice , indicates increased transmigration of MKs from the BM . Prolonged ( but not acute ) treatment with romiplostim decreased expression of GPIb-IX-V complex and Q9HCN6 , but not of GPIIbIIIa , and bleeding time increased in WT mice . Microcirculation was not altered by the increased number of large platelets in any of the assessed organs , but in Myh9(-/-) mice a much stronger increase in BM reticulin fibers was present after 4 weeks of romiplostim treatment vs WT mice . These data further encourage short-term use of thrombopoietic agents in patients with P35579 -RDs ; however , myelofibrosis has to be considered as a potential severe adverse effect during longer treatment . Reduction of GPIbIX/ Q9HCN6 expression by romiplostim requires further studies . Q8N1N2 -spectrum antioxidant therapy featuring astaxanthin coupled with lipoprivic strategies and salsalate for management of non-alcoholic fatty liver disease . Owing to the worldwide epidemic of obesity , and the popularity of diets rich in sugar and saturated fat , nonalcoholic fatty liver disease ( NAFLD ) is increasingly common ; it is usually associated with insulin resistance , and may be considered a component of the metabolic syndrome . The pathologies which can complicate hepatic steatosis -- steatohepatitis , cirrhosis , and hepatic cancer -- appear to result from an interaction of hepatic lipid overload and hepatic oxidative stress . It is therefore proposed that comprehensive regimens which effectively target each of these precipitating factors should achieve the best therapeutic benefit in NAFLD . Appropriate weight loss , and a diet low in saturated fat , glycemic index , and added sugars , should decrease hepatic lipid load . Measures which enhance adipocyte insulin sensitivity -- such as pioglitazone , astaxanthin , and spirulina -- may also be helpful in this regard , as may agents that boost hepatocyte capacity for fatty acid oxidation , such as metformin , carnitine , hydroxycitrate , long-chain omega-3 fats , and glycine . Astaxanthin and spirulina appear to have considerable potential for controlling the oxidative stress associated with NAFLD - the former because it may help to prevent the mitochondrial damage that renders mitochondria a key source of superoxide in this syndrome , the latter because it is exceptionally rich in phycocyanobilin , a phytochemical inhibitor of NAPDH oxidase . Other antioxidants which show some promise in this syndrome include high-dose folate , lipoic acid , melatonin , DB06151 , vitamin E , and taurine . Finally , treatment with salsalate , an inhibitor of O15111 -beta , has potential for blunting the adverse impact of hepatic steatosis on oxidative stress and inflammation . Extracellular glutamate level and DB01221 receptor subunit expression in mouse olfactory bulb following nanoparticle-rich diesel exhaust exposure . In this present study , we aimed to investigate the extracellular glutamate level and memory function-related gene expression in the mouse olfactory bulb after exposure of the animals to nanoparticle-rich diesel exhaust ( NRDE ) with or without bacterial cell wall component . Lipoteichoic acid ( P01374 ) , a cell wall component derived from Staphylococcus aureus , was used to induce systemic inflammation . Male BALB/c mice were exposed to clean air ( particle concentration , 4.58 microg/m(3) ) or NRDE ( 148.86 microg/m(3) ) 5 h per day on 5 consecutive days of the week for 4 wk with or without weekly intraperitoneal injection of P01374 . We examined the extracellular glutamate levels in the olfactory bulb using in vivo microdialysis and high-performance liquid chromatography assay . Then , we collected the olfactory bulb to examine the expression of N-methyl-D-aspartate ( DB01221 ) receptor subunits ( Q9UHB4 , Q12879 , and Q13224 ) and calcium/calmodulin-dependent protein kinase ( CaMK ) IV and cyclic AMP response element binding protein ( CREB ) -1 using real-time reverse-transcription polymerase chain reaction ( RT-PCR ) . NRDE and/or P01374 caused significantly increased extracellular glutamate levels in the olfactory bulb of mice . Moreover , the exposure of mice to NRDE upregulates Q9UHB4 , Q12879 , Q13224 , and CaMKIV mRNAs in the olfactory bulb , while P01374 upregulates only Q13224 and P16220 mRNAs . These findings suggest that NRDE and P01374 cause glutamate-induced neurotoxicity separately and accompanied by changes in the expression of DB01221 receptor subunits and related kinase and transcription factor in the mouse olfactory bulb . This is the first study to show the correlation between glutamate toxicity and memory function-related gene expressions in the mouse olfactory bulb following exposure to NRDE . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . Could decitabine treatment impair memory functions in humans ? The role of epigenetic mechanisms in cognitive functions and neurological/psychiatric disorders has been studied in a number of studies recently . One of these mechanisms is DNA methylation , for which DNA methyltransferases ( P26358 ) are responsible . Decitabine , or DB01262 , is a cytosine-analog P26358 inhibitor and is used in the treatment of certain myelodysplastic syndromes ( P43034 ) subsets . Several studies address the role of DNA methylation and negative effects of decitabine on memory formation and consolidation in animals . We , therefore , hypothesize that standard decitabine treatment for P43034 in patients without dementia might cause learning and memory deficits . A clinical trial is proposed to test the hypothesis which could support the role of DNA methylation in cognitive abilities of humans . ADP receptors -- targets for developing antithrombotic agents . Platelet P2 receptors -- P47900 , Q9H244 , and P51575 -- constitute the means by which adenine nucleotides can activate platelets . Coactivation of the Galphaq-coupled P47900 and Galphai2-coupled Q9H244 receptors is necessary for ADP-mediated platelet activation , which forms the basis of using P2 antagonists as antithrombotic drugs . P47900 receptor antagonists inhibit platelet activation , while P47900 knockout mice show longer bleeding times than normal mice but few other problems ; however , its ubiquitous expression in other tissues renders P47900 questionable as an antithrombotic target . The Q9H244 receptor is expressed nearly exclusively in platelets and brain , making it an attractive antithrombotic target . Antagonists for the Q9H244 receptor have been developed that either require metabolic activation to covalently inhibit Q9H244 and are irreversible , or simply are competitive in nature and thus reversible . DB00208 and clopidogrel are irreversible Q9H244 antagonists and have been repeatedly proven as clinical antithrombotic agents . In addition , a recently reported Q9H244 antagonist , CS-747 , shows promise as a future antithrombotic drug . The AR-C series of compounds represent reversible Q9H244 antagonists and have been used extensively to characterize the function of Q9H244 in platelets . Clinical studies show that AR-C69931MX is as effective as clopidogrel ; furthermore , the combination of AR-C69931MX ( cangrelor ) and clopidogrel confers greater antagonism of Q9H244 than either antagonist alone . The P51575 receptor is a calcium channel that functions to potentiate agonist-induced platelet shape change , and its inhibition or loss has little if any effect on hemostasis . A combination of P47900 and Q9H244 antagonists may represent an additional course of antithrombotic treatment . Synergism of Q9HC29 and Q96P20 activators promotes a unique transcriptional profile in murine dendritic cells . NLRs are cytoplasmic proteins that sense cellular stress and intracellular damage resulting from pathogen uptake . To date , the role of NLRs has been studied using combinations of NLR and TLR agonists , but the interplay between two different NLRs remains uncharacterized . In this study , we employed microarrays to investigate in DCs the regulation of gene transcription mediated by activation of Q9HC29 and Q96P20 pathways using P16444 and MSU . P16444 and MSU co-stimulation of murine BMDCs up-regulated the expression of genes encoding molecules for antigen presentation and co-stimulation ( MHC class II , P33681 , P42081 ) , integrins ( P05106 , P06756 ) , cytokines ( IL-1α , IL-1β , P05231 , P60568 , Q9NPF7 , IL-12p40 ) , and chemokines ( P09341 , P19875 ) . Transcription of the cytokine genes induced by P16444 and MSU partially depended on Q9HC29 but was independent of Q96P20 . Finally , we showed that P27361 and c- P05412 activation increased upon P16444 and MSU co-stimulation . As a whole , the results indicate that two different NLR activators synergize at the transcriptional level , leading to unique differential expression of genes involved in the innate immune response . Solution structure and backbone dynamics of the catalytic domain of matrix metalloproteinase-2 complexed with a hydroxamic acid inhibitor . P08253 is a member of the matrix metalloproteinase family that has been implicated in tumor cell metastasis and angiogenesis . Here , we describe the solution structure of a catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) , determined by three-dimensional heteronuclear NMR spectroscopy . The catalytic domain , designated MMP-2C , has a short peptide linker replacing the internal fibronectin-domain insertion and is enzymatically active . Distance geometry-simulated annealing calculations yielded 14 converged structures with atomic root-mean-square deviations ( r.m.s.d. ) of 1.02 and 1.62 A from the mean coordinate positions for the backbone and for all heavy atoms , respectively , when 11 residues at the N-terminus are excluded . The structure has the same global fold as observed for other MMP catalytic domains and is similar to previously solved crystal structures of P08253 . Differences observed between the solution and the crystal structures , near the bottom of the S1 ' specificity loop , appear to be induced by the large inhibitor present in the solution structure . The MMP-2C solution structure is compared with P22894 crystal structure bound to the same inhibitor to highlight the differences especially in the S1 ' specificity loop . The finding provides a structural explanation for the selectivity between P08253 and P22894 that is achieved by large inhibitors . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Binding affinities for sulfonamide inhibitors with matrix metalloproteinase-2 using a linear response method . Due to their involvement in many pathological conditions , matrix metalloproteinases ( MMPs ) , are very attractive therapeutic targets . Our study focuses on one of them , P08253 , which is involved in tumor progression and metastasis . Recently , the solution structure of the catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) was published by Feng et al . Using the Hanessian group published binding affinity data and the structure published by Feng as a basis , we have built a binding affinity model by targeting the S(2) ' pocket of the enzyme with a set of nine alpha-N-sulfonylamino hydroxamic acid derivatives . Two binding geometries of each ligand have been generated corresponding to two binding modes denoted A and B , respectively , of which the first one is targeting the S(2) ' pocket and the second one the S(1) pocket . For the binding affinity model developed for mode A the computed activities show a rmsd of 0.583 kcal/mol as compared with the experimental data , and a correlation coefficient r(2) of 0.779 , while in the case of the binding mode B a rmsd of 0.834 kcal/mol and correlation coefficient r(2) of 0.500 , respectively , were obtained . In conclusion , our data suggest a higher probability for the DB00120 (76) gated S(2) ' open form pocket to accommodate the substituent alpha versus the wide solvent exposed S(1) subsite , probability which some research groups could have overlooked due to extensive use in their calculations of non revealing S(2) ' pocket open state crystallographic structures instead of NMR ones . Induction of CYP3A expression by dehydroepiandrosterone : involvement of the pregnane X receptor . DB01708 ( DB01708 ) is a steroid produced by the human adrenal gland . Administration of pharmacological doses of DB01708 to rats changes expression of many genes , including the cytochrome P450 family members CYP4A1 and CYP3A23 . It is known that induction of CYP4A expression by DB01708 requires the peroxisome proliferator-activated receptor alpha ( Q07869 (alpha) ) . In the current study , Q07869 (alpha)-null mice were used to examine the role of Q07869 (alpha) in expression of CYP3A . In wild-type mice , 150 mg/kg DB01708 -sulfate induced Cyp4a and Cyp3a11 mRNAs by 5- and 2-fold , respectively . Induction of Cyp4a expression by DB01708 -sulfate was not observed in Q07869 (alpha)-null mice , whereas induction of Cyp3a11 expression by DB01708 -sulfate was similar between genotypes . This suggests that Q07869 (alpha) is not involved in induction of Cyp3a11 expression by DB01708 . Because expression of CYP3A family members can be induced by activation of another member of the nuclear receptor superfamily , the pregnane X receptor ( O75469 ) , we examined the ability of DB01708 to activate O75469 . In transient transfection assays , DB01708 and its metabolites DB01524 ( ADIOL ) , DB01456 , and androst-4-ene-3,17-dione were activators of O75469 . Maximal induction of a O75469 -responsive reporter gene of approximately 3-fold was observed at concentrations of 50 to 100 microM , indicating that these steroids are relatively weak activators of O75469 . Human and murine O75469 exhibited different specificities for DB01708 and its metabolites . ADIOL activated reporter gene expression in the presence of murine but not human O75469 . Results of these studies suggest that the induction of rodent CYP3A expression upon treatment with high doses of DB01708 occurs through activation of O75469 . Eight common genetic variants associated with serum DHEAS levels suggest a key role in ageing mechanisms . DB01708 sulphate ( DHEAS ) is the most abundant circulating steroid secreted by adrenal glands -- yet its function is unknown . Its serum concentration declines significantly with increasing age , which has led to speculation that a relative DHEAS deficiency may contribute to the development of common age-related diseases or diminished longevity . We conducted a meta-analysis of genome-wide association data with 14,846 individuals and identified eight independent common SNPs associated with serum DHEAS concentrations . Genes at or near the identified loci include Q9Y2L8 ( rs11761528 ; p = 3.15 × 10(-36) ) , Q06520 ( rs2637125 ; p = 2.61 × 10(-19) ) , Q92747 ( rs740160 ; p = 1.56 × 10(-16) ) , Q9C037 ( rs17277546 ; p = 4.50 × 10(-11) ) , Q96LC9 ( rs7181230 ; p = 5.44 × 10(-11) ) , Q03014 ( rs2497306 ; p = 4.64 × 10(-9) ) , O43521 ( rs6738028 ; p = 1.72 × 10(-8) ) , and P11712 ( rs2185570 ; p = 2.29 × 10(-8) ) . These genes are associated with type 2 diabetes , lymphoma , actin filament assembly , drug and xenobiotic metabolism , and zinc finger proteins . Several SNPs were associated with changes in gene expression levels , and the related genes are connected to biological pathways linking DHEAS with ageing . This study provides much needed insight into the function of DHEAS . High-density genetic map of the P38398 region of chromosome 17q12-q21 . To facilitate the positional cloning of the breast-ovarian cancer gene P38398 , we constructed a high-density genetic map of the 8.3-cM interval between D17S250 and GIP on chromosome 17q12-q21 . Markers were mapped by linkage in the CEPH and in extended kindreds in our breast cancer series . The map comprises 33 ordered polymorphisms , including 12 genes and 21 anonymous markers , yielding an average of one polymorphism every 250 kb . Twenty-five of the markers are PCR-based systems . The order of polymorphic genes and markers is cen-D17S250-D17S518- P04626 - P10827 - P10276 -D17S80 - P13645 -[D17S800-D17S857]-GAS- D17S856-EDH17B-D17S855-D17S859-D17S858- [ ++ + P01298 -D17S78 ] -D17S183- P02730 -D17S579- D17S509- [ D17S508-D17S190 = D17S810 ] -D17S791- [ D17S181 = D17S806 ] -D17S797- P17509 - P05106 - [ D17S507 = GIP ] -qter . P38398 lies in the middle of the interval , between P10827 and D17S183 . Markers from this map can be used to determine whether cancer is linked to P38398 in families , to evaluate whether tumors have lost heterozygosity at loci in the region , and to identify probes for characterizing chromosomal rearrangements from patients and from tumors . DB05332 as a treatment for immune thrombocytopenia : a review . " Immune thrombocytopenia " ( ITP ) is an autoimmune disorder that leads to peripheral destruction , as well as a decreased production of platelets . ITP most commonly presents as mild mucocutaneous bleeding . Though it is rare , the leading cause of mortality in persons with ITP is intracranial hemorrhage and those that do not respond to therapy are at increased risk . Our understanding of the pathophysiology of ITP has evolved immensely , especially over the last 60 years . The discovery of the platelet-production stimulator , thrombopoietin ( P07202 ) , lent clarity to an earlier hypothesis that inhibition of platelet production at the level of the megakaryocyte , at least in part , accounts for thrombocytopenia in adults with ITP . This facilitated the development of P07202 -based therapies to treat ITP . P40238 agonists are one of the most recent treatments to enter the landscape . Original production of a recombinant human P07202 was halted after clinical trials revealed the untoward effect of autoantibodies to the recombinant human P07202 with cross-reactivity to endogenous P07202 . Next-step development focused on stimulation of the P07202 receptor with fewer immunogenic agents . Currently , two such thrombopoietin receptor agonists , romiplostim and eltrombopag , are licensed in the USA to treat thrombocytopenia in adults with persistent or chronic ITP . Ongoing research will assess their efficacy in other immune-mediated and nonimmune-mediated primary and secondary thrombocytopenias . Epigenetic changes by zebularine leading to enhanced differentiation of human promyelocytic leukemia NB4 and KG1 cells . Aberrant DNA methylation is a critical epigenetic process involved in gene expression of tumor cells . Diverse DNA methyltransferase inhibitors are being studied as potential anticancer drugs , and there is interest in developing novel and more effective DNMTIs . We evaluated zebularine , a stable and low-toxic cytidine analog , effects on human promyelocytic leukemia cell lines , NB4 and KG1 . DB03068 caused a dose- and time-dependent NB4 and KG1 cell growth inhibition , did not induce myeloid differentiation but triggered concentration-dependent apoptosis as manifested by procaspase-3 and P25116 cleavage and the occurrence of early apoptosis detected by Annexin-V-propidium iodide . DB03068 co-treatment with all-trans retinoic acid ( RA ) at pharmacological dose ( 1 μM for NB4 cells ) and higher ( 3 μM for KG1 cells ) increased granulocytic differentiation in both cell lines . Pretreatment for 24 or 48 h with zebularine before the treatment with different doses of RA alone or RA with histone deacetylase inhibitors , phenyl butyrate , and BML-210 , resulted in significant acceleration and enhancement of differentiation and cell cycle arrest at G0/1 . DB03068 alone or in sequential combination with RA decreased expression of P26358 , caused fast and time-dependent expression of pan-cadherin and partial demethylation of P12830 but not tumor suppressor p15 . When used in combination with RA , zebularine increased expression of both genes transcript and protein . DB03068 induced regional chromatin remodeling by local histone H4 acetylation and histone H3- P19013 methylation in promoter sites of methylated P12830 and also in the promoter of unmethylated P38936 as evidenced by chromatin immunoprecipitation assay . Our results extend the spectrum of zebularine effects and the evaluation its utility in acute myeloid leukemia therapy based on epigenetics . High loading dose of clopidogrel is unable to satisfactorily inhibit platelet reactivity in patients with glycoprotein IIIA gene polymorphism : a genetic substudy of PRAGUE-8 trial . The study aimed to assess the impact of nine polymorphisms of genes encoding platelet receptors , enzymes , and hemostatic factors on clopidogrel efficacy to inhibit platelet reactivity in patients with stable coronary artery disease undergoing elective coronary angiography either with or without ad hoc percutaneous coronary intervention . The study was performed as a genetic substudy of the PRAGUE-8 trial . Ninety-five patients pretreated with 600 mg clopidogrel at least 6 h prior to coronary angiography were tested . Baseline platelet reactivity to ADP was assessed before the drug was administered . DB00758 efficacy was tested again at 12 and 28 h after administration . Polymorphisms of platelet receptors , glycoprotein ( GP ) Ia ( 807C/T ) , Q9HCN6 ( 13254C/T ) , P05106 ( PlA1/PlA2 ) , P25116 ( IVSn-14A/T ) , Q9H244 ( 32C/T ) , Q9H244 ( H1/H2 ) haplotype , gene variations of cyclooxygenase-1 , Leiden , and factor II mutations were studied . Flow cytometric tests of vasodilator-stimulated phosphoprotein phosphorylation states were used as a measure of drug efficacy . None of the gene polymorphisms influenced baseline ADP-induced platelet reactivity significantly . Twenty-eight hours after drug administration , differences in suppression of ADP-induced platelet reactivity were observed between polymorphism-positive and polymorphism-negative patients . Inhibition of platelet reactivity , after 600 mg of clopidogrel , was significantly less in carriers of PlA2 ( P=0.009 ) for mean decrease in platelet reactivity index . The proportion of clopidogrel nonresponders ( platelet reactivity index > 50 % ) was apparently higher in PlA2 carriers in comparison with PlA1/PlA1 patients ( 54 vs. 24 % , P=0.082 ) . A 600 mg loading dose of clopidogrel failed to acceptably inhibit platelet reactivity in patients who were positive for the PlA2 polymorphism . Pharmacokinetic profiles of the novel P35354 selective inhibitor cimicoxib in dogs . DB05095 ( CX ) is a novel imidazole derivative that is a cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drug and the latest P35354 selective inhibitor to be released for veterinary use . Currently there is limited information available on the pharmacokinetic ( PK ) properties of CX . The aim of the current study was to evaluate the PK features of CX after administration of the recommended dose and after administration of a more variable dose rate in the form of the commercially available tablet . In addition , the effects of food intake on the PK properties were also evaluated . In the first study , five healthy Beagle dogs received 2mg/kg CX via the oral route following a period of fasting . The second study was conducted using six healthy Labrador retriever dogs which each received an 80 mg tablet ( approximate dose 1.95-2.5mg/kg ) using a crossover design , both in the fasted and fed condition . The plasma concentrations of CX were detected by a validated HPLC method . No adverse effects were observed in any dogs during the experiment . The results from the PK analysis were similar between the studies , regardless of precision of dose and fasted and fed conditions . The mean peak concentration of CX was 0.49 and 0.43 μg/mL under fasted and fed conditions , respectively . The mean half-life was about 3h after all treatments . In addition , simulated multiple dosing data revealed that time over minimal effective concentration was similar after 1.95 , 2.0 and 2.5mg/kg dose administrations . These findings suggest that slight variation from the recommended dose should not alter the therapeutic outcome . In addition , CX can be administered to fed dogs without significantly affecting blood levels . NO-donor P35354 inhibitors . New nitrooxy-substituted 1,5-diarylimidazoles endowed with P35354 inhibitory and vasodilator properties . A series of NO-donor diarylimidazoles derived from the lead compound DB05095 were synthesized and evaluated for their P35354 inhibitory activity and their stability in whole blood as well as for vasodilator properties . The products are partly transformed into the corresponding alcohols following 24-h incubation in whole blood . All of them display good P23219 / P35354 selectivity , but are less potent than the lead ; a molecular modeling study was carried out to investigate their binding mode . The compounds are also capable of relaxing rat aorta strips precontracted with phenylephrine with a NO-dependent mechanism ; this property could confer reduced cardiotoxicity with respect to traditional P35354 inhibitors . High concentration of antioxidants DB06151 and mitoquinone-Q induces intercellular adhesion molecule 1 and oxidative stress by increasing intracellular glutathione . In endothelial cells , the intracellular level of glutathione is depleted during offering protection against proinflammatory cytokine P01375 -induced oxidative stress . Administration of anti-inflammatory drugs , i.e. , DB06151 ( Q9C000 ) or mitoquinone-Q ( mito-Q ) in low concentrations in the human pulmonary aortic endothelial cells offered protection against depletion of reduced glutathione and oxidative stress mediated by P01375 . However , this study addressed that administration of Q9C000 or mito-Q in high concentrations resulted in a biphasic response by initiating an enhanced generation of both reduced glutathione and oxidized glutathione and enhanced production of reactive oxygen species , along with carbonylation and glutathionylation of the cellular proteins . This study further addressed that O15111 ( IKK ) , a phosphorylation-dependent regulator of NF-kappaB , plays an important regulatory role in the P01375 -mediated induction of the inflammatory cell surface molecule P05362 . Of the two catalytic subunits of IKK ( IKKalpha and IKKbeta ) , low concentrations of Q9C000 and mito-Q activated IKKalpha activity , thereby inhibiting the downstream NF-kappaB and P05362 induction by P01375 . High concentrations of Q9C000 and mito-Q instead caused glutathionylation of IKKalpha , thereby inhibiting its activity that in turn enhanced the downstream NF-kappaB activation and P05362 expression by P01375 . Thus , establishing IKKalpha as an anti-inflammatory molecule in endothelial cells is another focus of this study . This is the first report that describes a stressful situation in the endothelial cells created by excess of antioxidative and anti-inflammatory agents Q9C000 and mito-Q , resulting in the generation of reactive oxygen species , carbonylation and glutathionylation of cellular proteins , inhibition of IKKalpha activity , and up-regulation of ICAM-1expression . Clinical and histologic response to single-dose treatment of moderate to severe psoriasis with an anti- P33681 monoclonal antibody . Pathologic T-cell activation is implicated in psoriasis progression . P33681 , a costimulatory molecule involved in T-cell activation , likely plays a key role . DB04901 , an IgG(1) anti- P33681 antibody , was evaluated for safety , pharmacokinetics , and preliminary clinical activity in this open-label , single-dose , dose-escalating study in patients with moderate to severe chronic plaque psoriasis . Twenty-four patients received DB04901 ( 0.05 mg/kg , 0.25 mg/kg , 1 mg/kg , 5 mg/kg , 10 mg/kg , or 15 mg/kg ) . Psoriasis Area and Severity Index , Physician 's Global Psoriasis Assessment , and Psoriasis Severity Scale scores improved in the highest-dose groups . Average plaque thickness and plaque CD3+ and CD8+ T-cell counts decreased in the 10 mg/kg dose group . Adverse events were primarily mild , transient , constitutional symptoms ; the most common related events were mild asthenia ( 29 % of patients ) , chills ( 25 % ) , and headache ( 21 % ) . The serum half-life of DB04901 was approximately 13 days . A single dose of DB04901 appears to be safe and well tolerated and has promising clinical activity in psoriasis . Thrombin inhibits migration of human hepatic myofibroblasts . Several lines of data recently pointed out a role of the serine proteinase thrombin in liver fibrogenesis , but its mechanism of action is unknown . The aim of this study was to evaluate the effect of thrombin on the migration of human liver myofibroblasts . We show here that thrombin inhibits both basal migration and platelet-derived growth factor ( PDGF ) -BB-induced migration of myofibroblasts . By using a thrombin antagonist , a protease-activated receptor ( PAR ) -1 mimetic peptide , and a P25116 antibody , we show that this effect is dependent on the catalytic activity of thrombin and on P25116 activation . Thrombin 's effect on basal migration was dependent on cyclooxygenase 2 ( P35354 ) activation because it was blocked by the P35354 inhibitors NS-398 and nimesulide , and pharmacological studies showed that it was relayed through prostaglandin E(2) and its EP(2) receptor . On the other hand , thrombin-induced inhibition of DB00102 -induced migration was not dependent on P35354 . We show that thrombin inhibits PDGF-induced Akt-1 phosphorylation . This effect was consecutive to inhibition of PDGF-beta receptor activation through active dephosphorylation . Thus thrombin , through two distinct mechanisms , inhibits both basal- and DB00102 -induced migration of human hepatic liver myofibroblasts . The fine tuning of myofibroblast migration may be one of the mechanisms used by thrombin to regulate liver fibrogenesis . Cloning and analysis of the thrombopoietin-induced megakaryocyte-specific glycoprotein VI promoter and its regulation by P15976 , Fli-1 , and Sp1 . The exposure of collagen fibers at sites of vascular injury results in the adherence of platelets and their subsequent activation . The platelet collagen receptor glycoprotein ( GP ) (1) VI plays a crucial role in platelet activation and thrombus formation and decreased levels or defective Q9HCN6 may lead to excessive bleeding . In addition , elevated levels of collagen receptors may predispose individuals to coronary heart disease or strokes . Q9HCN6 expression is restricted to platelets and their precursor cell , the megakaryocyte . In this study we investigate the regulation of Q9HCN6 expression and show that thrombopoietin induces its expression in the megakaryocytic cell line UT-7/ P07202 . A 5'-region flanking the transcription start point of the Q9HCN6 gene was cloned ( -694 to +29 ) and we report that this putative Q9HCN6 promoter bestows megakaryocye-specific expression . Deletion analyses and site-directed mutagenesis identified Sp1(227) , GATA(177) , and Ets(48) sites as essential for Q9HCN6 expression . We show that transcription factors P15976 , Fli-1 , and Sp1 can bind to and activate this promoter . Finally , Q9HCN6 mRNA was detected only in megakaryocytic cell lines expressing both Fli-1 and P15976 , and we show that overexpression of Fli-1 in a stable cell line ( which expresses endogenous P15976 and Sp1 ) results in expression of the endogenous Q9HCN6 gene . Effects of thioacetamide-induced hepatic failure on the N-methyl-D-aspartate receptor complex in the rat cerebral cortex , striatum , and hippocampus . Binding of different ligands and expression of receptor subunit mRNAs . Hepatic encephalopathy ( HE ) is characterized by symptoms pointing at disturbances in glutamatergic neurotransmission in the brain , particularly in the striatum . The binding parameters of ligands specific for different recognition sites in the N-methyl-D-aspartate ( DB01221 ) receptor complex and the distribution of the receptor subunit mRNAs ( Q9UHB4 , Q12879 -D ) were assessed in rats with acute HE induced with a hepatotoxin , thioacetamide ( TAA ) . The binding of : 1 . L-[3H]glutamate ( DB01221 -displaceable ) ; 2 . [3H]dizocilpine and N-(1-[2-thienyl]-cyclohexyl) [3H]piperidine ( [3H] DB01520 ) ; and 3 . The coactivator site agonist [3H]glycine was assayed in purified membranes of the cerebral cortex , hippocampus , and striatum . In HE rats , Bmax of DB01221 -displaceable glutamate binding was increased in the cerebral cortex and hippocampus , but slightly decreased in the striatum . In this region , the binding affinity was also slightly increased . In HE , Bmax of [3H]dizocilpine binding was unchanged in the striatum and cerebral cortex , but substantially decreased in the hippocampus . Pretreatment with phorbol ester enhanced the binding of dizocilpine more in HE than in control rats . Bmax of [3H] DB01520 binding was decreased in the cerebral cortex and striatum , but increased in the hippocampus . The different responses of these two phencyclidine site antagonists to HE may be indicative of a conformational change within the ion channel and/or the presence of microdomains reacting differently to extrinsic factors . HE did not affect glycine binding , but potentiated the maximal stimulation of [3H]dizocilpine binding by glycine in the cerebral cortex . The results emphasize the brain region and domain specificity of the responses of the DB01221 receptor complex to HE . Intraplatelet signaling mechanisms of the priming effect of matrix metalloproteinase-2 on platelet aggregation . OBJECTIVE : Platelets contain and release some matrix metalloproteinases ( MMPs ) , enzymes involved in the degradation of extracellular matrix , and one of these ( P08253 ) exerts a proaggregatory effect . We explored the signal transduction mechanisms activated by P08253 in human blood platelets . METHODS AND RESULTS : Recombinant , human P08253 , added before stimulation with subthreshold doses of different agonists , potentiated platelet activation , calcium influx , IP3 formation , and pleckstrin phosphorylation . Wortmannin and LY29400 , two P19957 -K inhibitors , suppressed the potentiating effects of P08253 and preincubation with P08253 enhanced the thrombin-induced association of the p85alpha P19957 -K subunit with the cytoskeleton and increased the phosphorylation of P31749 . Protein tyrosine kinase inhibitors , Q96HU1 kinase inhibitors , P04054 inhibitors , cyclooxygenase inhibitors and antagonists of the P47900 and Q9H244 receptors did not affect the potentiating activity of P08253 on platelets . CONCLUSION : Our data show that P08253 , at a concentration released by activated platelets , facilitates platelet activation acting at the level of a second messenger system common to different agonists and related to the activation of P19957 -K . Platelet-released P08253 may contribute to platelet activation in vivo . Pharmacological effects of lipid-lowering drugs recapitulate with a larger amplitude the phenotypic effects of common variants within their target genes . BACKGROUND : A major expectation underlying the search for novel susceptibility genes for common diseases using genome-wide association studies ( GWAS ) is that these discoveries will lead to new drug targets . This claim has not been verified yet . Here , we tested the hypothesis that common single nucleotide polymorphisms ( SNPs ) within drug target genes are associated with the corresponding phenotypes , using a population-based GWAS dataset and lipid-lowering drugs as a test case . METHODS : We examined the association between 36 genotyped and 193 imputed SNPs within four lipid-lowering drug target genes ( P04035 , Q07869 , Q8TDS4 / Q8TDS4 and P11597 ) and four non-lipid drug target genes ( P12821 , P30556 , Q9H244 , and P51164 ) and lipid phenotypes , blood pressure , and coronary artery disease in 5635 adult participants of the Lausanne , Switzerland , CoLaus study , genotyped using the Affymetrix 500K SNP chip technology . RESULTS : The phenotypes associated with SNPs within drug target genes recapitulated to a certain extent the pharmacological effects of the drug . The amplitude of the SNP effect was about 10 times smaller than the pharmacological effect of the corresponding drug . In particular , several P11597 SNPs were associated with an elevation in HDL-cholesterol levels , yet a lower diastolic blood pressure , providing evidence that the blood pressure elevation induced by the P11597 inhibitor torcetrapib is more likely compound specific than class specific . CONCLUSION : Pharmacological modulation of lipid-lowering drug targets recapitulates , and markedly amplifies , the phenotypic effects of common SNPs within these target genes . This data provides indirect evidence that , with certain limitations , large-scale GWAS represent a new tool for the discovery and the development of innovative drugs .

MH_train_1281

MH_train_1281

MH_train_1281

interacts_with DB00266?

multiple_choice

The bone morphogenetic protein signaling pathway is upregulated in a mouse model of total parenteral nutrition . Total parenteral nutrition ( O15533 ) results in intestinal mucosal atrophic changes due to an absence of enteral nutrition ; however , the mechanisms responsible for this are not fully understood . It has been shown that bone morphogenetic protein ( BMP ) activation inhibits intestinal epithelial cell ( EC ) proliferation . Therefore , we hypothesized that the BMP pathway could be upregulated by O15533 . To address this , we randomly assigned mice to receive O15533 or to be enterally fed ( control ) for 7 d . Mucosal EC isolates were harvested from the entire length of small intestine for RNA and protein measurements . Q8N1N2 -thickness , mid-small bowel was processed for histological examination . O15533 increased the abundance of P12643 , P12644 , and Q13873 at the RNA and protein levels . Phosphorylation of Q15797 , Q99717 , and Smad8 also was greater in the O15533 group than in the control , which helped to confirm activation of this pathway . Interestingly , the O15533 and control groups did not differ in the mRNA expression of the extracellular soluble bmp antagonists , noggin , gremlin , chordin , or follistatin . Compared to the control group , the expression of c-Myc ( cellular myelocytomatosis ) mRNA was lower , whereas the level of P38936 ( P38936 /CIP1) was greater , in the O15533 group . Because the BMP family may function through suppression of Wnt-beta-catenin signaling , this pathway was also examined . mRNA expression of Wnt 3 , Wnt5a , and the Wnt receptor Lrp5 were lower in the O15533 group compared to controls . The results suggest that the BMP signaling pathway may be involved in the development of intestinal mucosal atrophy due to O15533 administration . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . Oxidative stress renders retinal pigment epithelial cells susceptible to complement-mediated injury . Uncontrolled activation of the alternative pathway of complement is thought to be associated with age-related macular degeneration ( AMD ) . The alternative pathway is continuously activated in the fluid phase , and tissue surfaces require continuous complement inhibition to prevent spontaneous autologous tissue injury . Here , we examined the effects of oxidative stress on the ability of immortalized human retinal pigment epithelial cells ( ARPE-19 ) to regulate complement activation on their cell surface . Combined treatment with H(2)O(2) ( to induce oxidative stress ) and complement-sufficient serum was found to disrupt the barrier function of stable ARPE-19 monolayers as determined by transepithelial resistance ( Q9NZ01 ) measurements . Neither treatment alone had any effect . Q9NZ01 reduction was correlated with increased cell surface deposition of P01024 , and could be prevented by using P10643 -depleted serum , an essential component of the terminal complement pathway . Treatment with H(2)O(2) reduced surface expression of the complement inhibitors P08174 , P08174 , and P13987 , and impaired regulation at the cell surface by factor H present within the serum . Combined treatment of the monolayers with H(2)O(2) and serum elicited polarized secretion of vascular epidermal growth factor ( P15692 ) . Both , secretion of P15692 and Q9NZ01 reduction could be attenuated using either an alternative pathway inhibitor or by blocking P15692 receptor-1/2 signaling . Regarded together , these studies demonstrate that oxidative stress reduces regulation of complement on the surface of ARPE-19 cells , increasing complement activation . This sublytic activation results in P15692 release , which mediates disruption of the cell monolayer . These findings link oxidative stress , complement activation , and apical P15692 release , which have all been associated with the pathogenesis of AMD . Anti-vascular endothelial growth factor therapies in ophthalmology : current use , controversies and the future . Use of anti-vascular endothelial growth factor ( P15692 ) therapies was introduced for the treatment of ocular disorders in 2005 . In the UK , the current licensed and NICE approved indications are for the treatment of neovascular age-related macular degeneration ( nAMD ) , diabetic macular oedema ( Q5VZB9 ) , macular oedema secondary to a retinal vein occlusion ( RVO ) and choroidal neovascularization in pathological myopia . These diagnoses alone account for two-thirds of the main causes of legally registrable visual impairment and blindness . DB01270 ( Lucentis® ; Genentech/Novartis ) , a drug specifically designed for intraocular use , is the primary licensed medication . Controversially however , clinicians have been using an unlicensed cheaper drug , bevacizumab ( Avastin® ; Genentech/Roche ) , originally designed for systemic administration , with a similar mode of action and shown to have a similar efficacy . However , there are fears of greater side effects with bevacizumab though studies have not been sufficiently powered to show statistical difference . In the current global economic climate , anti- P15692 treatment places huge financial and logistical pressure on already strained health care systems . DB00112 is considerably more cost effective than ranibizumab , and thus using bevacizumab would widen access to treatment particularly in developing countries . This licensing issue also places clinicians in a difficult medico-legal position especially in Europe , where doctors are duty bound to use a licensed drug for a particular indication if this is available . As the indications of anti- P15692 therapies expand and the cost of health care provision becomes more expensive , the controversies surrounding their use will inevitably become more important . BMP canonical Smad signaling through Q15797 and Q99717 is required for endochondral bone formation . Bone morphogenetic protein ( BMP ) signaling is required for endochondral bone formation . However , whether or not the effects of BMPs are mediated via canonical Smad pathways or through noncanonical pathways is unknown . In this study we have determined the role of receptor Smads 1 , 5 and 8 in chondrogenesis . Deletion of individual Smads results in viable and fertile mice . Combined loss of Smads 1 , 5 and 8 , however , results in severe chondrodysplasia . Q15797 /5(CKO) ( cartilage-specific knockout ) mutant mice are nearly identical to Q15797 /5(CKO);Smad8(-/-) mutants , indicating that Smads 1 and 5 have overlapping functions and are more important than Smad8 in cartilage . The Q15797 /5(CKO) phenotype is more severe than that of Q13485 (CKO) mice , challenging the dogma , at least in chondrocytes , that Q13485 is required to mediate Smad signaling through BMP pathways . The chondrodysplasia in Q15797 /5(CKO) mice is accompanied by imbalances in cross-talk between the BMP , FGF and Ihh/ P12272 pathways . We show that Ihh is a direct target of BMP pathways in chondrocytes , and that FGF exerts antagonistic effects on Ihh expression . Finally , we tested whether FGF exerts its antagonistic effects directly through Smad linker phosphorylation . The results support the alternative conclusion that the effects of FGFs on BMP signaling are indirect in vivo . Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 receptor ( IL13Rα2 ) that differs from the physiological P24394 /IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL-13-PE ) encoding a mutated human P35225 fused to Pseudomonas exotoxin ( mhIL-13-PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 that binds to the physiological P24394 /IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL-13-PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL-13-PE and two protein formulations , hIL-13-PE used in clinical trials ( DB05305 ) and a second-generation mhIL-13-PE . DB05305 doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL-13-PE led to ∼40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL-13-PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 include its short half-life , which demanded frequent or continued administration , and binding to P24394 /IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM . MicroRNA-155 modulates Treg and Th17 cells differentiation and Th17 cell function by targeting O15524 . MicroRNA ( miR ) -155 is a critical player in both innate and adaptive immune responses . It can influence P01730 (+) T cell lineage choice . To clarify the role of miR-155 in P01730 (+) CD25(+) regulatory T ( Treg ) /T helper (Th)17 cell differentiation and function , as well as the mechanism involved , we performed gain-and loss-of-function analysis by transfection pre-miR-155 and anti-miR-155 into purified P01730 (+) T cells . The results showed that miR-155 positively regulated both Treg and Th17 cell differentiation . It also induced the release of interleukin ( IL ) -17A by Th17 cells , but not the release of P22301 and transforming growth factor ( TGF ) -β1 by Treg cells . Furthermore , we found that miR-155 reacted through regulating Janus kinase/signal transducer and activator of transcription ( JAK/ P35610 ) rather than TGF-β/mothers against decapentaplegic homolog ( SMAD ) signaling pathway in the process of Treg and Th17 cells differentiation . This may because suppressors of cytokine signaling ( Q9NSE2 )1 , the important negative regulator of JAK/ P35610 signaling pathway , was the direct target of miR-155 in this process , but Q15796 and Q99717 were not . Therefore , we demonstrated that miR-155 enhanced Treg and Th17 cells differentiation and Q16552 production by targeting O15524 . Q02318 expression in gilthead sea bream ( Sparus auratus , L. ) : effects of calcitriol and parathyroid hormone-related protein . Little is known about vitamin D metabolism in fishes . Several reports have shown hydroxylase activities in various organs to produce vitamin D metabolites , but the enzymes involved have not been isolated or characterized . We isolated and characterized a renal mitochondrial hydroxylase , Q02318 , that governs vitamin D metabolism in gilthead sea bream , Sparus auratus . The enzyme is highly expressed in kidney and to a far lesser extent in liver . When treated with 25-hydroxy vitamin D or calcitriol , the kidney responded differentially and time dependently with Q02318 mRNA expression levels . This response substantiates a role for Q02318 in fish vitamin D metabolism . This notion is strengthened by upregulation of Q02318 in sea bream treated with parathyroid hormone-related protein ( P12272 ) , and suggests an original role for P12272 in calcitriol-regulated processes n fish similar to the role of PTH in mammalian vitamin D-dependent processes . Factor XIIIa binding to activated platelets is mediated through activation of glycoprotein IIb-IIIa . Stabilization of a clot is dependent on fibrin cross-linking mediated by the transglutaminase , factor XIIIa ( FXIIIa ) . In addition to fibrin stabilization , FXIIIa acts on a number of platelet-reactive proteins , including fibronectin and vitronectin , as well as the platelet proteins , glycoprotein ( GP ) IIb-IIIa , myosin , and actin . However , conditions inducing the platelet-activation dependent binding of FXIIIa have not been characterized nor have the sites mediating FXIIIa binding been identified . The generation of FXIIIa and consequent detection of FXIIIa on the platelet surface were compared with other thrombin-induced activation events ; the rate at which FXIIIa bound to activated platelets was much slower than platelet degranulation or fibrin(ogen) binding . Whereas platelets could be rapidly induced to express a functional receptor for FXIIIa , the rate of FXIIIa binding to platelets is limited by the rate of conversion of FXIII to FXIIIa . Immunoprecipitation of radiolabeled platelets using polyclonal anti-FXIII A-chain antibody identified two proteins corresponding to P08514 and P05106 . Preincubation of intact platelets with DB00054 , a monoclonal antibody that blocks the fibrinogen binding site , or GRGDSP peptide inhibited FXIIIa binding by about 95 % when measured by flow cytometry ; FXIIIa binding to purified P08514 -IIIa was also inhibited by DB00054 . The binding of FXIIIa to purified P08514 -IIIa was enhanced by the addition of fibrinogen , but not by that of fibronectin or thrombospondin , suggesting that FXIIIa also binds to fibrinogen associated with the complex . These observations suggest that activated platelets bearing FXIIIa may enhance stabilization of platelet-rich thrombi through surface-localized cross-linking events . The cloning and reintroduction into animal cells of a functional CAD gene , a dominant amplifiable genetic marker . Rodent cells resistant to DB03459 , a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional P27708 , overproduce CAD as a result of amplification of the CAD gene . We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector . Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long . We introduced the cloned genes into CAD-deficient Chinese hamster ovary ( CHO ) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids . We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of DB03459 following protoplast fusion . The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes . The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization . In two of these transformants , the genes are located in one X chromosome or in a chromosome resembling the X . In the third case , the genes are located in a small metacentric or rearranged chromosome . Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries . BACKGROUND : Atherosclerosis affects aorta , coronary , carotid , and iliac arteries most frequently than any other body vessel . There may be common molecular pathways sustaining this process . Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis . RESULTS : We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression . We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis . Caveolae and JAK/ P35610 pathways , and P06702 / P05109 interacting proteins are certainly involved in the development of vascular disease . We found that the system of caveolae is directly connected with genes that respond to hormone receptors , and indirectly with the apoptosis pathway . Cytokines , chemokines and growth factors released in the blood flux were investigated in parallel . High levels of RANTES , IL-1ra , MIP-1 alpha , MIP-1 beta , P60568 , P05112 , P05113 , P05231 , P13232 , Q16552 , DB00102 , P15692 and P01579 were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels . CONCLUSION : The pattern of cytokine and P06702 / P05109 up-regulation characterizes atherosclerosis as a proinflammatory disorder . Activation of the JAK/ P35610 pathway is confirmed by the up-regulation of P05231 , P42224 , Q00978 and Q13651 genes in coronary and carotid plaques . The functional network constructed in our research is an evidence of the central role of P35610 protein and the caveolae system to contribute to preserve the plaque . Moreover , Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype . Isolation and characterization of P62333 . A novel ATPase family component of the yeast 26 S proteasome . Using a genetic strategy designed to find proteins involved in the function of the Saccharomyces cerevisiae transcriptional activator GAL4 , we isolated mutants in two genes which rescue a class of gal4 activation domain mutants . One of these genes , P62195 , encodes a member of a large family of putative ATPases , the Conserved ATPase containing Domain ( CAD ) proteins ( also known as AAA proteins ) that are involved in a wide variety of cellular functions . Subsequently , P62195 was identified as a subunit of the 26 S proteasome . We have now cloned the gene defined by the second complementation group . P62333 encodes an essential 49-kDa protein that is also a member of the CAD family and is 43 % identical to P62195 . The mutation in sug2-1 , like that in sug1-1 , is found in the CAD near the highly conserved ATPase motif . We present biochemical and genetic evidence that P62333 is associated in vivo with P62195 and is a novel P27708 subunit of the 26 S proteasome . With its highly conserved mammalian homologs , human Q8NFH3 and ground squirrel CADp44 , P62333 defines a new class of proteasomal CAD proteins . Purification and characterization of mouse O15528 overproduced by an Escherichia coli system coexpressing molecular chaperonins GroEL/ES . The expression of mouse O15528 in Escherichia coli has been dramatically enhanced by coexpression of GroEL/ES . To reveal the enzymatic properties of O15528 , we measured its hydroxylation activity toward vitamin D3 and DB01436 ( 1alpha(OH)D3 ) in addition to the physiological substrate DB00146 . Surprisingly , O15528 converted vitamin D3 to 1alpha, DB00146 . Both 1alpha-hydroxylation activity toward vitamin D3 , and 25-hydroxylation activity toward 1alpha(OH)D3 were observed . The Km and Vmax values for 25-hydroxylation activity toward 1alpha(OH)D3 were estimated to be 1.7 microM and 0.51 mol/min/mol P450 , respectively , while those for 1alpha-hydroxylation activity toward DB00146 were 0.050 microM and 2.73 mol/min/mol P450 , respectively . Note that the substrate must be fixed in the opposite direction in the substrate-binding pocket of O15528 between 1alpha-hydroxylation and 25-hydroxylation . Based on these results and the fact that human Q02318 and Streptomyces CYP105A1 also convert vitamin D3 to 1alpha, DB00146 , 1alpha-hydroxylation , and 25-hydroxylation of vitamin D3 appear to be closely linked together . Effects of peroxisome proliferator-activated receptor-alpha and -gamma agonist , DB04971 , on diabetic complications in Zucker diabetic fatty rats . This study has investigated the effects of DB04971 , a peroxisome proliferator-activated receptor ( Q07869 ) -alpha and P37231 agonist , on the pathogenesis of diabetic complications in the Zucker diabetic fatty ( ZDF ) rats , a model of type 2 diabetes . Comparison is made with troglitazone , a P37231 agonist . The ZDF rats exhibited hyperglycaemia and hyperlipidaemia , and developed diabetic complications such as cataract , nephropathy , and neuropathy . Treatment with DB04971 from the prediabetic stage controlled glycaemia and lipidaemia , and prevented the development of diabetic complications . DB00197 was less effective in controlling serum cholesterol and neuropathy . ZDF rats developed diabetic osteopenia with reduced bone turnover , and this was prevented by DB04971 and troglitazone , possibly mediated by increased bone turnover and bone formation . Since DB04971 controlled glycaemia and lipidaemia in ZDF rats and prevented several diabetic complications , it is suggested that treatment with DB04971 , which activates both Q07869 and P37231 , could provide a valuable therapeutic approach against diabetic complications in type 2 diabetes . Peroxisome proliferator-activated receptor-gamma agonists increase vascular endothelial growth factor expression in human vascular smooth muscle cells . Vascular endothelial growth factor ( P15692 ) , expressed in a variety of mesenchymal cells including vascular smooth muscle cells ( VSMC ) , is a potent mitogen for endothelial cells , and is used clinically applied for ischemic disease of peripheral vessels . To determine whether peroxisome proliferator-activated receptor gamma ( PPARgamma ) regulates P15692 production in VSMC , we examined P15692 secretion from VSMC treated with Q07869 agonists . DB00197 increased P15692 secretion in a time- and dose-dependent manner ( 261 +/- 35 % with 25 mM of troglitazone for 24 h ) , and also increased levels of P15692 mRNA . P15692 secretion was also increased by other PPARgamma agonists , pioglitazone , LY171883 , and 15d-PGJ2 ( 224 +/- 17.1 % , 247 +/- 36.8 % and 171 +/- 7.8 % , respectively ) , but not the PPARgamma agonists bezafibrate and Wy14643 ( 85.2 +/- 1.5 % , 94.6 +/- 3.2 , respectively ) . Our findings suggest that thiazolidinediones might be useful for the therapeutic angiogenesis for ischemic artery disease . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . Inhibition of human steroid 5beta-reductase ( P51857 ) by finasteride and structure of the enzyme-inhibitor complex . The Delta(4)-3-ketosteroid functionality is present in nearly all steroid hormones apart from estrogens . The first step in functionalization of the A-ring is mediated in humans by steroid 5alpha- or 5beta-reductase . DB01216 is a mechanism-based inactivator of 5alpha-reductase type 2 with subnanomolar affinity and is widely used as a therapeutic for the treatment of benign prostatic hyperplasia . It is also used for androgen deprivation in hormone-dependent prostate carcinoma , and it has been examined as a chemopreventive agent in prostate cancer . The effect of finasteride on steroid 5beta-reductase ( P51857 ) has not been previously reported . We show that finasteride competitively inhibits P51857 with low micromolar affinity but does not act as a mechanism-based inactivator . The structure of the P51857 .NADP(+)*finasteride complex determined at 1.7 A resolution shows that it is not possible for NADPH to reduce the Delta(1-2)-ene of finasteride because the cofactor and steroid are not proximal to each other . The P01024 -ketone of finasteride accepts hydrogen bonds from the catalytic residues DB00135 -58 and DB00142 -120 in the active site of P51857 , providing an explanation for the competitive inhibition observed . This is the first reported structure of finasteride bound to an enzyme involved in steroid hormone metabolism . Synergistic antileukemic effects between DB06080 and chemotherapy involve downregulation of cell cycle-regulated genes and c-Mos-mediated MAPK pathway . Internal tandem duplications ( ITDs ) of fms-like tyrosine kinase 3 ( P36888 ) receptor play an important role in the pathogenesis of acute myeloid leukemia ( AML ) and represent an attractive therapeutic target . DB06080 has demonstrated potent effects in AML cells with P36888 -ITDs . Here , we provide further evidence that DB06080 treatment significantly downregulates cyclins D and E but increases the expression of P38936 and p27 . DB06080 induces apoptosis through downregulation of Bcl-xL and upregulation of Q16611 , P55957 and Q92934 . We also evaluate the combinations of DB06080 and chemotherapy . DB06080 demonstrates significant sequence-dependent synergism with cytarabine and doxorubicin in cell lines and primary leukemia samples . The optimal combination was validated in MV4-11 xenografts . Low-density array analysis revealed the synergistic interaction involved in downregulation of cell cycle and mitogen-activated protein kinase pathway genes . P24385 and c-Mos were the most significantly inhibited targets on both transcriptional and translational levels . Treatment with short hairpin RNAs targeting either P24385 or c-Mos further sensitized MV4-11 cells to DB06080 . These findings suggest that specific pathway genes were further targeted by adding chemotherapy and support the rationale of combination therapy . Thus , a clinical trial using sequence-dependent combination therapy with DB06080 in AML is warranted . DB00092 ( anti- P06729 ) causes a selective reduction in circulating effector memory T cells ( Tem ) and relative preservation of central memory T cells ( Tcm ) in psoriasis . BACKGROUND : DB00092 ( anti- P06729 ) biological therapy selectively targets effector memory T cells ( Tem ) in psoriasis vulgaris , a model Type 1 autoimmune disease . METHODS : Circulating leukocytes were phenotyped in patients receiving alefacept for moderate to severe psoriasis . RESULTS : In all patients , this treatment caused a preferential decrease in effector memory T cells ( P32248 - CD45RA- ) ( mean 63 % reduction ) for both P01730 + and CD8+ Tem , while central memory T cells ( Tcm ) ( P32248 +CD45RA- ) were less affected , and naïve T cells ( P32248 +CD45RA+ ) were relatively spared . Circulating CD8+ effector T cells and Type 1 T cells ( P01579 -producing ) were also significantly reduced . CONCLUSION : DB00092 causes a selective reduction in circulating effector memory T cells ( Tem ) and relative preservation of central memory T cells ( Tcm ) in psoriasis . Bone morphogenetic protein signalling in P01138 -stimulated PC12 cells . Bone morphogenetic proteins ( BMPs ) are shown to potentiate P01138 -induced neuronal differentiation in PC12 phaeochromocytoma cells grown on collagen under low-serum conditions . Whereas , cell bodies remained rounded in control medium or with only BMPs present , addition of P12644 or P22004 robustly increased the neuritogenic effect of P01138 within 2 days . P01138 -increased phosphorylation of Q8TCB0 (Erk1) and Q8NFH3 (Erk2) between 2 and 24h was unaffected by addition of P22004 . PC12 cells transfected with the SBE(4x)-luc reporter showed that P12644 significantly increased receptor-activated Smad activity . Expression of constitutively active BMP receptor ALK2 activating Q15797 and Q99717 resulted in a strong increase in the SBE(4x)-luc reporter response . Adding the inhibitory O15105 drastically reduced this signal . In contrast to wild-type ( wt ) Q99717 , a Q99717 variant lacking five Erk phosphorylation sites in the linker region ( designated Q99717 /5SA ) showed a strong background transcriptional activity . A fusion construct ( Gal4- Q99717 /5SA ) was also highly transcriptionally active . Addition of the MEK inhibitor U0126 to PC12 cells expressing Gal4- Q99717 /wt did not increase background transcriptional activity . However , upon activation by constitutively active ALK2 both Gal4- Q99717 /wt and Gal4- Q99717 /5SA strongly stimulated transcription . The data show that serine residues of the linker region of Q99717 reduce spontaneous transcriptional activity and that P01138 -activated Erk does not antagonise BMP signalling at this site . Hence , P01138 and BMP signals are likely to interact further downstream at the transcriptional level in neuronal differentiation of the PC12 cells .

MH_train_1282

MH_train_1282

MH_train_1282

interacts_with DB08820?

multiple_choice

Differential regulation of cystic fibrosis transmembrane conductance regulator by interferon gamma in mast cells and epithelial cells . P13569 ( P13569 ) is a P13569 in epithelial cells ; recently , we identified it in mast cells . Previous work that we confirmed showed that interferon gamma ( IFNgamma ) down-regulated P13569 expression in epithelial cells ( T84 ) , but by contrast , we found that IFNgamma up-regulated P13569 mRNA and protein expression in rat and human mast cells . IFNgamma up-regulation of P13569 in mast cells was inhibited by p38 and extracellular signal-regulated kinase ( P29323 ) kinase inhibitors but not a Janus tyrosine kinase (JAK)2 inhibitor , whereas in T84 cells IFNgamma-mediated down-regulation of P13569 was O60674 -dependent and P29323 - and p38-independent . Furthermore , IFNgamma down-regulation of P13569 in T84 epithelial cells was P42224 -dependent , but up-regulation of P13569 in mast cells was P42224 -independent . Thus , differential regulatory pathways of P13569 expression in mast cells and epithelial cells exist that depend upon either p38/ P29323 or JAK/ P35610 pathways , respectively . Surprisingly , IFNgamma treatment of mast cells inhibited Cl(-) efflux , in contrast to up-regulation of P13569 /mRNA and protein expression . However , down-regulation of Cl(-) flux correlated with IFNgamma-mediated inhibition of mediator secretion . This and other work suggests that the effect of IFNgamma on P13569 expression in mast cells is important for their function . Flip and flop splice variants of AMPA receptor subunits in the spinal cord of amyotrophic lateral sclerosis . Excitotoxicity mediated by AMPA receptors has been suggested to be implicated in the pathogenesis of amyotrophic lateral sclerosis ( P35858 ) . To investigate the relevance of AMPA receptors to motor neuron degeneration in P35858 , we evaluated the expression of mRNAs coding for flip and flop splice variants of AMPA receptor subunits ( P42261 to P48058 ) in the cervical segment of the spinal cord from control individuals and patients with P35858 using in situ hybridization histochemistry . Transcript mRNAs coding for flop variants were significantly decreased in the ventral horn of the spinal cord from patients with P35858 , whereas the mRNAs for flip variants were preserved . These findings suggest that the relative abundance of flip variants vs. flop variants is increased in spinal motor neurons of P35858 patients when compared to that of control individuals . Flip variants promote assemblies of slowly desensitizing AMPA receptors . These results imply that spinal motor neurons of P35858 patients possess more slowly desensitizing AMPA receptors than those of control individuals . This expression change of AMPA receptors in P35858 may account for vulnerability of motor neurons in this disease . DB11320 stimulates hydrogen peroxide production by bronchial epithelial cells via histamine H1 receptor and dual oxidase . Oxidative stress has been implicated in the pathogenesis of bronchial asthma . Besides granulocytes , the airway epithelium can produce large amounts of reactive oxygen species and can contribute to asthma-related oxidative stress . DB11320 is a major inflammatory mediator present in large quantities in asthmatic airways . Whether histamine triggers epithelium-derived oxidative stress is unknown . We therefore aimed at characterizing human airway epithelial H2O2 production stimulated by histamine . We found that air-liquid interface cultures of primary human bronchial epithelial cells ( BECs ) and an immortalized BEC model ( Cdk4/hTERT HBEC ) produce H2O2 in response to histamine . The main source of airway epithelial H2O2 is an NADPH dual oxidase , Duox1 . Out of the four histamine receptors ( P35367 - Q9H3N8 ) , P35367 has the highest expression in BECs and mediates the H2O2-producing effects of histamine . P05112 induces Duox1 gene and protein expression levels and enhances histamine-induced H2O2 production by epithelial cells . Using P29320 -293 cells expressing Duox1 or Duox2 and endogenous P35367 , histamine triggers an immediate intracellular calcium signal and H2O2 release . Overexpression of P35367 further increases the oxidative output of Duox-expressing P29320 -293 cells . Our observations show that BECs respond to histamine with Duox-mediated H2O2 production . These findings reveal a mechanism that could be an important contributor to oxidative stress characteristic of asthmatic airways , suggesting novel therapeutic targets for treating asthmatic airway disease . Drosophila Answers to Q13148 Proteinopathies . Initially implicated in the pathogenesis of P13569 and HIV-1 transcription , nuclear factor Q13148 was subsequently found to be involved in the origin and development of several neurodegenerative diseases . In 2006 , in fact , it was reported for the first time the cytoplasmic accumulation of Q13148 in ubiquitin-positive inclusions of P35858 and FTLD patients , suggesting the presence of a shared underlying mechanism for these diseases . Today , different animal models of Q13148 proteinopathies are available in rodents , nematodes , fishes , and flies . Although these models recapitulate several of the pathological features found in patients , the mechanisms underpinning the progressive neuronal loss observed in Q13148 proteinopathies remain to be characterized . Compared to other models , Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies . At present , the development of Q13148 -related Drosophila models has further strengthened the hypothesis that both Q13148 " loss-of-function " and " gain-of-function " mechanisms can contribute to disease . The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies , along with the information they have generated , towards a better understanding of the mechanisms underlying Q13148 proteinopathies . Recurrent acute and chronic pancreatitis in two brothers with familial chylomicronemia syndrome . The chylomicronemia syndrome is well recognized as a rare etiologic factor of acute pancreatitis ; however , whether hypertriglyceridemia can cause chronic pancreatitis ( CP ) remains unclear . We describe the long-time course of 2 brothers with the familial chylomicronemia syndrome caused by identical compound heterozygous mutations in the lipoprotein lipase ( P06858 ) gene with markedly reduced P06858 activity . Other etiologic factors were excluded , including mutations in the P07477 , P00995 , and P13569 gene . Although both brothers had recurrent acute pancreatitis and the same P06858 genotype , CP became evident in only one patient . Progression to CP was associated with a more severe disease course . Thus , the chylomicronemia syndrome may cause CP in the absence of other known causative factors , and similar to alcoholic and hereditary CP , a more severe disease course is associated with disease progression . Endogenous concentrations of ouabain act as a cofactor to stimulate fluid secretion and cyst growth of in vitro ADPKD models via DB02527 and P00533 -Src-MEK pathways . In autosomal-dominant polycystic kidney disease ( ADPKD ) , renal cysts develop by aberrant epithelial cell proliferation and transepithelial fluid secretion . We previously showed that ouabain increases proliferation of cultured human ADPKD cells via stimulation of the P01133 receptor ( P00533 ) -Src-MEK/ P29323 signaling pathway . We examined whether ouabain affects fluid secretion and in vitro cyst growth of human ADPKD cell monolayers , ADPKD cell microcysts cultured in a three-dimensional collagen matrix , and metanephric organ cultures from Pkd1(m1Bei) mice . Physiological concentrations of ouabain alone did not affect net transepithelial basal-to-apical fluid transport in ADPKD monolayers or growth of cultured ADPKD microcysts . In contrast , in the presence of forskolin or 8-bromo- DB02527 , ouabain significantly enhanced ADPKD fluid secretion and microcyst expansion . Ouabain exerted this effect by enhancing DB02527 -dependent Cl(-) secretion via the P13569 . Similarly , ouabain accelerated DB02527 -dependent cyst enlargement in Pkd1(m1Bei) mice metanephroi , with a more prominent response in homozygous than heterozygous mice . Ouabain had no effect on fluid secretion and cystogenesis of normal human kidney cells and caused only slight cystic dilations in wild-type mouse kidneys . The effects of ouabain in ADPKD cells and Pkd1(m1Bei) metanephroi were prevented by inhibitors of P00533 ( AG1478 ) , Src ( Q99463 ) , and MEK ( U0126 ) . Together , our results show that ouabain , used in physiological concentrations , has synergistic effects on DB02527 -mediated fluid secretion and cyst growth , via activation of the P00533 -Src-MEK pathway . These data provide important evidence for the role of ouabain as an endogenous hormone that exacerbates ADPKD cyst progression . Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . The effects of a cyclooxygenase-2 ( P35354 ) expression and inhibition on human uveal melanoma cell proliferation and macrophage nitric oxide production . BACKGROUND : P35354 ( P35354 ) expression has previously been identified in uveal melanoma although the biological role of P35354 in this intraocular malignancy has not been elucidated . This study aimed to investigate the effect of a P35354 inhibitor on the proliferation rate of human uveal melanoma cells , as well as its effect on the cytotoxic response of macrophages . METHODS : Human uveal melanoma cell lines were transfected to constitutively express P35354 and the proliferative rate of these cells using two different methods , with and without the addition of Amfenac , was measured . DB00435 production by macrophages was measured after exposure to melanoma-conditioned medium from both groups of cells as well as with and without Amfenac , the active metabolite of DB06802 . RESULTS : Cells transfected to express P35354 had a higher proliferation rate than those that did not . The addition of Amfenac significantly decreased the proliferation rate of all cell lines . DB00435 production by macrophages was inhibited by the addition of melanoma conditioned medium , the addition of Amfenac partially overcame this inhibition . CONCLUSION : Amfenac affected both P35354 transfected and non-transfected uveal melanoma cells in terms of their proliferation rates as well as their suppressive effects on macrophage cytotoxic activity . Mechanism of oral absorbent DB05269 in lipid abnormalities in experimental uremic rats . BACKGROUND : We have reported that oral sorbent DB05269 ( Q9NRA2 ) is effective in delaying the induction of dialysis in patients with chronic renal failure ( CRF ) because of its effect on lipid metabolism . To clarify the precise mechanism of Q9NRA2 in lipid abnormalities in CRF , we examined the effect of Q9NRA2 on plasma lipid profile , total bile acids ( TBA ) , and lipoprotein lipase ( P06858 ) activity in experimental uremic rats . METHODS : Uremic rats were prepared using male Wistar rats by ligating 5/6 of the renal artery . Uremic rats were randomly divided into two groups as follows : a control group in which rats were maintained on the standard diet and an Q9NRA2 group in which rats were maintained on a diet containing 5 g of Q9NRA2 per 100 g of standard diet for 10 weeks . Plasma P06858 activity was measured as free fatty acid ( FFA ) generation after intravenous administration of heparin . RESULTS : Plasma creatinine at 1.5 +/- 0.1 mg/dl was lower in the Q9NRA2 group than the 1.9 +/- 0.5 mg/ml level in the control group . Q9NRA2 significantly decreased plasma total cholesterol from 192 +/- 29 to 142 +/- 25 mg/dl , triglycerides from 198 +/- 71 to 99 +/- 38 mg/dl , and TBA from 19.6 +/- 2.6 mumol/liter to 8.8 +/- 3.5 mumol/ml . Plasma P06858 activity at 0.22 +/- 0.01 mumol FFA/min/hr was significantly higher in the Q9NRA2 group than 0.15 +/- 0.03 mumol FFA/min/hr in the control group . CONCLUSIONS : These results suggest that Q9NRA2 may improve plasma lipid abnormalities by binding to bile acids in the intestinal lumen and preventing their reabsorption and inhibiting the reduction of P06858 activity in experimental uremic rats . Interplay between inhibitory ferric and stimulatory curcumin regulates phosphorylation-dependent human cystic fibrosis transmembrane conductance regulator and ΔF508 activity . Curcumin potentiates cystic fibrosis transmembrane conductance regulator ( P13569 ) activation in an DB00171 -independent but phosphorylation-dependent manner . The underlying molecular mechanisms are unclear . Here , P29320 -293T cells cultured in an Fe(3+)-containing medium were transiently transfected with P13569 constructs , and the role of the inhibitory Fe(3+) bridge between intracellular loop 3 and the regulatory domain of P13569 in this pathway was investigated . The results showed that ethylenediaminetetraacetic acid ( DB00974 ) stimulated phosphorylation-dependent P13569 activation and the stimulation was suppressed by the deletion of the regulatory domain or the insertion of a C832A mutation that removes the Fe(3+)-binding interface . Furthermore , curcumin potentiation of P13569 was significantly weakened not only by Fe(3+)-insensitive mutations at the interface between the regulatory domain and intracellular loop 3 but also by N-ethylmaleimide or DB00974 pretreatment that removes Fe(3+) . More importantly , potentiation of P13569 was completely suppressed by sufficient Fe(3+) . Finally , the insertion of Fe(3+)-insensitive H950R/S768R increased the curcumin-independent activity of ΔF508 but weakened its curcumin potentiation . Thus , Fe(3+) homeostasis in epithelia may play a critical role in regulating P13569 activity , and targeting Fe(3+)-chelating potentiators may direct new therapies for cystic fibrosis . Novel structural features of CDK inhibition revealed by an ab initio computational method combined with dynamic simulations . The rational development of specific inhibitors for the approximately 500 protein kinases encoded in the human genome is impeded by a poor understanding of the structural basis for the activity and selectivity of small molecules that compete for DB00171 binding . Combining classical dynamic simulations with a novel ab initio computational approach linear-scalable to molecular interactions involving thousands of atoms , we have investigated the binding of five distinct inhibitors to the cyclin-dependent kinase P24941 . We report here that polarization and dynamic hydrogen bonding effects , so far undetected by crystallography , affect both their activity and selectivity . The effects arise from the specific solvation patterns of water molecules in the DB00171 binding pocket or the intermittent formation of hydrogen bonds during the dynamics of CDK/inhibitor interactions and explain the unexpectedly high potency of certain inhibitors such as 3-(3H-imidazol-4-ylmethylene)-5-methoxy-1,3-dihydro-indol-2-one ( DB03428 ) . The Lys89 residue in the DB00171 -binding pocket of P24941 is observed to form temporary hydrogen bonds with the three most potent inhibitors . This residue is replaced in P11802 by Thr89 , whose shorter side-chain can not form similar bonds , explaining the relative selectivity of the inhibitors for P24941 . Our results provide a generally applicable computational method for the analysis of biomolecular structures and reveal hitherto unrecognized features of the interaction between protein kinases and their inhibitors . Managing the underlying cause of cystic fibrosis : a future role for potentiators and correctors . Cystic fibrosis ( CF ) , a severe genetic disease , is caused by mutations that alter the structure and function of P13569 , a plasma membrane channel permeable to chloride and bicarbonate . Defective anion transport in CF irreversibly damages the lungs , pancreas , liver , and other organs . CF mutations cause loss of P13569 function in multiple ways . In particular , class 3 mutations such as p.Gly551Asp strongly decrease the time spent by P13569 in the open state ( gating defect ) . Instead , class 2 mutations impair the maturation of P13569 protein and its transport from the endoplasmic reticulum to the plasma membrane ( trafficking defect ) . The deletion of phenylalanine 508 ( p.Phe508del ) , the most frequent mutation among CF patients ( 70-90 % ) , destabilizes the P13569 protein , thus causing both a trafficking and a gating defect . These two defects can be overcome with drug-like molecules generically called correctors and potentiators , respectively . The potentiator Kalydeco™ ( also known as DB08820 or VX-770 ) , developed by Vertex Pharmaceuticals , has been recently approved by the US FDA and the European Medicines Agency ( P15941 ) for the treatment of CF patients carrying at least one P13569 allele with the p.Gly551Asp mutation ( 2-5 % of all patients ) . In contrast , the corrector VX-809 , which significantly improves p.Phe508del- P13569 trafficking in vitro , is still under study in clinical trials . Because of multiple defects caused by the p.Phe508del mutation , it is probable that rescue of the mutant protein will require combined treatment with correctors having different mechanisms of action . This review evaluates the status of experimental and clinical research in pharmacotherapy for the CF basic defect . Clinical development of eniluracil : current status . DB03516 is a potent inactivator of dihydropyrimidine dehydrogenase ( Q12882 ) , which is the first enzyme in the degradative pathway of systemically administered 5-fluorouracil ( DB00544 ) . Two completely oral regimens of eniluracil plus DB00544 are being evaluated in clinical trials : ( 1 ) a chronic schedule with both agents administered P55957 in a 10:1 ratio for 28 days of a 5-week course , and ( 2 ) a 5-day schedule of eniluracil once daily on days 1 through 7 and DB00544 once daily on days 2 through 6 . The clinical development of eniluracil is being pursued in several tumor types , including colorectal cancer , breast cancer , and pancreatic cancer . Response rates achieved in a phase II study of the chronic schedule of oral eniluracil/ DB00544 in patients with colorectal cancer compare favorably with those obtained in trials of intravenous DB00544 and leucovorin , while results from other trials are awaited . Safety analysis for the 28-day schedule has revealed a low incidence of severe toxicities , particularly as compared with standard DB00544 regimens . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . The P15941 -C oncoprotein binds to the BH3 domain of the pro-apoptotic Q07812 protein and blocks Q07812 function . The pro-apoptotic Q07812 protein contains a BH3 domain that is necessary for its dimerization and for activation of the intrinsic apoptotic pathway . The P15941 ( mucin 1 ) heterodimeric protein is overexpressed in diverse human carcinomas and blocks apoptosis in the response to stress . In this study , we demonstrate that the oncogenic P15941 -C subunit associates with Q07812 in human cancer cells . P15941 -C· Q07812 complexes are detectable in the cytoplasm and mitochondria and are induced by genotoxic and oxidative stress . The association between P15941 -C and Q07812 is supported by the demonstration that the P15941 -C cytoplasmic domain is sufficient for the interaction with Q07812 . The results further show that the P15941 -C cytoplasmic domain CQC motif binds directly to the Q07812 BH3 domain at DB00151 -62 . Consistent with binding to the Q07812 BH3 domain , P15941 -C blocked Q07812 dimerization in response to ( i ) truncated P55957 in vitro and ( ii ) treatment of cancer cells with DNA-damaging agents . In concert with these results , P15941 -C attenuated localization of Q07812 to mitochondria and the release of cytochrome c . These findings indicate that the P15941 -C oncoprotein binds directly to the Q07812 BH3 domain and thereby blocks Q07812 function in activating the mitochondrial death pathway . Immunohistochemical and Q5TCZ1 analysis of Q00987 and P11802 in a dedifferentiated extraskeletal osteosarcoma arising in the vastus lateralis muscle : differential diagnosis and diagnostic algorithm . Extraskeletal osteosarcoma is a rare neoplasia within the broad differential diagnostic spectrum of calcifying intramuscular lesions . We present a case of a slowly increasing mass within the left vastus lateralis muscle . At first presentation the patient showed a partially calcified well defined mass with a diameter of 5 cm and with no direct contact to the femur . A biopsy from the periphery revealed an ossifying lesion compatible with myositis ossificans . The patient returned 18 months later with the lesion having increased to a diameter of 25 cm . The resection specimen revealed a well delimitated tumor with a central core of partially necrotic neoplastic bone . Besides , histology showed high mitotic areas with pleomorphic spindle cells and regions with cartilaginous differentiation . Immunohistochemistry demonstrated : vimentin+ , P28906 - , desmin- , actin- , P15941 - and pancytokeratin- with focal S100 protein positivity and a Ki-67 index of 20 % . Comparative genomic hybridization ( CGH ) revealed a gain of chromosomal material on 12q ; Q5TCZ1 analyses for the P11802 and Q00987 region showed high level amplifications . Consequently , a high-grade dedifferentiated extraskeletal osteosarcoma was diagnosed . In conclusion , analysis of the Q00987 and P11802 status is a powerful and discriminating diagnostic tool to distinguish dedifferentiated extraskeletal osteosarcoma from other benign/malignant ossifying lesions in the skeletal muscle . Antihistamine effects on prefrontal cortex activity during working memory process in preschool children : a near-infrared spectroscopy ( NIRS ) study . P35367 antagonists ( antihistamines ) are widely used for the treatment of allergic disorders in young children . This study examined the effects of antihistamine on prefrontal cortex activity in preschool children using near-infrared spectroscopy ( NIRS ) , an emerging brain-imaging method suitable for psychological experiments , especially in young children . We examined the changes of oxygenated hemoglobin concentration in the prefrontal cortex while children performed a spatial working memory task , 3h after taking a first-generation antihistamine ( ketotifen ) , second-generation antihistamine ( epinastine ) , or placebo . Fifteen healthy preschool children ( mean age , 5.5 years ) participated . DB00920 significantly impaired behavioral performance and cortical activation at the lateral prefrontal cortex in the working memory task , compared with epinastine and placebo . There were no sedative effects on neural response or behavioral performance after epinastine administration . This paper demonstrates for the first time differential sedation effects of first- and second-generation antihistamines on brain hemodynamic response in young children . Also discussed is the utility of the NIRS technique in neuropsychopharmacological studies of children . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . PDE10 inhibition increases P42261 and CREB phosphorylation and improves spatial and recognition memories in a Huntington 's disease mouse model . Huntington 's disease ( HD ) causes motor disturbances , preceded by cognitive impairment , in patients and mouse models . We showed that increased hippocampal DB02527 -dependent protein kinase ( PKA ) signaling disrupts recognition and spatial memories in R6 HD mouse models . However , unchanged levels of hippocampal phosphorylated ( p ) DB02527 -responsive element-binding protein ( CREB ) suggested unaltered nuclear PKA activity in R6 mice . Here , we extend this finding by showing that nuclear pPKA catalytic subunit ( Thr197 ) and pPKA substrate levels were unaltered in the hippocampus of R6/1 mice . Phosphodiesterases ( PDEs ) play an important role in the regulation of PKA activity . Q9Y233 , a DB02527 /cGMP dual-substrate PDE , was reported to be restricted to the nuclear region in nonstriatal neurons . Using cell fractionation we confirmed that Q9Y233 was enriched in nuclear fractions , both in wild-type and R6/1 mice hippocampus , without differences in its levels or intracellular distribution between genotypes . We next investigated whether inhibition of PDE10 with papaverine could improve cognitive function in HD mice . DB01113 treatment improved spatial and object recognition memories in R6/1 mice , and significantly increased pGluA1 and pCREB levels in R6/1 mice hippocampus . DB01113 likely acted through the activation of the PKA pathway as the phosphorylation level of distinct cGMP-dependent kinase ( cGK ) substrates was not modified in either genotype . Moreover , hippocampal DB02527 , but not cGMP , levels were increased after acute papaverine injection . Our results show that inhibition of PDE10 improves cognition in R6 mice , at least in part through increased P42261 and CREB phosphorylation . Thus , PDE10 might be a good therapeutic target to improve cognitive impairment in HD . Receptor kinase inhibitors target NSCLC : two antibodies and a small-molecule MET inhibitor . Joining cetuximab , sorafenib , afatinib , intedanib , and crizotinib in phase III development for non-small cell lung cancer ( NSCLC ) are ramucirumab ( developed by ImClone , a subsidiary of Lilly ) , necitumumab ( developed by ImClone and Bristol-Myers Squibb ) , and tivantinib ( ARQ 197 , developed by ArQule and Daiichi Sankyo ) . DB09559 is a second-generation anti- P00533 monoclonal antibody ( mAb ) similar to cetuximab . Enrollment has been stopped in one of two necitumumab phase III trials because of safety concerns . DB05578 is an anti- P35968 mAb targeting the same pathway as bevacizumab . Although the phase II safety data for ramucirumab appear better than the data for necitumumab , fewer phase III data are available . Tivantinib is a highly selective , orally available MET tyrosine kinase inhibitor . MET is overexpressed in 61 % of NSCLC cases . Although tivantinib is the last of the three agents discussed here to enter phase III , its phase II results are the most robust .

MH_train_1283

MH_train_1283

MH_train_1283

interacts_with DB00877?

multiple_choice

Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5-methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 -dependent enzyme methionine synthase . Q99707 specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells . DB00877 induces Q8NHJ6 (high) Q8N423 (high) dendritic cells promoting a new immunoregulatory pathway . Q8NHJ6 (high) Q8N423 (high) dendritic cells ( DCs ) may cause anergy in P01730 (+)CD45RO(+)CD25(+) T cells transforming them into regulatory T cells ( Tregs ) . Here , we tested whether chronic exposure to rapamycin may modulate this immunoregulatory pathway in renal transplant recipients . Forty renal transplant patients with biopsy-proven chronic allograft nephropathy and receiving calcineurin inhibitors were randomly assigned to either calcineurin inhibitor dose reduction or withdrawal with rapamycin introduction . At conversion and 2 years thereafter , we measured the rapamycin effects on circulating DCs ( BDCA1/ Q8WTT0 and Q8NHJ6 / Q8N423 expression ) , P01730 (+)/CD25(high)/Foxp3(+) Tregs , CD8(+)/ P10747 (-) T cells , and the Th1/Th2 balance in graft biopsies . In rapamycin-treated patients , peripheral Q8WTT0 (+) cells were significantly increased along with Q8NHJ6 / Q8N423 (+) DCs . The number of circulating P01730 (+)/CD25(high)/Foxp3(+)/ P16410 (+) Tregs , CD8(+) P10747 (-) T cells , and P17693 serum levels were higher in the rapamycin-treated group . The number of Q8NHJ6 / Q8N423 (+) Q8WTT0 (+) DC was directly and significantly correlated with circulating Tregs and CD8(+) P10747 (-) T cells . Q8NHJ6 / Q8N423 expression was increased in kidney biopsies at the end of the study period along with a significant bias toward a Th2 response within the graft only in the rapamycin-treated patients . Thus , rapamycin induces the upregulation of Q8NHJ6 and Q8N423 on the DC surface , and this effect is associated with an increase in the number of Tregs and expansion of the CD8(+) P10747 (-) T cell population . This suggests that P42345 inhibition may promote a novel immunoregulatory pathway . Preclinical in vivo evaluation of rapamycin in human malignant peripheral nerve sheath explant xenograft . Neurofibromatosis type 1 ( P21359 ) patients are prone to the development of malignant tumors , the most common being Malignant Peripheral Nerve Sheath Tumor ( MPNST ) . P21359 -MPNST patients have an overall poor survival due to systemic metastasis . Currently , the management of MPNSTs includes surgery and radiation ; however , conventional chemotherapy is not very effective , underscoring the need for effective biologically-targeted therapies . Recently , the P21359 gene product , neurofibromin , was shown to negatively regulate the phosphoinositide-3-kinase ( PI3K ) /Protein Kinase-B ( Akt ) /mammalian Target Of DB00877 ( P42345 ) pathway , with loss of neurofibromin expression in established human MPNST cell lines associated with high levels of P42345 activity . We developed and characterized a human P21359 -MPNST explant grown subcutaneously in NOD-SCID mice , to evaluate the effect of the P42345 inhibitor rapamycin . We demonstrate that rapamycin significantly inhibited human P21359 -MPNST P42345 pathway activation and explant growth in vivo at doses as low as 1.0 mg/kg/day , without systemic toxicities . While rapamycin was effective at reducing P21359 -MPNST proliferation and angiogenesis , with decreased CyclinD1 and P15692 respectively , there was no increase in tumor apoptosis . DB00877 effectively decreased activation of S6 downstream of P42345 , but there was accompanied increased Akt activation . This study demonstrates the therapeutic potential and limitations of rapamycin in P21359 -associated , and likely sporadic , MPNSTs . The combination of rapamycin and MAPK inhibitors enhances the growth inhibitory effect on Nara-H cells . The inhibition of the mammalian target of rapamycin ( P42345 ) signaling pathway promotes the initiation of autophagy , and the mitogen-activated protein kinase ( MAPK ) /extracellular signal-regulated protein kinase ( P29323 ) is well known to induce autophagy . Autophagy is a self-defense mechanism of cancer cells that are subjected to antitumor agents , and blocking autophagy can trigger apoptosis . In the present study , we demonstrate that an P42345 inhibitor , rapamycin , induces autophagy in the Nara-H malignant fibrous histiocytoma ( Q9H334 ) cell line through the activation of P27361 /2 . DB00877 -induced apoptosis was enhanced following the inhibition of the MEK/ P29323 pathway . In the Nara-H cells , we examined the effects of rapamycin treatment on cell proliferation and on the phosphorylation of the P42345 pathway components and autophagy by western blot analysis . Furthermore , we examined the effects of rapamycin with or without the MEK inhibitor , U0126 , on the induction of apoptosis by using fluorescence microscopy . DB00877 inhibited Nara-H cell proliferation and decreased the phosphorylation of the P42345 pathway in the Nara-H cells . DB00877 induced the apoptosis of Nara-H cells , and this apoptosis was enhanced by U0126 . Simultaneously , phospho- P27361 /2 was activated by rapamycin . The present study demonstrates that rapamycin induces autophagy in Nara-H cells by activating the MEK/ P29323 signaling pathway , and the rapamycin-induced apoptosis can be enhanced by the MEK inhibitor , U0126 . These results suggest that self‑protective mechanisms involving P42345 inhibitors in Nara-H cells are prevented by the inhibition of the MEK/ P29323 pathway . The combination of an P42345 inhibitor ( e.g. , rapamycin ) and an MEK inhibitor ( e.g. , U0126 ) may offer effective treatment for Q9H334 , as this combination effectively activates apoptotic pathways . DB01780 signaling reveals 14-3-3 protein function as a novel step in left-right patterning during amphibian embryogenesis . To gain insight into the molecular mechanisms underlying the control of morphogenetic signals by H+ flux during embryogenesis , we tested DB01780 -A ( FC ) , a compound produced by the fungus Fusicoccum amygdali Del . In plant cells , FC complexes with 14-3-3 proteins to activate H+ pumping across the plasma membrane . It has long been thought that FC acts on higher plants only ; here , we show that exposing frog embryos to FC during early development specifically results in randomization of the asymmetry of the left-right ( LR ) axis ( heterotaxia ) . Biochemical and molecular-genetic evidence is presented that 14-3-3-family proteins are an obligate component of Xenopus FC receptors and that perturbation of 14-3-3 protein function results in heterotaxia . The subcellular localization of 14-3-3 mRNAs and proteins reveals novel cytoplasmic destinations , and a left-right asymmetry at the first cell division . Using gain-of-function and loss-of-function experiments , we show that P62258 protein is likely to be an endogenous and extremely early aspect of LR patterning . These data highlight a striking conservation of signaling pathways across kingdoms , suggest common mechanisms of polarity establishment between C. elegans and vertebrate embryos , and uncover a novel entry point into the pathway of left-right asymmetry determination . A longitudinal magnetization transfer imaging evaluation of brain injury in a macaque model of neuroAIDS . Magnetization transfer ( MT ) imaging has been explored in prior studies of HIV patients and showed the potential capacity to assess brain injury after HIV infection . In the present study , adult pig-tailed macaques were infected with a highly neuropathogenic virus SIVsmmFGb . MT imaging was exploited to examine the monkey brains before simian immunodeficiency virus ( SIV ) inoculation and 2 , 4 , 8 , 12 , 16 , and 20 weeks post-SIV inoculation . Blood samples were collected from each animal for monitoring P01730 (+) and CD8(+) T cells before each Q9BWK5 scan . The MT ratios ( Q99707 ) in several brain regions of interest were evaluated longitudinally . Significant reductions of Q99707 were observed in whole brain and selected regions of interest ( genu , splenium , thalamus , caudate , centrum semiovale , frontal white matter , frontal gray matter , and putamen ) in the SIV-infected monkeys , consistent with those reported previously in HIV patients . In particular , the longitudinal results indicate that abnormal Q99707 reduction can be detected as early as in 2 weeks and Q99707 may be more sensitive to the brain injury in cortical regions than in subcortical regions during acute SIV infection . In addition , Q99707 reduction in genu , centrum semiovale , and thalamus significantly correlated with the P01730 (+) T cell percentage decrease . Also , the Q99707 reduction in thalamus correlated with the CD8(+) T cell percentage elevation . Taken together , this study reported the longitudinal evolution of Q99707 in different brain regions during SIV infection and further validates previous findings in HIV patients . The preliminary results suggest that MT imaging could be a robust and sensitive approach to characterize the neurodegeneration after SIV or HIV infection . Decrease in transient receptor potential melastatin 6 mRNA stability caused by rapamycin in renal tubular epithelial cells . DB00877 , an inhibitor of mammalian target of rapamycin ( P42345 ) , is used in treatments for transplantation and cancer . DB00877 causes hypomagnesemia , although precisely how has not been examined . Here , we investigated the effect of rapamycin on the expression of transient receptor potential melastatin 6 ( Q9BX84 ) , a Mg2+ channel . DB00877 and LY-294002 , an inhibitor of phosphatidilinositol-3 kinase ( PI3K ) located upstream of P42345 , inhibited epidermal growth factor ( P01133 ) -induced expression of the Q9BX84 protein without affecting Q96QT4 expression in rat renal Q7Z2Y5 -52E epithelial cells . Both rapamycin and LY-294002 decreased P01133 -induced Mg2+ influx . U0126 , a MEK inhibitor , inhibited P01133 -induced increases in c-Fos , p- P29323 , and Q9BX84 levels . In contrast , neither rapamycin nor LY-294002 inhibited P01133 -induced increases in p- P29323 and c-Fos levels . P01133 increased p-Akt level , an effect inhibited by LY-294002 and 1L-6-hydroxymethyl-chiro-inositol2- [ ( R ) -2-O-methyl-3-O-octadecylcarbonate ] ( Akt inhibitor ) . Akt inhibitor decreased Q9BX84 level similar to rapamycin and LY-294002 . These results suggest that a PI3K/Akt/ P42345 pathway is involved in the regulation of Q9BX84 expression . DB00877 inhibited the P01133 -induced increase in Q9BX84 mRNA but did not inhibit human Q9BX84 promoter activity . In the presence of actinomycin D , a transcriptional inhibitor , rapamycin accelerated the decrease in Q9BX84 mRNA . DB00877 decreased the expression and activity of a luciferase linked with the 3'-untranslated region of human Q9BX84 mRNA . These results suggest that Q9BX84 expression is up-regulated by a PI3K/Akt/ P42345 pathway and rapamycin reduces Q9BX84 mRNA stability , resulting in a decrease in the reabsorption of Mg2+ . A novel role for Bruton 's tyrosine kinase in hepatocyte growth factor-mediated immunoregulation of dendritic cells . The limited success of dendritic cell ( DC ) -based immunotherapy in multiple myeloma is partly due to hepatocyte growth factor ( P14210 ) -induced DC dysfunction . From a therapeutic standpoint , it is important to understand the molecular events involved in inhibition of DC activation/maturation by P14210 . Because Bruton 's tyrosine kinase ( Btk ) negatively regulates maturation and immunostimulatory function of DCs , a role for Btk in P14210 -induced inhibition of both murine and human DCs was investigated . We demonstrate that Btk is a novel proximal component of P14210 -induced c-MET ( P08581 ) signaling . Following P14210 treatment , Btk binds to c-MET and becomes activated . Btk activation in turn blocks the NF-κB pathway and subsequent DC activation via the c-Src-PI3K-AKT-mammalian target of rapamycin ( P42345 ) pathway . Notably , Btk activation is necessary for P14210 -induced association of c-Src and PI3K with c-MET . Furthermore , we provide the first evidence that P14210 inhibits DC activation by inducing autocrine interleukin ( IL ) -10 secretion , which requires activation of Btk . Blocking activation of Btk and its downstream the c-Src-PI3K-AKT- P42345 pathway prevents P14210 -induced P22301 secretion by DCs . In addition , neutralization of P22301 secretion from DCs impaired the inhibitory effect of P14210 on DCs . Thus , our study identifies a novel role for Btk in P14210 -induced DC inhibition . DB00877 antagonizes P01375 induction of P19320 on endothelial cells by inhibiting mTORC2 . Recruitment of circulating leukocytes into inflamed tissues depends on adhesion molecules expressed by endothelial cells ( ECs ) . Here we report that rapamycin pretreatment reduced the ability of P01375 -treated ECs to capture T cells under conditions of venular flow . This functional change was caused by inhibition of P01375 -induced expression of vascular cell adhesion molecule-1 ( P19320 ) and could be mimicked by knockdown of mammalian target of rapamycin ( P42345 ) or rictor , but not raptor , implicating mTORC2 as the target of rapamycin for this effect . Mechanistically , mTORC2 acts through Akt to repress Raf1- Q02750 /2- P27361 /2 signaling , and inhibition of mTORC2 consequently results in hyperactivation of P27361 /2 . Increased P27361 /2 activity antagonizes P19320 expression by repressing P01375 induction of the transcription factor P10914 . Preventing activation of P27361 /2 reduced the ability of rapamycin to inhibit P01375 -induced P19320 expression . In vivo , rapamycin inhibited mTORC2 activity and potentiated activation of P27361 /2 . These changes correlated with reduced endothelial expression of P01375 -induced P19320 , which was restored via pharmacological inhibition of P27361 /2 . Functionally , rapamycin reduced infiltration of leukocytes into renal glomeruli , an effect which was partially reversed by inhibition of P27361 /2 . These data demonstrate a novel mechanism by which rapamycin modulates the ability of vascular endothelium to mediate inflammation and identifies endothelial mTORC2 as a potential therapeutic target . P30305 mediates rapamycin-induced oncogenic responses in cancer cells . Because the mammalian target of rapamycin ( P42345 ) pathway is commonly deregulated in human cancer , P42345 inhibitors , rapamycin and its derivatives , are being actively tested in cancer clinical trials . Clinical updates indicate that the anticancer effect of these drugs is limited , perhaps due to rapamycin-dependent induction of oncogenic cascades by an as yet unclear mechanism . As such , we investigated rapamycin-dependent phosphoproteomics and discovered that 250 phosphosites in 161 cellular proteins were sensitive to rapamycin . Among these , rapamycin regulated four kinases and four phosphatases . A siRNA-dependent screen of these proteins showed that AKT induction by rapamycin was attenuated by depleting cellular P30305 phosphatase . DB00877 induces the phosphorylation of P30305 at Serine375 , and mutating this site to DB00160 substantially reduced P30305 phosphatase activity . Additionally , expression of P30305 ( S375A ) inhibited the AKT activation by rapamycin , indicating that phosphorylation of P30305 is critical for P30305 activity and its ability to transduce rapamycin-induced oncogenic AKT activity . Importantly , we also found that P30305 depletion in various cancer cell lines enhanced the anticancer effect of rapamycin . Together , using rapamycin phosphoproteomics , we not only advance the global mechanistic understanding of the action of rapamycin but also show that P30305 may serve as a drug target for improving P42345 -targeted cancer therapies . DB08865 for the treatment of patients with advanced non-small cell lung cancer . DB08865 is a potent small-molecule inhibitor of Q9UM73 ( anaplastic lymphoma kinase ; Q9UM73 ) and hepatocyte growth factor receptor ( P08581 , proto-oncogene c- DB00134 ) . A range of tumors , including subsets of non-small cell lung cancer ( NSCLC ) , anaplastic large cell lymphoma and inflammatory myofibroblastic tumors harbor an Q9UM73 rearrangement that leads to oncogenic activation of Q9UM73 . DB08865 has demonstrated preclinical and clinical activity against such malignancies through inhibition of Q9UM73 , and patients harboring Q9UM73 - rearranged NSCLC have demonstrated high response rates and prolonged progression-free survival in phase I and II studies . In August 2011 , crizotinib was approved for the treatment of advanced Q9UM73 -positive NSCLC . 14-3-3 epsilon prevents G2/M transition of fertilized mouse eggs by binding with P30305 . BACKGROUND : The 14-3-3 ( YWHA ) proteins are highly conserved in higher eukaryotes , participate in various cellular signaling pathways including cell cycle regulation , development and growth . Our previous studies demonstrated that 14-3-3ε ( P62258 ) is responsible for maintaining prophase | arrest in mouse oocyte . However , roles of 14-3-3ε in the mitosis of fertilized mouse eggs have remained unclear . Here , we showed that 14-3-3ε interacts and cooperates with P30305 phosphorylated at Ser321 regulating G2/M transition of mitotic progress of fertilized mouse eggs . RESULTS : Disruption of 14-3-3ε expression by RNAi prevented normal G2/M transition by inhibition of Q13421 activity and leaded to the translocation of P30305 into the nucleus from the cytoplasm . Overexpression of 14-3-3ε-WT and unphosphorylatable P30305 mutant ( P30305 -S321A ) induced mitotic resumption in dbcAMP-arrested eggs . In addition , we examined endogenous and exogenous distribution of 14-3-3ε and P30305 . Endogenous 14-3-3ε and P30305 were co-localized primarily in the cytoplasm at the P55008 , S , early G2 and M phases whereas P30305 was found to accumulate in the nucleus at the late G2 phase . Upon coexpression with RFP-14-3-3ε , GFP- P30305 -WT and GFP- P30305 -S321A were predominantly cytoplasmic at early G2 phase and then GFP- P30305 -S321A moved to the nucleus whereas P30305 -WT signals were observed in the cytoplasm without nucleus accumulation at late G2 phase at presence of dbcAMP . CONCLUSIONS : Our data indicate that 14-3-3ε is required for the mitotic entry in the fertilized mouse eggs . 14-3-3ε is primarily responsible for sequestering the P30305 in cytoplasm and 14-3-3ε binding to P30305 -S321 phosphorylated by PKA induces mitotic arrest at one-cell stage by inactivation of Q13421 in fertilized mouse eggs . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Divergence in signaling pathways involved in promotion of cell viability mediated by P09038 , P01138 , and P01133 in PC12 cells . We employed a series of inhibitors of intracellular cascade to disclose the precise molecular mechanisms by which basic fibroblast growth factor ( P09038 ) promotes viability of PC12 cells and compared with nerve growth factor ( P01138 ) and epidermal growth factor ( P01133 ) . The Q02750 and 2 inhibitors , U0126 and PD98059 , significantly suppressed cell viability mediated by P09038 in a dose-dependent manner , and to a greater extent compared with P01133 and P01138 . The degree of MEK dependency for growth factor-mediated cell viability was estimated to be in the order of P09038 , P01133 , and P01138 . DB00877 strongly inhibited the effect of P01138 on cell viability , compared with P09038 and P01133 . The mechanisms of action of P01138 -mediated cell viability may depend largely on P08133 S6 kinase-related signal transduction pathways comparing to P09038 and P01133 . The present findings suggest that different signal transduction systems may be involved in the molecular mechanisms by which P09038 , P01138 , and P01133 mediate cell viability . DB08911 : first global approval . DB08911 is an orally bioavailable mitogen-activated protein kinase ( MAPK ) kinase ( MEK ) inhibitor with antineoplastic activity . The compound specifically binds to Q02750 and P36507 , resulting in inhibition of growth factor-mediated cell signalling and cellular proliferation in various cancers . Originally developed by Japan Tobacco , GlaxoSmithKline has licensed exclusive worldwide rights to the compound and conducted development in a number of different cancer types . DB08911 , as a monotherapy , has been approved in the US for the treatment of unresectable or metastatic malignant melanoma with P15056 V600E or V600K mutations , as detected by an FDA-approved test . The compound , as a monotherapy , has also been submitted for regulatory review in the EU for P15056 mutation-positive malignant melanoma , and is in phase III development in Europe , Argentina , Canada and Oceania . Phase II development is underway for pancreatic cancer , non-small cell lung cancer and relapsed or refractory leukaemias . GlaxoSmithKline is also developing trametinib for use in combination with dabrafenib in P15056 V600 mutation-positive metastatic cutaneous melanoma ; the combination is at the preregistration stage in the EU and a phase III clinical programme is underway worldwide . Phase II development for this combination is also underway in colorectal cancer . Several phase I trials have also been initiated to evaluate trametinib in combination with other drugs for the treatment of various solid tumours and haematological malignancies . A paediatric oral solution formulation has been assessed against the oral tablet formulation in a phase I trial . This article summarizes the milestones in the development of trametinib leading to this first approval for unresectable or metastatic P15056 mutation-positive malignant melanoma . The anti-androgen drug dutasteride renders triple negative breast cancer cells more sensitive to chemotherapy via inhibition of HIF-1α-/ P15692 -signaling . BACKGROUND : Triple negative breast cancer ( TNBC ) is characterized by lack of expression of both estrogen and progesterone receptor as well as lack of amplification of P04626 . Patients with TNBC carry an unfavorable prognosis compared to other breast cancer subtypes given that endocrine or P04626 targeted therapies are not effective , rendering chemotherapy the sole effective treatment option to date . Therefore , there is a high demand for additional novel treatment options . FINDINGS : We previously published a list of genes showing both higher gene expression rates in TNBC and , in addition , are known to encode targets of non-oncologic drugs . P18405 , which encodes the type-1 isoform of the steroid-5alpha-reductase , which is involved in androgen metabolism , was found to be one of these genes . DB01126 is a dual blocker of both the type-1 and type-2 isoform of P18405 and is indicated in the treatment of benign prostate hyperplasia . Treatment of TNBC cell lines with dutasteride was associated with a dose-dependent decrease in cell viability , altered protein expression of P15692 and HIF-1α and increased chemosensitivity . CONCLUSION : Our results demonstrate that the P18405 -corresponding anti-androgenic drug dutasteride might act as a combinatorial therapeutic option besides standard chemotherapy in highly aggressive TNBC . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Role of chronic inhibition of dopamine-metabolizing enzymes in the regulation of renal sodium and phosphate excretion in the rat remnant kidney . BACKGROUND/AIMS : The present study examined the effects of chronic selective or combined inhibition of type A monoamine oxidase ( MAO ) and catechol-O-methyltransferase ( P21964 ) on daily urinary excretion of dopamine and metabolites and on natriuresis and phosphaturia in 3/4 nephrectomized ( 3/4nx ) and Sham rats . METHODS : The 3/4nx and Sham rats were placed in metabolic cages and received the P21397 -selective inhibitor Ro-411049 ( 7.5 mg x kg(-1) bid ) and/or the P21964 -selective inhibitor DB03336 3-202 ( 30 mg x kg(-1) bid ) orally for 3 days during high sodium diet . RESULTS : Selective P21964 inhibition increased the urinary excretion of the deaminated metabolite ( 3,4-dihydroxyphenylacetic acid , DOPAC ) and decreased the urinary excretion of the methylated ( 3-methoxytyramine , 3-MT ) and deaminated plus methylated metabolite ( homovanillic acid , HVA ) in both groups . Selective P21397 inhibition increased the urinary excretion of 3-MT and reduced the urinary excretion of both DOPAC and HVA in either 3/4nx or Sham rats . Combined inhibition of P21397 and P21964 did not significantly change the urinary excretion of DOPAC and markedly decreased the urinary excretion of 3-MT and HVA in both groups . Selective or combined inhibition of P21397 and P21964 did not alter the daily urinary excretion of dopamine , sodium or phosphate in either 3/4nx or Sham rats . CONCLUSIONS : Chronic selective or combined inhibition of P21397 and P21964 is not of major importance in regulating the dopamine-dependent natriuresis and phosphaturia in either 3/4nx or Sham rats . Gateways to clinical trials . 11D10 , 9vPnC-MnCc ; DB00051 , DB00718 , DB00092 , ALN-RSV01 , AME-133 , Q99217 -317 , DB00992 , Amlodipine besylate/atorvastatin calcium , Anisodamine , Anti- P05113 receptor antibody , Apremilast , Aripiprazole , Atacicept , DB01072 sulfate , Atrasentan ; DB04975 , DB00112 , BIBW-2992 , DB04853 , BMS-387032 ; cAC10 , Caldaret hydrate , CD-NP , DB04918 medocaril , Celivarone fumarate , DB08904 , Cholesteryl hydrophobized polysaccharide-Her2 protein complex , Choline fenofibrate , Cilengitide , Cinaciguat , Curcumin , Custirsen sodium , Cypher , CYT-6091 ; Dalcetrapib , Deforolimus , Desvenlafaxine succinate , DB05297 , DP6-001 ; E-7010 , E75 , Ecogramostim , P01133 -P64K , EnvPro , Enzastaurin hydrochloride , DB01175 oxalate , DB00973 , DB00973 /simvastatin ; DB05076 ; Gefitinib , Golimumab , Green tea catechins , GTI-2040 , GW-406381 ; HPV16 E6 E7 , HPV-16/18 AS04 , HPV-6/11/16/18 ; ICC-1132 , Immune globulin intravenous ( human ) , DB05039 , Intranasal insulin ; Kahalalide F ; Lactobacillus rhamnosus , Laromustine , Laropiprant , GTI-2040 ; MAb 3H1 , DB06612 , Mifamurtide , Milataxel , DB05313 ; Nebicapone , DB01280 , Neuradiab ; Oncolytic HSV ; PCV7 , PHX-1149 , Pimecrolimus , DB06813 , Pramiconazole ; DB01270 , Reolysin , DB06372 , Rolofylline , DB06176 ; S-32865 , Shigella dysenteriae 1 vaccine ; Taranabant , Taxus , DB05657 ; Ustekinumab ; Vitespen ; Zileuton , Zycure . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually . Cl- DB05511 enhances P01375 -α release in peritoneal macrophages stimulated with LPS . P0DMS8 ( A3R ) belongs to the Gi/Gq-coupled receptor family , that leads to the intracellular DB02527 reduction and intracellular calcium increase , respectively . A3R is widely expressed and it can play a crucial role in many patho-physiological conditions , including inflammation . Here we investigate the effect of Cl- DB05511 , A3R agonist , on the production of P01375 -α . We found that Cl- DB05511 enhances LPS-induced P01375 -α release in peritoneal macrophages . This effect is reduced by MRS1191 , A3R antagonist and by forskolin , activator of adenylyl cyclase . pIκBα increased in LPS+Cl- DB05511 -treated macrophages , while total IκB kinase-β ( IKKβ ) reduced . Indeed , p65NF-κB nuclear translocation increased in cells treated with LPS+Cl- DB05511 . Moreover , IMD 0354 , IKKβ inhibitor , significantly abrogated the effect of Cl- DB05511 on P01375 -α release . Inhibition of protein kinase C ( PKC ) significantly reduced Cl- DB05511 -induced P01375 -α release in LPS-stimulated macrophages . Furthermore , LY-294002 , PI3K inhibitor , reduced the P01375 -α production enhanced by Cl- DB05511 , although the phosphorylation status of Akt did not change in cells treated with LPS+Cl- DB05511 than LPS alone . In summary , these data show that Cl- DB05511 is able to enhance P01375 -α production in LPS-treated macrophages in an NF-κB- dependent manner .

MH_train_1284

MH_train_1284

MH_train_1284

interacts_with DB08815?

multiple_choice

[ DB02424 administration reduces the number of P07900 -positive germ cells in the mouse embryo : preliminary results ] . 5 mg of DB02424 , an inhibitor of stress protein P07900 which express on mammalian germ cells , were administered to E8 pregnant mice . E17 embryos were removed , and a quantitative analysis of HSP90-immunoreactive cells in the gonad was performed , in comparison to control embryos . First , we observed that the number of germ cells is lower in male than in female embryos , as well in control and experimental embryos . External features of experimental and control embryos did not display any difference . Embryos exposed to geldanamycin exhibit a significant decrease of immunoreactive germ cells . In two embryos , we observed a group of ectopic immunoreactive cells in the pelvic area . We conclude that geldanamycin inhibits germ cells migration , and suggest that this inhibition can lead to ectopic germ cell populations , similar to teratomas . Q8N0V5 V ( Mgat5 ) -mediated N-glycosylation negatively regulates Th1 cytokine production by T cells . The differentiation of naive P01730 (+) T cells into either proinflammatory Th1 or proallergic Th2 cells strongly influences autoimmunity , allergy , and tumor immune surveillance . We previously demonstrated that beta1,6GlcNAc-branched complex-type ( Q8N0V5 V ( Mgat5 ) ) N-glycans on TCR are bound to galectins , an interaction that reduces TCR signaling by opposing agonist-induced TCR clustering at the immune synapse . Mgat5(-/-) mice display late-onset spontaneous autoimmune disease and enhanced resistance to tumor progression and metastasis . In this study we examined the role of beta1,6GlcNAc N-glycan expression in Th1/Th2 cytokine production and differentiation . beta1,6GlcNAc N-glycan expression is enhanced by TCR stimulation independent of cell division and declines at the end of the stimulation cycle . DB00075 -activated splenocytes and naive T cells from Mgat5(-/-) mice produce more P01579 and less P05112 compared with wild-type cells , the latter resulting in the loss of P05112 -dependent down-regulation of IL-4Ralpha . DB02034 , an inhibitor of Q16706 , blocked beta1,6GlcNAc N-glycan expression and caused a similar increase in P01579 production by T cells from humans and mice , but no additional enhancement in Mgat5(-/-) T cells . Mgat5 deficiency did not alter P01579 / P05112 production by polarized Th1 cells , but caused an approximately 10-fold increase in P01579 production by polarized Th2 cells . These data indicate that negative regulation of TCR signaling by beta1,6GlcNAc N-glycans promotes development of Th2 over Th1 responses , enhances polarization of Th2 cells , and suggests a mechanism for the increased autoimmune disease susceptibility observed in Mgat5(-/-) mice . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Leukocytosis and Mobilization of P28906 + Hematopoietic Progenitor Cells by DB06809 , a P61073 Antagonist . Stromal cell-derived factor-1 ( P48061 / P48061 ) plays a key regulatory role in the trafficking of hematopoietic cells . DB06809 is a specific antagonist of the binding of P48061 to its receptor , P61073 . This phase I study assessed the hematological effects , pharmacokinetics , and safety of administration of DB06809 to 32 healthy volunteers , including its ability to mobilize P28906 + hematopoietic progenitor cells . A generalized leukocytosis occurred after a single subcutaneous injection of DB06809 ( 80 microg/kg ) resulting in a maximum white blood cell count of 19.49 +/- 1.27 x 103/microL ( mean +/- SEM ) at 6 hours . No changes were observed in erythrocyte or platelet counts . Circulating P28906 + cells increased 5-fold after administration of DB06809 at 80 mug/kg and 15.5-fold in response to DB06809 at 240 mug/kg , both at 9 hours after injection . Myeloid progenitor cells-colony forming unit granulocytemacrophage ( CFU-GM ) ; CFU-granulocyte , eosinophil , monocyte , megakaryocyte ( CFU-GEMM ) ; and burst forming units-erythroid showed similar increases in mobilization to the blood with increasing doses of DB06809 . The mobilized cells were in a slow or noncycling state as determined by in vitro high specific activity of 3H-thymidine . Pharmacokinetic studies showed a near linear increase in peak drug levels with increasing doses and nearly complete elimination of the drug by 24 hours . DB06809 was well tolerated with only mild and transient toxicities ( injection site erythema , headache , paresthesia , nausea , and abdominal distension ) observed . These observations suggest that DB06809 may be a clinically useful agent for hematopoietic progenitor cell mobilization . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) 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 ) -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 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 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 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- 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 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . DB00939 sodium is an inhibitor of both the P09917 and cyclooxygenase pathways of the arachidonic acid cascade in vitro . DB00939 sodium was compared to other nonsteroidal antiinflammatory drugs in terms of its potency to inhibit the formation of 5-HETE and LTB4 in human leukocytes and the formation of prostaglandin E2 in bovine seminal vesicles as measures of its ability to inhibit the P09917 and cyclooxygenase pathways of the arachidonic acid cascade . DB00939 sodium was about 2-4 times less potent than BW-755C in inhibiting P09917 enzyme activity and three times more potent than benoxaprofen , while naproxen , ibuprofen , and indomethacin showed IC50 greater than 100 microM . DB00939 sodium and indomethacin were the most potent inhibitors of the formation of DB00917 in bovine seminal vesicles followed by ibuprofen , naproxen , and benoxaprofen in this order . DB00939 sodium , like BW-755C , can be considered a dual inhibitor of P09917 and cyclooxygenase pathways of arachidonic acid cascade . This finding may explain in part the antiinflammatory activity of meclofenamate sodium . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Effects of lurasidone on executive function in common marmosets . Cognitive impairment is one of the major symptoms of schizophrenia , and is considered largely due to dysfunctions in the prefrontal cortex ( P27918 ) . DB08815 , a novel atypical antipsychotic agent with high binding affinity for dopamine D2 , serotonin P34969 , 5- Q13049 and P08908 receptors has been reported to have superior efficacy in rodents ' models of cognitive impairment . However , the beneficial effect of lurasidone on cognitive impairment has not been evaluated in non-human primates . In this study , we investigated the effect of lurasidone on executive function , which is one of the cognitive domains , in common marmosets and compared the results to those of other antipsychotics . The effects of lurasidone , haloperidol , olanzapine , risperidone , quetiapine and clozapine on executive function were evaluated in naïve marmosets using the object retrieval with detours ( ORD ) task . Before drug treatment , marmosets ' success rates in the easy trial of the test were almost 90 % . However , maximum success in the difficult trial of the task reached only 50 % after 8 days of training . DB00502 , olanzapine and risperidone decreased correct performance even in the easy trial of the task . All drugs , except lurasidone , impaired success rate in the difficult trial . On the other hand , lurasidone dose-dependently increased marmosets ' success rates in the difficult trial with significant effect at 10mg/kg . In conclusion , we have shown in this study that lurasidone , unlike conventional antipsychotics , improves cognition associated with executive function in common marmosets . These findings suggest that lurasidone would be more useful for treatment of schizophrenia cognitive impairment than other antipsychotics . Chemical ablation of androgen receptor in prostate cancer cells by the histone deacetylase inhibitor LAQ824 . P10275 plays a critical role in the development of primary as well as advanced hormone-refractory prostate cancer . Therefore , ablation of androgen receptor from prostate cancer cells is an interesting concept for developing a new therapy not only for androgen-dependent prostate cancer but also for metastatic hormone-refractory prostate cancer , for which there is no effective treatment available . We report here that LAQ824 , a cinnamyl hydroxamatic acid histone deacetylase inhibitor currently in human clinical trials , effectively depleted androgen receptor in prostate cancer cells at nanomolar concentrations . LAQ824 seemed capable of depleting both the mutant and wild-type androgen receptors in either androgen-dependent and androgen-independent prostate cancer cells . Although LAQ824 may exert its effect through multiple mechanisms , several lines of evidence suggest that inactivation of the heat shock protein-90 ( Hsp90 ) molecular chaperone is involved in LAQ824-induced androgen receptor depletion . Besides androgen receptor , LAQ824 reduced the level of Hsp90 client proteins HER-2 ( ErbB2 ) , Akt/ P31749 , and P04049 in LNCaP cells . Another Hsp90 inhibitor , 17-allyamino-17-demethoxygeldanamycin ( 17- P29372 ) , also induced androgen receptor diminution . LAQ824 induced Hsp90 acetylation in LNCaP cells , which resulted in inhibition of its DB00171 -binding activity , dissociation of Hsp90-androgen receptor complex , and proteasome-mediated degradation of androgen receptor . Consequently , LAQ824 blocked androgen-induced prostate-specific antigen production in LNCaP cells . LAQ824 effectively inhibited cell proliferation and induced apoptosis of these prostate cancer cells . These results reveal that LAQ824 is a potent agent for depletion of androgen receptor and a potential new drug for prostate cancer . Role of serotonin in the regulation of interferon-gamma production by human natural killer cells . Monocytes , recovered directly from peripheral blood by counter-current centrifugal elutriation ( CCE ) , were shown to provide two regulatory signals for induction of interferon-gamma ( P01579 ) in natural killer ( NK ) cells in response to interleukin-2 ( P60568 ) : an upregulating signal and an inhibitory signal . The inhibitory signal was time-dependent , irreversible , and operating on a pretranslational level , as indicated by the inability of enriched NK cells to accumulate P01579 mRNA in the presence of elutriated monocytes . Monocyte-induced inhibition of P01579 production was abrogated by the biogenic amine serotonin , acting via the 5-hydroxytryptamine , or serotonin ( P08908 ) , subset of serotonin receptors ( 5-HTR ) . Thereby , serotonin effectively promoted P01579 production in the presence of monocytes . We conclude that serotonergic P08908 receptors transduce signals that are required for NK cells to produce P01579 in response to P60568 . Novel combination treatments targeting chronic myeloid leukemia stem cells . Chronic myeloid leukemia ( CML ) is currently considered incurable in most patients . Stem cell transplantation , an accepted curative option for which extensive experience has been gained , is limited by high morbidity and mortality rates , particularly in older patients . Tyrosine kinase inhibitors targeting P11274 - P00519 are widely used and induce remission in a high proportion of patients , but resistance and incomplete response to these agents portends eventual relapse and disease progression . Although P11274 - P00519 inhibitors eradicate most CML cells , they are largely ineffective against the reservoir of quiescent leukemic stem cells ( LSCs ) . Thus a strong medical need exists for therapies that effectively eradicate LSCs and is currently a focus of extensive research . To date , evidence obtained from in vitro studies , animal models , and clinical CML specimens suggests that an effective approach may be to partner existing P11274 - P00519 inhibitors with compounds targeting key stem cell molecular effectors , including Wnt/β-catenin , hedgehog pathway components , histone deacetylase ( HDAC ) , transforming growth factor-β ( TGF-β ) , O60674 , promyelocytic leukemia protein , and arachidonate P09917 ( P09917 ) . Novel combinations may sensitize LSCs to P11274 - P00519 inhibitors , thereby overcoming resistance and creating the possibility of improving disease outcome beyond the current standard of care . Vitamin D deficiency modulates Graves ' hyperthyroidism induced in BALB/c mice by thyrotropin receptor immunization . DB00024 receptor ( P16473 ) antibodies and hyperthyroidism are induced by immunizing mice with adenovirus encoding the P16473 or its A-subunit . Depleting regulatory T cells ( Treg ) exacerbates thyrotoxicosis in susceptible BALB/c mice and induces hyperthyroidism in normally resistant C57BL/6 mice . Vitamin D plays an important role in immunity ; high dietary vitamin D intake suppresses ( and low intake enhances ) adaptive immune responses . Vitamin D-induced immunosuppression may enhance Treg . Therefore , we hypothesized that decreased vitamin D intake would mimic Treg depletion and enhance hyperthyroidism induced by A-subunit adenovirus immunization . BALB/c mice had a reduced ability vs. C57BL/6 mice to generate the active metabolite of vitamin D ( 1,25-dihydroxyvitamin D3 ) . Vitamin D deficiency induced subtle immune changes in BALB/c ( not C57BL/6 ) mice . Compared with mice fed regular chow , vitamin D-deprived BALB/c mice had fewer splenic B cells and decreased interferon-gamma responses to mitogen and lacked memory T-cell responses to A-subunit protein . However , vitamin D deficiency did not alter P16473 antibody responses measured by ELISA , DB00024 binding inhibition , or DB02527 generation from P16473 -expressing cells . Unexpectedly , compared with vitamin D-sufficient mice , vitamin D-deficient BALB/c mice had lower preimmunization T(4) levels and developed persistent hyperthyroidism . This difference was unrelated to the immunological changes between vitamin D-deficient or -sufficient animals . Previously , we found that different chromosomes or loci confer susceptibility to P16473 antibody induction vs. thyroid function . Our present studies provide evidence that an environmental factor , vitamin D , has only minor effects on induced immunity to the P16473 but directly affects thyroid function in mice . Characterization of the P40933 niche in primary and secondary lymphoid organs in vivo . P40933 is a cytokine critical for development , maintenance , and response of T cells , natural killer ( NK ) cells , NK T cells , and dendritic cells . However , the identity and distribution of P40933 -expressing cells in lymphoid organs are not well understood . To address these questions , we established and analyzed P40933 - P27918 knock-in mice . We found that P40933 was highly expressed in thymic medulla , and medullary thymic epithelial cells with high MHC class II expression were the major source of P40933 . In bone marrow , P40933 was detected primarily in P19320 (+)PDGFRβ(+)CD31(-)Sca-1(-) stromal cells , which corresponded to previously described P48061 -abundant reticular cells . In lymph nodes , P40933 -expressing cells were mainly distributed in the T-cell zone and medulla . P40933 was expressed in some fibroblastic reticular cells and gp38(-)CD31(-) double-negative stromal cells in the T-cell zone . Blood endothelial cells , including all high endothelial venules , also expressed high P40933 levels in lymph nodes , whereas lymphatic endothelial cells ( LECs ) lacked P40933 expression . In spleen , P40933 was expressed in P19320 (+) stromal cells , where its expression increased as mice aged . Finally , P40933 expression in blood and LECs of peripheral lymphoid organs significantly increased in LPS-induced inflammation . Overall , we have identified and characterized several P40933 -expressing cells in primary and secondary lymphoid organs , providing a unique perspective of P40933 niche in immune microenvironment . This study also suggests that some stromal cells express P13232 and P40933 differentially and suggests a way to functionally classify different stromal cell subsets . Molecular sampling of the allosteric binding pocket of the DB00024 receptor provides discriminative pharmacophores for antagonist and agonists . The P16473 ( thyrotropin receptor ) is activated endogenously by the large hormone thyrotropin and activated pathologically by auto-antibodies . Both activate and bind at the extracellular domain . Recently , SMLs ( small-molecule ligands ) have been identified , which bind in an allosteric binding pocket within the transmembrane domain . Modelling driven site-directed mutagenesis of amino acids lining this pocket led to the delineation of activation and inactivation sensitive residues . Modified residues showing CAMs ( constitutively activating mutations ) indicate signalling-sensitive positions and mark potential trigger points for agonists . Silencing mutations lead to an impairment of basal activity and mark contact points for antagonists . Mapping these residues on to a structural model of P16473 indicates locations where an SML may switch the receptor to an inactive or active conformation . In the present article , we report the effects of SMLs on these signalling-sensitive amino acids at the P16473 . Surprisingly , the antagonistic effect of SML compound 52 was reversed to an agonistic effect , when tested at the P62158 Y667A . Switching agonism to antagonism and the reverse by changing either SMLs or residues covering the binding pocket provides detailed knowledge about discriminative pharmacophores . It prepares the basis for rational optimization of new high-affinity antagonists to interfere with the pathogenic activation of the P16473 . Distribution of polyamines and their biosynthetic enzymes in intestinal adaptation . P11926 ( ODC ) and the polyamines have been shown to be important for growth processes in the intestinal mucosa . The highest activity of ODC is found in the differentiated , nonproliferating villus-tip cells rather than in the rapidly proliferating undifferentiated crypt cells . During poststarvation refeeding and lactation , we now show that increases in ODC activity paralleled the time course of mucosal hyperplasia and thymidine incorporation . Increases in ODC ( threefold ) were similar in villus and crypt cells , and the villus-crypt gradient of decreasing ODC activity ( 40:1 ) was maintained . The activity of the other polyamine biosynthetic enzyme , S-adenosylmethionine decarboxylase ( P18827 ) , was highest in the crypt cells in the basal state and increased throughout the entire villus-crypt axis during refeeding and lactation , preserving a villus-crypt gradient opposite to that of ODC . During hyperplasia , all three polyamines increased . DB01917 was highest in the villus-tip cells , paralleling ODC activity , whereas spermidine and spermine were highest in the crypt cells and paralleled the distribution of P18827 activity . Thus P18827 activity and spermidine and spermine content may play a more important role than ODC and putrescine in regulation of intestinal mucosal proliferation . It is also possible that the threefold increases in the low levels of ODC in the crypt cells are adequate to trigger cell proliferation , whereas the higher ODC levels in villus cells may represent an association with the differentiation of the enterocytes . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Polymorphisms of the HTR1a allele are linked to frontal brain electrical asymmetry . Polymorphic variations in genes related to serotonin synthesis , transport , recognition , or degradation may convey subtle changes in serotonin system architecture that may place an individual at risk for psychopathology when faced with life stressors . The relationship between three key serotonin alleles and frontal brain electrical asymmetry , a putative endophenotype of depression , was examined . Risk alleles were hypothesized to predict relatively greater right frontal brain activity regardless of current clinical state . A sample of 313 college-age individuals , spanning a range of depressive severity from no symptomotology to clinically meaningful levels , participated . Resting encephalographic ( EEG ) activity was recorded from 64 scalp sites on four occasions separated by at least 24h ( two 8-min recording sessions occurring at each occasion ) . Alpha power asymmetry scores between homologous sites were calculated for each session and then averaged to form a trait metric of asymmetry for each pair . PCR based genotyping was conducted for the HTR1a , HTR2a , and HTTLPR genes . Variations in the HTR1a gene were related to trait EEG asymmetry , regardless of any history of depression . Compared to subjects with at least one non-risk allele , subjects with homozygous P08908 risk alleles had significantly greater relative right frontal activity at sites P08709 / P00451 , P12259 /F6 , and F1/F2 . In conclusion , variation in HTR1a can influence trait level brain activity , which may ultimately be indicative of risk for psychopathology . Biological production of monoethanolamine by engineered Pseudomonas putida P28222 . Pseudomonas putida P28222 was engineered for the production of monoethanolamine ( MEA ) from glucose via the decarboxylation of the central metabolite L-serine , which is catalyzed by the enzyme L-serine decarboxylase ( P18827 ) . The host was first evaluated for its tolerance towards MEA as well as its endogenous ability to degrade this alkanolamine . Growth inhibition was observed at MEA concentrations above 100 mM , but growth was never completely arrested even at 750 mM of MEA . P. putida P28222 was able to catabolize MEA in the absence of ammonia , but deletion of the eutBC genes that encode ethanolamine ammonia-lyase ( EAL ) enzyme sufficed to eliminate this capacity . For the biological production of MEA , the sdc genes from Arabidopsis thaliana ( full-length and a truncated version ) and Volvox carteri were expressed in P. putida P28222 . From 20 mM of glucose , negligible amounts of MEA were produced by P. putida P28222 ΔeutBC expressing the sdc genes from A. thaliana and V. carteri . However , 0.07 mmol of MEA was obtained per g of cell dry weight of P. putida P28222 ΔeutBC expressing the truncated variant of the A. thaliana P18827 . When the medium was supplemented with L-serine ( 30 mM ) , MEA production increased to 1.25 mmol MEA g⁻¹ CDW , demonstrating that L-serine availability was limiting MEA production . Phosphorylation regulates the activity of the eEF-2-specific Ca(2+)- and calmodulin-dependent protein kinase III . The activity of the eukaryotic elongation factor 2 ( eEF-2 ) -specific Ca(2+)- and calmodulin-dependent protein kinase III ( P62158 PK III ) is regulated by phosphorylation . The kinase can be inactivated by treatment with alkaline phosphatase and subsequently reactivated by endogenous protein kinase . This kinase can be substituted for by the catalytic subunit of DB02527 -dependent protein kinase but not by casein kinase II . The purified kinase preparation contains only one protein as judged by gel electrophoresis . This protein has a molecular mass of approximately 90 kDa and an isoelectric point of 5.2 . Reactivation of the O00418 is associated with the phosphorylation of this protein . The amino acid sequence obtained from the 90-kDa protein reveals substantial homology with that of murine heat shock protein 86 ( P07900 ) a member of the P08238 -family . Conventional preparations of P08238 contain an inactive O00418 that could be activated after dephosphorylation and phosphorylation by the catalytic subunit of DB02527 -dependent protein kinase . The effects of early and late administration of inhibitors of inducible nitric oxide synthase in a thioacetamide-induced model of acute hepatic failure in the rat . BACKGROUND/AIMS : DB00435 ( NO ) is a pivotal mediator of inflammation . Its role in acute hepatic failure ( P00451 ) is controversial . We investigated the role of NO , and the hypothesis that inhibition of inducible NO synthase ( P35228 ) activity would improve outcome in liver failure in rats , using the P35228 inhibitors L-NAME and aminoguanidine ( Q99217 ) . METHODS : P00451 was induced by two intraperitoneal injections of thioacetamide ( TAA ) . Seven groups ( n=10 ) were studied . Group I : TAA alone . Groups II , III and IV were additionally pre-treated with the NO precursor L-arginine ( 300 mg/kg i.p. ) , or P35228 inhibitors Q99217 ( 100 mg/kg s.c. ) , or N(G)-nitro-L-arginine methyl ester ( L-NAME ) ( 100 mg/kg s.c. ) for 5 days , respectively . Groups V , VI and VII received L-arginine , Q99217 or L-NAME commencing immediately after TAA administration . Clinical and biochemical parameters were assessed serially , and mortality investigated in further similar cohorts for each regime . RESULTS : Q99217 , pre-treatment but not post-treatment , significantly improved outcome including mortality ( 10 vs. 70 % , P < 0.005 ) . The less selective P35228 inhibitor L-NAME was not beneficial . DB00125 pre-and post-treatment , and P35228 inhibition post-treatment , worsened clinical parameters of TAA-induced liver failure . CONCLUSIONS : Administration of the P35228 inhibitor Q99217 prior to insult reduces the severity of damage and improves mortality .

MH_train_1285

MH_train_1285

MH_train_1285

interacts_with DB00215?

multiple_choice

NOS Inhibition Modulates Immune Polarization and Improves Radiation-Induced Tumor Growth Delay . DB00435 synthases ( NOS ) are important mediators of progrowth signaling in tumor cells , as they regulate angiogenesis , immune response , and immune-mediated wound healing . Ionizing radiation ( IR ) is also an immune modulator and inducer of wound response . We hypothesized that radiation therapeutic efficacy could be improved by targeting NOS following tumor irradiation . Herein , we show enhanced radiation-induced ( 10 Gy ) tumor growth delay in a syngeneic model ( C3H ) but not immunosuppressed ( Nu/Nu ) squamous cell carcinoma tumor-bearing mice treated post-IR with the constitutive NOS inhibitor N(G)-nitro-l-arginine methyl ester ( L-NAME ) . These results suggest a requirement of T cells for improved radiation tumor response . In support of this observation , tumor irradiation induced a rapid increase in the immunosuppressive Th2 cytokine P22301 , which was abated by post-IR administration of L-NAME . In vivo suppression of P22301 using an antisense P22301 morpholino also extended the tumor growth delay induced by radiation in a manner similar to L-NAME . Further examination of this mechanism in cultured Jurkat T cells revealed L-NAME suppression of IR-induced P22301 expression , which reaccumulated in the presence of exogenous NO donor . In addition to L-NAME , the guanylyl cyclase inhibitors ODQ and thrombospondin-1 also abated IR-induced P22301 expression in Jurkat T cells and Q14201 -1 macrophages , which further suggests that the immunosuppressive effects involve P29474 . Moreover , cytotoxic Th1 cytokines , including P60568 , IL12p40 , and IFNγ , as well as activated CD8(+) T cells were elevated in tumors receiving post-IR L-NAME . Together , these results suggest that post-IR NOS inhibition improves radiation tumor response via Th1 immune polarization within the tumor microenvironment . Oleanolic acid induces relaxation and calcium-independent release of endothelium-derived nitric oxide . BACKGROUND AND PURPOSE : The present study investigated the mechanisms by which oleanolic acid , a component of olive oil , increases release of nitric oxide ( NO ) . EXPERIMENTAL APPROACH : Measurements of isometric tension , NO concentration , or endothelial cell calcium were made in rat isolated mesenteric arteries . Immunoblotting for endothelial NOS ( P29474 ) and Akt kinase were performed in primary cultures of human umbilical vein endothelial cells ( HUVECs ) . KEY RESULTS : Oleanolic acid ( 3-30 microM ) evoked endothelium-dependent relaxations in noradrenaline-contracted rat superior and small mesenteric arteries . In rat superior mesenteric arteries , oleanolic acid induced simultaneous increases in NO concentration and relaxation , and these responses were inhibited by an inhibitor of NOS , asymmetric DB01686 ( 300 microM ) and by the NO scavenger , oxyhaemoglobin ( 10 microM ) . Oleanolic acid-evoked NO increases were not reduced in Ca(2+)-free solution and in the presence of an inhibitor of endoplasmic reticulum calcium-ATPase , thapsigargin ( 1 microM ) . Oleanolic acid evoked relaxation without changes in endothelial cell calcium , but decreased smooth muscle calcium in arterial segments . Oleanolic acid failed to increase calcium in HUVECs , but increased time-dependently phosphorylation of Akt kinase at DB00133 (473) ( Akt- DB00133 (473) ) and P29474 at DB00133 (1177) ( P29474 - DB00133 (1177) ) , which was attenuated by inhibitors of phosphoinositide-3-kinase . CONCLUSIONS AND IMPLICATIONS : This study provides direct evidence that a component of olive oil , oleanolic acid , activated endothelium-dependent release of NO and decreased smooth muscle cell calcium followed by relaxation . The oleanolic acid-evoked endothelium-derived NO release was independent of endothelial cell calcium and involved phosphoinositide-3-kinase-dependent phosphorylation of Akt- DB00133 (473) followed by phosphorylation of P29474 - DB00133 (1177) . Serotonin and vasopressin interact in the hypothalamus to control communicative behavior . The present study investigated whether serotonin ( 5-HT ) agonists could inhibit the ability of arginine-vasopressin ( AVP ) to induce a form of scent marking called flank marking by their actions in the medial preoptic-anterior hypothalamus ( MPOA-AH ) . DOI , a 5- Q13049 ,2B,2C receptor agonist , did not inhibit AVP-induced flank marking , but mCPP a 5- Q13049 antagonist and P41595 ,2C agonist inhibited AVP-induced flank marking . In addition , the finding that 8-OH-DPAT , CGS-12066A and SC53116 also inhibited AVP-induced flank marking suggests that 5-HT could also inhibit flank marking by acting through P08908 , P34969 , P28222 and/or Q13639 receptor subtypes . These data support the hypothesis that 5-HT acts within the MPOA-AH to inhibit the ability of AVP to induce flank marking . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . Green tea polyphenol extract attenuates inflammation in interleukin-2-deficient mice , a model of autoimmunity . Green tea polyphenols ( GrTP ) have been previously shown to decrease endotoxin-induced tumor necrosis factor-alpha production and lethality in mice . Our present studies demonstrate that GrTP inhibit inflammatory responses and may be useful in treating chronic inflammatory states , such as inflammatory bowel disease . In this preliminary study , we examined whether GrTP decrease disease activity in interleukin-2-deficient ( P60568 (-/-) mice . Eight-week old P60568 (-/-) C57BL/6J mice who were fed nonpurified diet were randomly assigned to receive water with GrTP ( 5 g/L ) or water alone ( control ) for up to 6 wk . After 1 wk , explant colon and lamina propria lymphocyte ( P06858 ) cultures were established from a subgroup of mice and supernatants collected . Culture supernatants from GrTP-treated mice showed decreased spontaneous interferon-gamma and tumor necrosis factor-alpha secretion compared with that of controls . At 6 wk , the GrTP group had less severe colitis as demonstrated by lower histologic scores and wet colon weights . This was associated with lower plasma levels of serum amyloid A , increased weight gain and improved hematocrits . These results show that GrTP attenuated inflammation in P60568 (-/-) mice and suggest a role for GrTP in treating chronic inflammatory diseases such as inflammatory bowel disease . Q9BVG9 : high efficiency for synthesizing phosphatidylserine containing docosahexaenoic acid . DB00144 ( PS ) , the major anionic phospholipid in eukaryotic cell membranes , is synthesized by the integral membrane enzymes PS synthase 1 ( PSS1 ) and 2 ( Q9BVG9 ) . Q9BVG9 is highly expressed in specific tissues , such as brain and testis , where docosahexaenoic acid ( DB01708 , 22:6n-3 ) is also highly enriched . The purpose of this work was to characterize the hydrocarbon-chain preference of Q9BVG9 to gain insight on the specialized role of Q9BVG9 in PS accumulation in the DB01708 -abundant tissues . Flag-tagged Q9BVG9 was expressed in P29320 cells and immunopurified in a functionally active form . Purified Q9BVG9 utilized both PE plasmalogen and diacyl PE as substrates . Nevertheless , the latter was six times better utilized , indicating the importance of an ester linkage at the sn-1 position . Although no sn-1 fatty acyl preference was noted , Q9BVG9 exhibited significant preference toward DB01708 compared with 18:1 or 20:4 at the sn-2 position . Preferential production of DB01708 -containing PS ( DB01708 -PS ) was consistently observed with Q9BVG9 purified from a variety of cell lines as well as with microsomes from mutant cells in which PS synthesis relies primarily on Q9BVG9 . These findings suggest that Q9BVG9 may play a key role in PS accumulation in brain and testis through high activity toward DB01708 -containing substrates that are abundant in these tissues . A clinical trial with chimeric monoclonal antibody DB05304 and low dose interleukin-2 pulsing scheme for advanced renal cell carcinoma . PURPOSE : DB05304 is a chimeric monoclonal antibody that binds to carbonic anhydrase IX( Q16790 /MN) , which is present on greater than 95 % of RCCs of the clear cell subtype . The suggested working mechanism of DB05304 is by ADCC . Because the number of activated ADCC effector cells can be increased by a low dose interleukin-2 pulsing schedule , a multicenter study was initiated to investigate whether DB05304 combined with LD- P60568 could lead to an improved clinical outcome in patients with progressive RCC . MATERIALS AND METHODS : A total of 35 patients with progressive clear cell RCC received weekly infusions of DB05304 for 11 weeks combined with a daily LD- P60568 regimen . Patients were monitored longitudinally for ADCC capacity . Radiological assessment of metastatic lesions was performed at week 16 and regularly until disease progression . RESULTS : A durable clinical benefit was achieved in 8 of 35 patients ( 23 % ) , including 3 with a partial response and 5 with stabilization at 24 weeks or greater . Mean survival was 22 months . In general treatment was well tolerated with little toxicity . The number of effector cells increased during treatment but lytic capacity per cell did not increase . ADCC and clinical outcome did not appear to correlate . CONCLUSIONS : DB05304 combined with LD- P60568 in patients with metastatic RCC is safe and well tolerated . With a substantial clinical benefit and a median survival of 22 months in patients with metastatic RCC who have progressive disease at study entry combination therapy showed increased overall survival compared to DB05304 monotherapy . Survival was at least similar to that of currently used cytokine regimens but with a favorable toxicity profile . DB06288 promotes cognitive flexibility in rats : the role of P34969 receptors . The antagonism of P34969 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug , amisulpride . It has been previously demonstrated that the selective P34969 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task ( ASST ) . Therefore , the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility . The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound 's action at P34969 receptors . Rats repeatedly exposed ( 1 h daily for 7 days ) to restraint stress demonstrated impaired performance on the extra-dimensional ( ED ) set-shifting stage of the ASST . DB06288 ( 3 mg/kg ) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group . The P34969 receptor agonist , AS19 ( 10 mg/kg ) , abolished the pro-cognitive efficacy of amisulpride ( 3 mg/kg ) . The present study suggests that the antagonism of P34969 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride . These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . Role of serotonin in the regulation of interferon-gamma production by human natural killer cells . Monocytes , recovered directly from peripheral blood by counter-current centrifugal elutriation ( CCE ) , were shown to provide two regulatory signals for induction of interferon-gamma ( P01579 ) in natural killer ( NK ) cells in response to interleukin-2 ( P60568 ) : an upregulating signal and an inhibitory signal . The inhibitory signal was time-dependent , irreversible , and operating on a pretranslational level , as indicated by the inability of enriched NK cells to accumulate P01579 mRNA in the presence of elutriated monocytes . Monocyte-induced inhibition of P01579 production was abrogated by the biogenic amine serotonin , acting via the 5-hydroxytryptamine , or serotonin ( P08908 ) , subset of serotonin receptors ( 5-HTR ) . Thereby , serotonin effectively promoted P01579 production in the presence of monocytes . We conclude that serotonergic P08908 receptors transduce signals that are required for NK cells to produce P01579 in response to P60568 . β2 integrin induces TCRζ-Syk-phospholipase C-γ phosphorylation and paxillin-dependent granule polarization in human NK cells . Cytotoxic lymphocytes kill target cells through polarized release of the content of lytic granules at the immunological synapse . In human NK cells , signals for granule polarization and for degranulation can be uncoupled : Binding of β(2) integrin LFA-1 to ICAM is sufficient to induce polarization but not degranulation , whereas CD16 binding to IgG triggers unpolarized degranulation . In this study , we investigated the basis for this difference . P60568 -expanded human NK cells were stimulated by incubation with plate-bound ligands of LFA-1 ( P05362 ) and CD16 ( human IgG ) . Surprisingly , LFA-1 elicited signals similar to those induced by CD16 , including tyrosine phosphorylation of the TCR ζ-chain , tyrosine kinase Syk , and phospholipase C-γ . Whereas CD16 activated Ca(2+) mobilization and O43561 phosphorylation , LFA-1 did not , but induced strong Pyk2 and paxillin phosphorylation . LFA-1-dependent granule polarization was blocked by inhibition of Syk , phospholipase C-γ , and protein kinase C , as well as by paxillin knockdown . Therefore , common signals triggered by CD16 and LFA-1 bifurcate to provide independent control of Ca(2+)-dependent degranulation and paxillin-dependent granule polarization . Renal changes induced by a cyclooxygenase-2 inhibitor during normal and low sodium intake . P35354 ( P35354 ) has been identified in renal tissues under normal conditions , with its expression enhanced during sodium restriction . To evaluate the role of P35354 -derived metabolites in the regulation of renal function , we infused a selective inhibitor ( nimesulide ) in anesthetized dogs with normal or low sodium intake . The renal effects elicited by nimesulide and a non-isozyme-specific inhibitor ( meclofenamate ) were compared during normal sodium intake . In ex vivo assays , meclofenamate , but not nimesulide , prevented the platelet aggregation elicited by arachidonic acid . During normal sodium intake , nimesulide infusion ( n=6 ) had no effects on arterial pressure or renal hemodynamics but did reduce urinary sodium excretion , urine flow rate , and fractional lithium excretion . In contrast , nimesulide administration increased arterial pressure and decreased renal blood flow , urine flow rate , and fractional lithium excretion during low sodium intake ( n=6 ) . P35354 inhibition reduced urinary prostaglandin E(2) excretion in both groups but did not modify plasma renin activity in dogs with low ( 8.1+/-1.1 ng angiotensin I. mL(-1). h(-1) ) or normal ( 1.8+/-0.4 ng angiotensin I. mL(-1). h(-1) ) sodium intake . DB00939 infusion in dogs with normal sodium intake ( n=8 ) induced a greater renal hemodynamic effect than nimesulide infusion . These results suggest that P35354 -derived metabolites ( 1 ) are involved in the regulation of sodium excretion in dogs with normal sodium intake , ( 2 ) play an important role in the regulation of renal hemodynamic and excretory function in dogs with low sodium intake , and ( 3 ) are not involved in the maintenance of the high renin levels during a long-term decrease in sodium intake . P60568 inhibits DB01221 receptor-mediated currents directly and may differentially affect subtypes . Using whole-cell patch-clamp recordings , this study investigated the effects of interleukin-2 ( P60568 ) on N-methyl-d-aspartate ( DB01221 ) receptor-mediated currents ( I( DB01221 ) ) in rat cultured hippocampal neurons and human embryonic kidney ( P29320 ) 293 cells expressing recombinant DB01221 receptors . We found that P60568 ( 0.01-1ng/ml ) immediately and significantly decreased peak I( DB01221 ) in cultured neurons . Interestingly , the peak I( DB01221 ) induced in P29320 293 cells was also inhibited by P60568 . We also found that P60568 differentially decreased the peak amplitudes of Q12879 - and Q13224 -containing DB01221 receptor-mediated currents ( I( Q12879 ) and I( Q13224 ) ) by 54+/-5 % and 30+/-4 % , respectively . These results provide new evidence that P60568 induces rapid inhibition of peak currents of DB01221 receptor-mediated responses with possible Q9UHB4 / Q12879 and Q9UHB4 / Q13224 subtype-differentiation , and suggest that the inhibition is mediated by direct interaction between P60568 and DB01221 receptors . F-actin involvement in guinea pig sperm motility . Sperm motility is a must for natural fertilization to occur . During their travel through the epididymis , mammalian spermatozoa gradually acquire the ability to move . This is accomplished through a sliding movement of the outer doublet microtubules of the axoneme which is energized by the dynein ATPase . Within its complex structure , the mammalian sperm flagellum contains F-actin and thus , we decided to test in the guinea pig sperm flagellum the role of F-actin in motility . During maturation , capacitation , and the acrosome reaction , a gradual decrease of the relative concentration of F-actin was observed . Motility increased as spermatozoa became able to fertilize . P06396 , phalloidin , and KI inhibited sperm motility . P06396 canceled sperm motility within 20 min of treatment while 0.6 M KI had immediate effects . Phalloidin diminished hyperactive sperm motility slightly . All three compounds significantly increased the relative concentration of F-actin . Latrunculins are conventional drugs that destabilize the F-actin cytoskeleton . DB02621 ( O43561 A ) did not affect sperm motility ; but significantly increased F-actin relative concentration . The results suggested that in guinea pig spermatozoa , randomly severing F-actin filaments inhibits flagellar motility ; while end filament alteration does not . Thus , specific filament regions seem to be important for sperm motility . Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV-1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE3-tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e.g. , all-trans-retinoic acid ( RA ) , 13-cis-retinoic acid ( 13- DB00982 ) , arotinoid acid ( DB02877 ) and m-carboxy-arotinoid acid ( m-carboxy- DB02877 , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 ( 24 nM ) greater than P10826 ( 4.0 nM ) greater than P13631 ( 1.3 nM ) , while the ED50 values for DB02877 and 13- DB00982 decreased in the order of P10276 ( 6.5 nM , 190 nM ) greater than P13631 ( 2.3 nM , 140 nM ) greater than P10826 ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy- DB02877 , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 greater than P10826 greater than P13631 . In vivo effects of a combined P28222 receptor/ P31645 antagonist in experimental pulmonary hypertension . AIMS : A mechanism for co-operation between the serotonin ( 5-hydroxytryptamine , 5-HT ) transporter and P28222 receptor in mediating pulmonary artery vasoconstriction and proliferation of pulmonary artery smooth muscle cells has been demonstrated in vitro . Here we determine , for the first time , the in vivo effects of a combined P28222 receptor/serotonin transporter antagonist ( LY393558 ) with respect to the development of pulmonary arterial hypertension ( PAH ) and its in vitro effects in human pulmonary artery smooth muscle cells ( hPASMCs ) derived from idiopathic PAH ( IPAH ) patients . METHODS AND RESULTS : We determined the effects of LY393558 as well as a selective serotonin transporter inhibitor , citalopram , on right ventricular pressure , right ventricular hypertrophy , and pulmonary vascular remodelling in wildtype mice and mice over-expressing serotonin transporter ( P31645 + mice ) before and after hypoxic exposure . We also compared their effectiveness at reversing PAH in P31645 + mice and hypoxic mice . Further , we examined the proliferative response to serotonin in IPAH hPASMCs . We also clarified the pharmacology of serotonin-induced vasoconstriction and P28222 receptor/serotonin transporter interactions in mouse isolated pulmonary artery . DB00215 had a moderate effect at preventing and reversing experimental PAH in vivo whereas LY393558 was more effective . LY393558 was more effective than citalopram at reversing serotonin-induced proliferation in IPAH hPASMCs . There is synergy between P28222 receptor and serotonin transporter inhibitors against serotonin-induced vasoconstriction in mouse pulmonary arteries . CONCLUSION : P28222 receptor and serotonin transporter inhibition are effective at preventing and reversing experimental PAH and serotonin-induced proliferation of PASMCs derived from IPAH patients . Targeting both the serotonin transporter and P28222 receptor may be a novel therapeutic approach to PAH . Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . Mechanism of oral absorbent DB05269 in lipid abnormalities in experimental uremic rats . BACKGROUND : We have reported that oral sorbent DB05269 ( Q9NRA2 ) is effective in delaying the induction of dialysis in patients with chronic renal failure ( CRF ) because of its effect on lipid metabolism . To clarify the precise mechanism of Q9NRA2 in lipid abnormalities in CRF , we examined the effect of Q9NRA2 on plasma lipid profile , total bile acids ( TBA ) , and lipoprotein lipase ( P06858 ) activity in experimental uremic rats . METHODS : Uremic rats were prepared using male Wistar rats by ligating 5/6 of the renal artery . Uremic rats were randomly divided into two groups as follows : a control group in which rats were maintained on the standard diet and an Q9NRA2 group in which rats were maintained on a diet containing 5 g of Q9NRA2 per 100 g of standard diet for 10 weeks . Plasma P06858 activity was measured as free fatty acid ( FFA ) generation after intravenous administration of heparin . RESULTS : Plasma creatinine at 1.5 +/- 0.1 mg/dl was lower in the Q9NRA2 group than the 1.9 +/- 0.5 mg/ml level in the control group . Q9NRA2 significantly decreased plasma total cholesterol from 192 +/- 29 to 142 +/- 25 mg/dl , triglycerides from 198 +/- 71 to 99 +/- 38 mg/dl , and TBA from 19.6 +/- 2.6 mumol/liter to 8.8 +/- 3.5 mumol/ml . Plasma P06858 activity at 0.22 +/- 0.01 mumol FFA/min/hr was significantly higher in the Q9NRA2 group than 0.15 +/- 0.03 mumol FFA/min/hr in the control group . CONCLUSIONS : These results suggest that Q9NRA2 may improve plasma lipid abnormalities by binding to bile acids in the intestinal lumen and preventing their reabsorption and inhibiting the reduction of P06858 activity in experimental uremic rats . Prevention of acute and chronic allograft rejection by a novel retinoic acid receptor-alpha-selective agonist . To investigate the involvement of retinoic acid receptor ( RAR ) -alpha in allograft rejection , we investigated the effect of a novel selective agonist to the receptor , ER-38925 , in a mouse cardiac allograft model . Prophylactic treatment with ER-38925 inhibited the acute rejection of the mouse cardiac allograft ( BALB/c --> C3H/HeN ) at 0.3 and 3 mg/kg , and its effect was enhanced in combination with tacrolimus . In this model , ER-38925 remarkably inhibited cytotoxic T lymphocyte induction and alloantigen-stimulated production of cytokines , i.e. P60568 , IL-12 and P01579 . In the chronic rejection model , combined treatment with tacrolimus and ER-38925 reduced the grade and incidence of arteriosclerosis in the cardiac allografts significantly more potently than tacrolimus monotherapy . ER-38925 inhibited the proliferation of rat aortic smooth muscle cells stimulated in vitro , presumably through the induction of a cyclin-dependent kinase inhibitor , p27(kip-1) . Those results provide a rationale for using P10276 agonists as immunosuppressants in human organ transplantation .

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P35354 predicts adverse effects of tamoxifen : a possible mechanism of role for nuclear P04626 in breast cancer patients . P35354 ( P35354 ) is associated with breast tumour progression . Clinical and molecular studies implicate human epidermal growth factor receptor 2 ( P04626 ) in the regulation of P35354 expression . Recent reports raise the possibility that P04626 could mediate these effects through direct transcriptional mechanisms . The relationship between P04626 and P35354 was investigated in a cohort of breast cancer patients with or without endocrine treatment . A tissue microarray comprising tumours from 560 patients with 10-year follow-up was analysed for P04626 , P27361 /2 , polyoma enhancer activator 3 ( P43268 ) and P35354 expression . Subcellular localisation of P04626 was assessed by immunofluorescence and confocal microscopy . Expression of markers examined was analysed in relation to classic clinicopathological parameters and disease-free survival in the presence and absence of tamoxifen . P35354 expression associated with both membranous and nuclear expression of P04626 ( P=0.0033 and P < 0.00001 respectively ) . No association was detected between P35354 and either P27361 /2 or P43268 ( P=0.7 and P=0.3 respectively ) . None of the markers were found to be independently prognostic . Membrane P04626 , nuclear P04626 and P35354 , however , were all found to predict poor disease-free survival in patients on endocrine treatment ( P=0.0017 , P=0.0003 and P=0.0202 respectively ) . Moreover , patients who were positive for P35354 predicted adverse effects of tamoxifen ( P=0.0427 ) . These clinical ex vivo data are consistent with molecular observations that P04626 can regulate P35354 expression through direct transcriptional mechanisms . P35354 expression correlates with disease progression on endocrine treatment . This study supports a role for P35354 as a predictor of adverse effects of tamoxifen in breast cancer patients . Generation and phenotypic characterization of new human ovarian cancer cell lines with the identification of antigens potentially recognizable by HLA-restricted cytotoxic T cells . This study describes a simple method for long-term establishment of human ovarian tumor lines and prediction of T-cell epitopes that could be potentially useful in the generation of tumor-specific cytotoxic T lymphocytes ( CTLs ) . Nine ovarian tumor lines ( INT.Ov ) were generated from solid primary or metastatic tumors as well as from ascitic fluid . Notably all lines expressed HLA class I , intercellular adhesion molecule-1 ( P05362 ) , polymorphic epithelial mucin ( P15941 ) and cytokeratin ( CK ) , but not HLA class II , P33681 .1 ( P33681 ) or Q13072 . While of the 9 lines tested 4 ( INT.Ov1 , 2 , 5 and 6 ) expressed the folate receptor ( P15328 ) and 6 ( INT.Ov1 , 2 , 5 , 6 , 7 and 9 ) expressed the epidermal growth factor receptor ( P00533 ) ; MAGE-1 and p185HER-2/neu were only found in 2 lines ( INT.Ov1 and 2 ) and Q13065 expression in 1 line ( INT.Ov2 ) . The identification of class I MHC ligands and T-cell epitopes within protein antigens was achieved by applying several theoretical methods including : 1 ) similarity or homology searches to MHCPEP ; 2 ) BIMAS and 3 ) artificial neural network-based predictions of proteins MAGE , GAGE , P00533 , p185HER-2/neu and P15328 expressed in INT.Ov lines . Because of the high frequency of expression of some of these proteins in ovarian cancer and the ability to determine HLA binding peptides efficiently , it is expected that after appropriate screening , a large cohort of ovarian cancer patients may become candidates to receive peptide-based vaccines . Identification of a variant in P35968 associated with serum P35968 and pharmacodynamics of DB06589 . PURPOSE : P15692 receptor ( VEGFR ) kinases are important drug targets in oncology that affect function of systemic endothelial cells . To discover genetic markers that affect VEGFR inhibitor pharmacodynamics , we performed a genome-wide association study of serum soluble vascular P35968 concentrations [ sVEGFR2 ] , a pharmacodynamic biomarker for P35968 inhibitors . EXPERIMENTAL DESIGN : We conducted a genome-wide association study ( GWAS ) of [ sVEGFR2 ] in 736 healthy Old Order Amish volunteers . Gene variants identified from the GWAS were genotyped serially in a cohort of 128 patients with advanced solid tumor with baseline [ sVEGFR2 ] measurements , and in 121 patients with renal carcinoma with [ sVEGFR2 ] measured before and during pazopanib therapy . RESULTS : rs34231037 ( C482R ) in P35968 , the gene encoding sVEGFR2 was found to be highly associated with [ sVEGFR2 ] , explaining 23 % of the variance ( P = 2.7 × 10(-37) ) . Association of rs34231037 with [ sVEGFR2 ] was replicated in 128 patients with cancer with comparable effect size ( P = 0.025 ) . Furthermore , rs34231037 was a significant predictor of changes in [ sVEGFR2 ] in response to pazopanib ( P = 0.01 ) . CONCLUSION : Our findings suggest that genome-wide analysis of phenotypes in healthy populations can expedite identification of candidate pharmacogenetic markers . Genotyping for germline variants in P35968 may have clinical utility in identifying patients with cancer with unusual sensitivity to effects of P35968 kinase inhibitors . Heat-shock transcription factor Q00613 has a critical role in human epidermal growth factor receptor-2-induced cellular transformation and tumorigenesis . The heat-shock transcription factor Q00613 was recently shown to have a key role in the development of tumors associated with activation of Ras or inactivation of p53 . Here , we show that Q00613 is required for the cell transformation and tumorigenesis induced by the human epidermal growth factor receptor-2 ( P04626 ) oncogene responsible for aggressive breast tumors . Upon expression of P04626 , untransformed human mammary epithelial MCF-10A cells underwent neoplastic transformation , formed foci in culture and tumors in nude mouse xenografts . However , expression of P04626 in MCF-10A cells with knockdown of Q00613 did not cause either foci formation or tumor growth in xenografts . The antitumorigenic effect of downregulation of Q00613 was associated with P04626 -induced accumulation of the cyclin-dependent kinase inhibitor P38936 and decrease in the mitotic regulator survivin , which resulted in growth inhibition and cell senescence . In fact , either knockout of P38936 or overexpression of survivin alleviated these effects of Q00613 knockdown . The proliferation of certain human P04626 -positive breast cancer lines also requires Q00613 , as its knockdown led to upregulation of P38936 and/or decrease in survivin , precipitating growth arrest . Similar effects were observed with a small-molecular-weight inhibitor of the heat-shock response NZ28 . The effects of Q00613 knockdown on the growth arrest and senescence of P04626 -expressing cells were associated with downregulation of heat-shock protein (Hsp)72 and Hsp27 . Therefore , Q00613 is critical for proliferation of P04626 -expressing cells , most likely because it maintains the levels of HSPs , which in turn control regulators of senescence P38936 and survivin . Does reduction of circulating prostaglandin E2 reduce fetal hypothalamic-pituitary-adrenal axis activity ? OBJECTIVE : Placental production of prostaglandin E2 ( DB00917 ) increases in fetal sheep as term approaches . It has been suggested that placental DB00917 may act as a hormone to activate the fetal hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . Alternatively , we have proposed that local generation of prostaglandins in the fetal brain and/or pituitary might play a more prominent role in the stimulation of fetal adrenocorticotropin ( DB01285 ) secretion . We performed the present experiments to test the hypothesis that the elevated concentrations of DB00917 in fetal plasma do not tonically stimulate fetal DB01285 secretion . METHODS : We studied chronically catheterized late-gestation fetal sheep . Selective inhibitors of prostaglandin synthase-1 and -2 ( P23219 and P35354 ) were injected intravenously . Fetal blood pressure and heart rate were monitored continuously , and circulating concentrations of DB00917 , DB01285 , and cortisol were measured by specific immunoassay . RESULTS : Injection of vehicle did not have an effect on circulating levels of DB00917 or DB01285 , but it did have a mild stimulatory effect on cortisol . The selective P35354 inhibitor , DB04743 ( Cayman Chemical , Ann Arbor , MI ) , significantly decreased plasma DB00917 concentrations . The selective P23219 inhibitor , DB02709 ( Cayman Chemical ) , produced smaller decreases in plasma DB00917 concentrations and significantly increased mean arterial blood pressure . Neither inhibitor significantly altered plasma DB01285 or cortisol concentrations . CONCLUSION : These results demonstrate that reduction of circulating DB00917 concentrations in response to intravenous injection of P23219 and P35354 inhibitors does not reduce fetal Q9Y251 axis activity . We conclude that DB00917 in late-gestation ovine fetal plasma does not tonically stimulate fetal DB01285 secretion . Crosstalk between PDGF and P05019 receptors in rat liver myofibroblasts : implication for liver fibrogenesis . P05019 ( P05019 ) and platelet-derived growth factor ( PDGF ) have been identified as significant mitogens for liver myofibroblasts ( LMFs ) , one of the cell populations playing a role in liver fibrogenesis . In the present work , we aimed to elucidate a possible interaction between PDGF receptor ( P09619 ) and P08069 ( IGF-IR ) signaling in LMFs . Among different rat liver cells , P09619 alpha- and beta-subunits were mainly expressed in hepatic stellate cells and LMFs , and were upregulated during their in vitro cultivation . In LMFs , DB00102 ( 10 ng/ml ) stimulated DNA synthesis approximately two-fold and this effect was similar to that of P05019 . P05019 and DB00102 differentially affected IGF-IR and P09619 signaling . High concentrations of P05019 decreased levels of IGF-IR and P35568 and inhibited the expression and activation of PDGFRalpha . DB00102 prevented P05019 -induced downregulation of the IGF-IR , but did not affect expression of its cognate receptor subunits . Transphosphorylation of P09619 and IGF-IR was not observed . PDGF effectively activated terminal Q96HU1 kinases , P19957 kinase and Akt kinase , whereas P05019 demonstrated weaker effects . PLCgamma(1) was phosphorylated only in response to PDGF , but not to P05019 . In rat LMFs , blockade of the IGF-IR via inhibition of the IGF-IR kinase completely abrogated IGF- and PDGF-induced mitogenesis and the ability of PDGF to phosphorylate PLCgamma(1) . In conclusion , the presented data demonstrate that the P09619 signaling requires a functional IGF-IR and that DB00102 stabilizes the IGF-IR function through preventing the P05019 -induced downregulation of the IGF-IR . These interactions might be relevant in vivo for the fibroproliferative response during liver injury . DB05595 , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC(50) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 did not block membrane binding of radiolabeled folic acid , 5-methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 had a minimal effect on the cytoplasmic accumulation of 5-methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC(50) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Platelet-derived growth factor ( PDGF ) -C neutralization reveals differential roles of PDGF receptors in liver and kidney fibrosis . Platelet-derived growth factors ( PDGF ) are key mediators of organ fibrosis . We investigated whether Q9NRA1 (-/-) mice or mice treated with neutralizing Q9NRA1 antibodies are protected from bile duct ligation-induced liver fibrosis , and we compared the effects with those of Q9NRA1 deficiency or neutralization on kidney fibrosis induced by unilateral ureteral obstruction . Unexpectedly , and in contrast to kidney fibrosis , Q9NRA1 deficiency or antagonism did not protect from liver fibrosis or functional liver impairment . Furthermore , the hepatic infiltration of monocytes/macrophages/dendritic cells and chemokine mRNA expression ( CC chemokine ligand [ DB00833 ]5 , P13500 , and CC chemokine receptor 2 [ P41597 ] ) remained unchanged . Transcript expression of PDGF ligands increased in both liver and kidney fibrosis and was not affected by neutralization of Q9NRA1 . In kidney fibrosis , Q9NRA1 deficiency or antagonism led to reduced expression and signaling of PDGF-receptor ( R ) -α- and P09619 -β-chains . In contrast , in liver fibrosis there was either no difference ( Q9NRA1 (-/-) mice ) or even an upregulation of P09619 -β and signaling ( anti- Q9NRA1 group ) . Finally , in vitro studies in portal myofibroblasts pointed to a predominant role of PDGF-B and Q9GZP0 signaling in liver fibrosis . In conclusion , our study revealed significant differences between kidney and liver fibrosis in that Q9NRA1 mediates kidney fibrosis , whereas antagonism of Q9NRA1 in liver fibrosis appears to be counteracted by significant upregulation and increased P09619 -β signaling . Q9NRA1 antagonism , therefore , may not be effective to treat liver fibrosis . Effect of HI-6 on cytokines production after immunity stimulation by DB05299 in a mouse model . OBJECTIVES : HI-6 or asoxime in some sources is an antidotum for nerve agents . In recent experiments , implication of HI-6 in immunity response was proved ; however , the issue was not studied in details . In this experiment , role of cytokines in HI-6 impact on immunity was searched . DESIG N : BALB/c mice were exposed to saline , HI-6 in a dose 1-100 mg/kg and/or 1 DB05299 ( KLH ) 1 mg/kg . Mice were sacrificed 21 days after experiment beginning and interleukins ( IL ) 1 , 2 , 4 , 6 were determined by Enzyme Linked Immunosorbent Assay ( ELISA ) . RESULTS : The animals had no pathological manifestation . From the tested cytokines , no significant alteration was found for the IL-1 , P05112 and P05231 . P60568 was significantly increased in a dose response manner . CONCLUSIONS : The experimental data well correlates with the previous work where HI-6 caused increase of antibodies production . HI-6 is suitable to be used as an adjuvant whenever immunity should be pharmacologically altered . Discovery of DB05130 , a Potent , Selective , and Orally Bioavailable hCCR2 Antagonist . We report the identification of 13 ( DB05130 ) as a potent human P41597 ( hCCR2 ) antagonist . DB05130 exhibited an IC50 of 3.7 nM in antagonism of monocyte chemoattractant protein-1 binding to hCCR2 , an IC50 of 4.7 nM in antagonism of chemotaxis activity , an IC50 of 84 μM in inhibition of the hERG potassium current , a free fraction of 58 % in protein binding , high selectivity over other chemokine receptors and G-protein-coupled receptors , and acceptable oral bioavailability in rodents and primates . In human clinical trials , DB05130 exhibited a pharmacokinetic profile suitable for once-a-day dosing ( T 1/2 = 15 h ) . Synergistic proapoptotic effects of the two tyrosine kinase inhibitors pazopanib and lapatinib on multiple carcinoma cell lines . DB06589 and lapatinib are two tyrosine kinase inhibitors that have been designed to inhibit the P15692 tyrosine kinase receptors 1 , 2 and 3 ( pazopanib ) , and the P00533 and P04626 receptors in a dual manner ( lapatinib ) . DB06589 has also been reported to mediate inhibitory effect on a selected panel of additional tyrosine kinases such as P09619 and c-kit . Here , we report that pazopanib and lapatinib act synergistically to induce apoptosis of A549 non-small-cell lung cancer cells . Systematic assessment of the kinome revealed that both pazopanib and lapatinib inhibited dozens of different tyrosine kinases and that their combination could suppress the activity of some tyrosine kinases ( such as c- DB00134 ) that were not or only partially affected by either of the two agents alone . We also found that pazopanib and lapatinib induced selective changes in the transcriptome of A549 cells , some of which were specific for the combination of both agents . Analysis of a panel of unrelated human carcinoma cell lines revealed a signature of 52 genes whose up- or downregulation reflected the combined action of pazopanib and lapatinib . Indeed , pazopanib and lapatinib exerted synergistic cytotoxic effects on several distinct non-small-cell lung cancer cells as well as on unrelated carcinomas . Altogether , these results support the contention that combinations of tyrosine kinase inhibitors should be evaluated for synergistic antitumor effects . Such combinations may lead to a ' collapse ' of pro-survival signal transduction pathways that leads to apoptotic cell death . DB01268 : a newly approved small-molecule inhibitor of angiogenesis . The advent of targeted therapies has allowed treatment to be directed at signaling pathways integral to tumor growth and survival . DB01268 ( SU11248 , sunitinib malate ; Pfizer Inc. , New York , NY , USA ) is a novel oral small-molecule multitargeted receptor tyrosine kinase inhibitor that has demonstrated direct antitumor activity and antiangiogenic action . It targets the vascular endothelial growth factor receptor ( VEGFR ) , platelet derived growth factor receptor ( P09619 ) , stem-cell factor receptor and Fms-like tyrosine kinase receptor 3 receptor tyrosine-kinases . In January 2006 , sunitinib malate was granted approval by the U.S . Food and Drug Administration for the treatment of gastrointestinal stromal tumor after disease progression on , or intolerance to , imatinib mesylate , as well as for the treatment of metastatic renal cell cancer . This review will discuss the development of sunitinib , particularly in acute myeloid leukemia , imatinib-resistant gastrointestinal stromal tumors and renal cell cancer . The review will also discuss ongoing trials with sunitinib in other malignancies such as neuroendocrine tumors and breast cancer , as well as its potential future development in combination therapy with other agents and in other malignancies . Preclinical pharmacokinetics of DB01270 ( rhuFabV2 ) after a single intravitreal administration . PURPOSE : DB01270 ( DB01270 ; Lucentis , Genentech , South San Francisco , CA ) is a humanized monoclonal antibody fragment designed to bind all forms of P15692 , thereby blocking vessel permeability and angiogenesis in neovascular age-related macular degeneration . This study evaluated the pharmacokinetic ( PK ) and serum bioavailability of ranibizumab after a single intravitreal ( ITV ) or intravenous ( IV ) dose in cynomolgus monkeys . METHODS : Monkeys received ranibizumab as either a bilateral ITV dose ( 500 or 2000 microg/eye ; n = 6/group ) or a single IV dose ( 1000 or 4000 microg/animal ; n = 4/group ) . After ITV administration , ranibizumab concentrations were measured in several ocular compartments and in serum for 10 days and , after IV administration , for 48 hours . Pharmacokinetic parameters were estimated by compartmental and noncompartmental methods . RESULTS : DB01270 cleared in parallel from all ocular compartments , with a terminal half-life of approximately 3 days . It distributed rapidly to the retina ( 6-24 hours ) , and concentrations were approximately one third that in the vitreous . After ITV injection , bioavailability ( F ) was 50 % to 60 % . Serum concentrations were very low , reflecting wider distribution and faster clearance when ranibizumab reached the serum . After IV administration , the terminal half-life was approximately 0.5 day . CONCLUSIONS : This study demonstrates that ranibizumab has a PK profile that is favorable for its clinical use in treating neovascular AMD by monthly ITV injection . Nuclear factor-kappaB enhances ErbB2-induced mammary tumorigenesis and neoangiogenesis in vivo . The ( P04626 /Neu ) ErbB2 oncogene is commonly overexpressed in human breast cancer and is sufficient for mammary tumorigenesis in transgenic mice . Nuclear factor ( NF ) -kappaB activity is increased in both human and murine breast tumors . The immune response to mammary tumorigenesis may regulate tumor progression . The role of endogenous mammary epithelial cell NF-kappaB had not previously been determined in immune-competent animals . Furthermore , the role of the NF-kappaB components , p50 and p65 , in tumor growth was not known . Herein , the expression of a stabilized form of the NF-kappaB-inhibiting P25963 protein ( IkappaBalphaSR ) in breast tumor cell lines that express oncogenic ErbB2 inhibited DNA synthesis and growth in both two- and three-dimensional cultures . Either NF-kappaB inhibition or selective silencing of p50 or p65 led to a loss of contact-independent tumor growth in vitro . IkappaBalphaSR reversed the features of the oncogene-induced phenotype under three-dimensional growth conditions . The NF-kappaB blockade inhibited ErbB2-induced mammary tumor growth in both immune-competent and immune-deficient mice . These findings were associated with both reduced tumor microvascular density and a reduction in the amount of vascular endothelial growth factor . The expression of IkappaBalphaSR in breast cancer tumors inhibited angiogenesis . Thus , mammary epithelial cell NF-kappaB activity enhances ErbB2-mediated mammary tumorigenesis in vivo by promoting both growth and survival signaling via the promotion of tumor vasculogenesis . DB04998 inhibits activation of nuclear factor-kappaB ( NF-kappaB ) by forming a complex with NF-kappaB essential modulator ( Q9Y6K9 ) and nucleolin . DB04998 , also known as DB04998 , is an experimental anticancer drug that recently entered human clinical trials . It is a member of a novel class of antiproliferative agents known as G-rich oligonucleotides ( P09341 ) , which are non-antisense , guanosine-rich phosphodiester oligodeoxynucleotides that form stable G-quadruplex structures . The biological activity of GROs results from their binding to specific cellular proteins as aptamers . One important target protein of GROs has been previously identified as nucleolin , a multifunctional protein expressed at high levels by cancer cells . Here , we report that DB04998 also associates with nuclear factor-kappaB ( NF-kappaB ) essential modulator ( Q9Y6K9 ) , which is a regulatory subunit of the inhibitor of kappaB ( IkappaB ) kinase ( IKK ) complex , and also called IKKgamma . In the classic NF-kappaB pathway , the IKK complex is required for phosphorylation of P25963 and subsequent activation of the transcription factor NF-kappaB . We found that treatment of cancer cells with DB04998 inhibits IKK activity and reduces phosphorylation of P25963 in response to tumor necrosis factor-alpha stimulation . Using a reporter gene assay , we showed that DB04998 blocks both tumor necrosis factor-alpha-induced and constitutive NF-kappaB activity in human cancer cell lines derived from cervical , prostate , breast , and lung carcinomas . In addition , we showed that , in DB04998 -treated cancer cells , Q9Y6K9 is coprecipitated by nucleolin , indicating that both proteins are present in the same complex . Our studies suggest that abrogation of NF-kappaB activity may contribute to the anticancer effects of DB04998 and that nucleolin may play a previously unknown role in regulating the NF-kappaB pathway . Rapid purification of a new humanized single-chain Fv antibody/human interleukin-2 fusion protein reactive against P04626 receptor . Human embryonic kidney 293 cells were transfected with plasmid pcDNA-H520C9scFv-rhIL-2 containing a chimeric cDNA encoding the humanized 520C9 scFv/recombinant human P60568 fusion protein ( H520C9scFv-rhIL-2 ) . The transfected cells in plateau growing phase were cultured in serum-free medium for three days . The supernatant was collected , concentrated and purified using an affinity column packed with CNBr-activated Sepharose 4B coupled with anti-rhIL-2 mouse monoclonal antibody . The purified fusion protein was analyzed by ELISA , SDS-PAGE and Western blot . The fusion protein showed only one band in both silver stained electrophoresis gel and Western blot developed by ECL chemiluminescence system . Its molecular weight was confirmed to be about 45 kD . This fusion protein possessed binding specificity against p185 positive SKOV3 and B16/neu cells , and it might stimulate P60568 -dependent CTLL-2 cell proliferation as well . DB02424 attenuates 3‑nitropropionic acid‑induced apoptosis and JNK activation through the expression of HSP 70 in striatal cells . Although selective striatal cell death is a characteristic hallmark in the pathogenesis of Huntington 's disease ( HD ) , the underlying mechanism of striatal susceptibility remains to be clarified . Heat shock proteins ( HSPs ) have been reported to suppress the aggregate formation of mutant huntingtin and concurrent striatal cell death . In a previous study , we observed that heat shock transcription factor 1 ( Q00613 ) , a major transcription factor of HSPs , significantly attenuated 3‑nitropropionic acid (3NP)‑induced reactive oxygen species ( ROS ) production and apoptosis through the expression of HSP 70 in striatal cells . To investigate the differential roles of HSPs in 3NP‑induced striatal cell death , the effect of geldanamycin ( GA ) , an P08238 inhibitor , was examined in 3NP‑stimulated striatal cells . GA significantly attenuated 3NP‑induced striatal apoptosis and ROS production with an increased expression of HSP 70 . Triptolide ( TL ) , an HSP 70 inhibitor , abolished GA‑mediated protective effects in 3NP‑stimulated striatal cells . To understand the underlying mechanism by which GA‑mediated HSP 70 protects striatal cells against 3NP stimulation , the involvement of various signaling pathways was examined . GA significantly attenuated 3NP‑induced c‑Jun N‑terminal kinase ( JNK ) phosphorylation and subsequent c‑Jun phosphorylation in striatal cells . Taken together , the present study demonstrated that GA exhibits protective properties against 3NP‑induced apoptosis and JNK activation via the induction of HSP 70 in striatal cells , suggesting that expression of HSP 70 may be a valuable therapeutic target in the treatment of HD .

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interacts_with DB00563?

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DB00945 antagonizes the cytotoxic effect of methotrexate in lung cancer cells . DB00563 ( MTX ) has been widely used for the treatment of cancer and rheumatoid arthritis ( RA ) . DB00945 ( ASA ) is a non-selective cyclooxygenase ( P36551 ) inhibitor that contributes to the treatment of inflammatory conditions such as RA . It has been observed that the antitumor effect of ASA can be attributed to inhibition of cell cycle progression , induction of apoptosis and inhibition of angiogenesis . In the present study , we revealed that the treatment with a combination of MTX and ASA resulted in antagonism of the cytotoxic effect as demonstrated by P50991 and colony formation assays . ASA alleviated the MTX-mediated S phase accumulation and recovered the P55008 phase . MTX-mediated accumulation of the S phase marker cyclin A was also alleviated by ASA . Notably , FAS protein levels were upregulated by MTX in A549 cells . The antagonism of MTX efficacy caused by ASA was accompanied by altered expression of caspase-3 , Bcl-2 and FAS but not dihydrofolate reductase ( P00374 ) . This suggests that the alteration of caspase-3 , Bcl-2 and FAS was involved in the antagonism between ASA and MTX . Exogenously added folic acid reversed the MTX-mediated P00374 inhibition following either MTX or MTX + ASA treatments . Most importantly , we demonstrated for the first time that the commonly used non-steroidal anti-inflammatory drug for headache ASA and possibly other P23219 /2 inhibitors can produce a strong antagonistic effect on the growth inhibition of lung cancer cells when administered in combination with MTX . The clinical implication of our finding is obvious , i.e. , the clinical efficacy of MTX therapy can be compromised by ASA and their concomitant use should be avoided . The blockage of Ras/ P29323 pathway augments the sensitivity of SphK1 inhibitor P12755 II in human hepatoma HepG2 cells . The treatment of hepatocellular carcinoma ( HCC ) remains a challenge and the future of cancer therapy will incorporate rational combinations directed to molecular targets that cooperate to drive critical pro-survival signaling . Q9NYA1 ( SphK1 ) has been shown to regulate various processes important for cancer progression . Given the up-regulated expression of SphK1 in response to the silence of N-ras and other interactions between Ras/ P29323 and SphK1 , it was speculated that combined inhibition of Ras/ P29323 and SphK1 would create enhanced antitumor effects . Experimental results showed that dual blockage of N-ras/ P29323 and SphK1 resulted in enhanced growth inhibitions in human hepatoma cells . Similarly , Q02750 /2 Inhibitor U0126 potentiated P12755 II-induced apoptosis in hepatoma HepG2 cells , consistently with the further attenuation of Akt/ P29323 /NF-κB signaling pathway . It was also shown that the combination of P12755 II and U0126 further attenuated the migration of hepatoma HepG2 cells via Q05397 /MLC-2 signaling pathway . Taken together , the dual inhibition of SphK1 and Ras/ P29323 pathway resulted in enhanced effects , which might be an effective therapeutic approach for the treatment of HCC . DB00563 and cytarabine inhibit progression of human lymphoma in NOD/SCID mice carrying a mutant dihydrofolate reductase and cytidine deaminase fusion gene . An SFG-based retroviral bicistronic vector containing a double-mutant dihydrofolate reductase-cytidine deaminase fusion cDNA ( F/S P00374 -CD ) with IRES-eGFP confers resistance to both methotrexate ( MTX ) and cytarabine ( ara-C ) . Two weeks after transplantation with marrow transduced with either a fusion or a control gene ( eGFP-IRES-NeoR ) , human lymphoma ( P12755 -DLCL-1 ) cells were injected sc into the flanks of nonobese diabetic/severe combined immune deficiency mice . In mock-transplanted mice , maximal tolerated dose ( MTD ) of posttransplant MTX/ara-C ( 15/10 mg/kg/day , x3 ) was unable to control tumor growth . Transfer of the fusion gene allowed doses of MTX/ara-C ( 25/15 mg/kg/day , x4 ) twofold higher than the MTD to be tolerated . The tumor burden defined the efficiency of posttransplant chemotherapy ; early treatment , 48 h after tumor inoculation , provided tumor-free survival , while starting treatment after having palpable tumor growth ( 7 days ) delayed tumor growth a median time of 28 days . In addition , the early treated group had higher gene expression in peripheral blood and marrow cells than the late treated group ( P < 0.05 ) , suggesting that early treatment allowed for enrichment of transduced marrow progenitors . These results encourage clinical studies using this retroviral fusion gene construct . Q8N0V5 V ( Mgat5 ) -mediated N-glycosylation negatively regulates Th1 cytokine production by T cells . The differentiation of naive P01730 (+) T cells into either proinflammatory Th1 or proallergic Th2 cells strongly influences autoimmunity , allergy , and tumor immune surveillance . We previously demonstrated that beta1,6GlcNAc-branched complex-type ( Q8N0V5 V ( Mgat5 ) ) N-glycans on TCR are bound to galectins , an interaction that reduces TCR signaling by opposing agonist-induced TCR clustering at the immune synapse . Mgat5(-/-) mice display late-onset spontaneous autoimmune disease and enhanced resistance to tumor progression and metastasis . In this study we examined the role of beta1,6GlcNAc N-glycan expression in Th1/Th2 cytokine production and differentiation . beta1,6GlcNAc N-glycan expression is enhanced by TCR stimulation independent of cell division and declines at the end of the stimulation cycle . DB00075 -activated splenocytes and naive T cells from Mgat5(-/-) mice produce more P01579 and less P05112 compared with wild-type cells , the latter resulting in the loss of P05112 -dependent down-regulation of IL-4Ralpha . DB02034 , an inhibitor of Q16706 , blocked beta1,6GlcNAc N-glycan expression and caused a similar increase in P01579 production by T cells from humans and mice , but no additional enhancement in Mgat5(-/-) T cells . Mgat5 deficiency did not alter P01579 / P05112 production by polarized Th1 cells , but caused an approximately 10-fold increase in P01579 production by polarized Th2 cells . These data indicate that negative regulation of TCR signaling by beta1,6GlcNAc N-glycans promotes development of Th2 over Th1 responses , enhances polarization of Th2 cells , and suggests a mechanism for the increased autoimmune disease susceptibility observed in Mgat5(-/-) mice . Changes in laboratory results for cancer patients treated with interleukin-2 . The systemic administration of interleukin-2 ( P60568 ) can lead to significant antitumor responses in some patients with metastatic cancer in whom standard therapy has failed . A limitation of this immunotherapy is the toxicity associated with P60568 infusion . To assess toxicity , we determined aspartate aminotransferase ( Q9NRA2 ; EC 2.6.1.1 ) , alanine aminotransferase ( ALT ; EC 2.6.1.2 ) , gamma-glutamyltransferase ( P19440 ; EC 2.3.2.2 ) , lactate dehydrogenase ( LD ; EC 1.1.1.27 ) , alkaline phosphatase ( ALP ; EC 3.1.3.1 ) , creatine kinase ( CK ; EC 2.7.3.2 ) , total bilirubin ( TBI ) , direct bilirubin ( DBI ) , creatinine , urea nitrogen , and P02741 in serum from 21 patients before and during five consecutive days of P60568 treatment . Ten patients were followed for an additional five days after the end of P60568 therapy . The P60568 infusion caused liver toxicity and prerenal azotemia , as evidenced by significant increases ( P less than 0.05 ) of all analytes except CK by day 1 . There was a progressive increase in the results ( except CK ) for these tests until P60568 treatment was stopped . Seven tests related to liver function ( Q9NRA2 , ALT , P19440 , LD , ALP , DBI , and TBI ) showed increases , but the test results indicated significant improvement and moved toward the baseline value five days after the end of P60568 therapy . Concentrations of creatinine and urea nitrogen in serum were normal three days after the cessation of P60568 therapy . Gamma-glutamyl transpeptidase and glutathione biosynthesis in non-tumorigenic and tumorigenic rat liver oval cell lines . Glutathione synthesis and growth properties were studied in the gamma-glutamyl transpeptidase( P19440 )-negative , non-tumorigenic rat liver oval cell line OC/CDE22 , and in its P19440 -positive , tumorigenic counterpart line M22 . DB03408 synthetase ( GGCS ) activities were comparable . Growth rates of M22 cells exceeded those of OC/CDE22 cells at non-limiting and limiting exogenous cysteine concentrations . A monoclonal antibody ( Ab 5F10 ) that inhibits the transpeptidatic but not the hydrolytic activity of P19440 did not affect the growth rates of OC/CDE22 , and decreased those of M22 to the OC/CDE22 level . In DB00143 -depleted M22 , but not in OC/CDE22 cells , the rate and extent of DB00143 repletion with exogenous cysteine and glutamine exceeded those obtained with exogenous cysteine and glutamate . With Ab 5F10 , repletion with cysteine/glutamine was similar to that obtained with cysteine/glutamate . Repletion with exogenous DB00143 occurred only in M22 cells , and was abolished by the P19440 inhibitor acivicin . Repletion with gamma-glutamylcysteine ( GGC ) in OC/CDE22 was resistant to acivicin whereas that in M22 was inhibited by acivicin . Repletion with exogenous DB00143 or cysteinylglycine ( CG ) required aminopeptidase activity and was lower than that obtained with cysteine . Unless reduced , CG disulfide did not support DB00143 repletion . The findings are compatible with the notions that ( i ) P19440 -catalyzed transpeptidation was largely responsible for the growth advantage of M22 cells at limiting cysteine concentration , and for their high DB00143 content via the formation of GGC from a gamma-glutamyl donor ( glutamine ) and cyst(e)ine , and ( ii ) aminopeptidase/dipeptidase activity is rate-limiting in DB00143 repletion when DB00143 or CG serve as cysteine sources . A comparative study of the antipyretic effects of indomethacin and dipyrone in rats . OBJECTIVE : Compare the antipyretic effects of dipyrone and indomethacin . MATERIALS AND METHODS : Fever was induced in rats by i. v. LPS or i . c . v. interleukins ( IL ) , prostaglandins ( PG ) , arachidonic acid ( AA ) , pre-formed pyrogenic factor ( PFPF ) , tumour necrosis factor-alpha ( P01375 ) or corticotrophin releasing hormone ( P06850 ) . DB04817 and indomethacin were administered i.p. , arginine vasopressin V1-receptor antagonist , d(CH2)5 DB00135 (Me)AVP , into the ventral septal area . Cyclooxygenase ( P23219 /-2 ) blocking activity was assessed in transfected COS-7 cells . P06850 release from isolated hypothalami was determined by ELISA . RESULTS : Indomethacin or dipyrone reduced LPS , IL-1beta , P05231 or P01375 induced fever and P06850 release from rat hypothalamus . Only dipyrone inhibited P10145 , PFPF or PGF2alpha fever . Only indomethacin inhibited fever induced by AA or IL-1beta , plus AA . Neither antipyretic affected fever caused by DB00917 or P06850 . d(CH2)5Tyr(Me)AVP only blocked antipyresis induced by indomethacin . DB04817 at a very high concentration ( 10 mM ) inhibited only P23219 , while indomethacin ( 0.1 microM ) blocked P23219 and P35354 in COS-7 cells . CONCLUSION : The antipyretic effect of dipyrone differs from that of indomethacin in that it does not depend on AVP release or inhibition of PG synthesis . Enhanced therapeutic effects of doxorubicin and paclitaxel in combination with liposome-entrapped ends-modified raf antisense oligonucleotide against human prostate , lung and breast tumor models . P04049 protein kinase plays an important role in cell growth , proliferation and cell survival . We have previously described the use of liposome-entrapped antisense raf oligonucleotide ( DB04973 ) to inhibit P04049 expression resulting in tumor growth inhibition and radiosensitization . The present study was undertaken to evaluate the chemosensitization effects of DB04973 in combination with doxorubicin or paclitaxel on a panel of human tumor xenografts . DB04973 ( 25.0 mg/kg i.v. x 10 ) displayed significant antitumor activity ( P < 0.05 ) when administered as a single agent in prostate ( PC-3 ) , lung ( A549 ) and breast ( MDA-MB 231 ) carcinoma models . Doxorubicin ( 1.0-4.0 mg/kg i.v. per week x 3 ) and paclitaxel ( 1.0-4.0 mg/kg i.v. on alternate days x 3 ) were administered as single agents at non-toxic doses that led to only minimal to moderate antitumor activity . However , a combination of DB04973 with doxorubicin or paclitaxel led to significantly enhanced antitumor activity in all the tumor types tested ( PC-3 , P < 0.03 ; A549 , P < 0.035 ; MDA-MB 231 , P < 0.045 ) as compared with DB04973 alone or chemotherapeutic agents alone treated groups . This effect of chemosensitization appeared to be sequence-specific because a mismatch control oligonucleotide continued to show significant tumor growth . Additionally , no inhibition in P04049 expression in MDA-MB 231 tumor tissue was observed with mismatch oligonucleotide treatment . On the other hand , DB04973 treatment led to > 75 % inhibition of P04049 expression in tumor tissue . These preclinical observations support the use of DB04973 in combination with chemotherapeutic agents to improve the treatment of human cancers . SAR and in vivo evaluation of 4-aryl-2-aminoalkylpyrimidines as potent and selective O60674 ( O60674 ) inhibitors . We report the discovery of a series of 4-aryl-2-aminoalkylpyrimidine derivatives as potent and selective O60674 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 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 ) it was advanced into clinical trials . Neurokinin type-1 receptor antagonist inhibits enhancement of T cell functions by DB05875 in normal and neuromanipulated capsaicin-treated rats . Substance P ( SP ) plays a major role in the regulation of the interaction between immune and nervous systems . SP administration stimulates Con A-induced proliferation of spleen and peripheral blood lymphocytes from normal and neonatally capsaicin treated rats , which correlated with enhanced P60568 production and expression of activation antigens such as P60568 receptor alpha chain ( CD25 ) and RT1B MHC class II molecule . Moreover , SP markedly increased the percentage of P06127 + and P01730 + T lymphocytes in the peripheral blood of capsaicin-treated rats . Concomitant administration of SP with the non-peptide Neurokinin-1 receptor ( P25103 ) antagonist SR140333 completely inhibited the SP-mediated augmentation of Con A-induced PBL proliferation and P60568 production as well as of P01730 + CD25+ and P01730 + RT1B+ T cell numbers in normal and capsaicin-treated rats . DB05790 also blocked the increased percentage of peripheral blood P01730 + T cells induced by SP in capsaicin-treated rats . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . Long-term exposure to methotrexate induces immunophenotypic changes , decreased methotrexate uptake and increased dihydrofolate gene copy number in Jurkat T cells . DB00563 ( MTX ) treatment of rheumatoid arthritis may require increasing doses to maintain clinical efficacy . An overall plateau of clinical response is reached after only six months of treatment . To study the immunologic , biochemical and genetic effects of MTX on T cells , the Jurkat T cell line was made MTX-resistant by serial addition of methotrexate sodium into culture medium . Cells proliferated and divided successfully in MTX concentrations ranging to 15 microM . MTX resistance of Jurkat T cells in vitro was accompanied by significantly ( P < 0.05 ) decreased expression of P06729 , CD3 , P01730 , P10747 , and Q07108 , P60568 production , and MTX uptake assessed by cell association or disassociation of 3[H]-MTX or fluoresceinated MTX ( FMTX ) , respectively . In addition , there was P00374 gene amplification and increased levels of P00374 in all resistant cell lines . Both permanent and transient phenotypic changes developed in resultant cell lines exposed to increasing concentrations of MTX in vitro . Expression of P01730 and CD25 and sensitivity to MTX returned to near-parental levels after removal of MTX from culture medium , whereas expression of P27487 and MTX uptake were significantly increased . Expression of P06729 , CD3 , Q07108 and P60568 production as well as the P00374 levels did not return to the parental phenotype after removal from MTX . We conclude that MTX-cultured cells express depressed levels of cell-surface markers vital for T cell function and activation . The return of enhancement of these cell-surface markers critical to T cell activation suggests a possible mechanism for the severe flares experienced by rheumatoid arthritis patients when drug treatment is discontinued . Absolute bioavailability and effect of formulation change , food , or elevated pH with rabeprazole on cobimetinib absorption in healthy subjects . DB05239 is a potent and highly selective inhibitor of Q02750 /2 . Since cobimetinib exhibited absorption variability in cancer patients , a series of single-dose studies in healthy subjects were conducted to determine absolute bioavailability and elucidate potential effects of formulation , food , and elevated gastric pH on cobimetinib bioavailability . Three crossover trials were performed with a 20 mg cobimetinib oral dose : absolute bioavailability using a 2 mg intravenous infusion ( n = 13 ) , relative bioavailability of tablets versus capsules and food effect ( n = 20 ) , and drug interaction with a proton pump inhibitor ( 20 mg of rabeprazole daily for 5 days prior to cobimetinib administration ; n = 20 ) . Absolute bioavailability of cobimetinib was 46.2 % ( 24.2 , CV % ) , likely due to metabolism rather than incomplete absorption . The mean systemic clearance of cobimetinib was low ( 11.7 L/h [ 28.2 , CV % ] ) . Administration of cobimetinib tablets with a high-fat meal delayed drug absorption ( prolonged tmax ) but had no statistically significant effect on cobimetinib exposure ( Cmax and AUC0-∞ ) . Tablet and capsule formulations of cobimetinib showed comparable exposures . DB05239 exhibited delayed absorption ( tmax ) in the presence of rabeprazole , with no statistically significant effects on drug exposure ( Cmax and AUC0-∞ ) in the fasted state . In conclusion , cobimetinib oral absorption was not affected by change in formulation , food , or elevated gastric pH . Molecular basis of differential resistance to cycloguanil and pyrimethamine in Plasmodium falciparum malaria . DB01131 and pyrimethamine are antifolate drugs with distinct chemical structures that are used commonly in the prophylaxis and treatment of Plasmodium falciparum malaria . Clinical reports and field studies have suggested that some parasites refractory to proguanil can be treated with pyrimethamine , and vice versa . Analysis of the P. falciparum dihydrofolate reductase ( P00374 ) from different parasites reveals the structural basis for differential susceptibility to these antifolate drugs . Parasites harboring a pair of point mutations from Ala-16 to DB00161 -16 and from DB00133 -108 to DB00156 -108 are resistant to cycloguanil ( the active metabolite of proguanil ) but not to pyrimethamine . A single DB00174 -108 mutation , on the other hand , confers resistance to pyrimethamine with only a moderate decrease in susceptibility to cycloguanil . Significant cross-resistance to both drugs occurs in parasites having mutations that include DB00133 -108 ---- DB00174 -108 and DB00167 -164 ---- DB00149 -164 . These results reflect the distinct structures of pyrimethamine and cycloguanil and suggest fine differences in binding within the active site cavity of P00374 . Alternative inhibitors , used alone or in combination , may be effective against some strains of cycloguanil- or pyrimethamine-resistant malaria . Development of a yeast protein fragment complementation assay ( DB11245 ) system using dihydrofolate reductase ( P00374 ) with specific additives . A yeast protein fragment complementation assay ( DB11245 ) system based on dihydrofolate reductase ( P00374 ) is difficult to be operated because it is not as sensitive to trimethoprim ( P54849 ) as the system using a prokaryotic microorganism . Here , the DB11245 system using P00374 , specific inhibitors , and a substrate in the yeast Saccharomyces cerevisiae was newly developed . As a model , the human oncoprotein Ras and the Ras-binding domain ( RBD ) of P04049 were individually and genetically fused to P00374 fragment , and each genetic construct was coexpressed under the control of the P22466 promoter . An interaction between Ras and RBD could be evaluated on the basis of cell proliferation . To establish the experimental conditions for the yeast DB11245 system based on the P00374 reconstitution , we examined yeast host strains and the concentration of inhibitory additives to prevent endogenous P00374 activity , namely , P54849 and sulfanilamide , and the substrate of P00374 , namely , folic acid . The transformant harboring wild-type Ras or its variants showed positive interaction signals , and the order of interactions for combination corresponded to the results of other in vitro assays . Moreover , combinatorial mutated Ras-binding domains were constructed , and the interaction of RBDs with Ras using this yeast DB11245 system was examined . As a result , various types of mutated clone for RBD were obtained . These demonstrations suggest that the yeast DB11245 system based on P00374 can be one of good , convenient , and inexpensive tools for investigating eukaryotic protein-protein interactions in vivo . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . Passive smoke effects on cough and airways in young guinea pigs : role of brainstem DB05875 . Children raised with extended exposure to environmental tobacco smoke ( ETS ) experience increased cough and wheeze . This study was designed to determine whether extended ETS exposure enhances citric acid-induced cough and bronchoconstriction in young guinea pigs via a neurokinin-1 ( NK-1 ) receptor mechanism at the first central synapse of lung afferent neurons , the nucleus tractus solitarius . Guinea pigs were exposed to ETS from 1 to 6 weeks of age . At 5 weeks of age , guide cannulae were implanted bilaterally in the medial nucleus tractus solitarius at a site that produced apnea in response to the glutamate agonist D,L-homocysteic acid . At 6 weeks of age , either vehicle or a P25103 antagonist , DB05790 , was injected into the nucleus tractus solitarius of the conscious guinea pigs who were then exposed to citric acid aerosol . ETS exposure significantly enhanced citric acid-induced cough by 56 % and maximal Penh ( a measure of airway obstruction ) by 43 % , effects that were attenuated by the P25103 antagonist in the nucleus tractus solitarius . We conclude that in young guinea pigs extended exposure to ETS increases citric acid-induced cough and bronchoconstriction in part by an P25103 mechanism in the nucleus tractus solitarius . Light and X-ray scattering show decorin to be a dimer in solution . P07585 is a widely distributed member of the extracellular matrix small leucine-rich repeat glycoprotein/proteoglycan family . For investigation of its physical properties , decorin from two sources ( young steer skin and a recombinant adenovirus ) was used . The first sample was extracted into 7 m urea and purified , while the second was isolated from medium conditioned by 293A cells infected with adenovirus and purified without chaotropes . The only chemical differences detected between these materials were a slightly shorter glycosaminoglycan chain and the retention of the propeptide on the latter . Circular dichroism spectra of the two samples were virtually identical , showing a high proportion of beta-sheet and beta-turn and little alpha-helix . The protein cores were completely denatured in 2.25 m guanidine HCl ( GdnHCl ) but recovered their secondary structure on removal of chaotrope . Light scattering of material eluted from gel-filtration columns in DB03754 -buffered saline , pH 7.0 , gave molecular mass values of 165 +/- 1 kDa and 84.6 +/- 4 kDa for intact decorin and the glycoprotein core produced by digestion with chondroitin ABC lyase , respectively . Intact recombinant prodecorin had a mass of 148 +/- 18 kDa . These values , which are double those estimated from SDS gel electrophoresis or from the known sequences and compositions , were halved in 2.5 m GdnHCl . Data from solution x-ray scattering of intact decorin and its core in DB03754 -buffered saline are consistent with a dimeric particle whose protein component has a radius of gyration of 31.6 +/- 0.4 A , a maximum diameter of 98 +/- 5 A , and approximates two intertwined C shapes . DB01373 efflux activity of plasma membrane Ca2+ ATPase-4 ( P23634 ) mediates cell cycle progression in vascular smooth muscle cells . We explored the role played by plasma membrane calcium ATPase-4 ( P23634 ) and its alternative splice variants in the cell cycle of vascular smooth muscle cells ( VSMC ) . A novel variant ( PMCA4e ) was discovered . Quantitative real-time-PCR-quantified P23634 splice variant proportions differed in specific organs . The PMCA4a:4b ratio in uninjured carotid arteries ( ∼1:1 ) was significantly reduced by wire denudation injury ( to ∼1:3 ) by modulation of alternative splicing , as confirmed by novel antibodies against PMCA4a/e and PMCA4b . Laser capture microdissection localized this shift to the media and adventitia . Primary carotid VSMC from P23634 knock-out ( P4KO ) mice showed impaired [(3)H]thymidine incorporation and P55008 phase arrest as compared with wild type ( P4WT ) . Electroporation of expression constructs encoding PMCA4a , PMCA4b , and a PMCA4b mutant lacking PDZ binding rescued this phenotype of P4KO cells , whereas a mutant with only 10 % of normal Ca(2+) efflux activity could not . Microarray of early P55008 -synchronized VSMC showed 39-fold higher Rgs16 ( NFAT ( nuclear factor of activated T-cells ) target ; MAPK inhibitor ) and 69-fold higher P07585 ( P55008 arrest marker ) expression in P4KO versus P4WT . Validation by Western blot also revealed decreased levels of P12004 D1 and Q12968 in P4KO . Microarrays of P4KO VSMC rescued by PMCA4a or PMCA4b expression showed reversal of perturbed Rgs16 , P07585 , and Q12968 expression levels . However , PMCA4a rescue caused a 44-fold reduction in AP-2β , a known anti-proliferative transcription factor , whereas PMCA4b rescue resulted in a 50-fold reduction in p15 ( P12004 D1/Cdk4 inhibitor ) . We conclude that Ca(2+) efflux activity of P23634 underlies P55008 progression in VSMC and that PMCA4a and PMCA4b differentially regulate specific downstream mediators . JAK- P35610 and JAK-PI3K-mTORC1 pathways regulate telomerase transcriptionally and posttranslationally in ATL cells . Adult T-cell leukemia ( ATL ) is a heterogeneous tumor that is resistant to chemotherapy . Telomerase activity plays a critical role in tumorigenesis and is associated with the prognosis of ATL patients . Interleukin ( IL ) -2 commonly promotes tumor growth in chronic ATL cells . The signaling pathways involved in P60568 -regulated telomerase activation were studied in ATL cells derived from chronic ATL patients . P60568 challenge enhanced tyrosine phosphorylation of Janus-activated kinase (JAK)1-3 and P42229 , and induced P23458 and O60674 to associate with P42229 in P60568 -dependent ATL cells . Chromatin immunoprecipitation assays revealed that P42229 directly bound to the human telomerase reverse transcriptase ( hTERT ) promoter . P42229 short interfering RNA inhibited hTERT transcription in P60568 -stimulated ATL cells . Inhibitors of PI3K , HSP90 , and P42345 reduced P60568 -induced hTERT mRNA , protein expression , and telomerase activity . AKT , HSP90 , P42345 , S6 kinase , and hTERT immunoprecipitate from P60568 -stimulated cells contained telomerase activity , suggesting that hTERT directly interacts with , and is regulated by , these proteins . Binding of the p85 regulatory subunit of PI3K to O60674 was enhanced in an P60568 -dependent manner , indicating that O60674 propagates activation signals from the P60568 receptor and links hTERT activation to both the P42229 and PI3K pathways . Finally , P60568 -induced activation of telomerase and P42229 was observed in primary leukemic cells . These results indicate that P60568 stimulation induces hTERT activation through the JAK/ P35610 pathway and the JAK/PI3K/AKT/HSP90/mTORC1 pathway in P60568 -responsive ATL cells . These signaling proteins represent novel and promising molecular therapeutic targets for P60568 -dependent ATL .

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P00797 angiotensin system modulates P42345 pathway through AT2R in HIVAN . P42345 ( P42345 ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 and p70S6K . DB09026 , a renin inhibitor attenuated phosphorylation of both P42345 and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 ) phosphorylation of p70S6K in a dose dependent manner . HIV/ P04629 also displayed enhanced phosphorylation of both P42345 and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 and p70S6K . These findings indicate that HIV stimulates P42345 pathway in HIVAN through the activation of renin angiotensin system via AT2R . Environment-related adaptive changes of gut commensal microbiota do not alter colonic toll-like receptors but modulate the local expression of sensory-related systems in rats . Pathogenic and protective roles have been attributed to gut commensal microbiota ( GCM ) in gastrointestinal inflammatory and functional disorders . We have shown that the adaptation to a new environment implies specific changes in the composition of GCM . Here we assessed if environment-related adaptive changes of GCM modulate the expression of colonic Toll-like receptors ( TLRs ) and sensory-related systems in rats . Adult male SD rats were maintained under different environmental conditions : barrier-breed-and-maintained , barrier-breed adapted to conventional conditions or conventional-breed-and-maintained . Fluorescent in situ hybridization and real-time quantitative PCR ( qPCR ) were used to characterize luminal ceco-colonic microbiota . Colonic expression of O60603 , O00206 , O60602 , and Q9NYK1 , cannabinoid receptors ( P21554 /CB2 ) , μ-opioid receptor ( MOR ) , transient receptor potential vanilloid ( Q8NER1 , Q8NET8 , and Q9HBA0 ) , protease-activated receptor 2 ( P55085 ) , and calcitonin gene-related peptide were quantified by RT-qPCR . P21554 , CB2 and MOR expression , was evaluated also by immunohistochemistry . In rats , housing-related environmental conditions induce specific changes of GCM , without impact on the expression of TLR-dependent bacterial recognition systems . Expression of sensory-related markers ( MOR , Q8NET8 , P55085 , and CB2 ) decreased with the adaptation to a conventional environment , correlating with changes in Bacteroides spp. , Lactobacillus spp. , and Bifidobacterium spp. counts . This suggests an interaction between GCM and visceral sensory mechanisms , which might be part of the mechanisms underlying the beneficial effects of some bacterial groups on functional and inflammatory gastrointestinal disorders . DB08810 protects against ethanol-induced gastric mucosal injury in rats : role of 5-hydroxytryptamine , prostaglandins and sulfhydryl compounds . This study was designed to determine the gastroprotective properties of cinitapride ( CNT ) , a novel prokinetic benzamide derivative agonist of Q13639 and 5-HT1 receptors and 5-HT2 antagonist , on mucosal injury produced by 50 % ( v/v ) ethanol . Results were compared with those for 5-hydroxytryptamine ( 5-HT : 10 mg kg-1 ) . The possible involvements of gastric mucus secretion , endogenous prostaglandins ( PGs ) and sulfhydryl compounds ( SH ) in the protection mediated by CNT were also examined . Intraperitoneal administration of CNT ( 0.50 and 1 mg kg-1 ) , 30 min before ethanol , significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus . CNT ( 1 mg kg-1 ) also produced a significant increase in gastric mucosal levels of DB00917 , but did not induce any significant changes in SH values . On the contrary , pretreatment with 5-HT worsened ethanol-induced erosions , however , did not affect gastric mucus secretion , glycoprotein content or DB00917 levels , although the non-protein SH fraction was significantly decreased . The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism . We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . P62158 interacts with angiotensin-converting enzyme-2 ( Q9BYF1 ) and inhibits shedding of its ectodomain . P12821 -2 ( Q9BYF1 ) is a regulatory protein of the renin-angiotensin system ( DB01367 ) and a receptor for the causative agent of severe-acute respiratory syndrome ( P49591 ) , the P49591 -coronavirus . We have previously shown that Q9BYF1 can be shed from the cell surface in response to phorbol esters by a process involving P01375 converting enzyme ( P78536 ; P78536 ) . In this study , we demonstrate that inhibitors of calmodulin also stimulate shedding of the Q9BYF1 ectodomain , a process at least partially mediated by a metalloproteinase . We also show that calmodulin associates with Q9BYF1 and that this interaction is decreased by calmodulin inhibitors . The effect of DB05804 and allopregnanolone sulfate on the binding of [(3)H]ifenprodil to the N-methyl-d-aspartate receptor in rat frontal cortex membrane . Neurosteroids have been shown to modulate the N-methyl-d-aspartate ( DB01221 ) receptor function . DB01708 sulfate ( DHEAS ) is shown to participate in memory and learning processes as well as preventing glutamate neurotoxicity in hippocampus . In this study we have focused on the modulatory effect of neurosteroids on ifenprodil binding to the Q13224 subunit of the DB01221 receptor . We show that DHEAS and allopregnanolone sulfate ( ALLOPREGS ) exert different effects on the [(3)H]ifenprodil binding at 10 , 30 or 100 nM , corresponding to physiological concentrations . The effects include changes in the ifenprodil displacement curve , changing it from a one-site fit into a two-site fit leaving B(max) , K(d) and K(off) unaffected . Our results indicate that DHEAS and ALLOPREGS induce an allosteric modulation of the DB01221 receptor , an observation that might contribute to the understanding of the effects of these neurosteroids . DB09026 : An orally active renin inhibitor . P00797 inhibitors are antihypertensive drugs that block the first step in the renin-angiotensin system . Their mechanism of action differs from that of the angiotensin-converting enzyme inhibitors and angiotensin-receptor antagonists , but like these drugs , renin inhibitors interrupt the negative feedback effects of angiotensin II on renin secretion . The renin-angiotensin-aldosterone system ( RAAS ) has long been recognized to play a significant role in hypertension pathophysiology . Certain agents that modify the RAAS can control blood pressure and improve cardiovascular outcomes . Optimization of this compound by Novartis led to the development of aliskiren - the only direct renin inhibitor which is clinically used as an antihypertensive drug . DB09026 is the first of a new class of antihypertensive agents . DB09026 is a new renin inhibitor of a novel structural class that has recently been shown to be efficacious in hypertensive patients after once-daily oral dosing . In short-term studies , it was effective in lowering blood pressure either alone or in combination with valsartan and hydrochlorothiazide , and had a low incidence of serious adverse effects . It was approved by the Food and Drug Administration in 2007 for the use as a monotherapy or in combination with other antihypertensives . Greater reductions in blood pressure have been achieved when aliskiren was used in combination with hydrochlorothiazide or an angiotensin-receptor blocker . The most common adverse effects reported in clinical trials were headache , fatigue , dizziness , diarrhea , and nasopharyngitis . DB09026 has not been studied in patients with moderate renal dysfunction ; as an RAAS-acting drug , it should be prescribed for such patients only with caution . Local renin-angiotensin II systems , angiotensin-converting enzyme and its homologue Q9BYF1 : their potential role in the pathogenesis of chronic obstructive pulmonary diseases , pulmonary hypertension and acute respiratory distress syndrome . P00797 -angiotensin II-aldosterone axis has long been known as a regulator of blood pressure and fluid homeostasis . Yet , local renin-angiotensin II systems have been discovered and novel actions of angiotensin II ( AngII ) have emerged among which its ability to act as a immunomodulator and profibrotic molecule . The enzyme responsible for its synthesis , Angiotensin-converting-enzyme ( P12821 ) , is present in high concentrations in lung tissue . In the present paper , we review data from studies of the past decade that implicate AngII and functional polymorphisms of the P12821 gene that increase P12821 activity with increased susceptibility for asthma and chronic obstructive pulmonary disease ( P48444 ) and for pulmonary hypertension . Moreover , drugs that inhibit the synthesis of AngII ( P12821 inhibitors ) or that antagonize its actions on its receptors ( Angiotensin II receptor blockers -ARBs ) have been shown to provide beneficial effects . Another recent discovery reviewed is the presence of a homologue of P12821 , Q9BYF1 , which cleaves a single amino acid from AngII and forms a heptapeptide with vasodilatory actions , Ang 1-7 . The balance between P12821 and Q9BYF1 is crucial for controlling AngII levels . P12821 and Q9BYF1 also appear to modify the severity of Acute Respiratory Distress Syndrome ( ARDS ) , with Q9BYF1 playing a protective role . Finally , mention is made to the recent discovery of Q9BYF1 as a receptor for the P49591 Corona Virus . The universal field hypothesis of catatonia and neuroleptic malignant syndrome . Catatonia and neuroleptic malignant syndrome ( Q5H8A3 ) are uncommon disorders that can be life-threatening . Many researchers consider them as clinically divergent entities ; however , they share similar and overlapping literature on causative agents , phenomenology , and treatment response . This hypothesis considers both disorders as a single entity that result from variable combinations of the following : 1 ) gamma-aminobutyric acid ( GABA ) hypoactivity at the GABAA receptor ; 2 ) dopamine hypoactivity at the D2 receptor ; 3 ) serotonin hyperactivity at the P08908 receptor and hypoactivity at the 5- Q13049 receptor ; and 4 ) glutamate hypoactivity at the N-methyl-D-aspartate ( NDMA ) receptor . In this paper , evidence to support this hypothesis is limited to retrospective human studies of catatonia and Q5H8A3 . The four components of the hypothesis are : 1 ) GABAA agonists have been shown to alleviate catatonia and Q5H8A3 ; 2 ) D2 antagonism is proportional to the relative likelihood of Q5H8A3 and catatonia ; 3 ) P08908 agonism with 5- Q13049 antagonism is implicated in catatonia and Q5H8A3 ; 4 ) DB01221 receptor antagonists , such as phencyclidine and ketamine , reduce glutamate transmission . This hypothesis proposes that it is the interaction of these systems that prediposes , initiates , and maintains the twin syndromes of catatonia and Q5H8A3 . Pharmacological modulation of myocardial tumor necrosis factor alpha production by phosphodiesterase inhibitors . Phosphodiesterase ( PDE ) inhibitors are used as therapeutic agents for management of congestive heart failure . PDE inhibitors are potent inotropic and vasodilator drugs , which have also been shown to inhibit tumor necrosis factor alpha ( P01375 ) production . P01375 is a pleiotropic cytokine that has the ability to produce cardiac depressant and other cardiovascular effects in many disease conditions . P01375 levels are elevated in patients with chronic congestive heart failure , and it is possible that P01375 may play a role in this condition . The effects of PDE inhibitors on P01375 secretion from rat heart were evaluated in this study . Rat left ventricle was minced and incubated for 4 hr with various PDE inhibitors , and the amount of P01375 secretion was evaluated by cytotoxicity assay . Ro-20 , 1724 , etazolate , amrinone , milrinone and pentoxifylline inhibited unstimulated P01375 production , with IC50 values of 1.87 , 2.07 , 13.9 , 153 and 201 microM , respectively . Lipopolysaccharide-induced P01375 secretion from rat left ventricle was also evaluated in this study . DB01427 , milrinone and pentoxifylline inhibited lipopolysaccharide-induced P01375 secretion , with IC50 values of 14.8 , 81.6 and 748 microM , respectively , whereas Ro-2D , 1724 and etazolate had no effect on lipopolysaccharide-induced P01375 secretion . These results demonstrated that P01375 was secreted from rat left ventricle after 4 hr and different pharmacological manipulations were able to inhibit the secretion of P01375 from left ventricle . These initial pharmacological results may provide an important tool for further investigation into the beneficial effects of PDE inhibitors in congestive heart failure or other conditions where P01375 levels are elevated . Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress . Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis . Because oxidative stress contributes to atherosclerosis , we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon . Bovine aortic endothelial cells were exposed to static , laminar ( 15 dyn/cm2 ) , and oscillatory shear stress ( +/-15 dyn/cm2 ) . Oscillatory shear increased superoxide ( O2.- ) production by more than threefold over static and laminar conditions as detected using electron spin resonance ( P03372 ) . This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources . DB05262 also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence . DB02134 -dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress . This was associated with decreased xanthine dehydrogenase ( P47989 ) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase ( XO ) to P47989 . We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase . These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity . Transfection of these cells with p47phox restored XO protein levels . Finally , in bovine aortic endothelial cells , prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress . These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress . Novel 3,4-diarylpyrazolines as potent cannabinoid P21554 receptor antagonists with lower lipophilicity . Novel 3,4-diarylpyrazolines 1 as potent P21554 receptor antagonists with lipophilicity lower than that of DB05077 are described . The key change is the replacement of the arylsulfonyl group in the original series by a dialkylaminosulfonyl moiety . The absolute configuration ( 4S ) of eutomer 24 was established by X-ray diffraction analysis and 24 showed a close molecular fit with rimonabant in a P21554 receptor-based model . Compound 17 exhibited the highest P21554 receptor affinity ( Ki = 24 nM ) in this series , as well as very potent P21554 antagonistic activity ( pA2 = 8.8 ) and a high P21554 /CB2 subtype selectivity ( approximately 147-fold ) . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . Induction of G2 arrest and binding to cyclophilin A are independent phenotypes of human immunodeficiency virus type 1 Vpr . P62937 ( CypA ) is a member of a family of cellular proteins that share a peptidyl prolyl cis-trans isomerase ( PPIase ) activity . CypA was previously reported to be required for the biochemical stability and function ( specifically , induction of G2 arrest ) of the human immunodeficiency virus type 1 ( HIV-1 ) protein R ( Vpr ) . In the present study , we examine the role of the Vpr-CypA interaction on Vpr-induced G2 arrest . We find that Vpr coimmunoprecipitates with CypA and that this interaction is disrupted by substitution of proline-35 of Vpr as well as incubation with the CypA inhibitor cyclosporine A ( DB00091 ) . Surprisingly , the presence of CypA or its binding to Vpr is dispensable for the ability of Vpr to induce G2 arrest . Vpr expression in CypA-/- cells leads to induction of G2 arrest in a manner that is indistinguishable from that in CypA+ cells . DB00091 abolished CypA-Vpr binding but had no effect on induction of G2 arrest or Vpr steady-state levels . In view of these results , we propose that the interaction with CypA is independent of the ability of Vpr to induce cell cycle arrest . The interaction between Vpr and CypA is intriguing , and further studies should examine its potential effects on other functions of Vpr . Interactome mapping of the phosphatidylinositol 3-kinase-mammalian target of rapamycin pathway identifies deformed epidermal autoregulatory factor-1 as a new glycogen synthase kinase-3 interactor . The phosphatidylinositol 3-kinase-mammalian target of rapamycin ( PI3K- P42345 ) pathway plays pivotal roles in cell survival , growth , and proliferation downstream of growth factors . Its perturbations are associated with cancer progression , type 2 diabetes , and neurological disorders . To better understand the mechanisms of action and regulation of this pathway , we initiated a large scale yeast two-hybrid screen for 33 components of the PI3K- P42345 pathway . Identification of 67 new interactions was followed by validation by co-affinity purification and exhaustive literature curation of existing information . We provide a nearly complete , functionally annotated interactome of 802 interactions for the PI3K- P42345 pathway . Our screen revealed a predominant place for glycogen synthase kinase-3 ( GSK3 ) A and B and the AMP-activated protein kinase . In particular , we identified the deformed epidermal autoregulatory factor-1 ( O75398 ) transcription factor as an interactor and in vitro substrate of P49840 and P49841 . Moreover , GSK3 inhibitors increased O75398 transcriptional activity on the P08908 serotonin receptor promoter . We propose that O75398 may represent a therapeutic target of lithium and other GSK3 inhibitors used in bipolar disease and depression . SynGAP regulates protein synthesis and homeostatic synaptic plasticity in developing cortical networks . Disrupting the balance between excitatory and inhibitory neurotransmission in the developing brain has been causally linked with intellectual disability ( ID ) and autism spectrum disorders ( P51689 ) . Excitatory synapse strength is regulated in the central nervous system by controlling the number of postsynaptic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors ( AMPARs ) . De novo genetic mutations of the synaptic P20936 ( SynGAP ) are associated with ID and P51689 . SynGAP is enriched at excitatory synapses and genetic suppression of SynGAP increases excitatory synaptic strength . However , exactly how SynGAP acts to maintain synaptic AMPAR content is unclear . We show here that SynGAP limits excitatory synaptic strength , in part , by suppressing protein synthesis in cortical neurons . The data presented here from in vitro , rat and mouse cortical networks , demonstrate that regulation of translation by SynGAP involves P29323 , P42345 , and the small Q15382 . Furthermore , these data show that Q13224 -containing NMDARs and the cognitive kinase CaMKII act upstream of SynGAP and that this signaling cascade is required for proper translation-dependent homeostatic synaptic plasticity of excitatory synapses in developing cortical networks . DB08911 , a novel MEK kinase inhibitor , suppresses lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -α production and endotoxin shock . Lipopolysaccharide ( LPS ) , one of the most prominent pathogen-associated molecular patterns ( PAMPs ) , activates macrophages , causing release of toxic cytokines ( i.e. tumor necrosis factor ( P01375 ) -α ) that may provoke inflammation and endotoxin shock . Here , we tested the potential role of trametinib , a novel and highly potent MAPK/ P29323 kinase ( MEK ) inhibitor , against LPS-induced P01375 -α response in monocytes , and analyzed the underlying mechanisms . We showed that trametinib , at nM concentrations , dramatically inhibited LPS-induced P01375 -α mRNA expression and protein secretion in transformed ( RAW 264.7 cells ) and primary murine macrophages . In ex-vivo cultured human peripheral blood mononuclear cells ( PBMCs ) , this MEK inhibitor similarly suppressed P01375 -α production by LPS . For the mechanism study , we found that trametinib blocked LPS-induced MEK- P29323 activation in above monocytes , which accounted for the defective P01375 -α response . Macrophages or PBMCs treated with a traditional MEK inhibitor PD98059 or infected with Q02750 /2-shRNA lentivirus exhibited a similar defect as trametinib , and nullified the activity of trametinib . On the other hand , introducing a constitutively-active ( CA ) P27361 restored P01375 -α production by LPS in the presence of trametinib . In vivo , mice administrated with trametinib produced low levels of P01375 -α after LPS stimulation , and these mice were protected from LPS-induced endotoxin shock . Together , these results show that trametinib inhibits LPS-induced P01375 -α expression and endotoxin shock probably through blocking MEK- P29323 signaling . The danger signal P04271 integrates pathogen- and danger-sensing pathways to restrain inflammation . Humans inhale hundreds of Aspergillus conidia without adverse consequences . Powerful protective mechanisms may ensure prompt control of the pathogen and inflammation . Here we reveal a previously unknown mechanism by which the danger molecule P04271 integrates pathogen- and danger-sensing pathways to restrain inflammation . Upon forming complexes with O60603 ligands , P04271 inhibited O60603 via RAGE , through a paracrine epithelial cells/neutrophil circuit that restrained pathogen-induced inflammation . However , upon binding to nucleic acids , P04271 activated intracellular TLRs eventually resolve danger-induced inflammation via transcriptional inhibition of P04271 . Thus , the spatiotemporal regulation of TLRs and RAGE by P04271 provides evidence for an evolving braking circuit in infection whereby an endogenous danger protects against pathogen-induced inflammation and a pathogen-sensing mechanism resolves danger-induced inflammation . Proof of concept study to assess fetal gene expression in amniotic fluid by nanoarray PCR . Microarray analysis of cell-free RNA in amniotic fluid ( AF ) supernatant has revealed differential fetal gene expression as a function of gestational age and karyotype . Once informative genes are identified , research moves to a more focused platform such as quantitative reverse transcriptase-PCR . Standardized NanoArray PCR ( P60880 ) is a recently developed gene profiling technology that enables the measurement of transcripts from samples containing reduced quantities or degraded nucleic acids . We used a previously developed P60880 gene panel as proof of concept to determine whether fetal functional gene expression could be ascertained from AF supernatant . RNA was extracted and converted to cDNA from 19 AF supernatant samples of euploid fetuses between 15 to 20 weeks of gestation , and transcript abundance of 21 genes was measured . Statistically significant differences in expression , as a function of advancing gestational age , were observed for 5 of 21 genes . P08758 , P08236 , and P62937 showed decreasing gene expression over time , whereas O15234 and O43296 showed increasing gene expression over time . Statistically significantly increased expression of P42345 and P52630 was seen in female compared with male fetuses . This study demonstrates the feasibility of focused fetal gene expression analysis using P60880 technology . In the future , this technique could be optimized to examine specific genes instrumental in fetal organ system function , which could be a useful addition to prenatal care . Counteraction between angiotensin II and angiotensin-(1-7) via activating angiotensin type I and Mas receptor on rat renal mesangial cells . In the updated concept of renin-angiotensin system ( DB01367 ) , it contains the angiotensin converting enzyme ( P12821 ) -angiotensin ( Ang ) II-angtiogensin type 1 receptor ( AT1 ) axis and the angiotensin-converting enzyme-related carboxypeptidase ( Q9BYF1 ) -Ang-(1-7)-Mas axis . The former axis has been well demonstrated performing the vasoconstrictive , proliferative and pro-inflammatory functions by activation of AT1 receptors , while the later new identified axis is considered counterbalancing the effects of the former . The present study is aimed at observing the interaction between Ang-(1-7) and Ang II on cultured rat renal mesangial cells ( MCs ) . RT-PCR , Western blot and immunofluorescent staining and confocal microscopy results showed that both AT1 and Mas receptor were co-distributed in rat renal MCs . Ang-(1-7) showed similar effects on Ang II in cultured MCs that stimulated phosphorylated extracellular signal-regulated kinase ( P29323 )1/2 phosphorylation and transforms growth factor-β1 synthesis , and cell proliferation and extracellular matrix synthesis . Co-treatment of the cell with Ang-(1-7) and Ang II , Ang-(1-7) counteracted AngII-induced effects in a concentration dependent manner , but failed to alter the changes induced by endothelin-1 . The stimulating effect of Ang II was mediated by AT1 receptor while all the effects of Ang-(1-7) were blocked by Mas receptor antagonist A-779 , but not by AT1 receptor antagonist losartan or P50052 receptor antagonist PD123319 . These results suggest that Ang-(1-7) and Ang II specifically interact with each other on rat renal MCs via activation of their specific receptors , Mas and AT1 receptor respectively . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . Investigation of Q13639 receptors in bronchial hyperresponsiveness in cigarette smoke-exposed mice . BACKGROUND : Chronic obstructive pulmonary disease ( P48444 ) arises from an interaction between genetic host factors and environmental exposures ( mainly cigarette smoke ( CS ) ) . Genome Wide Association studies have demonstrated that genetic variations in the gene encoding 5-hydroxytryptamine 4 receptors ( 5-HT(4)R ) , Q13639 , were associated with measures of airway obstruction and with P48444 . We hypothesised that 5-HT(4) receptors , in addition to 5-HT2AR and muscarinic receptors , contribute to the pathogenesis of P48444 by facilitating cholinergic bronchoconstriction . METHODS : The levels of pulmonary 5-HT(4)R mRNA were measured in CS-exposed mice by qRT-PCR . We investigated the effect of CS exposure on bronchial hyperresponsiveness ( BHR ) to 5-HT and evaluated the contribution of 5-HT2AR , muscarinic receptors and 5-HT(4)R in the response to 5-HT by using the corresponding antagonists and 5-HT(4)R knockout ( KO ) mice . RESULTS : The 5-HT(4)R mRNA levels were significantly elevated upon acute ( 3 days ) , subacute ( 4 weeks ) and chronic ( 24 weeks ) CS exposure . Both acute and subacute CS exposure significantly increased BHR to 5-HT . Antagonism of 5-HT2AR abolished the CS-induced BHR to 5-HT , and antagonism of muscarinic receptors significantly reduced the response to 5-HT . However , pre-treatment with GR113808 , a specific 5-HT(4)R antagonist , did not alter the response to 5-HT in CS-exposed mice . Accordingly , the CS-induced BHR to 5-HT was not different between wild-type and 5-HT(4)R KO mice . CONCLUSION : CS increased the levels of 5-HT(4)R mRNA in the lungs , concomitantly with bronchial responsiveness to 5-HT . Our in vivo data using pharmacologic and genetic approaches suggest that 5-HT(4) receptors are not involved in the BHR to 5-HT in CS-exposed mice . Could treatment with arundic acid ( DB05343 ) increase vulnerability for depression ? Arundic acid ( DB05343 ) is believed to be neuroprotective because of its actions on glia cells ; i.e. , its inhibitory effects on the synthesis of a calcium-binding protein P04271 . DB05343 is undergoing clinical trials for the treatment of patients with stroke and Alzheimer 's disease . Recent clinical studies point to a pervasive comorbidity of depression with stroke and Alzheimer 's disease . Previously , P04271 has been implicated in the pathobiological mechanisms of depression . Preclinical studies have shown that antidepressant treatment significantly increases brain P04271 . Here we hypothesize that available data that link P04271 with depression , along with the proposed inhibitory action of DB05343 on P04271 synthesis , indicate that this compound could increase vulnerability for depression in patients at risk for this disorder , and we propose that evaluation of patients with stroke and Alzheimer 's disease for the presence of depression should be routine in clinical trials employing DB05343 . Although it may be open for discussion whether the neuroprotective effects of DB05343 are exclusively due to its inhibition of P04271 synthesis , the latter action of DB05343 warrants studies of the effects of this drug in the pathobiology of depression .

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interacts_with DB01050?

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Initial characterization of the glutamate-cysteine ligase modifier subunit Gclm(-/-) knockout mouse . Novel model system for a severely compromised oxidative stress response . Glutamate-cysteine ligase ( GCL ) is the rate-limiting enzyme in the DB00143 biosynthesis pathway . In higher eukaryotes , this enzyme is a heterodimer comprising a catalytic subunit ( P48506 ) and a modifier subunit ( P48507 ) , which change the catalytic characteristics of the holoenzyme . To define the cellular function of P48507 , we disrupted the mouse Gclm gene to create a null allele . Gclm(-/-) mice are viable and fertile and have no overt phenotype . In liver , lung , pancreas , erythrocytes , and plasma , however , DB00143 levels in Gclm(-/-) mice were 9-16 % of that in Gclm(+/+) littermates . DB00151 levels in Gclm(-/-) mice were 9 , 35 , and 40 % of that in Gclm(+/+) mice in kidney , pancreas , and plasma , respectively , but remained unchanged in the liver and erythrocytes . Comparing the hepatic GCL holoenzyme with P48506 in the genetic absence of P48507 , we found the latter had an approximately 2-fold increase in K(m) for glutamate and a dramatically enhanced sensitivity to DB00143 inhibition . The major decrease in DB00143 , combined with diminished GCL activity , rendered Gclm(-/-) fetal fibroblasts strikingly more sensitive to chemical oxidants such as H(2)O(2) . We conclude that the Gclm(-/-) mouse represents a model of chronic DB00143 depletion that will be very useful in evaluating the role of the P48507 subunit and DB00143 in numerous pathophysiological conditions as well as in environmental toxicity associated with oxidant insult . DB00055 differentially regulates both viability and differentiation of osteoblasts mediated by bisphosphonates . DB00055 ( P25054 ) is a cytoprotective anticoagulant that can promote cutaneous healing . We examined the effect of P25054 on viability and differentiation of the osteoblastic line , MG63 , in the presence and absence of bisphosphonates ( BPs ) . Osteoblasts were cultured and treated for 24 or 48 h with DB00630 ( Aln ) , DB00399 ( DB00399 ) or DB00282 ( Pam ) at concentrations ranging from 10(-4) to 10(-6) M . Cell differentiation was measured using type 1 collagen production , Alizarin red staining and alkaline phosphatase activity , whereas cell viability was assessed using MTT and crystal violet assays . All three BPs induced MG63 cell death in a dose- and time-dependent manner . Pam- and DB00399 -related cell death was prevented by P25054 treatment ; however , cell death induced by Aln was accelerated by P25054 . P25054 induced MG63 cell differentiation that was enhanced by Aln , but inhibited by Pam or DB00399 . Q9UNN8 ( Q9UNN8 ) was expressed by MG63 cells and mediated the protective effect of P25054 on DB00399 -induced viability . In summary , we have demonstrated that ( 1 ) P25054 favorably regulates MG63 viability and differentiation toward bone growth , ( 2 ) P25054 differentially regulates the effects of specific BPs and ( 3 ) at least part of the effects of P25054 is mediated through Q9UNN8 . These findings highlight the potential importance of the PC pathway in bone physiology and provide strong evidence that P25054 may influence bone cells and has potential to be a therapeutic drug for bone regeneration , depending on concurrent BP treatment . Compound FLZ inhibits lipopolysaccharide-induced inflammatory effects via down-regulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways in RAW264.7 macrophages . AIM : The aim of this study was to investigate the effect of the squamosamide derivative FLZ ( N-2-(4-hydroxy-phenyl)-ethyl-2-(2,5-dimethoxy-phenyl)-3-(3-methoxy-4-hydroxy-phenyl)-acrylamide ) on lipopolysaccharide ( LPS ) -induced inflammatory mediator production and the underlying mechanism in RAW264.7 macrophages . METHODS : RAW264.7 cells were preincubated with non-toxic concentrations of compound FLZ ( 1 , 5 , and 10 micromol/L ) for 30 min and then stimulated with 10 microg/L LPS . The production of nitric oxide ( NO ) , the expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase 2 ( P35354 ) , and the activation of nuclear factor kappa-B ( NF-kappaB ) and mitogen-activated protein kinase ( MAPK ) pathways were examined . RESULTS : FLZ significantly inhibited the LPS-induced production of NO , as well as the expression of P35228 and P35354 at both the RNA and the protein levels in RAW264.7 cells . The LPS-induced increase in the DNA binding activity of NF-kappaB and activator protein 1 ( AP-1 ) , the nuclear translocation of NF-kappaB p65 , the degradation of the inhibitory kappaBalpha protein ( P25963 ) and the phosphorylation of P25963 , O15111 ( IKK ) alpha/beta , c-Jun NH(2)-terminal kinase ( JNK ) and p38 MAPKs were all suppressed by FLZ . However , the phosphorylation of extracellular signal-regulated kinase ( P29323 ) was not affected . Further study revealed that FLZ inhibited the phosphorylation of transforming growth factor-beta ( TGF-beta ) -activated kinase 1 ( TAK1 ) , which is an upstream signaling molecule required for IKKalpha/beta , JNK and p38 activation . CONCLUSION : FLZ inhibited the LPS-induced production of inflammatory mediators at least partly through the downregulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways . DB05007 -- a multitargeted tyrosine kinase inhibitor : results of a phase II study in subjects with non-small cell lung cancer who have progressed after responding to treatment with either gefitinib or erlotinib . INTRODUCTION : Although patients with non-small cell lung cancer ( NSCLC ) whose tumors harbor epidermal growth factor receptor ( P00533 ) activating mutations commonly experience significant regressions when treated with erlotinib or gefitinib , they uniformly develop resistance to these agents . The secondary P00533 T790M mutation is found in 50 % of patients with acquired resistance . Herein , we studied DB05007 , an oral small molecule inhibitor of multiple receptor tyrosine kinases , including P00533 , P35968 , P04626 , and EphB4 , in NSCLC patients known or suspected of having tumors harboring T790M . METHODS : Eligible patients included those with relapsed or recurrent advanced NSCLC who progressed after ≥12 weeks of stable disease or response to erlotinib or gefitinib and/or those patients with a documented P00533 T790M . DB05007 300 mg was administered once daily . The primary end point was objective response rate . Pretreatment plasma samples were collected for mutation testing of circulating tumor DNA . RESULTS : Forty-one patients were enrolled ; 33 were evaluable for efficacy . One partial response was observed ( response rate 3 % and 90 % confidence interval , 0 % to 14 % ) . Of patients whose tumors harbored T790M , 67 % ( 8/12 ) had progression of disease as best response compared with 14 % ( 3/21 ) of those without this mutation . Plasma samples from 40 patients were available for mutation testing , 14 ( 35 % ) of which were found to have P00533 mutations . CONCLUSIONS : The 3 % response rate observed did not meet the prespecified threshold to recommend further study of DB05007 in patients who develop acquired resistance to erlotinib or gefitinib . Patients with T790M had a significantly worse progression-free survival . Q9P0J0 , a cell death regulatory gene product , is a subunit of bovine mitochondrial DB00157 :ubiquinone oxidoreductase ( complex I ) . The sequences of 42 subunits of DB00157 :ubiquinone oxidoreductase ( complex I ) from bovine heart mitochondria have been described previously . Seven are encoded by mitochondrial DNA , whereas the remaining 35 are nuclear gene products imported into the organelle from the cytoplasm . An additional protein , which does not correspond to any previously known subunit of the complex I assembly , has now been detected . Denaturing gels of subcomplex Ilambda , the hydrophilic arm of complex I , clearly show a hitherto unidentified band , which was digested with trypsin and subjected to mass-spectrometric analysis to provide several peptide sequences , used in cDNA cloning and sequencing . Measurement of the intact protein mass indicated that the N terminus is acetylated . The new complex I subunit ( B16.6 ) is the bovine homolog of Q9P0J0 , the product of a cell death regulatory gene induced by interferon-beta and retinoic acid , thus providing a new link between the mitochondrion and its electron-transport chain and apoptotic cell death . Vanadate induction of NF-kappaB involves O15111 beta and P45985 in macrophages . The present studies investigated the signaling pathways of vanadate , a vanadium ion with +5 oxidation state , to activate NF-kappaB transcription factor , a pivotal regulator of inflammatory responses . Treatment of macrophages with vanadate results in the activation of both NF-kappaB and c-Jun N-terminal kinase ( JNK ) . The activity of a recently identified cellular kinase , O15111 -beta ( IKKbeta ) , was significantly elevated concomitant with the increased degradation of P25963 and enhanced NF-kappaB activity in cells exposed to vanadate . To determine whether the IKK pathway and JNK pathway are interconnected or bifurcate upon vanadate stimulation , cells were transfected with either a kinase inactive form of IKKbeta or a kinase inactive form of P45985 ( P45985 ) . Inactive IKKbeta was able to block vanadate-induced degradation of P25963 , yet it was unable to influence the activation of JNK by vanadate . Conversely , blockage of JNK activation by transfection of a kinase-inactive form of P45985 resulted in partially inhibition of vanadate-induced P25963 degradation . Both vanadate-induced degradation of P25963 and activation of JNK were potently inhibited by pretreatment of cells with DB06151 or dimercaprol . These results demonstrate that early activation of stress kinases or change of cellular redox states plays a key role in vanadate-induced activation of NF-kappaB and JNK . [ Effect of NF-kappaB on inhibition of non-small cell lung cancer cell cyclooxygenase-2 by brucine ] . OBJECTIVE : To study the molecular mechanism of cyclooxygenase-2 ( P35354 ) , one of effective ingredient of brucine , in inducing non-small cell lung cancer cell apoptosis . METHOD : P35354 promoter , transcription factor deletion mutants and P35354 mRNA 3'-UTR-containing report plasmids were transfected with Renillia to non-small cell lung cancer A549 cell , in order to detect the activity of report gene luciferase and minimum cis-acting element of P35354 promoter inhibited by brucine . The influence of brucine on IkappaB phosphorylation and the nuclear translocation of p65 were detected by immunoblotting assay . RESULT : Brucine significantly suppressed LPS-induced P35354 promoter activation , but revealed minor impact on P35354 mRNA stability . NF-kappaB in the vicinity of P35354 promoter-262 was an important cis-acting element of brucine for inhibiting the activity of P35354 promoter . Brucine was found to inhibit the phosphorylation of P25963 as well as the nuclear translocation of p65 . CONCLUSION : Brucine can improve A549 cells apoptosis by inhibiting the activity of NF-kappaB and the subsequent P35354 gene expression . Comparative effects of azimilide and ambasilide on the human ether-a-go-go-related gene ( Q12809 ) potassium channel . OBJECTIVE : To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related ( Q12809 ) channel . METHODS : Q12809 was stably transfected into Chinese hamster ovary ( CHO- P04264 ) cells and currents were measured using a whole cell , voltage-clamp technique . RESULTS : DB04957 had a ' dual effect ' , inhibiting current at voltage steps above -40 mV and augmenting current at -40 and -50 mV . Tail current inhibition following a step to +30 mV did not vary with temperature ( IC(50) 610 nM at 22 degrees C and 560 nM at 37 degrees C ) . The agonist effect at -50 mV was concentration-dependent and correlated with a hyperpolarizing shift in the V(1/2) of activation ( r=0.98 , P < 0.05 ) . Time constants of inactivation were faster and there was a -10 mV shift in the V(1/2) of steady state inactivation suggestive of open and inactivated state binding . By comparison , ambasilide inhibited Q12809 channels with lower potency ( IC(50) 3.6 microM ) , in a voltage- and time-dependent but frequency-independent manner ( 0.03-1 Hz ) . Ambasilide had no effect on activation or inactivation gating but prolonged both fast and slow components of deactivation consistent with unbinding from the open state . The net effect of both drugs was similar during a voltage ramp which simulated a cardiac action potential . CONCLUSIONS : Inhibition of Q12809 channels by azimilide and ambasilide exhibits a similar time and voltage-dependence . While both exhibit affinity for the open state , azimilide also binds to inactivated channels . Identification of flavopiridol analogues that selectively inhibit positive transcription elongation factor ( P-TEFb ) and block HIV-1 replication . The positive transcription elongation factor ( P-TEFb ; P50750 /cyclin T1 ) regulates RNA polymerase II-dependent transcription of cellular and integrated viral genes . It is an essential cofactor for HIV-1 Tat transactivation , and selective inhibition of P-TEFb blocks HIV-1 replication without affecting cellular transcription ; this indicates that P-TEFb could be a potential target for developing anti-HIV-1 therapeutics . DB03496 , a small molecule CDK inhibitor , blocks HIV-1 Tat transactivation and viral replication by inhibiting P-TEFb kinase activity , but it is highly cytotoxic . In the search for selective and less cytotoxic P-TEFb inhibitors , we prepared a series of flavopiridol analogues and evaluated their kinase inhibitory activity against P-TEFb and P24941 /cyclin A , and tested their cellular antiviral potency and cytotoxicity . We identified several analogues that selectively inhibit P-TEFb kinase activity in vitro and show antiviral potency comparable to that of flavopiridol , but with significantly reduced cytotoxicity . These compounds are valuable molecular probes for understanding P-TEFb-regulated cellular and HIV-1 gene transcription and provide potential anti-HIV-1 therapeutics . Q9P0J0 , a death-regulatory gene product , suppresses Stat3 activity via functional interaction . P40763 ( Stat3 ) is a latent cytoplasmic transcription factor that can be activated by cytokines and growth factors . Stat3 plays important roles in cell growth , anti-apoptosis and cell transformation , and is constitutively active in various cancers . We examined its potential regulators by yeast two-hybrid screening . Q9P0J0 , a gene product related to interferon-beta- and retinoic acid-induced cancer cell death , was identified and demonstrated to interact with Stat3 in various cell types . The interaction is specific for Stat3 , but not for Stat1 and Stat5a . The interaction regions in both proteins were mapped , and the cellular localization of the interaction was examined . Q9P0J0 itself co-localizes with mitochondrial markers , and forms aggregates at the perinulear region with co-expressed Stat3 , which inhibits Stat3 nuclear translocation stimulated by epidermal growth factor ( P01133 ) . Q9P0J0 represses Stat3 transcriptional activity and its target gene expression , and also suppresses cell growth in Src-transformed cells and a Stat3-expressing cell line . Our data suggest that Q9P0J0 is a novel negative regulator of Stat3 . Effects of aspirin on atherosclerosis and the cyclooxygenase-2 expression in atherosclerotic rabbits . BACKGROUND : Atherosclerosis is a complex vascular inflammatory disease . DB00945 is a mainstay in the prevention of vascular complications of atherosclerosis . In this study , the effectiveness of aspirin in suppressing atherosclerosis and the inflammation process was evaluated in rabbits fed with a high fat diet . METHODS : Eighteen male New Zealand rabbits were randomly divided into 3 groups : control group , untreated cholesterol-fed group , aspirin treated cholesterol-fed group , which were fed for 12 weeks . After 12 weeks , the aorta was harvested for pathologic morphology observation . Immunohistochemical analysis of cyclooxygenase-2 ( P35354 ) , macrophage and vascular smooth muscle cell ( VSMC ) was performed . The statistical analysis was performed by the statistical program SPSS10.0 . RESULTS : The aorta plaque/intima size ( P/I ) by pathologic morphology observation was 0 % , ( 59.6 +/- 13.7 ) % and ( 36.3 +/- 16.5 ) % in the control , untreated cholesterol-fed group and aspirin treated group , respectively . The maximum plaque thickness , the degree of artery stenosis and the proportion of the intimal circumference occupied by atheroma of the 3 groups were significantly different from each other ( P < 0.01 ) . The expression of P35354 and macrophage in plaque of the aspirin treated group were decreased compared with that in untreated cholesterol-fed group . However , no difference was found in the expression of VSMC between the aspirin treated and the untreated cholesterol-fed group . CONCLUSION : The mechanism of atherosclerosis suppression by aspirin in cholesterol-fed rabbits is related to the inhibition of P35354 expression together with the reduced inflammation followed by , but not related to the hypolipidemic effects . DB01892 is a dual inhibitor of cyclooxygenase-1 and P09917 . The acylphloroglucinol derivative hyperforin is the major lipophilic constituent in the herb Hypericum perforatum ( St. John 's wort ) . The aim of the present study was to investigate if hyperforin as well as extracts of H. perforatum can suppresses the activities of P09917 ( P09917 ) and cyclooxygenases ( P36551 ) , key enzymes in the formation of proinflammatory eicosanoids from arachidonic acid ( AA ) . In freshly isolated human polymorphonuclear leukocytes stimulated with Ca(2+) ionophore A23187 , hyperforin inhibited P09917 product formation with IC(50) values of about 1-2 microM , in the absence or presence of exogenous AA ( 20 microM ) , respectively , being almost equipotent to the well-documented P09917 inhibitor zileuton ( IC(50) = 0.5-1 microM ) . Experiments with purified human P09917 demonstrate that hyperforin is a direct P09917 inhibitor ( IC(50) approximately 90 nM ) , acting in an uncompetitive fashion . In thrombin- or ionophore-stimulated human platelets , hyperforin suppressed P23219 product ( 12(S)-hydroxyheptadecatrienoic acid ) formation with an IC(50) of 0.3 and 3 microM , respectively , being about 3- to 18-fold more potent than aspirin . At similar concentrations , hyperforin suppressed P23219 activity in platelets in presence of exogenous AA ( 20 microM ) as well as in cell-free systems . DB01892 could not interfere with P35354 product formation and did not significantly inhibit 12- or 15-LO in platelets or leukocytes , respectively . We conclude that hyperforin acts as a dual inhibitor of P09917 and P23219 in intact cells as well as on the catalytic activity of the crude enzymes , suggesting therapeutic potential in inflammatory and allergic diseases connected to eicosanoids . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Nrf2 is a critical modulator of the innate immune response in a model of uveitis . Uveitis is an inflammatory condition that can lead to blindness . It is therefore important to understand the pathophysiology against which to develop targeted therapy . Herein , we tested whether the oxidant-responsive transcription factor Nrf2 is involved in regulating the innate immune response and oxidative damage in the LPS uveitis model . As shown by dihydroethidium staining , intraperitoneally injected LPS increased reactive oxygen species in the retina and iris-ciliary body of Nrf2+/+ and Nrf2-/- mice . After LPS injection , P05362 , P05231 , P01375 , P35354 , P35228 , and P13500 mRNAs were increased more in the retina and iris-ciliary body of Nrf2-/- than in those of Nrf2+/+ mice . NQO-1 and P48507 , two Nrf2-responsive antioxidant enzymes , had reduced expression in Nrf2+/+ retinas after LPS injection , but no change in expression in Nrf2-/- mice . The number of FITC-Con A-labeled leukocytes adherent to the retinal vascular endothelium increased after LPS treatment in both Nrf2+/+ and Nrf2-/- mice compared to control injections , with more adherent leukocytes in Nrf2-/- than in Nrf2+/+ mice . Pretreatment with the Nrf2 activator 1-(2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl)imidazole increased antioxidant gene expression in the retina , reduced inflammatory mediator expression , and reduced leukocyte adherence to retinal vasculature after LPS treatment in Nrf2+/+ mice , but had no effect on Nrf2-/- mice . Treatment targeting the Nrf2 pathway may be a new therapy for uveitis . Paradoxical effects of resveratrol on the two prostaglandin H synthases . Prostaglandin H synthase ( PGHS ) is the primary enzyme responsible for the biosynthesis of prostaglandins and thromboxanes . Of the two isoenzymes of PGHS , P23219 is constitutively expressed and P35354 is inducible by mitogens or other inflammatory stimuli . Constitutive expression of P35354 in neoplastic tissues has been implicated in carcinogenesis . DB02709 , a lignan , was recently shown to be an anticarcinogen that selectively inhibits P23219 . In vitro experiments to resolve these seemingly paradoxical observations revealed that resveratrol is not only an inhibitor of P23219 but also is an activator of P35354 . DB02709 non-competitively inhibited P23219 with a P04264 of 26 +/- 2 microM but enhanced the P35354 activity nearly twofold . Additionally , resveratrol did not serve as a reducing co-substrate for the peroxidase activities of either enzyme despite being an easily oxidizable phenolic compound . DB02709 inhibited the peroxidase activity of P23219 ( IC50 = 15 microM ) better than that of P35354 ( IC50 = > 200 microM ) . Inhibition of the perxidase activity but not the cyclooxygenase activity of P35354 resulted in the production of PGG2 from arachidonic acid . A plausible relationship between these observation and the anticarcinogenic activity of resveratrol is discussed . Evidence for colorectal cancer cell specificity of aspirin effects on NF kappa B signalling and apoptosis . Epidemiological evidence indicates that non-steroidal anti-inflammatory drugs ( NSAIDs ) protect against colorectal cancer ( CRC ) to a greater degree than other non-gastrointestinal cancers , but the molecular basis for this difference is unknown . We previously reported that aspirin induces signal-specific I kappa B alpha degradation followed by NF kappa B nuclear translocation in CRC cells , and that this mechanism contributes substantially to aspirin-induced apoptosis . Here , we explored the hypothesis that cell-type specific effects on NF kappa B signalling are responsible for the observed differences in protection by aspirin against CRC compared to breast and gynaecological cancers . We also assessed whether P35354 expression , mutation status of adenomatous polyposis coli ( P25054 ) , beta-catenin , p53 , or DNA mismatch repair ( P22897 ) genes in CRC lines influenced aspirin-induced effects . We found that aspirin induced concentration-dependent I kappa B alpha degradation , NF kappa B nuclear translocation and apoptosis in all CRC lines studied . However , there was no such effect on the other cancer cell types , indicating a considerable degree of cell-type specificity . The lack of effect on NF kappa B signalling , paralleled by absence of an apoptotic response to aspirin in non-CRC lines , strongly suggests a molecular rationale for the particular protective effect of NSAIDs against CRC . Effects on NF kappa B and apoptosis were observed irrespective of P35354 expression , or mutation status in P25054 , beta-catenin , p53 and DNA P22897 genes , underscoring the generality of the aspirin effect on NF kappa B in CRC cells . These findings raise the possibility of cell-type specific targets for the development of novel chemopreventive agents . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE .

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DB06612 for severe eosinophilic asthma . In this large ( 616 patients ) , double-blind , placebo-controlled , dose-ranging study of mepolizumab ( a monoclonal antibody that blocks P05113 binding to its receptor ) , patients were given placebo , 75- , 250- or 750-mg mepolizumab by intravenous infusion every 4 weeks for 1 year . Exacerbation rates at all doses were 50 % less than those in the placebo group . There were no changes in any other asthma measures ( symptoms , quality of life or lung function ) . This may be a useful advance for a subgroup of severe asthma with frequent exacerbations and persistent eosinophilia , which may be about half of severe asthmatics . More information on patient selection and cost-benefit will be required . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . Purification and characterization of a high molecular weight histone deacetylase complex ( Q92769 ) of maize embryos . The dynamic state of core histone acetylation is maintained by histone acetyltransferases and deacetylases . In germinating maize embryos , four nuclear histone deacetylases can be distinguished . From a chromatin fraction prepared at 72 h after start of embryo germination , we have purified the nuclear histone deacetylase Q92769 to homogeneity . Using a sequence of chromatographic steps , we achieved the purification of an enzymatically active high molecular weight protein complex with an apparent molecular mass of 400 kDa , as determined by gel filtration chromatography . The purified enzyme was characterized in terms of enzymatic and kinetic properties , and sensitivity to several histone deacetylase inhibitors . In SDS-polyacrylamide gels , Q92769 split into three polypeptides of 45 , 42 , and 39 kDa , suggesting that the native enzyme is a multimer-protein complex . Electrophoresis under nondenaturing conditions in combination with second dimension SDS-gel electrophoresis indicated that all three protein components of the Q92769 complex were enzymatically active . Polyclonal antibodies against each of the three polypeptides were raised in rabbits . Each antiserum reacted with all three polypeptides on Western blots , suggesting that P29466 , Q8NFH3 , and p39 are highly homologous . This homology was confirmed by amino acid sequencing of peptides generated from each of the three Q92769 components . Induction of O15534 mRNA expression in immortalized gonadotropes by gonadotropin-releasing hormone ( DB00644 ) : involvement of protein kinase C and Q96HU1 kinase signaling . The initiation and maintenance of reproductive function in mammals is critically dependent on the pulsatile secretion of gonadotropin-releasing hormone ( DB00644 ) . This peptide drives the pulsatile release of DB00094 and LH from the pituitary pars distalis via signaling pathways that are activated by the type I P30968 ( P30968 ) . Recently , a microarray analysis study reported that a number of genes , including mPer1 , are induced by DB00644 in immortalized gonadotrope cells . In view of these data , we have begun to analyze in detail the signaling pathways mediating the action of DB00644 on mPer1 expression in these cells . Using quantitative real-time polymprose cho read ( PCR ) , we could confirm that exposure of immortalized gonadotropes ( LbetaT2 cells ) to the DB00644 analog , buserelin , markedly induces mPer1 ( but not mPer2 ) expression . Consistent with P30968 signaling via the protein kinase ( PK ) -C pathway , exposure of the cells to phorbol 12,13-dibutyrate rapidly elevates both mPer1 and LHbeta subunit mRNA levels , while pharmacological inhibition of PKC prevents the mPer1 and LHbeta response to buserelin . As DB00644 is known to regulate gonadotropin synthesis via activation of Q8NFH3 /44 mitogen-activated protein kinase ( MAPK ) signaling pathways , we then examined the involvement of this pathway in regulating mPer1 expression in gonadotropes . Our data reveal that DB00644 -induced mPer1 expression is blocked following acute exposure to a MAPK kinase inhibitor . Although the involvement of this signaling mechanism in the regulation of mPer1 is known in neurons , e.g. , in the suprachiasmatic nuclei , the induction of mPer1 in gonadotropes represents a novel mechanism of DB00644 signaling , whose functional significance is still under investigation . Expression of genes involved in the embryo-maternal interaction in the early-pregnant canine uterus . Although there is no acute luteolytic mechanism in the absence of pregnancy in the bitch , a precise and well-timed embryo-maternal interaction seems to be required for the initiation and maintenance of gestation . As only limited information is available about these processes in dogs , in this study , the uterine expression of possible decidualization markers was investigated during the pre-implantation stage ( days 10-12 ) of pregnancy and in the corresponding nonpregnant controls . In addition , the expression of selected genes associated with blastocyst development and/or implantation was investigated in embryos flushed from the uteri of bitches used for this study ( unhatched and hatched blastocysts ) . There was an upregulated expression of prolactin receptor ( P16471 ) and P01344 observed pre-implantation . The expression of PRL and of IGF1 was unaffected , and neither was the expression of progesterone- or estrogen receptor β ( Q92731 ) . In contrast , ( P03372 ) levels were elevated during early pregnancy . Prostaglandin ( PG ) -system revealed upregulated expression of DB00917 -synthase and its receptors , PTGER2 and P35408 , and of the PG-transporter . Elevated levels of P42330 mRNA , but not the protein itself , were noted . Expression of prostaglandin-endoperoxide synthase 2 ( P35354 ) remained unaffected . Most of the transcripts were predominantly localized to the uterine epithelial cells , myometrium and , to a lesser extent , to the uterine stroma . O14684 ( O14684 ) mRNA was abundantly expressed in both groups of embryos and appeared higher in the hatched ones . The expression level of P01344 mRNA appeared higher than that of IGF1 mRNA in hatched embryos . In unhatched embryos IGF1 , P01344 , and P35354 mRNA levels were below the detection limit . Electrostatic steering at acetylcholine binding sites . The electrostatic environments near the acetylcholine binding sites on the nicotinic acetylcholine receptor ( nAChR ) and acetylcholinesterase were measured by diffusion-enhanced fluorescence energy transfer ( DEFET ) to determine the influence of long-range electrostatic interactions on ligand binding kinetics and net binding energy . Changes in DEFET from variously charged Tb3+ -chelates revealed net potentials of -20 mV at the nAChR agonist sites and -14 mV at the entrance to the P22303 active site , in physiological ionic strength conditions . The potential at the alphadelta-binding site of the nAChR was determined independently in the presence of DB01199 to be -14 mV ; the calculated potential at the alphagamma-site was approximately threefold stronger than at the alphadelta-site . By determining the local potential in increasing ionic strength , Debye-Hückel theory predicted that the potentials near the nAChR agonist binding sites are constituted by one to three charges in close proximity to the binding site . Examination of the binding kinetics of the fluorescent acetylcholine analog dansyl- P13671 -choline at ionic strengths from 12.5 to 400 mM revealed a twofold decrease in association rate . Debye-Hückel analysis of the kinetics revealed a similar charge distribution as seen by changes in the potentials . To determine whether the experimentally determined potentials are reflected by continuum electrostatics calculations , solutions to the nonlinear Poisson-Boltzmann equation were used to compute the potentials expected from DEFET measurements from high-resolution models of the nAChR and P22303 . These calculations are in good agreement with the DEFET measurements for P22303 and for the alphagamma-site of the nAChR . We conclude that long-range electrostatic interactions contribute -0.3 and -1 kcal/mol to the binding energy at the nAChR alphadelta- and alphagamma-sites due to an increase in association rates . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . Leukotriene C4 production during hypoxic pulmonary vasoconstriction in isolated rat lungs . Leukotriene inhibitors preferentially inhibit hypoxic pulmonary vasoconstriction in isolated rat lungs . If lipoxygenase products are involved in the hypoxic pressor response they might be produced during acute alveolar hypoxia and a leukotriene inhibitor should block both the vasoconstriction and leukotriene production that occurs in response to hypoxia . We investigated in isolated blood perfused rat lungs whether leukotriene C4 ( LTC4 ) could be recovered from whole lung lavage fluid during ongoing hypoxic vasoconstriction . Lung lavage from individual rats had slow reacting substance ( SRS ) -like myotropic activity by guinea pig ileum bioassay . Pooled lavage ( 10 lungs ) as analyzed by reverse phase high performance liquid chromatography had an ultraviolet absorbing component at the retention time for LTC4 . At radioimmunoassay , and SRS myotropic activity by bioassay . LTC4 was not found during normoxic ventilation , during normoxic ventilation after a hypoxic pressor response , or during vasoconstriction elicited by DB00761 . DB00711 citrate , a leukotriene synthesis blocker , concomitantly inhibited the hypoxic vasoconstriction and LTC4 production . Thus P09917 products may play a role in the sequence of events leading to hypoxic pulmonary vasoconstriction . Thromboxane A(2) generation , in the absence of platelet P23219 activity , in patients with and without atherothrombotic myocardial infarction . BACKGROUND : DB00945 's therapeutic action is via inhibition of platelet cyclooxygenase 1 ( P23219 ) thromboxane A2 ( TxA2 ) production . The aim of this study was to evaluate TxA2 production , in the absence of platelet P23219 activity , in coronary atherosclerotic heart disease patients with and without atherothrombotic myocardial infarction ( MI ) . METHODS AND RESULTS : TxA2 production , in the absence of platelet P23219 activity , was evaluated in 44 patients taking aspirin on 3 commercially available assays that detect metabolites of TxA2 in the urine . Two assays measure urine 11-dehydro-thromboxane B2 ( TxB2 ) alone and 1 measures urine 11-dehydro-TxB2 plus 11-dehydro-2,3-dinor-TxB2 . Platelet P23219 inhibition was confirmed on < 10 % platelet aggregation in response to ≥1 mmol/L arachidonic acid . Median urine 11-dehydro-TxB2 was no different in those with and without a diagnosis of atherothrombotic MI ( 325 vs. 311 pg/mg creatinine , P=0.59 via polyclonal ELISA ) and ( 312 vs. 244 pg/mg creatinine , P=0.11 via LC-MS/MS ) . Median urine 11-dehydro-TxB2 plus 11-dehydro-2,3-dinor-TxB2 , however , was higher in those with vs. those without a diagnosis of atherothrombotic MI ( 1,035 vs. 606 pg/mg creatinine , P=0.03 via monoclonal ELISA ) . CONCLUSIONS : Differences in TxA2 production , in the absence of platelet P23219 activity , between those with vs. without atherothrombotic MI were not observed when TxA2 generation was assessed on 11-dehydro-TxB2 production alone ( polyclonal ELISA or LC-MS/MS ) , but differences were observed when TxA2 generation was assessed using 11-dehydro-TxB2 plus 11-dehydro-2,3-dinor-TxB2 ( monoclonal ELISA ) . These findings highlight important differences between different commercially available assays for TxA2 generation and suggest that 11-dehydro-2,3-dinor-TxB2 may be critical to the biology of atherothrombosis . Roles of P09917 activating protein in cell proliferation and apoptosis . 5-Lipoxygenase activating protein ( P20292 ) functions as a facilitator of P09917 ( 5- P28300 ) activity . However , on the basis of the induction of apoptosis by the P20292 inhibitor MK886 in cells lacking 5- P28300 , it is possible that this fatty acid-binding protein has other activities . This study was designed to examine potential roles of P20292 in apoptosis and cell proliferation . Overexpression of P20292 protein ( 2.2-fold ) was achieved by stable transfection of P08700 -dependent murine prolymphoid progenitor cells ( FL5.12 ) with a construct expressing the cDNA under a CMV promoter . The overexpressed protein was localized to nuclear membranes as with endogenous P20292 . The initial growth rate of P20292 -transfected cells was greater than that of control cells . After 48 h , when cell density had increased , the growth rate of P20292 -transfected cells declined substantially and there and there was a decrease in viability relative to control transfected cells . The P20292 -transfected cells were also more susceptible to withdrawal of P08700 than were control cells . There was , however , no difference between P20292 and control cells in their susceptibility to MK886 , NDGA , or etoposide during the log growth phase . Overexpression of P20292 did not alter Bcl-xL protein expression , but did decrease Bax protein and somewhat increased P23219 and P35354 mRNA levels . The failure of increased P20292 to alter susceptibility to MK886 provides further support to the concept that this agent induces apoptosis by mechanisms unrelated to P20292 . The data also suggest that P20292 can affect cell proliferation . CD8 T cell-specific downregulation of histone hyperacetylation and gene activation of the P05112 gene locus by ROG , repressor of GATA . Chromatin remodeling of type 2 cytokine gene loci occurs during differentiation of naive P01730 and CD8 T cells into type 2 helper ( Th2 ) and cytotoxic ( Tc2 ) T cells . P05112 production and histone hyperacetylation in P05112 -associated nucleosomes in developing Tc2 cells were significantly lower than those of Th2 cells ; however , cytokine production and histone hyperacetylation of P05113 and P35225 genes were equivalent . Developing Tc2 cells expressed lower P23771 levels and dramatically increased levels of repressor of GATA ( ROG ) . A ROG response element in the P35225 gene exon 4 displayed Tc2-specific binding of ROG , Q13547 , and Q92769 and exhibited repression of P05112 gene activation . Thus , ROG may confer CD8 T cell-specific repression of histone hyperacetylation and activation of the P05112 gene locus . Biosensor analysis of dynamics of interleukin 5 receptor subunit beta(c) interaction with P05113 : Q01344 (alpha) complexes . To gain insight into P05113 receptor subunit recruitment mechanism , and in particular the experimentally elusive pathway for assembly of signaling subunit beta(c) , we constructed a soluble beta(c) ectodomain ( s(beta)(c) ) and developed an optical biosensor assay to measure its binding kinetics . Functionally active s(beta)(c) was anchored via a C-terminal DB00117 tag to immobilized anti- DB00117 monoclonal antibodies on the sensor surface . Using this surface , we quantitated for the first time direct binding of s(beta)(c) to Q01344 (alpha) complexed to either wild-type or single-chain P05113 . Binding was much weaker if at all with either R(alpha) or P05113 alone . Kinetic evaluation revealed a moderate affinity ( 0. P35326 microM ) and relatively fast off rate for the s(beta)(c) interaction with P05113 :R(alpha) complexes . The data support a model in which beta(c) recruitment occurs with preformed P05113 :R(alpha) complex . Dissociation kinetics analysis suggests that the P05113 -alpha-beta(c) complex is relatively short-lived . Overall , this study solidifies a model of sequential recruitment of receptor subunits by P05113 , provides a novel biosensor binding assay of beta(c) recruitment dynamics , and sets the stage for more advanced characterization of the roles of structural elements within R(alpha) , beta(c) , and cytokines of the P05113 / P08700 /GM- P04141 family in receptor recruitment and activation . Benzo[b]thiophene-based histone deacetylase inhibitors . Benzo[b]thienyl hydroxamic acids , a novel class of histone deacetylase ( HDAC ) inhibitors , were identified via a targeted screen of small molecule hydroxamic acids . Various substitutions were explored in the P01031 - and P13671 -positions of the benzo[b]thiophene core to characterize SAR and develop optimal inhibitors . It was determined that substitution at the P13671 -position of the benzo[b]thiophene core with a three-atom spacer yielded optimal Q13547 inhibition and anti-proliferative activity in murine erythroleukemia ( SC-9 ) cells . Trichostatin A induces P09917 promoter activity and mRNA expression via inhibition of histone deacetylase 2 and 3 . The P09917 ( P09917 ) is the key enzyme in the formation of leukotrienes . We have previously shown that the histone deacetylase ( HDAC ) inhibitor trichostatin A ( P32119 ) activates P09917 transcription via recruitment of Sp1 , Sp3 and RNA polymerase II to the proximal promoter . To identify the HDACs involved in the regulation of P09917 promoter activity isoform-specific HDAC inhibitors were applied . P09917 promoter activity and mRNA expression were up-regulated by the class I HDAC inhibitors apicidin and MS-275 but not by class II inhibitors . Knockdown of HDAC 1 , 2 and 3 revealed that Q92769 and O15379 but not Q13547 is involved in the up-regulation of P09917 mRNA expression . To analyse the chromatin modifications at the P09917 promoter associated with HDAC inhibition , the time course of P09917 mRNA induction by trichostatin A was investigated and the concomitant changes in histone modifications at the P09917 promoter in HL-60 , U937 and Mono Mac6 cells were determined . Chromatin immunoprecipitation analysis revealed that trichostatin A increases acetylation of histones H3 and H4 at the P09917 core promoter in HL-60 and U937 cells whereas no significant changes were observed in Mono Mac6 cells . The appearance of H3 and H4 acetylation preceded the P09917 mRNA induction whereas in all three cell lines , induction of P09917 mRNA expression correlated with histone H3 lysine 4 trimethylation ( H3K4me3 ) , a marker for transcriptional activity of gene promoters . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . The effects of 5-aza-2'- deoxycytidine and trichostatin A on gene expression and DNA methylation status in cloned bovine blastocysts . We previously found that treatment of both donor cells and early cloned embryos with combination of DB01262 ( 5-aza-dC ) and trichostatin A ( P32119 ) significantly improve the in vitro and full-term development of nuclear transfer ( NT ) bovine embryos . To investigate how this treatment improved the epigenetic reprogramming of somatic cell nuclei , we compared the expression levels of DNA methylation- , chromatin structure- , and development-related genes in in vitro fertilized ( IVF group ) , NT ( C-NT group ) , and 5-aza-dC and P32119 -treated NT ( T-NT group ) single blastocyst using quantitative real-time PCR . We also compared the DNA methylation status of satellite I among three groups using bisulfite sequencing analysis and combined bisulfite restriction analysis ( COBRA ) . There were significantly lower levels of P26358 , DNMT3b , Q92769 , and P01344 transcripts in T-NT blastocysts than in C-NT blastocysts , whereas the relative abundance of Q01860 and P48431 mRNA was significantly increased in T-NT blastocysts compared to C-NT blastocysts . In addition , the treatment also reduced the DNA methylation levels of NT blastocysts on satellite I sequence . It is likely that P32119 may act synergistically with 5-aza-dC to exert such modifications in gene expression and DNA methylation , subsequently enhancing developmental potential ( in vitro and full-term ) of treated cloned embryos . Effects of a novel P08908 receptor agonist , E4424 , on gastric adherent mucus levels following restraint stress in rats . Several novel arylpiperazine serotonin 1A receptor agonists , developed as anxiolytics , have antisecretory and gastroprotective effects in rats . E4424 ( 2-¿4-[4-(4-chloropyrazol-1-yl)butyl]-1-piperazinyl ¿pyrimidine ; DB04970 dihydrochloride ) , has potent anti-gastric secretory and antiulcer effects . Preliminary data indicated an enhancing effect of E4424 on gastric mucus that may underlie its gastroprotective actions . We therefore tested the effects of acute and chronic administration of E4424 and of a reference P08908 receptor agonist , 8-OHDPAT [ 8-hydroxy-2-(di-n-propylamino)tetralin ] , on gastric mucus levels in rats subjected to cold-restraint stress , a procedure associated with depletion of gastric mucus and the development of mucosal injury . Acute oral administration of E4424 increased adherent mucus levels by 12 % , 11 % , and 13 % , relative to controls . Chronic E4424 significantly increased gastric mucus relative to controls ( 69 % increase ) . Acute oral treatment with 8-OHDPAT did not affect gastric mucus level . Acute intraperitoneal 8-OHDPAT slightly increased mucus levels . Chronic twice per day 8-OHDPAT did not affect mucus levels ; however , chronic once per day treatment with 8-OHDPAT significantly elevated gastric mucus levels at the highest doses used . For E4424 , there is a strong correlation between reduction of gastric mucosal injury and increase in gastric mucus level , suggesting that the action of E4424 on glandular mucus levels is an important mechanism underlying its gastroprotective effects . P04150 recruitment of histone deacetylase 2 inhibits interleukin-1beta-induced histone H4 acetylation on lysines 8 and 12 . We have investigated the ability of dexamethasone to regulate interleukin-1beta ( IL-1beta ) -induced gene expression , histone acetyltransferase ( O60235 ) and histone deacetylase ( HDAC ) activity . Low concentrations of dexamethasone ( 10(-10) M ) repress IL-1beta-stimulated granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) expression and fail to stimulate secretory leukocyte proteinase inhibitor expression . Dexamethasone ( 10(-7) M ) and IL-1beta ( 1 ng/ml ) both stimulated O60235 activity but showed a different pattern of histone H4 acetylation . Dexamethasone targeted lysines P13647 and P08779 , whereas IL-1beta targeted K8 and K12 . Low concentrations of dexamethasone ( 10(-10) M ) , which do not transactivate , repressed IL-1beta-stimulated K8 and K12 acetylation . Using chromatin immunoprecipitation assays , we show that dexamethasone inhibits IL-1beta-enhanced acetylated K8-associated GM- P04141 promoter enrichment in a concentration-dependent manner . Neither IL-1beta nor dexamethasone elicited any GM- P04141 promoter association at acetylated P13647 residues . Furthermore , we show that GR acts both as a direct inhibitor of CREB binding protein ( CBP ) -associated O60235 activity and also by recruiting Q92769 to the p65-CBP O60235 complex . This action does not involve de novo synthesis of HDAC protein or altered expression of CBP or p300/CBP-associated factor . This mechanism for glucocorticoid repression is novel and establishes that inhibition of histone acetylation is an additional level of control of inflammatory gene expression . This further suggests that pharmacological manipulation of of specific histone acetylation status is a potentially useful approach for the treatment of inflammatory diseases . Progesterone together with estrogen attenuates homologous upregulation of gonadotropin-releasing hormone receptor mRNA in primary cultured rat pituitary cells . In a previous study , we clearly demonstrated that an application of gonadotropin-releasing hormone ( DB00644 ) to cultured rat pituitary cells increased the expression of P30968 ( P30968 ) mRNA through transcriptional activation of P30968 gene rather than suppression of the turnover rate of P30968 mRNA . Along with DB00644 , gonadal steroids seem to be an important regulator for P30968 expression in the pituitary gland . Recent in vivo studies reported that an application of gonadal steroids to gonadectomized animals modulated P30968 mRNA expression in the pituitary gland . However , it has not been clearly understood whether steroids may act directly at the pituitary or indirectly via modulation of hypothalamic DB00644 release . Therefore , we assessed the effects of estrogen and progesterone on P30968 mRNA expression in primary cultured female rat pituitary cells . Neither estradiol nor progesterone modulates the basal expression of P30968 mRNA in primary cultured pituitary cells . When cultured pituitary cells were exposed to different doses of estradiol in combination with DB00644 ( 0.2 nM ) , the DB00644 -stimulated increment of P30968 mRNA expression was not significantly changed by estradiol at any given doses . However , when different doses of progesterone were added to primary cultured pituitary cells in combination with DB00644 ( 0.2 nM ) , DB00644 -induced increases in P30968 mRNA levels were reduced in a dose-related manner , showing a significant reduction at 100 nM progesterone . Furthermore , the addition of estradiol reinforced the suppressive effect of progesterone on the homologous upregulation of P30968 mRNA expression . Collectively , our results clearly demonstrated that progesterone directly attenuates the homologous upregulation of P30968 mRNA expression at the pituitary level , and that estradiol potentiates the effect of progesterone . P06401 modulator for emergency contraception : a randomized controlled trial . OBJECTIVE : Compare the efficacy and adverse effects of DB05366 , a new progesterone receptor modulator , to levonorgestrel for emergency contraception . METHODS : We performed a randomized , double-blinded noninferiority trial , enrolling healthy women seeking emergency contraception within 72 hours of unprotected intercourse . Participants were randomly assigned to receive a single dose of 50 mg of DB05366 , plus a placebo 12 hours later or two doses of 0.75 mg of levonorgestrel taken 12 hours apart . Follow-up was scheduled 5 to 7 days after the expected onset of the next menstrual period . Posttreatment pregnancy was established by a positive urine test at follow-up and confirmed by quantitative serum beta-hCG . Daily diaries were used from the time of emergency contraception use until next menses to record adverse effects and sexual activity . RESULTS : Product efficacy was evaluable in 775 of DB05366 users and 774 of levonorgestrel users . Pregnancies occurred in 7 ( 0.9 % , 95 % confidence interval 0. P35326 .6 % ) and 13 ( 1.7 % , 95 % confidence interval 0.8-2.6 % ) women , respectively . Based on the estimated cycle day of unprotected intercourse , 85 % and 69 % of anticipated pregnancies , respectively , were averted . Nausea was reported by a somewhat greater percentage of DB05366 than levonorgestrel users ( 29 % compared with 24 % , P=.03 ) , but the distribution of other adverse effects was similar in both groups . Women in both groups experienced considerable variation in menstrual cycle length as compared with their reported individual normal cycle lengths . CONCLUSION : DB05366 is at least as effective as levonorgestrel in preventing pregnancies after unprotected intercourse and has a similar side effect profile . LEVEL OF EVIDENCE : I .

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interacts_with DB08907?

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A set of consensus mammalian mediator subunits identified by multidimensional protein identification technology . The Mediator is a multiprotein transcriptional coactivator that is expressed ubiquitously in eukaryotes from yeast to mammals and is required for induction of RNA polymerase II ( pol II ) transcription by DNA binding transcription factors . In the work described here , we exploit multidimensional protein identification technology ( MudPIT ) to carry out a proteomic analysis of the subunit composition of the mammalian Mediator complex . By comparing MudPIT data sets obtained from six independent Mediator preparations immunoaffinity purified through their Nut2 ( Q9BTT4 ) , Med25 ( Q9NWA0 ) , Intersex ( Q9NX70 ) , A0JLT2 ( A0JLT2 ) , AK007855 ( Q9H204 ) , or CRSP70 ( O95402 ) subunits , we identify a set of consensus mammalian Mediator subunits . In addition , we identify as Mediator-associated proteins the P49336 -like cyclin-dependent kinase CDK11 and the Q9UHV7 -like Q71F56 protein ( Q71F56 ) , which is mutated in patients with the congenital heart defect transposition of the great arteries ( TGA ) . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Are human anti-idiotypic anti-OC125 antibodies formed after immunization with the anti- Q8WXI7 antibody DB04964 ? The two monoclonal antibodies ( MAb ) OC125 and DB04964 both recognize antigenic determinants on the cancer antigen 125 ( Q8WXI7 ) molecule . The aim of the present study was to clarify how far antibodies formed by patients treated with DB04964 may cross-react with idiotopes of OC125 . Serum samples from 15 ovarian cancer patients treated with DB04964 were tested for the presence of anti-idiotypic anti-OC125 antibodies ; 4 of these patients previously had received OC125 F(ab')2 fragments . Six patients treated only with DB04964 and all patients pretreated with OC125 fragments developed a considerable increase of human anti-mouse anti-bodies after DB04964 infusion , indicating an immune response to antibody infusion . However , none of the patients treated only with DB04964 developed detectable levels of anti-idiotypic anti-OC125 antibodies , whereas in 2 patients pretreated with OC125 fragments , the pre-existing concentration of anti-idiotypic anti-OC125 antibodies further increased after DB04964 infusion . The binding of these newly formed antibodies to OC125 was not inhibited by the DB04964 , but about 70 % of the binding was inhibited by the Q8WXI7 antigen . The present data suggest that the idiotopes expressed on the antibodies DB04964 and OC125 , respectively , are completely different . Thus , anti-idiotypic anti- DB04964 antibodies may not cross-react with OC125 . A greater number of cases is needed to clarify how far the increase of anti-idiotypic anti-OC125 antibodies observed in 2 patients pretreated with OC125 fragments really is due to DB04964 infusion . Coactivators and corepressors of NF-kappaB in IkappaB alpha gene promoter . In this study , we investigated recruitment of coactivators ( Q15788 , P12931 -2 , and Q9Y6Q9 ) and corepressors ( Q13547 , Q92769 , O15379 , Q9Y618 , and NCoR ) to the IkappaB alpha gene promoter after NF-kappaB activation by tumor necrosis factor-alpha . Our data from chromatin immunoprecipitation assay suggest that coactivators and corepressors are simultaneously recruited to the promoter , and their binding to the promoter DNA is oscillated in HEK293 cells . Q15788 , P12931 -2 , and Q9Y6Q9 all enhanced IkappaB alpha transcription . However , the interaction of each coactivator with the promoter exhibited different patterns . After tumor necrosis factor-alpha treatment , Q15788 signal was increased gradually , but P12931 -2 signal was reduced immediately , suggesting replacement of P12931 -2 by Q15788 . Q9Y6Q9 signal was increased at 30 min , reduced at 60 min , and then increased again at 120 min , suggesting an oscillation of Q9Y6Q9 . The corepressors were recruited to the promoter together with the coactivators . The binding pattern suggests that the corepressor proteins formed two types of corepressor complexes , Q9Y618 - Q13547 and NCoR- O15379 . The two complexes exhibited a switch at 30 and 60 min . The functions of cofactors were confirmed by gene overexpression and RNA interference-mediated gene knockdown . These data suggest that gene transactivation by the transcription factor NF-kappaB is subject to the regulation of a dynamic balance between the coactivators and corepressors . This model may represent a mechanism for integration of extracellular signals into a precise control of gene transcription . Differential effects of gonadotropin-releasing hormone I and II on the urokinase-type plasminogen activator/plasminogen activator inhibitor system in human decidual stromal cells in vitro . To date , the factors capable of regulating the coordinate expression of the urokinase-type plasminogen activator ( uPA ) and its endogenous inhibitor , plasminogen activator inhibitor ( P05121 ) , at the maternal-fetal interface remain poorly characterized . In these studies we examined the ability of the classical form of gonadotropin-releasing hormone ( DB00644 ) I and the second , mammalian form of this hormone , DB00644 II , to regulate uPA and P05121 mRNA and protein expression levels in cultures of stromal cells isolated from first trimester decidual tissues using quantitative competitive-PCR and ELISA , respectively . DB00644 I and DB00644 II increased uPA mRNA and protein expression levels in these primary cell cultures in a dose- and time-dependent manner . In contrast , DB00644 I increased , whereas DB00644 II decreased P05121 mRNA and protein expression levels in these cells . DB00050 , a P30968 antagonist , inhibited the regulatory effects of DB00644 I , but not DB00644 II , on uPA and P05121 expression levels in these decidual stromal cell cultures . Taken together , these observations suggest that DB00644 I and DB00644 II differentially regulate the balance between uPA and P05121 expression levels in the human decidua , possibly via distinct receptor-mediated signaling pathways . HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing P16671 -dependent cholesteryl ester accumulation in macrophages . Protease inhibitors decrease the viral load in HIV patients , however the patients develop hypertriglyceridemia , hypercholesterolemia , and atherosclerosis . It has been assumed that protease inhibitor-dependent increases in atherosclerosis are secondary to the dyslipidemia . Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of P16671 and the accumulation of cholesteryl esters . The use of P16671 -blocking antibodies , a P16671 morpholino , and monocytes isolated from P16671 null mice demonstrated that protease inhibitor-induced increases in cholesteryl esters were dependent on P16671 upregulation . These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating P16671 and cholesteryl ester accumulation independent of dyslipidemia . Studies with P01130 null mice demonstrated that low doses of protease inhibitors induce an increase in the level of P16671 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids . Furthermore , the lack of P16671 protected the animals from protease inhibitor-induced atherosclerosis . Finally , ritonavir increased P37231 and P16671 mRNA levels in a PKC- and P37231 -dependent manner . We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of P16671 and the subsequent accumulation of sterol in macrophages . Discovery of ( 2E ) -3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide ( DB05223 ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 enzyme and COLO 205 cellular IC(50) ) , liver microsomal stability ( t(1/2) ) , cytochrome P450 inhibitory ( 3A4 IC(50) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT-116 , PC-3 , A2780 , MV4-11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials . Role of steroid receptor coactivators in glucocorticoid and transforming growth factor beta regulation of plasminogen activator inhibitor gene expression . TGFbeta is a major regulator of extracellular matrix deposition and a potent inducer of type-1 plasminogen activator inhibitor ( P05121 ) gene expression . We have reported that liganded glucocorticoid receptor ( GR ) represses TGFbeta transactivation of P05121 in Hep3B human hepatoma cells and that it interacts functionally and physically with the C-terminal activation domain of P84022 , a mediator of TGFbeta signaling . The ligand binding domain of GR is required for GR-mediated transrepression , but the GR DNA binding domain and activation function 1 domains are not . We report here that overexpression of steroid receptor coactivator-1 ( Q15788 ) and GR-interacting protein-1 ( Q9Y3R0 ) enhanced repression by liganded GR , and by a GR mutant defective in repression . Surprisingly , Q15788 and Q9Y3R0 also enhanced TGFbeta-induced activation from the TGFbeta-responsive sequence of the P05121 gene by a GR-independent mechanism . Coimmunoprecipitation and mammalian one-hybrid experiments demonstrated that Q15788 and Q9Y3R0 interact physically with endogenous P84022 and functionally with the C-terminal domain of P84022 to directly enhance transcription . Thus , the GR coactivators , Q15788 and Q9Y3R0 , act as both corepressors of the glucocorticoid repression of P05121 gene transcription , and coactivators of TGFbeta-induced activation of the P05121 promoter . Identification and functional signature of genes regulated by structurally different P00519 kinase inhibitors . Dasatinib is an DB00171 -competitive , multi-targeted P12931 and P00519 kinase inhibitor that can bind P11274 - P00519 in both the active and inactive conformations . From a clinical standpoint , dasatinib is particularly attractive because it has been shown to induce hematologic and cytogenetic responses in imatinib-resistant chronic myeloid leukemia patients . The fact because the combination of imatinib and dasatinib shows the additive/synergistic growth inhibition on wild-type Q92817 P11274 - P00519 -expressing cells , we reasoned that these P00519 kinase inhibitors might induce the different molecular pathways . To address this question , we used DNA microarrays to identify genes whose transcription was altered by imatinib and dasatinib . K562 cells were cultured with imatinib or dasatinib for 16 h , and gene expression data were obtained from three independent microarray hybridizations . Almost all of the imatinib- and dasatinib-responsive genes appeared to be similarly increased or decreased in K562 cells ; however , small subsets of genes were identified as selectively altered expression by either imatinib or dasatinib . The distinct genes that are selectively modulated by dasatinib are cyclin-dependent kinase 2 ( P24941 ) and P49336 , which had a maximal reduction of < 5-fold in microarray screen . To assess the functional importance of dasatinib regulated genes , we used RNA interference to determine whether reduction of P24941 and P49336 affected the growth inhibition . K562 and TF-1BCR- P00519 cells , pretreated with P24941 or P49336 small interfering RNA , showed additive growth inhibition with imatinib , but not with dasatinib . These findings demonstrate that the additive/synergistic growth inhibition by imatinib and dasatinib may be mediated in part by P24941 and P49336 . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Estradiol increases cell growth in human astrocytoma cell lines through ERα activation and its interaction with Q15788 and Q9Y6Q9 coactivators . Estradiol ( E2 ) regulates several cellular functions through the interaction with estrogen receptor subtypes , ERα and ERβ , which present different functional and regulation properties . ER subtypes have been identified in human astrocytomas , the most common and aggressive primary brain tumors . We studied the role of ER subtypes in cell growth of two human astrocytoma cell lines derived from tumors of different evolution grades : U373 and D54 ( grades III and IV , respectively ) . E2 significantly increased the number of cells in both lines and the co-administration with an ER antagonist ( ICI 182 , 780 ) significantly blocked E2 effects . ERα was the predominant subtype in both cell lines . E2 and ICI 182 , 780 down-regulated ERα expression . The number of U373 and D54 cells significantly increased after PPT ( ERα agonist ) treatment but not after DPN ( ERβ agonist ) one . To determine the role of Q15788 and Q9Y6Q9 coactivators in ERα induced cell growth , we silenced them with RNA interference . Coactivator silencing blocked the increase in cell number induced by PPT . The content of proteins involved in proliferation and metastasis was also determined after PPT treatment . Western blot analysis showed that in U373 cells the content of PR isoforms ( PR-A and PR-B ) , P00533 , P15692 and cyclin D1 increased after PPT treatment while in D54 cells only the content of P00533 was increased . Our results demonstrate that E2 induces cell growth of human astrocytoma cell lines through ERα and its interaction with Q15788 and Q9Y6Q9 and also suggest differential roles of ERα on cell growth depending on astrocytoma grade . Novel mechanism of steroid action in skin through glucocorticoid receptor monomers . Glucocorticoids ( GCs ) , important regulators of epidermal growth , differentiation , and homeostasis , are used extensively in the treatment of skin diseases . Using keratin gene expression as a paradigm of epidermal physiology and pathology , we have developed a model system to study the molecular mechanism of GCs action in skin . Here we describe a novel mechanism of suppression of transcription by the glucocorticoid receptor ( GR ) that represents an example of customizing a device for transcriptional regulation to target a specific group of genes within the target tissue , in our case , epidermis . We have shown that GCs repress the expression of the basal-cell-specific keratins P13647 and P02533 and disease-associated keratins K6 , P08779 , and Q04695 but not the differentiation-specific keratins K3 and P13645 or the simple epithelium-specific keratins K8 , P05783 , and P08727 . We have identified the negative recognition elements ( nGREs ) in all five regulated keratin gene promoters . Detailed footprinting revealed that the function of nGREs is to instruct the GR to bind as four monomers . Furthermore , using cotransfection and antisense technology we have found that , unlike Q15788 and Q9Y3R0 , which are not involved in the GR complex that suppresses keratin genes , histone acetyltransferase and CBP are . In addition , we have found that GR , independently from GREs , blocks the induction of keratin gene expression by P05412 . We conclude that GR suppresses keratin gene expression through two independent mechanisms : directly , through interactions of keratin nGREs with four GR monomers , as well as indirectly , by blocking the P05412 induction of keratin gene expression . P15121 inhibitor fidarestat attenuates leukocyte-endothelial interactions in experimental diabetic rat retina in vivo . PURPOSE : Dysregulation of the polyol pathway has been implicated as a major cause of diabetic retinopathy . The aldose reductase inhibitor fidarestat was recently reported to prevent retinal oxidative stress and overexpression of vascular endothelial growth factor ( P15692 ) protein in diabetic rats . In this study , we investigated the effect of fidarestat on leukocyte-endothelial cell interactions in an in vivo experimental model for diabetic retina . MATERIALS AND METHODS : Diabetes was induced in six-week-old male Long-Evans rats by intraperitoneal injection of streptozotocin ( Q11206 ) ( 75 mg/kg ) . The rats were divided into four experimental groups : non-diabetic control rats , untreated diabetic rats , and diabetic rats treated with a low ( 4 mg/kg/day ) or high ( 16 mg/kg/day ) oral dose of fidarestat . After four weeks of treatment , accumulated leukocytes in the retina were counted in vivo by acridine orange digital fluorography . Intercellular adhesion molecule-1 ( P05362 ) and P15692 -164 mRNA levels in the retina were analyzed using the quantitative reverse transcription-polymerase chain reaction . P05362 protein expression in the retina was investigated by immunohistochemistry . RESULTS : DB02021 treatment significantly decreased concentrations of sorbitol and fructose in the retinas of Q11206 -induced diabetic rats . Leukocyte accumulation in the retinas of fidarestat-treated rats was significantly less than in the untreated diabetic group ( P < 0.01 ) . DB02021 treatment significantly reduced the expression P05362 mRNA , but not P15692 -164 mRNA , in the retina of diabetic rats . Immunohistochemical study also revealed the suppressive effect of fidarestat on expression of P05362 . CONCLUSIONS : Oral administration of fidarestat attenuated leukocyte accumulation in the retina of Q11206 induced-diabetic rats , suggesting that fidarestat may have a therapeutic role in preventing the progression of diabetic retinopathy . P04150 interacting protein-1 restores glucocorticoid responsiveness in steroid-resistant airway structural cells . Glucocorticoid ( GC ) insensitivity represents a profound challenge in managing patients with asthma . The mutual inhibition of transcriptional activity between GC receptor ( GR ) and other regulators is one of the mechanisms contributing to GC resistance in asthma . We recently reported that interferon regulatory factor ( Q969Q1 ) -1 is a novel transcription factor that promotes GC insensitivity in human airway smooth muscle ( P17405 ) cells by interfering with GR signaling ( Tliba et al. , Am J Respir Cell Mol Biol 2008;38:463-472 ) . Here , we sought to determine whether the inhibition of GR function by P10914 involves its interaction with the transcriptional co-regulator GR-interacting protein 1 ( Q9Y3R0 ) , a known GR transcriptional co-activator . We here found that siRNA-mediated Q9Y3R0 depletion attenuated P10914 -dependent transcription of the luciferase reporter construct and the mRNA expression of an P10914 -dependent gene , P28907 . In parallel experiments , Q9Y3R0 silencing significantly reduced GR-mediated transactivation activities . Co-immunoprecipitation and Q86UG4 pull-down assays showed that Q9Y3R0 , through its repression domain , physically interacts with P10914 identifying Q9Y3R0 as a bona fide transcriptional co-activator for P10914 . Interestingly , the previously reported inhibition of GR-mediated transactivation activities by either P01375 and P01579 treatment or P10914 overexpression was fully reversed by increasing cellular levels of Q9Y3R0 . Together , these data suggest that the cellular accumulation of P10914 may represent a potential molecular mechanism mediating altered cellular response to GC through the depletion of Q9Y3R0 from the GR transcriptional regulatory complexes . Immunophenotyping of human HT29 colon cancer cell primary tumours and their metastases in severe combined immunodeficient mice . The immunophenotype of HT29 human colon cancer cells implanted into severe combined immunodeficient mice was assessed in primary tumours and their metastases in the lungs using an indirect immunohistochemical method . After primary tumours were surgically removed , the metastases were given time to develop , thus paralleling the clinical situation . While vimentin was negative in both primary and secondary tumours , P12830 was present as membrane-bound labelling in the primary tumours only . Whereas the markers p53 , Q86YT6 , P12004 and P06731 were consistently positive in both primary and metastatic tumours , P16070 variant 6 and Q8WXI7 were negative in metastases but positive in the primary tumours . There was a significant increase in the percentage of cells labelled for p53 in the primary tumours compared with the metastases . For the proliferation markers , there was no significant difference in labelling between primary tumours and metastases for Q86YT6 . Of the cytokeratins examined , CK 20 gave the strongest and most consistent reaction in both primary and secondary tumours . The results indicate that , for certain immunohistochemical markers , results are the same in both primary tumours and metastases . Hence , in these cases , antigens that are expressed on the primary tumour as well as on the metastases can serve as target molecules for immunologically based forms of treatment of metastases . Current and future pharmacologic options for the management of patients unable to achieve low-density lipoprotein-cholesterol goals with statins . Low-density lipoprotein-cholesterol ( LDL-C ) lowering is the mainstay of the current treatment guidelines in the management of cardiovascular risk . P04035 inhibitors ( statins ) are currently the most effective LDL-C-lowering drugs . However , a substantial number of patients do not reach treatment targets with statins . Therefore , an unmet medical need exists for lipid-lowering drugs with novel mechanisms of action to reach the recommended cholesterol target levels , either by monotherapy or combination therapy . Upregulation of the P01130 with squalene synthase inhibitors has shown promising results in animal studies but the clinical development of the lead compound lapaquistat ( DB05317 ) has recently been discontinued . DB00973 combined with statins allowed significantly more patients to reach their LDL-C targets . Other inhibitors of intestinal cholesterol absorption such as disodium ascorbyl phytostanol phosphate ( DB05449 ) and bile acid transport inhibitors have shown positive results in early development trials , whereas the prospect of acyl coenzyme A : cholesterol acyltransferase inhibition in cardiovascular prevention is dire . Selective inhibition of messenger RNA ( mRNA ) by antisense oligonucleotides is a new approach to modify cholesterol levels . The inhibition of apolipoprotein B mRNA is in advanced development and mipomersen sodium ( ISIS 301012 ) has shown striking results in phase II studies both as monotherapy as well as in combination with statins . Structural mechanism of the Pan- P11274 - P00519 inhibitor ponatinib ( DB08901 ) : lessons for overcoming kinase inhibitor resistance . The P11274 - P00519 inhibitor imatinib has revolutionized the treatment of chronic myeloid leukemia . However , drug resistance caused by kinase domain mutations has necessitated the development of new mutation-resistant inhibitors , most recently against the T315I gatekeeper residue mutation . DB08901 ( DB08901 ) inhibits both native and mutant P11274 - P00519 , including T315I , acting as a pan- P11274 - P00519 inhibitor . Here , we undertook a combined crystallographic and structure-activity relationship analysis on ponatinib to understand this unique profile . While the ethynyl linker is a key inhibitor functionality that interacts with the gatekeeper , virtually all other components of ponatinib play an essential role in its T315I inhibitory activity . The extensive network of optimized molecular contacts found in the DFG-out binding mode leads to high potency and renders binding less susceptible to disruption by single point mutations . The inhibitory mechanism exemplified by ponatinib may have broad relevance to designing inhibitors against other kinases with mutated gatekeeper residues . Ras signaling mechanisms underlying impaired GluR1-dependent plasticity associated with fragile X syndrome . Fragile X syndrome , caused by the loss of Q06787 gene function and loss of fragile X mental retardation protein ( Q06787 ) , is the most commonly inherited form of mental retardation . The syndrome is characterized by associative learning deficits , reduced risk of cancer , dendritic spine dysmorphogenesis , and facial dysmorphism . However , the molecular mechanism that links loss of function of Q06787 to the learning disability remains unclear . Here , we report an examination of small GTPase Ras signaling and synaptic AMPA receptor ( AMPA-R ) trafficking in cultured slices and intact brains of wild-type and Q06787 knock-out mice . In Q06787 knock-out mice , synaptic delivery of GluR1- , but not GluR2L- and P48058 -containing AMPA-Rs is impaired , resulting in a selective loss of GluR1-dependent long-term synaptic potentiation ( LTP ) . Although Ras activity is upregulated , its downstream MEK ( extracellular signal-regulated kinase kinase ) - P29323 ( extracellular signal-regulated kinase ) signaling appears normal , and phosphoinositide 3-kinase ( PI3K ) -protein kinase B ( P31749 ; or Akt ) signaling is compromised in Q06787 knock-out mice . Enhancing Ras-PI3K- P31749 signaling restores synaptic delivery of GluR1-containing AMPA-Rs and normal LTP in Q06787 knock-out mice . These results suggest aberrant Ras signaling as a novel mechanism for fragile X syndrome and indicate manipulating Ras-PI3K- P31749 signaling to be a potentially effective approach for treating patients with fragile X syndrome . The transcriptional coactivator DRIP/mediator complex is involved in vitamin D receptor function and regulates keratinocyte proliferation and differentiation . Mediator is a multisubunit coactivator complex that facilitates transcription of nuclear receptors . We investigated the role of the mediator complex as a coactivator for vitamin D receptor ( P11473 ) in keratinocytes . Using P11473 affinity beads , the vitamin D receptor interacting protein ( DRIP ) /mediator complex was purified from primary keratinocytes , and its subunit composition was determined by mass spectrometry . The complex included core subunits , such as Q15648 /MED1 ( MED1 ) , that directly binds to P11473 . Additional subunits were identified that are components of the RNA polymerase II complex . The functions of different mediator components were investigated by silencing its subunits . The core subunit MED1 facilitates P11473 activity and regulating keratinocyte proliferation and differentiation . A newly described subunit Q13503 also has a role in promoting keratinocyte proliferation and differentiation , whereas Q9BTT4 has an inhibitory role . Blocking MED1/ Q13503 expression caused hyperproliferation of keratinocytes , accompanied by increases in mRNA expression of the cell cycle regulator cyclin D1 and/or glioma-associated oncogene homolog . Blocking MED1 or Q13503 expression also resulted in defects in calcium-induced keratinocyte differentiation , as indicated by decreased expression of differentiation markers and decreased translocation of P12830 to the membrane . These results show that keratinocytes use the transcriptional coactivator mediator to regulate P11473 functions and control keratinocyte proliferation and differentiation . DB01093 -treated HL60 cells : a model of neutrophil-like cells mainly expressing Q07343 subtype . The human promyelocytic HL60 cells acquired a neutrophilic phenotype after a 7- to 10-day DB01093 treatment . Fc gammaRII was up-regulated . Fc gammaRI was also up-regulated by an additional P01579 treatment . These cells are able to produce O2*- by NADPH oxidase activation in the presence of immune complexes or phorbol-12-myristate-13-acetate ( PMA ) . A change of their DB05876 subtype profile was also observed : Q07343 was the predominant isoenzyme , Q08499 was down-regulated and P27815 was no longer detectable . Additionally , the more NADPH oxidase was activated by PMA , the less P27815 was expressed , suggesting that NADPH oxidase activity could be used as a surrogate marker of P27815 down-regulation . DB01954 and DB03849 ( cilomilast ) , two selective DB05876 inhibitors , dose-dependently inhibited receptor-coupled activation of superoxide . These results suggest that Q07343 is the main subtype involved in regulating superoxide induced by Fc gammaRs activation . Furthermore , these cells , expressing almost exclusively Q07343 subtype , could be useful to identify selective Q07343 inhibitors . Discrimination between agonists and antagonists by the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid-selective glutamate receptor . A mutation analysis of the ligand-binding domain of P48058 subunit . The crystal structures of the ligand-binding core of the agonist complexes of the glutamate receptor-B ( P42262 ) subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid ( AMPA ) -selective glutamate receptor indicate that the distal anionic group of agonist molecules are stabilized by interactions with an N-terminal region of an alpha-helix ( helix F ) in the lobe 2 ( " domain 2 , " Armstrong , N. , and Gouaux , E. ( 2000 ) Neuron 28 , 165-181 ) of the two-lobed ligand-binding domain . We used site-directed mutagenesis to further analyze the role of this region in the recognition of both agonists and antagonists by the AMPA receptor . Wild-type and mutated versions of the ligand-binding domain of P48058 were expressed in insect cells as secreted soluble polypeptides and subjected to binding assays using [(3)H]AMPA , an agonist , and [(3)H]Ro 48-8587 ( 9-imidazol-1-yl-8-nitro-2,3,5,6-tetrahydro[1,2,4]triazolo[1,5-c] quinazoline-2,5-dione ) , a high affinity AMPA receptor antagonist , as radioligands . Single alanine substitutions at residues DB00149 -672 and DB00156 -677 severely affected the affinities for all agonists , as seen in ligand competition assays , whereas similar mutations at residues DB00128 -673 , DB00133 -674 , DB00145 -675 , DB00133 -676 , and Lys-678 selectively affected the binding affinities of one or two of the agonists . In striking contrast , the binding affinities of [(3)H]Ro 48-8587 and of another competitive antagonist , DB03759 , were not affected by any of these alanine mutations , suggesting the absence of critical side-chain interactions . Together with ligand docking experiments , our results indicate a selective engagement of the side chains of the helix F region in agonist binding , and suggest that conformational changes involving this region may play a critical role in receptor activation . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 .

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interacts_with DB01197?

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The thioredoxin reductase inhibitor auranofin triggers apoptosis through a Bax/Bak-dependent process that involves peroxiredoxin 3 oxidation . P30044 ( TrxR ) is a key selenoprotein antioxidant enzyme and a potential target for anti-cancer drugs . One potent inhibitor of TrxR is the gold ( I ) compound auranofin , which can trigger mitochondrial-dependent apoptosis pathways . The exact mechanism of apoptosis induction by auranofin is not yet clear , but there are indications that mitochondrial oxidative stress is a central event . We assessed the redox state of the peroxiredoxins ( Prxs ) in Jurkat T-lymphoma cells treated with auranofin , and found that mitochondrial Prx3 was considerably more sensitive to oxidation than the cytosolic Prx1 and 2 , indicating selective mitochondrial stress . Prx3 oxidation was detected at apoptotic doses of auranofin in several cell types , and occurred before other mitochondrial events including cytochrome c release and mitochondrial depolarisation . DB00995 was also able to sensitise U937 cells to P01375 -mediated apoptosis . DB00995 -induced apoptosis was effectively blocked by the overexpression of Bcl-2 , and Bax/Bak deficient mouse embryonic fibroblasts were also resistant to apoptosis , indicating a central role for the pro-apoptotic proteins of this family in auranofin-triggered apoptosis . DB00995 exposure inhibited the proliferation of apoptosis-resistant cells , and at higher doses of auranofin could cause cell death through necrosis . We conclude that auranofin induces apoptosis in cells through a Bax/Bak-dependent mechanism associated with selective disruption of mitochondrial redox homeostasis in conjunction with oxidation of Prx3 . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . Combination of DB05210 and gefitinib induces apoptosis of triple-negative breast cancer cells through the PI3K/AKT- P42345 pathway . To investigate the apoptotic mechanism of triple-negative breast cancer ( TNBC ) cells induced by gefitinib and PI3K inhibitor DB05210 . MDA-MB-231 , MDA-MB-436 , and MCF-7 cells were incubated with 0.1 μmol/l gefitinib , 1 μmol/l gefitinib , 10 μmol/l gefitinib , 1 μmol/l DB05210 , 0.1 μmol/l gefitinib+1 μmol/l DB05210 , 1 μmol/l gefitinib+1 μmol/l DB05210 , and 10 μmol/l gefitinib+1 μmol/l DB05210 . Then , cell viability and survival were determined using an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide ( MTT ) assay and Hoechst staining . The apoptosis-related factors and phosphoinositide-3-kinase/protein kinase B , the mammalian target of rapamycin ( PI3K/AKT- P42345 ) signaling pathway-related factors were detected by western blot . For TNBC cells , cell viability or survival was not significantly inhibited by gefitinib or DB05210 alone ; however , marked cell apoptosis was noted in the gefitinib and DB05210 combination groups , and this effect was dose dependent . Also , the expressions of apoptosis markers , such as cleaved caspase-3 , Bcl-2/Bax , were altered by the gefitinib and DB05210 combination . Moreover , phosphorylated AKT ( p-AKT ) and 70 kDa ribosomal protein S6-kinase ( p-p70S6K ) were also inhibited by the gefitinib and DB05210 combination , which may be responsible for the apoptosis . Gefitinib combined with DB05210 could induce cell apoptosis in TNBC cells and this effect was mediated through the P00533 -PI3K-AKT- P42345 -p70S6K pathway . Our studies have set the stage for future clinical trials of TNBC therapy by the combination of gefitinib and DB05210 . Development of a sensitive , accurate and robust liquid chromatography/mass spectrometric method for profiling of angiotensin peptides in plasma and its application for atherosclerotic mice . Quantification of angiotensin ( Ang ) peptides in biological matrices is a challenge due to their low picomolar ( pM ) concentration and poor analytical performance of current methods . This work aimed to select an optimal strategy for liquid chromatography/mass spectrometry ( LC/MS ) quantification of major angiotensins in plasma of wild type and atherosclerotic mice . Optimal LC/MS set-up for Ang quantification was chosen , based on analytical performance , from : nanoflow/orbitrap , nanoflow/triple quadrupole and preconcentration nanoflow/triple quadrupole . The best LC/MS configuration ( preconcentration nanoflow/triple quadrupole ) was validated and used for measurement of angiotensins ( Ang I , II , III , IV and ( 1-7 ) ) in plasma of 6-month-old atherosclerotic apolipoprotein E/ P01130 double knock-outs ( ApoE/ P01130 ( -- / -- ) ) and wild type C57BL/6J ( WT ) mice . The method established for Ang quantification was selective , accurate and highly sensitive with LLOQ of 5pgmL(-1) . The peak area intra-day precisions for Ang II and Ang-(1-7) were in the range 3.0-5.1 and 3.5-5.8 , respectively , with corresponding accuracy of 95.4-103.5 % and 95.6-106.3 % . Plasma angiotensin profile was substantially modified in ApoE/ P01130 knock-out mice with increase in concentration of Ang II from 37.6±21.3pgmL(-1) in WT to 200.2±47.6pgmL(-1) . Concentrations of Ang I , III and IV were also increased 3-10 fold in ApoE/ P01130 ( -- / -- ) mice while that of Ang-(1-7) was unchanged . We conclude that the method developed could be effectively used for accurate , comprehensive profiling of angiotensin peptides in mouse plasma . We identified substantial changes in renin-angiotensin system in a genetic mouse model of atherosclerosis consistent with the overactivation of angiotensin converting enzyme ( P12821 ) and the impairment of Q9BYF1 . [ The effect of blood pressure-reducing therapy with captopril on tubular marker excretion in type-1 diabetics with nephropathy ] . A prospective open clinical trial was carried out with 23 hypertensive type I diabetics ( 13 men , ten women , mean age 49 +/- 9.1 years , duration of diabetes 18 +/- 9.1 years ) with early nephropathy . Glomerular and tubular renal function and metabolic parameters were monitored during 8 months ' treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , captopril , in addition to previous antihypertensive treatment with one or more drugs . Blood pressure control tended to improve on captopril ( systolic pressures 152 +/- 13 vs 140 +/- 13 mm Hg , P < 0.05 ; diastolic pressures 89 +/- 10 vs 87 +/- 10 mm Hg , not significant ) . Proteinuria ( > 0.5 g/24 hours ) fell into the microalbuminuria range ( albumin excretion 2-20 mg/mmol creatinine ) in four out of 13 patients , and microalbuminuria disappeared in four out of ten patients . Urinary levels of the brush border enzyme O60502 ( NAG ) , a marker of tubular dysfunction , were initially raised and fell significantly after 8 months ' treatment with captopril ( 20.3 +/- 14.4 vs 8.8 +/- 8.1 U/g creatinine ; P < 0.01 ) . DB01197 did not affect metabolic control ( HbA1 , total , HDL and LDL cholesterol , triglycerides , apolipoproteins A1 and B ) or the insulin dosage . These results show that long-term treatment with captopril may favourably influence both albumin excretion and NAG activity , a marker of tubular dysfunction , in type I diabetics with nephropathy . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . Epigenetic regulation of angiotensin-converting enzyme 2 ( Q9BYF1 ) by Q96EB6 under conditions of cell energy stress . Q9BYF1 ( angiotensin-converting enzyme 2 ) counterbalances the actions of P12821 ( angiotensin-converting enzyme ) by metabolizing its catalytic product , the vasoactive and fibrogenic peptide AngII ( angiotensin II ) , into Ang-(1-7) [ angiotensin-(1-7) ] . Enhanced Q9BYF1 expression may be protective in diabetes , cardiovascular disease and cancer . However , relatively little is known about the specific physiological factors regulating Q9BYF1 expression . In the present paper , we show , by Western blotting and qPCR ( quantitative real-time PCR ) , that Q9BYF1 expression is increased under conditions of cell stress , including hypoxic conditions , IL (interleukin)-1β treatment and treatment with the AMP mimic AICAR ( 5-amino-4-imidazolecarboxamide riboside ) . The NAD+-dependent deacetylase Q96EB6 ( silent information regulator T1 ) was found to be up-regulated after AICAR treatment but , conversely , was down-regulated after IL-1β treatment . ChIP analysis demonstrated that Q96EB6 bound to the Q9BYF1 promoter and that binding was increased after AICAR treatment , but decreased after IL-1β treatment . Inhibition of Q96EB6 activity ablated the AICAR-induced increase in Q9BYF1 . In conclusion , we have established that the expression of the Q9BYF1 transcript is controlled by the activity of Q96EB6 under conditions of energy stress . Antioxidant activity of oak ( Quercus ) leaves infusions against free radicals and their cardioprotective potential . The aim of present study was to evaluate antioxidant capacity and cardioprotective potential of leaves infusions and partially purified fractions of Quercus sideroxyla and Q. eduardii ( red oaks ) and Q. resinosa ( white oak ) . Consumption of polyphenol-rich beverages derived from plants , such as oak may represent a beneficial diet in terms of cardiovascular protection . Infusions from Oak leaves were obtained and probed for total phenolics by Folin-Ciocalteu , DPPH and hydroxyl radicals scavenging by DPPH test and Deoxy-D-ribose method , the antioxidant capacity was evaluated by P42345 and ORAC tests , inhibitions of Low Density Lipoproteins ( LDL ) oxidation and Angiotensin Converting Enzyme ( P12821 ) activity were measured . A HPLC analysis was performed by HPLC-MS . Bioactive polyphenols such as gallic and ellagic acids , catechin , quercetin and derivatives : naringenin and naringin were detected in Quercus infusions . A distinctive HPLC profile was observed among the red and white oak samples . Q. resinosa infusions have exhibited the highest antioxidant activity in comparison with the other species , although in the inhibition of LDL oxidation no differences were observed . In the inhibition of the P12821 , Q. resinosa was more effective ( IC50 , 18 ppm ) than Q. sideroxyla , showing same effect as the control DB01197 . From the results it is possible to postulate that not only chelating activity is important in these infusions , especially in Q. resinosa . DB06699 : a novel gonadotropin-releasing hormone blocker for the treatment of prostate cancer . Androgen deprivation therapy with gonadotropin releasing-hormone ( DB00644 ) receptor agonists provides the mainstay of endocrine treatment for advanced prostate cancer . Although effective , DB00644 agonists induce an initial testosterone surge , which can cause painful and potentially dangerous clinical flare . DB06699 is a novel P30968 blocker that provides immediate , profound and sustained testosterone reduction , without an initial surge . In a Phase III trial , degarelix and leuprolide showed similar long-term efficacy in maintaining testosterone levels of 0.5 ng/ml or less over 1 year , and induced significantly faster testosterone and prostate-specific antigen suppression . DB06699 was well tolerated ; the most common side effects were mild/moderate injection-site reactions and hot flashes . Findings to date suggest that degarelix may make an important contribution to the treatment of prostate cancer . DB08816 as an alternative in clopidogrel-associated neutropenia . DB00945 in combination with platelet Q9H244 receptor blocker has become the mainstay antiplatelet treatment strategy for the prevention of stent thrombosis . DB00208 was the first widely used Q9H244 receptor blockers , but clopidogrel has mostly replaced the use of ticlopidine due to its more favorable adverse event profile on bone marrow . However , when clopidogrel induced bone marrow toxicity occurs , little is known about the efficacy and safety of alternative treatments , and thus , in these cases , medical decisions may be very difficult . We report a case of clopidogrel-induced severe neutropenia in a patient treated with coronary stent and safety of alternative treatment with ticagrelor . Lichen planus secondary complications associated with the use of biologic therapy for rheumatoid arthritis . OBJECTIVES : Biologic therapy such as DB00005 , which is a tumor necrosis factor alpha ( P01375 -α ) inhibitor , has been extensively used as election therapy in rheumatoid arthritis . The purpose of this case presentation was to inform about the possibility that lichen planus lesions could potentially become complicated by secondary infections in patients treated with DB00005 . Furthermore , we aimed at analyzing if the complication of the cutaneous lesion was coincidental or it was due to the immunosuppressive systemic therapy , and whether the infected lesion would respond to antibiotic therapy . CASE SUMMARY : The patient was a 59-year-old woman with rheumatoid arthritis and that have had lichen planus lesions for approximately 25 years . Only recently , she had been received immunosuppressive therapy ( DB00005 and DB00563 ) . Further on , the lichen planus flared up with a secondary infection determined by a DB01603 -sensitive Staphylococcus aureus . Uncommon myocardial complications were also characteristic of this case . RESULTS : While a case report described already the appearance of lichen planus following DB00005 therapy ( Battistella M et al. , 2008 ) , the possibility that the lesion could become secondary complicated following this therapy was never reported before , according to our knowledge . Additionally , we describe in this case the interplay between DB00005 therapy and hypertrophic cardiomyopathy . CONCLUSIONS : Our case is not a lichen planus induced by DB00005 , but it is aggravated and secondary infected with DB01603 -sensitive Staphylococcus during the therapy . The additional cardiac complication ( hypertrophic cardiomyopathy ) may represent solely an evolutive sign of rheumatoid arthritis and therefore not influenced by DB00005 . All-trans retinoic acid inhibits the increases in fibronectin and P05121 induced by TGF-beta1 and Ang II in rat mesangial cells . AIM : To investigate the effect of all-trans RA ( atRA ) on the increases in plasminogen activator inhibitor-1 ( P05121 ) and fibronectin that are induced by transforming growth factor-beta1 ( TGF-beta1 ) and angiotensin II ( Ang II ) in cultured rat glomerular mesangial cells . METHODS : Subconfluent glomerular mesangial cells were serum-starved for 48 h and pretreated with atRA with subsequent stimulation of TGF-beta1 and Ang II . Protein expressions of cell-associated fibronectin and P05121 in glomerular mesangial cells were evaluated by Western blot analysis . mRNA expression of RA receptors in glomerular mesangial cells was examined by RT-PCR . RESULTS : Retinoic acid receptor-alpha , -gamma ( P10276 , -gamma ) and retinoid X receptor-alpha , -beta , -gamma ( RXR-alpha , -beta , -gamma ) mRNA were expressed in rat glomerular mesangial cells . atRA pretreatment effectively reduced fibronectin expression in glomerular mesangial cells stimulated with TGF-beta 1 or Ang II for 48 h . TGF-beta 1 stimulated P05121 expression reached a maximum at 5 h . atRA did n't affect the early ( 5 h ) P05121 induction by TGF-beta 1 , but markedly attenuated the sustained ( 48 h ) P05121 induction . atRA also decreased the prolonged effect of Ang II on P05121 expression . CONCLUSION : These results indicate that atRA inhibits the increases in fibronectin that are induced by TGF-beta1 and Ang II in cultured glomerular mesangial cells . The data also suggest that this effect of atRA is associated with a change in P05121 levels . Structural basis for substrate fatty acyl chain specificity : crystal structure of human very-long-chain acyl- DB01992 dehydrogenase . Very-long-chain acyl- DB01992 dehydrogenase ( P49748 ) is a member of the family of acyl- DB01992 dehydrogenases ( ACADs ) . Unlike the other ACADs , which are soluble homotetramers , P49748 is a homodimer associated with the mitochondrial membrane . P49748 also possesses an additional 180 residues in the C terminus that are not present in the other ACADs . We have determined the crystal structure of P49748 complexed with myristoyl- DB01992 , obtained by co-crystallization , to 1.91-A resolution . The overall fold of the N-terminal approximately 400 residues of P49748 is similar to that of the soluble ACADs including medium-chain acyl- DB01992 dehydrogenase ( P11310 ) . The novel C-terminal domain forms an alpha-helical bundle that is positioned perpendicular to the two N-terminal helical domains . The fatty acyl moiety of the bound substrate/product is deeply imbedded inside the protein ; however , the adenosine pyrophosphate portion of the C14- DB01992 ligand is disordered because of partial hydrolysis of the thioester bond and high mobility of the DB01992 moiety . The location of DB00142 -422 with respect to the P06681 - P01024 of the bound ligand and DB03147 confirms DB00142 -422 to be the catalytic base . In P11310 , Gln-95 and DB00142 -99 form the base of the substrate binding cavity . In P49748 , these residues are glycines ( DB00145 -175 and DB00145 -178 ) , allowing the binding channel to extend for an additional 12A and permitting substrate acyl chain lengths as long as 24 carbons to bind . P49748 deficiency is among the more common defects of mitochondrial beta-oxidation and , if left undiagnosed , can be fatal . This structure allows us to gain insight into how a variant P49748 genotype results in a clinical phenotype . DB02709 neuroprotection in a chronic mouse model of multiple sclerosis . DB02709 is a naturally occurring polyphenol that activates Q96EB6 , an NAD-dependent deacetylase . DB05073 , a pharmaceutical formulation of resveratrol with enhanced systemic absorption , prevents neuronal loss without suppressing inflammation in mice with relapsing experimental autoimmune encephalomyelitis ( EAE ) , a model of multiple sclerosis ( MS ) . In contrast , resveratrol has been reported to suppress inflammation in chronic EAE , although neuroprotective effects were not evaluated . The current studies examine potential neuroprotective and immunomodulatory effects of resveratrol in chronic EAE induced by immunization with myelin oligodendroglial glycoprotein peptide in C57/Bl6 mice . Effects of two distinct formulations of resveratrol administered daily orally were compared . DB02709 delayed the onset of EAE compared to vehicle-treated EAE mice , but did not prevent or alter the phenotype of inflammation in spinal cords or optic nerves . Significant neuroprotective effects were observed , with higher numbers of retinal ganglion cells found in eyes of resveratrol-treated EAE mice with optic nerve inflammation . Results demonstrate that resveratrol prevents neuronal loss in this chronic demyelinating disease model , similar to its effects in relapsing EAE . Differences in immunosuppression compared with prior studies suggest that immunomodulatory effects may be limited and may depend on specific immunization parameters or timing of treatment . Importantly , neuroprotective effects can occur without immunosuppression , suggesting a potential additive benefit of resveratrol in combination with anti-inflammatory therapies for MS . Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals . In diabetic P01130 -/- mice , an ectopic P12643 -Msx2 gene regulatory program is upregulated in association with vascular calcification . We verified the procalcific actions of aortic Msx2 expression in vivo . CMV-Msx2 transgenic ( CMV-Msx2Tg(+) ) mice expressed 3-fold higher levels of aortic Msx2 than nontransgenic littermates . On high-fat diets , CMV-Msx2Tg(+) mice exhibited marked cardiovascular calcification involving aortic and coronary tunica media . This corresponded to regions of Msx2 immunoreactivity in adjacent adventitial myofibroblasts , suggesting a potential paracrine osteogenic signal . To better understand Msx2-regulated calcification , we studied actions in 10T1/2 cells . We found that conditioned media from Msx2-transduced 10T1/2 cells ( Msx2-CM ) is both pro-osteogenic and adipostatic ; these features are characteristic of Wnt signaling . Msx2-CM stimulated Wnt-dependent TCF/LEF transcription , and Msx2-transduced cells exhibited increased nuclear beta-catenin localization with concomitant alkaline phosphatase induction . Msx2 upregulated Wnt3a and Wnt7a but downregulated expression of the canonical inhibitor Dkk1 . Dkk1 treatment reversed osteogenic and adipostatic actions of Msx2 . DB06285 , a Q03431 agonist that inhibits murine vascular calcification , suppressed vascular P12643 -Msx2-Wnt signaling . Analyses of CMV-Msx2Tg(+) mice confirmed that Msx2 suppresses aortic Dkk1 and upregulates vascular Wnts ; moreover , TOPGAL(+) ( Wnt reporter ) ; CMV-Msx2Tg(+) mice exhibited augmented aortic LacZ expression . Thus , Msx2-expressing cells elaborated an osteogenic milieu that promotes vascular calcification in part via paracrine Wnt signals . Involvement of retinoic acid receptor alpha in the stimulation of tissue-type plasminogen-activator gene expression in human endothelial cells . Retinoids stimulate tissue-type plasminogen-activator ( t-PA ) gene expression in human endothelial cells , and are likely to do so by binding to one or more nuclear retinoid receptors . The present study was initiated to identify the retinoid receptor(s) involved in this process . Expression and regulation of retinoic acid receptors ( RARs ) and retinoid X receptors ( RXRs ) were analyzed by Northern-blot analysis of total or poly(A)-rich RNA prepared from cultured human umbilical vein endothelial cells ( HUVEC ) . Prior to any exposure to retinoids , HUVEC express two transcripts for P10276 ( 3.6 kb and 2.8 kb ) , and low levels of transcripts for P10826 ( 3.4 kb and 3.2 kb ) and P13631 ( 3.3 kb and 3.1 kb ) . Two RXR subtypes were identified , RXR-alpha ( 4.8 kb ) and , at a much lower concentration , RXR-beta ( 2.4 kb ) ; no evidence for the presence of RXR-gamma was found . Furthermore , HUVEC express cellular retinol-binding protein I ( P09455 ) and cellular retinoic-acid-binding protein I ( P29762 ) mRNA . Exposure of HUVEC to 1 microM retinoic acid or the DB04942 , Ch55 , led to the induction of the two P10826 mRNAs , RXR-alpha mRNA and P09455 mRNA , whereas the expression of the other receptor and P29762 transcripts did not change appreciably . Using retinoid analogues that bind preferentially to one of the RAR or RXR subtypes , we found evidence that P10276 is involved in the retinoid-induced t-PA expression in HUVEC . This conclusion was strengthened by experiments in which blocking of P10276 with a specific P10276 antagonist , Ro 41-5253 , was demonstrated to suppress the induction of t-PA by retinoids . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth .

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5-hydroxytryptamine stimulates phosphorylation of Q8TCB0 / Q8NFH3 mitogen-activated protein kinase activation in bovine aortic endothelial cell cultures . 5-Hydroxytryptamine ( 5-HT ) is sequestered and released by endothelial cells , acts as an endothelial cell mitogen , promotes the release of nitric oxide ( NO ) , and has been associated with the Q8TCB0 / Q8NFH3 mitogen-activated protein kinase ( MAPK ) cascade . NO also acts as a cell mitogen and promotes signals that culminate in the phosphorylation of MAPK . The aim of this study was to test whether endothelial 5-HT receptors stimulate dual ( tyrosyl- and threonyl- ) phosphorylation of MAPK through a mitogen-activated protein kinase kinase-1 ( MEK-1 ) and P29474 -dependent pathway in bovine aortic endothelial cells ( BAECs ) . As shown by Western blot analysis , 5-HT and the P28222 -selective agonist 5-nonyloxytryptamine ( 5-NOT ) stimulate time- and concentration-dependent ( 0.001-10 microM ) phosphorylation of MAPK in these cells . The agonist-stimulated phosphorylation of MAPK was blocked by the 5-HT1b-receptor antagonist isamoltane ( 0.01-10 p3M ) and the MEK-1 inhibitor PD 098059 ( [ 2-(2'-amino-3'-methoxy-phenyl)-oxanaphthalen-4-one ] ; 0.01-10 microM¿ . The P29474 inhibitor L-N(omega)-iminoethyl-L-ornithine ( L-NIO ; 0.01-10 microM ) failed to block the 1 microM 5-NOT-stimulated responses . Our findings suggest that the 5-HT receptors ( specifically P28222 ) mediate signals to MEK-1 and subsequently to MAPK through an P29474 -independent pathway in BAECs . Xaliproden ( SR57746A ) induces P08908 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 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 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 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . 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 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . DB00013 induces survival or pro-apoptotic signals in human mesangial cells depending on the apoptotic stimulus . Deregulated apoptosis of MCs ( mesangial cells ) is associated with a number of kidney diseases including end-stage diabetic nephropathy . Cell death by apoptosis is a tightly orchestrated event , whose mechanisms are not completely defined . In the present study we show that the uPA ( urokinase-type plasminogen activator ) / Q03405 ( uPA receptor ) system can initiate both cell survival and pro-apoptotic signals in human MCs in response to different apoptotic stimuli . uPA abrogated MC apoptosis induced by serum withdrawal conditions and enhanced apoptosis initiated in MCs by high glucose . Effects of uPA were independent of its proteolytic activity and required Q03405 for both pro- and anti-apoptotic effects . Studies on the Q03405 interactome provide evidence that the opposing effects of uPA were directed via different Q03405 -interacting transmembrane partners . Exposure of MCs to RGD ( DB00125 - DB00145 - DB00128 ) peptide led to abrogation of the anti-apoptotic effect of uPA , which implies involvement of integrins in this process . A pro-apoptotic effect of uPA under high-glucose conditions was mediated via association of Q03405 and the cation-independent M6P ( mannose-6-phosphate ) / P11717 ( insulin-like growth factor 2 receptor ) . Both receptors were co-precipitated and co-localized in MCs . Studies on the underlying signalling indicate that the P27361 /2 ( extracellular-signal-regulated kinase 1/2 ) , Akt and Q92934 ( Bcl-2/Bcl-X(L)-antagonist , causing cell death ) protein were involved in regulation of apoptosis by uPA in MCs . P11717 mediated Q92934 perinuclear localization during apoptosis initiated by uPA and high glucose . In conclusion , we provide evidence that , in MCs , the uPA/ Q03405 system regulates survival/apoptosis processes in a stimulus-specific fashion via a mitochondria-dependent mechanism and that Q92934 protein serves as a downstream molecule . Guanylate cyclase C-mediated antinociceptive effects of linaclotide in rodent models of visceral pain . BACKGROUND DB08890 is a novel , orally administered investigational drug currently in clinical development for the treatment of constipation-predominant irritable bowel syndrome ( IBS-C ) and chronic idiopathic constipation . Visceral hyperalgesia is a major pathophysiological mechanism in IBS-C . Therefore , we investigated the anti-nociceptive properties of linaclotide in rodent models of inflammatory and non-inflammatory visceral pain and determined whether these pharmacological effects are linked to the activation of guanylate cyclase C ( P25092 ) . METHODS Orally administered linaclotide was evaluated in non-inflammatory acute partial restraint stress ( PRS ) and acute water avoidance stress ( P42768 ) models in Wistar rats , and in a trinitrobenzene sulfonic acid ( TNBS ) -induced inflammatory model in Wistar rats and P25092 null mice . KEY RESULTS In TNBS-induced colonic allodynia , linaclotide significantly decreased the number of abdominal contractions in response to colorectal distension without affecting the colonic wall elasticity change in response to distending pressures after TNBS . However , linaclotide had no effect on visceral sensitivity under basal conditions . In addition , linaclotide significantly decreased colonic hypersensitivity in the PRS and P42768 models . In wild type ( wt ) and P25092 null mice , the instillation of TNBS induced similar hyperalgesia and allodynia . However , in post-inflammatory conditions linaclotide significantly reduced hypersensitivity only in wt mice , but not in P25092 null mice . CONCLUSIONS & INFERENCES These findings indicate that linaclotide has potent anti-nociceptive effects in several mechanistically different rodent models of visceral hypersensitivity and that these pharmacological properties of linaclotide are exerted through the activation of the P25092 receptor . Therefore , linaclotide may be capable of decreasing abdominal pain in patients suffering from IBS-C . Targeting ornithine decarboxylase in Myc-induced lymphomagenesis prevents tumor formation . Checkpoints that control Myc-mediated proliferation and apoptosis are bypassed during tumorigenesis . Genes encoding polyamine biosynthetic enzymes are overexpressed in B cells from E mu-Myc transgenic mice . Here , we report that disabling one of these Myc targets , P11926 ( Odc ) , abolishes Myc-induced suppression of the Cdk inhibitors P38936 (Cip1) and p27(Kip1) , thereby impairing Myc 's proliferative , but not apoptotic , response . Moreover , lymphoma development was markedly delayed in E mu-Myc;Odc(+/-) transgenic mice and in E mu-Myc mice treated with the Odc inhibitor difluoromethylornithine ( DB06243 ) . Strikingly , tumors ultimately arising in E mu-Myc;Odc(+/-) transgenics lacked deletions of Arf , suggesting that targeting Odc forces other routes of transformation . Therefore , Odc is a critical Myc transcription target that regulates checkpoints that guard against tumorigenesis and is an effective target for cancer chemoprevention . DB01780 signaling reveals 14-3-3 protein function as a novel step in left-right patterning during amphibian embryogenesis . To gain insight into the molecular mechanisms underlying the control of morphogenetic signals by H+ flux during embryogenesis , we tested DB01780 -A ( FC ) , a compound produced by the fungus Fusicoccum amygdali Del . In plant cells , FC complexes with 14-3-3 proteins to activate H+ pumping across the plasma membrane . It has long been thought that FC acts on higher plants only ; here , we show that exposing frog embryos to FC during early development specifically results in randomization of the asymmetry of the left-right ( LR ) axis ( heterotaxia ) . Biochemical and molecular-genetic evidence is presented that 14-3-3-family proteins are an obligate component of Xenopus FC receptors and that perturbation of 14-3-3 protein function results in heterotaxia . The subcellular localization of 14-3-3 mRNAs and proteins reveals novel cytoplasmic destinations , and a left-right asymmetry at the first cell division . Using gain-of-function and loss-of-function experiments , we show that P62258 protein is likely to be an endogenous and extremely early aspect of LR patterning . These data highlight a striking conservation of signaling pathways across kingdoms , suggest common mechanisms of polarity establishment between C. elegans and vertebrate embryos , and uncover a novel entry point into the pathway of left-right asymmetry determination . [ Effect of urotensin II on secretion of adrenomedullin from human vascular endothelial cells ] . OBJECTIVE : To study the effect of human urotensin II ( HU II ) on secretion of adrenomedullin ( P35318 ) from human vascular endothelial cells ( Q15223 ) and its mechanism . METHODS : In cultured Q15223 , different concentrations of HUII were used to stimulate the P35318 secretion from Q15223 , and the inhibitors of different signal transduction pathway were used to investigate their effects on P35318 secretion . The contents of P35318 in medium were determined by radio immunoassay . RESULTS : HUII stimulated secretion of P35318 from Q15223 in a time-dependent and concentration-dependent manner . The contents of P35318 in the experiment groups were changed compared with that in control group ( P < 0.05 ) . The increase of P35318 could be inhibited by inhibitor of extracellular signal-regulated protein kinase ( PD(98059) ) , inhibitor of O75791 kinase ( SB(202190) ) , inhibitor of calmodulin ( W(7) ) and inhibitor of Ca(2+) ( nicardipine ) ( P < 0.05 ) . The inhibition ratio in those groups was 68 % , 78 % , 24 % and 25 % respectively . But the inhibitor of Calcineurin ( CaN ) and inhibitor of protein kinase C ( H(7) ) had no influence on the secretion of P35318 from Q15223 ( P > 0.05 ) . CONCLUSION : The stimulated effect of HUII on the P35318 secretion from Q15223 may be mediated by Ca(2+) , ERKs , P62158 -PK and O75791 signal transduction pathways . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Potential opposite roles of the extracellular signal-regulated kinase ( P29323 ) pathway in autism spectrum and bipolar disorders . Signal transduction from the synapse to the nucleus subsequently involves transient increases in synaptic Ca2+ , activation of P62158 kinases , activation of the GTPase Ras , activation of the P29323 mitogen-activated protein kinase pathway , and finally GSK3 inhibition and CREB-activation . Genetic studies in autism have identified mutations and copy number variations in a number of genes involved in this synapse to nucleus signaling path . In particular , a gain of function mutation in the Q13936 gene , deletions and disruption of the Q96PV0 gene , a copy number variation encompassing the P27361 gene and a duplication of P62258 indicate that in a subset of autism patients the P29323 cascade is inappropriately activated . Predicted functional consequences of this hyperactivation would be an increase in complexity of the dendritic tree , and via inhibition of GSK3 , a delayed circadian phase . The latter effect indeed fits the frequent sleep disturbances observed in autistic patients . Interestingly , the sleep disturbances in bipolar disorder patients are frequently characterized as phase advanced . A selective evaluation of genetic mutations in bipolar patients indicates that the activity of the P29323 cascade , at least in a subset of patients , presumably is hypoactive . Thus , with respect to the P29323 pathway , autism and bipolar disorder seem each other 's counter pole . Can P35354 inhibitor-induced increase in cardiovascular disease risk be modified by essential fatty acids ? Selective P35354 inhibitors increase the risk of myocardial infarction and stroke . This has been attributed to their ability to inhibit endothelial P35354 derived prostacyclin ( DB01240 ) but not platelet P23219 derived thromboxane A2 ( TXA2 ) . On the other hand , aspirin blocks both P23219 and P35354 enzymes without decreasing DB01240 but blocks TXA2 synthesis that explains its beneficial action in the prevention of coronary heart disease ( Q8NE62 ) . The inhibitory action of aspirin on P23219 and P35354 enzymes enhances the tissue concentrations of dihomo-gamma-linolenic acid ( DB00154 ) , arachidonic acid , eicosapentaenoic acid ( EPA ) , and docosahexaenoic acid ( DB01708 ) . These fatty acids form precursors to PGE1 , DB01240 , PGI3 , lipoxins ( LXs ) , and resolvins that have anti-inflammatory actions . In contrast , increase in the concentrations of DB00154 , AA , EPA , and DB01708 is much less with specific P35354 inhibitors since they do not block the formation of eicosanoids through P23219 pathway . P35354 inhibitors interfere with the formation of LXs and resolvins that have neuroprotective and cardioprotective actions . EPA and DB01240 have anti-arrhythmic action . EPA , DB01708 , and AA augment eNO formation that prevents atherosclerosis . This suggests that P35354 inhibitors increase cardiovascular and stroke risk by interfering with the formation of eNO , DB01240 , LXs , and resolvins and implies that combining EFAs with P35354 inhibitors could prevent these complications . Mechanism of inhibition of the P42262 AMPA receptor channel opening by talampanel and its enantiomer : the stereochemistry of the 4-methyl group on the diazepine ring of 2,3-benzodiazepine derivatives . Stereoselectivity of 2,3-benzodiazepine compounds provides a unique way for the design of stereoisomers as more selective and more potent inhibitors as drug candidates for treatment of the neurological diseases involving excessive activity of AMPA receptors . Here we investigate a pair of enantiomers known as DB04982 and its ( + ) counterpart about their mechanism of inhibition and selectivity toward four AMPA receptor subunits or P42261 -4 . We show that DB04982 is the eutomer with the endismic ratio being 14 for the closed-channel and 10 for the open-channel state of P42262 . Kinetic evidence supports that DB04982 is a noncompetitive inhibitor and it binds to the same site for those 2,3-benzodiazepine compounds with the C-4 methyl group on the diazepine ring . This site , which we term as the " M " site , recognizes preferentially those 2,3-benzodiazepine compounds with the C-4 methyl group being in the R configuration , as in the chemical structure of DB04982 . Given that DB04982 inhibits P42261 and P42262 , but is virtually ineffective on the P42263 and P48058 AMPA receptor subunits , we hypothesize that the " M " site(s) on P42261 and P42262 to which DB04982 binds is different from that on P42263 and P48058 . If the molecular properties of the AMPA receptors and DB04982 are used for selecting an inhibitor as a single drug candidate for controlling the activity of all AMPA receptors in vivo , DB04982 is not ideal . Our results further suggest that addition of longer acyl groups to the N-3 position should produce more potent 2,3-benzodiazepine inhibitors for the " M " site . Differential actions of vasopeptidase inhibition versus angiotensin-converting enzyme inhibition on diuretic therapy in experimental congestive heart failure . BACKGROUND : DB00886 ( OMA ) , a vasopeptidase inhibitor , simultaneously inhibits angiotensin-converting enzyme ( P12821 ) and neutral endopeptidase , which degrades vasodilatory factors ( eg , P35318 ) and natriuretic peptides . Based on the beneficial cardiorenal and humoral properties of the natriuretic peptides , we hypothesized that an acute vasopeptidase inhibitor with or without diuretic would result in more favorable cardiorenal and hormonal actions than P12821 inhibition plus diuretic ( ACEI+D ) in congestive heart failure . METHODS AND RESULTS : We compared the actions of OMA alone and with diuretic ( OMA+D ) to ACEI+D in a model of pacing-induced congestive heart failure . OMA+D decreased pulmonary arterial and pulmonary capillary wedge pressures to a greater level than OMA alone or ACEI+D . Glomerular filtration rate was lower with ACEI+D than with either OMA group . Plasma renin activity and aldosterone immediately increased with ACEI+D , whereas OMA+D resulted in higher plasma renin activity and a delayed increase in aldosterone . OMA alone did not increase plasma renin activity and aldosterone , but resulted in a sustained increase in plasma adrenomedullin , with higher urinary atrial natriuretic peptide , adrenomedullin , and cGMP excretions than with ACEI+D . CONCLUSIONS : Acute administration of OMA with or without diuretic results in more favorable cardiorenal and humoral responses in experimental congestive heart failure than does ACEI+D . There is no acute activation of renin and aldosterone with OMA alone such as occurs with ACEI+D and OMA+D . Thus , OMA with or without a diuretic possesses beneficial cardiorenal and humoral actions comparable to those observed with ACEI+D that can be explained by potentiation of natriuretic peptides . Changes of several brain receptor complexes in the cerebral cortex of patients with Alzheimer disease : probable new potential pharmaceutical targets . Although Alzheimer disease ( AD ) has been linked to defects in major brain receptors , studies thus far have been limited to the determination of receptor subunits or specific ligand binding studies . However , the availability of current technology enables the determination and quantification of brain receptor complexes . Thus , we examined levels of native receptor complexes in the brains of patients with AD . Cortical tissue was obtained from control subjects ( n = 12 females and 12 males ) and patients with AD ( n = 12 females and 12 males ) within a 3-h postmortem time period . The tissues were kept frozen until further biochemical analyses . Membrane proteins were extracted and subsequently enriched by ultracentrifugation using a sucrose gradient . Membrane proteins were then electrophoresed onto native gels and immunoblotted using antibodies against individual brain receptors . We found that the levels were comparable for complexes containing GluR2 , GluR3 and P48058 as well as P08908 . Moreover , the levels of complexes containing muscarinic AChR M1 , Q9UHB4 and GluR1 were significantly increased in male patients with AD . Nicotinic AChRs 4 and 7 as well as dopaminergic receptors D1 and D2 were also increased in males and females with AD . These findings reveal a pattern of altered receptor complex levels that may contribute to the deterioration of the concerted activity of these receptors and thus result in cognitive deficits observed in patients with AD . It should be emphasised that receptor complexes function as working units rather than individual subunits . Thus , the receptor deficits identified may be relevant for the design of experimental therapies . Therefore , specific pharmacological modulation of these receptors is within the pharmaceutical repertoire . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . Dual effects of histone deacetylase inhibition by trichostatin A on endothelial nitric oxide synthase expression in endothelial cells . Inhibition of histone deacetylases by trichostatin A ( P32119 ) has pleiotropic effects on gene expression . We demonstrated that at low dose ( 0.1 microg ) P32119 increased the P29474 mRNA levels , which was followed by a time- and dose-dependent down-regulation . Cycloheximide , a protein synthesis inhibitor , completely abolished P32119 -induced decrease in P29474 expression , indicating that new protein synthesis is required for the inhibiting effect . DB06693 -- an inhibitor P04035 and geranylgeranylation reaction dose-dependently antagonized P32119 -induced reduction . This mevastatin-mediated antagonism was completely abolished by geranylgeranylpyrophosphate , suggesting that geranylgeranyl modification is needed to activate the P29474 mRNA destabilizing factor -- a mechanism responsible for statin-mediated P29474 upregulation . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients . Transcriptional regulation of artemin is related to neurite outgrowth and actin polymerization in mature Q86YR7 neurons . Q5T4W7 is a member of the glial cell line-derived neurotrophic factor ( P39905 ) family of ligands that helps to ensure the survival of sensory neurons . We used an in vitro isolated dorsal root ganglia model to study the effects of artemin on the adult rat neuronal system and investigate differentially regulated genes . We found that 285 genes were differentially transcribed by artemin after 3 h of treatment , including genes related to cell adhesion and actin polymerization . A series of genes involved in the regulation of actin dynamics , including coronin , Myr 5 , P42768 interacting protein , cofilin , drebrin and dynamin were down-regulated by artemin , suggesting that it plays a previously undefined role in the regulation of actin polymerization and synaptic vesicle movement . Q5T4W7 also down-regulated the expression of genes related to cell adhesion and matrix assembly , including biglycan , plectin , nestin , neuronatin and the neuron-glia- P62158 -related cell adhesion molecule , which is functionally relevant to neurite elongation in Q86YR7 neurons . Q5T4W7 resulted in increases in total neurite length and branching of the Q86YR7 neurons . Also artemin caused an increase of synaptic vesicle clustering . Our results showed that the inhibition of DNA methylation suppressed the artemin-dependent neurite growth , suggesting that the genetic regulation could be relevant to neurite elongation in mature Q86YR7 . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process .

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What future for combination therapies ? For most patients who require lipid-lowering treatment , statin monotherapy is the appropriate treatment . However , in those patients where statin monotherapy does not produce optimal lipid levels , the combination of a statin with niacin , a bile acid sequestrant , a fibric acid derivative , a cholesterol absorption inhibitor or a fish oil preparation may provide improved control . The choice of combination therapy depends upon the patient 's lipid profile and tolerability of the medication . Combination of a statin with niacin , a bile acid sequestrant or ezetimibe , a cholesterol absorption inhibitor , should be considered for patients with very high low-density lipoprotein cholesterol ( LDL-C ) levels , while combination with either a fibric acid derivative or a fish oil should be considered for patients with high LDL-C and high triglyceride levels . A number of new lipid-lowering agents are currently in development , including cholesteryl ester transfer protein ( P11597 ) inhibitors , acyl coenzyme A : cholesterol acyltransferase ( ACAT ) inhibitors , ileal bile acid transport ( Q12908 ) inhibitors , microsomal triglyceride transfer protein ( P55157 ) inhibitors and dual peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma agonists . Introduction of these novel therapies will provide opportunities for developing different combination strategies that may help to optimise lipid profiles in patients who are currently difficult to treat . The introduction of new combinations will require careful study to ensure that the risks of drug interactions and adverse events are minimised . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Down-regulation of hepatic cytochrome P450 enzymes in rats with trinitrobenzene sulfonic acid-induced colitis . Hepatic cytochrome P450 ( P450 ) enzymes are down-regulated during inflammation . In this study , an animal model of inflammatory bowel disease was subjected to characterization of hepatic P450 expression under inflammatory conditions . Rats were treated intracolonically with 100 mg/kg trinitrobenzene sulfonic acid ( TNBS ) dissolved in 30 % ethanol , and homogenates of colonic mucosa and hepatic microsomes of the rats were prepared . The colitis was accompanied by appearance of higher levels of portal endotoxin , interleukin-6 , and nitric oxide metabolites and decreases in contents and activities for hepatic CYP3A2 , CYP2C11 , and , to a lesser extent , P05177 and P05181 . DB04743 , a preferential P35354 inhibitor , protected rats with TNBS-induced colitis ( TNBS-colitis ) against the down-regulation of hepatic CYP3A2 . DB00781 , which neutralizes endotoxin , curcumin , which has anti-inflammatory properties , and gadolinium chloride , which inactivates macrophages , attenuated the down-regulation of CYP3A2 . Similar effects were observed in other P450s such as CYP2C11 , but the agents were less effective in attenuating the down-regulation . Our data suggest that endogenous substances leaked from damaged colon in the rats with TNBS-colitis activate Kupffer cells , leading to down-regulation of hepatic P450s with differential susceptibility to the inflammatory stimuli . The colitis model , instead of exogenous administration of lipopolysaccharide or cytokines , could be applied to the study on mechanisms for altered hepatic P450 expression and other liver functions under mild inflammatory conditions . Q9UBK8 Q9UBK8 66A > G has no effect on total homocysteine , folate , and DB00115 concentrations in renal transplant patients . The association of variants of the gene encoding methionine synthase reductase ( Q9UBK8 ) with hyperhomocysteinemia , folate and Vitamin B(12) status in kidney graft recipients is unknown . We examined two mutations in Q9UBK8 in a cross-sectional study of 733 kidney graft recipients . The allele frequency of Q9UBK8 66G was 0.55 . 369 patients ( 50.3 % ) were heterozygous and 219 patients ( 29.9 % ) were homozygous for the mutation . None of the patients showed the 997C > G mutation . The allelic variants of Q9UBK8 66A > G showed no significant association with total homocysteine ( tHcy ) levels , both in univariate analyses , and in a multivariate model controlling for age , gender , body mass index , renal function , time since transplantation , underlying kidney disease , as well as the P42898 677C > T/1298A > C genotypes . Similarly , no significant associations between the Q9UBK8 66A > Ggenotypes and plasma folate or Vitamin B(12) levels were found . In conclusion , Q9UBK8 66A > G has no major effect on tHcy , folate , or Vitamin B(12) plasma concentrations in kidney graft recipients . Cyclooxygenase isozymes are expressed in human myeloma cells but not involved in anti-proliferative effect of cyclooxygenase inhibitors . Considering possible tumorigenic activity of cyclooxygenase ( P36551 ) isozymes in myeloma , we examined expression levels of P23219 and -2 in seven human myeloma cell lines ( Q5SW96 -77 , IM-9 , RPMI-8226 , HPC , HS-Sultan , TSPC-1 , and U-266 ) . As analyzed by reverse transcriptase-polymerase chain reaction ( RT-PCR ) , all the cell lines constitutively expressed P23219 , while P35354 levels markedly varied among different cell lines . Induction of P35354 by phorbol ester was observed in RPMI-8226 and HPC cells . In contrast , P35354 was constitutively expressed in Q5SW96 -77 and IM-9 cells . Moreover , the high expression level of P35354 protein in Q5SW96 -77 cells was verified by Western blotting . Intact cells of Q5SW96 -77 converted 14C-labeled arachidonic acid to prostaglandin E2 , F2alpha , and D2 , and this activity was dose-dependently inhibited by selective P35354 inhibitors ( SC-58125 and NS-398 ) , a non-selective P36551 inhibitor ( indomethacin ) , and relatively high concentrations of a selective P23219 inhibitor ( SC-560 ) . These P36551 inhibitors also suppressed the proliferation of Q5SW96 -77 cells , but significant suppression was seen only at 100 microM , a much higher concentration than those sufficient for the P36551 inhibition . Moreover , proliferation of the myeloma cells lacking P35354 was also suppressed by 100 microM of SC-58125 . These results suggested that the anti-proliferative effect of the P36551 inhibitors is independent of the inhibition of P35354 . JTT-705 blocks cell proliferation and angiogenesis through p38 kinase/p27(kip1) and Ras/ P38936 (waf1) pathways . The excessive proliferation and migration of vascular smooth muscle cells ( SMCs ) participate in the growth and instability of atherosclerotic plaque . We examined the direct role of a newly developed chemical inhibitor of cholesteryl ester transfer protein , JTT-705 , on SMC proliferation and angiogenesis in endothelial cells ( ECs ) . JTT-705 inhibited human coronary artery SMC proliferation . JTT-705 induced the phosphorylation of p38 mitogen-activated protein kinase ( MAPK ) and extracellular-signal-regulated kinases ( P29323 ) in SMCs . In addition , the anti-proliferative effects of JTT-705 in SMCs were blocked by p38 MAPK inhibitor . JTT-705 induced the upregulation of p- P38936 (waf1) , and this effect was blocked by dominant-negative Ras ( N17 ) , but not by inhibitors of p38 MAPK or P29323 . In addition , JTT-705 also induced the upregulation of p27(kip1) , and this effect was blocked by p38 MAPK inhibitor . Interestingly , culture medium from JTT-705-treated SMCs blocked human coronary artery EC tube formation in an in vitro model of angiogenesis indirectly via a decrease in vascular endothelial growth factor ( P15692 ) from SMCs and directly via an anti-proliferative effect in ECs . JTT-705 blocked the proliferation of SMCs through the activation of p38 kinase/p27(kip1) and Ras/ P38936 (waf1) pathways , and simultaneously blocked EC tube formation associated with a decrease in P15692 production from SMCs and an anti-proliferative effect in ECs . Our results indicate that JTT-705 may induce a direct anti-atherogenic effect in addition to its inhibitory effect of P11597 activity . Isoenzyme-specific cyclooxygenase inhibitors : a whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6 . NSAIDs inhibit the conversion of arachidonic acid into DB03866 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase ( P36551 ) . Two genetically distinct isoforms have been discovered , P23219 and P35354 . While P23219 is thought to account for homeostatic amounts of eicosanoids , P35354 is induced during inflammation leading to pathologic amounts of eicosanoids . Since NSAIDs inhibit both P36551 isoforms , antiinflammatory drug research has refocused to discovering P35354 inhibitors that do not inhibit P23219 . For this purpose , we have developed a whole cell assay system using the human erythroleukemic cell line HEL as a source for P23219 and the human monocytic cell line Mono Mac 6 as a source for P35354 . Mono Mac 6 cells express high amounts of P35354 upon stimulation with lipopolysaccharide ( LPS ) in the absence of any detectable P23219 protein . On the other hand , we find HEL cells to naturally express P23219 protein , but not P35354 . Testing of a panel of NSAIDs as well as some P35354 specific inhibitors showed that this assay system is suitable for identifying compounds that selectively inhibit either P23219 or P35354 . This test system offers the advantage of assessing P23219 and P35354 inhibitors within the human species , within a similar test set-up , and circumvents the need for tedious purification of either platelets or peripheral blood monocytes . Ca2+ response of rat mesangial cells to DB00171 analogues . The aim of this investigation was to characterise the effects of DB00171 analogues and UTP on the single cell intracellular Ca2+ concentration ( [Ca2+]i ) in cultured rat mesangial cells . Typically , there were two phases in the Ca2+ response to the agonists , an initial fast transient peak and a subsequent slower decline , or plateau , phase . For the peak amplitude in [Ca2+]i the agonists had about equal effect . But when taking in consideration the percentage of responding cells and the integrated Ca2+ response over 1 min , the order of efficacy of nucleotide agonists ( 100 microM ) was UTP = DB00171 > ATPgammaS > ADP = 2MeS- DB00171 ( 2-methylthio- DB00171 ) . DB00640 , AMP and beta,gamma-Me- DB00171 ( 100 microM ) had no effect . DB04786 ( 100 microM ) and reactive blue ( 50 microM ) decreased the number of responding cells . Removing Ca2+ from the bath diminished neither the peak in [Ca2+]i nor the percentage of responding cells , but the average [Ca2+]i increase in 1 min was significantly reduced . The results indicate that P41231 receptors are present in rat mesangial cells but it can not be excluded that there are receptors distinct from P41231 which also mediate a rise in [Ca2+]i . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . Novel function of histamine signaling in hyperlipidemia-induced atherosclerosis : DB11320 H1 receptors protect and H2 receptors accelerate atherosclerosis . DB11320 is not only essential for acute inflammatory reactions , but it also participates in a chronic inflammatory disorder . We generated apolipoprotein E ( apoE ) and histamine receptors ( HHRs ) , including the major H1 and H2 receptors ( P35367 , P25021 ) double knockout mice ( DKO ) to clarify the role of HHRs in hyperlipidemia-induced atherosclerosis , in which apoE-KO and DKO mice were fed a high cholesterol diet . We found that pronounced hyperlipidemia-induced atherosclerotic progression occurred in P35367 /apoE-DKO mice , but in P25021 /apoE-DKO mice less atherosclerosis , despite pro-atherogenic serum cholesterol levels compared with apoE-KO mice . Furthermore , the increased expressions of scavenger receptors ( SRs ) , such as SR-A , P16671 and lectin-like oxidized low-density lipoprotein receptor 1 ( P78380 ) , nuclear factor-kappa B ( NFκB ) , monocyte chemoattractant protein ( P13500 ) , matrix metalloproteinases ( MMPs ) or liver X receptor ( LXR ) -related inflammatory signaling factors , were consistent with the pro-atherogenic phenotype of P25021 /apoE-DKO mice . We hypothesize that histamine/ P35367 and P25021 signaling has conflicting innate functions , inflammatory/atherogenic and anti-inflammatory/anti-atherogenic actions , and that there are innate links between histamine signaling and hyperlipidemia-induced atherosclerosis , independently of serum cholesterol metabolism . Specific histamine signaling blockers , in particular , P25021 blockers , are a possible novel therapeutic target for hyperlipidemia-induced atherosclerosis . Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . Splenic autonomic denervation increases inflammatory status but does not aggravate atherosclerotic lesion development . The brain plays a prominent role in the regulation of inflammation . Immune cells are under control of the so-called cholinergic anti-inflammatory reflex , mainly acting via autonomic innervation of the spleen . Activation of this reflex inhibits the secretion of proinflammatory cytokines and may reduce the development of atherosclerosis . Therefore , the aim of this study was to evaluate the effects of selective parasympathetic ( Px ) and sympathetic ( Sx ) denervation of the spleen on inflammatory status and atherosclerotic lesion development . Female P02649 *3-Leiden. P11597 mice , a well-established model for human-like lipid metabolism and atherosclerosis , were fed a cholesterol-containing Western-type diet for 4 wk after which they were subdivided into three groups receiving either splenic Px , splenic Sx , or sham surgery . The mice were subsequently challenged with the same diet for an additional 15 wk . Selective Px increased leukocyte counts ( i.e. , dendritic cells , B cells , and T cells ) in the spleen and increased gene expression of proinflammatory cytokines in the liver and peritoneal leukocytes compared with Sx and sham surgery . Both Px and Sx increased circulating proinflammatory cytokines IL-1β and P05231 . However , the increased proinflammatory status in denervated mice did not affect atherosclerotic lesion size or lesion composition . CONCLUSION : Predominantly selective Px of the spleen enhances the inflammatory status , which , however , does not aggravate diet-induced atherosclerotic lesion development . DB08827 . Aegerion Pharmaceuticals is developing lomitapide , a small-molecule , microsomal triglyceride transfer protein ( P55157 ) inhibitor , for the treatment of both familial and primary hypercholesterolemia . Oral , once-daily lomitapide will be targeted at patients resistant to P04035 inhibitors ( statins ) either due to abnormalities in liver function or to discontinuation because of muscle pain . An oral formulation of lomitapide is in phase III development for homozygous familial hypercholesterolemia ( hyperlipoproteinemia type IIa ) in the US , Canada , Italy , and South Africa . This review discusses the key development milestones and therapeutic trials of this drug . [ P02656 , P11597 , beta-fibrinogen and P42898 are genetic determinants of carotid intim-media thickness ( Stanislas cohort ) ] . The purpose of this study was to examine the relationship between carotid intima-media thickness ( CIMT ) interindividual variability and 16 polymorphisms of 11 genes associated with cardiovascular risk factors ( genes among lipid and homocysteine metabolisms , blood viscosity , platelet aggregation , leukocyte adhesion and renin-angiotensin system ) . CIMT was measured by high resolution B mode ultrasonography in an healthy population of 77 men and 84 women , aged 35-54 years and selected from a French cohort : the Stanislas cohort . The polymorphisms studied were genotyped by a multilocus approach . Statistical analysis were done by Q9UNW9 after adjustment of CIMT for age , BMI and smoking and by multiple regression analyses . No association was found with P04114 Thr71 DB00167 , P02656 -482C/T , -455T/C , GpIIIa P1A , AT1R 1166A/C , AGT Met235Thr , P35520 Ile278Thr , P16581 98G/T and P16581 Ser128Arg , polymorphism neither in men nor in women . Although , in women we found always no association for the P02656 3206T/G , 3175C/G , 1100C/T , the P11597 Ile405Val , the P42898 677C/T and the fibrinogen -455G/A polymorphism 's , in men these polymorphism 's were associated with CIMT variability ( 0.01 < or = p < or = 0.05 ) . The most interesting finding was that altogether these genes in men were able to explain a considerable part , 20.6 % , of CIMT variability . Therefore , our study gives a new opportunity to understand CIMT variability . Intraretinal lipid transport is dependent on high density lipoprotein-like particles and class B scavenger receptors . PURPOSE : In our companion paper we demonstrated that circulating lipoproteins enter the retina via the retinal pigment epithelium ( Q96AT9 ) and possibly Müller cells . In order to understand how these lipids are transported within the retina , expression and localization of the main proteins known to be involved in systemic lipid transport was determined . METHODS : Expression of O95477 , apoA1 ( the major HDL protein ) , Q8WTV0 , SR- Q15878 , P16671 , lecithin:cholesterol acyltransferase ( P04180 ) , and cholesteryl ester transfer protein ( P11597 ) was determined by reverse transcriptase polymerase chain reaction ( RT-PCR ) and immunoblots . Localization was determined by immunohistochemistry using fresh monkey vibrotome sections and imaged by confocal microscopy . RESULTS : O95477 and apoA1 were localized to the ganglion cell layer , retinal pigment epithelium ( Q96AT9 ) , and rod photoreceptor inner segments . ApoA1 was also observed associated with rod photoreceptor outer segments , presumably localized to the interphotoreceptor matrix ( IPM ) . The scavenger receptors Q8WTV0 and SR- Q15878 localized mainly to the ganglion cell layer and photoreceptor outer segments ; in the latter they appear to be associated with microtubules . P04180 and P11597 localized mainly to the IPM . CONCLUSIONS : The presence and specific localization of these well-known lipid transport proteins suggest that the retina employs an internal lipid transport mechanism that involves processing and maturation of HDL-like particles . P08183 , Q9UNQ0 , and P60484 determine the response of glioblastoma to temozolomide and ABT-888 therapy . PURPOSE : Little is known about the optimal clinical use of ABT-888 ( veliparib ) for treatment of glioblastoma . ABT-888 is a PARP inhibitor undergoing extensive clinical evaluation in glioblastoma , because it may synergize with the standard-of-care temozolomide ( DB00853 ) . We have elucidated important factors controlling ABT-888 efficacy in glioblastoma . EXPERIMENTAL DESIGN : We used genetically engineered spontaneous glioblastoma mouse models and allograft models that were orthotopically transplanted into wild-type ( WT ) and Abcb1/Abcg2-deficient ( KO ) recipients . RESULTS : ABT-888/ DB00853 is not efficacious against p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR allografts in wild-type recipients , indicating inherent resistance . Abcb1/Abcg2 mediated efflux of ABT-888 at the blood-brain barrier ( BBB ) causes a 5-fold reduction of ABT-888 brain penetration ( P < 0.0001 ) that was fully reversible by elacridar . Efficacy studies in WT and KO recipients and/or concomitant elacridar demonstrate that Abcb1/Abcg2 at the BBB and in tumor cells impair DB00853 /ABT-888 combination treatment efficacy . DB04881 also markedly improved DB00853 /ABT-888 combination treatment in the spontaneous p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR glioblastoma model . Importantly , ABT-888 does enhance DB00853 efficacy in Pten deficient glioblastoma allografts and spontaneous tumors , even in Abcb1/Abcg2 proficient wild-type mice . Loss of P60484 occurs frequently in glioblastoma ( 36 % ) and in silico analysis on patient with glioblastoma samples revealed that it is associated with a worse overall survival ( 310 days vs. 620 days , n = 117 ) . CONCLUSIONS : The potential of ABT-888 in glioblastoma can best be demonstrated in patients with P60484 null tumors . Therefore , clinical trials with ABT-888 should evaluate these patients as a separate group . Importantly , inhibition of P08183 and Q9UNQ0 ( by elacridar ) may improve the efficacy of DB00853 /ABT-888 therapy in all glioblastoma patients . DB09036 : first global approval . The anti-interleukin-6 ( P05231 ) chimeric monoclonal antibody siltuximab is the first drug to be approved for the treatment of multicentric Castleman 's disease ( O95822 ) in the US and European union ( EU ) , having gained approval under the FDA priority review program in the US and from an accelerated assessment and recommendation by the Committee for Medicinal Products for Human Use ( CHMP ) in the EU . Development of the drug is continuing in smoldering multiple myeloma . This article summarizes the milestones in the development of siltuximab leading to this first approval for O95822 . DB00112 ( avastin ) does not harm retinal function after intravitreal injection as shown by electroretinography in adult mice . PURPOSE : Scavenging of P15692 by specific antibodies is a promising way to treat ocular conditions connected with neovascularization . Intravitreal injections of DB00112 ( bevacizumab ) are performed frequently as a treatment of such conditions . In this study , the authors examine whether the retinal function in wild-type mice is affected by an intravitreal injection of DB00112 . METHODS : Electroretinography was performed in four different experimental groups of wild-type C57BL/6 mice before treatment and 1 , 4 , 12 , and 25 days afterwards . The first group was injected intravitreally with BSS , the second one received injections of a vehicle solution , and the third group was injected with the commercial DB00112 solution . In a fourth group , sham surgery was performed . Immunohistochemistry was performed in some eyes to evaluate penetration of the bevacizumab molecule through the retina . RESULTS : In all four groups , a similar behavior of the ERG parameters could be detected . One day after the injections , the amplitudes showed a clear decrease . Later on , they recovered gradually . No difference could be seen between eyes injected with DB00112 or vehicle solution . DB00112 immunoreactivity was already present in the whole retina half an hour after the intravitreal injection and was not detectable 25 days later . Moreover , binding of bevacizumab to endogenous mouse P15692 could be shown . CONCLUSIONS : Based on the electroretinographic findings , the authors conclude that bevacizumab does not have any toxic effects on the mouse retina and its function . The bevacizumab molecule penetrates the retina quickly . Therefore , it can act safely and very quickly , also in deeper retinal layers after its injection . Comparative transcriptional network modeling of three Q07869 -α/γ co-agonists reveals distinct metabolic gene signatures in primary human hepatocytes . AIMS : To compare the molecular and biologic signatures of a balanced dual peroxisome proliferator-activated receptor ( Q07869 ) -α/γ agonist , aleglitazar , with tesaglitazar ( a dual Q07869 -α/γ agonist ) or a combination of pioglitazone ( Pio ; Q07869 -γ agonist ) and fenofibrate ( Feno ; Q07869 -α agonist ) in human hepatocytes . METHODS AND RESULTS : Gene expression microarray profiles were obtained from primary human hepatocytes treated with EC(50)-aligned low , medium and high concentrations of the three treatments . A systems biology approach , Causal Network Modeling , was used to model the data to infer upstream molecular mechanisms that may explain the observed changes in gene expression . DB08915 , tesaglitazar and Pio/Feno each induced unique transcriptional signatures , despite comparable core Q07869 signaling . Although all treatments inferred qualitatively similar Q07869 -α signaling , aleglitazar was inferred to have greater effects on high- and low-density lipoprotein cholesterol levels than tesaglitazar and Pio/Feno , due to a greater number of gene expression changes in pathways related to high-density and low-density lipoprotein metabolism . Distinct transcriptional and biologic signatures were also inferred for stress responses , which appeared to be less affected by aleglitazar than the comparators . In particular , Pio/Feno was inferred to increase Q16236 activity , a key component of the stress response pathway , while aleglitazar had no significant effect . All treatments were inferred to decrease proliferative signaling . CONCLUSIONS : DB08915 induces transcriptional signatures related to lipid parameters and stress responses that are unique from other dual Q07869 -α/γ treatments . This may underlie observed favorable changes in lipid profiles in animal and clinical studies with aleglitazar and suggests a differentiated gene profile compared with other dual Q07869 -α/γ agonist treatments . The Q96P20 inflammasome is activated by nanoparticles through DB00171 , ADP and adenosine . The NLR pyrin domain containing 3 ( Q96P20 ) inflammasome is a major component of the innate immune system , but its mechanism of activation by a wide range of molecules remains largely unknown . Widely used nano-sized inorganic metal oxides such as silica dioxide ( nano-SiO2 ) and titanium dioxide ( nano-TiO2 ) activate the Q96P20 inflammasome in macrophages similarly to silica or asbestos micro-sized particles . By investigating towards the molecular mechanisms of inflammasome activation in response to nanoparticles , we show here that active adenosine triphosphate ( DB00171 ) release and subsequent DB00171 , adenosine diphosphate ( ADP ) and adenosine receptor signalling are required for inflammasome activation . Nano-SiO2 or nano-TiO2 caused a significant increase in P47900 , P41231 , A2A and/or A2B receptor expression , whereas the Q99572 receptor was downregulated . Interestingly , IL-1β secretion in response to nanoparticles is increased by enhanced DB00171 and ADP hydrolysis , whereas it is decreased by adenosine degradation or selective A2A or A2B receptor inhibition . Downstream of these receptors , our results show that nanoparticles activate the Q96P20 inflammasome via activation of P98160 -InsP3 and/or inhibition of adenylate cyclase ( ADCY ) - DB02527 pathways . Finally , a high dose of adenosine triggers inflammasome activation and IL-1β secretion through adenosine cellular uptake by nucleotide transporters and by its subsequent transformation in DB00171 by adenosine kinase . In summary , we show for the first time that extracellular adenosine activates the Q96P20 inflammasome by two ways : by interacting with adenosine receptors at nanomolar/micromolar concentrations and through cellular uptake by equilibrative nucleoside transporters at millimolar concentrations . These findings provide new molecular insights on the mechanisms of Q96P20 inflammasome activation and new therapeutic strategies to control inflammation . Reduction of histamine H1 receptor binding induced by high-fat diet can be prevented by DB01708 and dietary fiber in specific brain areas of male rats . High-fat ( HF ) diet and obesity are risk factors for a number of mental health problems including depression , cognitive dysfunction , dementia , and neurodegenerative diseases . DB11320 H1 receptors ( H1Rs ) are involved in many of these conditions . This study examined P35367 receptor binding density in the brain of male rats fed a high-saturated fat ( HF ) diet , as well as the effect of docosahexaenoic acid ( DB01708 ) , galacto-oligosaccharide ( GOS ) and resistant starch ( RS ) supplementation of HF diet . Alterations of P35367 expression in the post-mortem rat brain were detected by [ (3)H ] - DB06691 binding autoradiography . We found that HF diet significantly decreased P35367 binding densities in the substantia nigra ( SN ) , caudate putamen ( CPu ) , hypothalamic arcuate nucleus ( Arc ) , ventral tegmental area ( VTA ) , piriform cortex ( Pir ) and primary motor cortex ( M1 ) , compared with low-fat fed rats , and the suppression of receptor binding density ranged from 31 % to 48 % . Interestingly , supplementing the HF diet with 0.5 % n-3 polyunsaturated docosahexaenoic acid ( DB01708 ) prevented reduction of P35367 binding densities in the SN and CPu . Addition of galacto-oligosaccharide ( GOS ) and resistant starch ( RS ) to the diet blunted HF induced reduction of P35367 ligand binding in the SN and Pir , respectively . In conclusion this study showed that HF diet can alter P35367 binding densities in various brain regions , and many of these changes can be prevented by adding DB01708 , GOS or RS to the diet . Knockdown of Q03426 does not lead to changes in Q96P20 expression or activation . BACKGROUND : Mutations in the Mevalonate Kinase gene ( Q03426 ) are causes of a rare autoinflammatory disease : Mevalonate Kinase Deficiency and its more acute manifestation , Mevalonic Aciduria . The latter is characterized , among other features , by neuroinflammation , developmental delay and ataxia , due to failed cerebellar development or neuronal death through chronic inflammation . Pathogenesis of neuroinflammation in Mevalonate Kinase Deficiency and Mevalonic Aciduria has not yet been completely clarified , however different research groups have been suggesting the inflammasome complex as the key factor in the disease development . A strategy to mimic this disease is blocking the mevalonate pathway , using P04035 inhibitors ( Statins ) , while knock-out mice for Mevalonate Kinase are non-vital and their hemyzygous ( i.e only one copy of gene preserved ) littermate display almost no pathological features . FINDINGS : We sought to generate a murine cellular model closely resembling the pathogenic conditions found in vivo , by direct silencing of Mevalonate Kinase gene . Knockdown of Mevalonate Kinase in a murine microglial cellular model ( BV-2 cells ) results in neither augmented Q96P20 expression nor increase of apoptosis . On the contrary , statin treatment of BV-2 cells produces an increase both in Mevalonate Kinase and Q96P20 expression . CONCLUSIONS : MKD deficiency could be due or affected by protein accumulation leading to Q96P20 activation , opening novel questions about strategies to tackle this disease . Tyrosine phosphorylation of Q92835 promotes its proteasomal degradation . OBJECTIVE : The activity of the SH2-containing-phosphatidylinositol-5'-phosphatase ( Q92835 , also known as Q92835 ) , a critical hematopoietic-restricted negative regulator of the P19957 kinase ( PI3K ) pathway , is regulated in large part via its protein levels . We sought to determine the mechanism(s) involved in its downregulation by P11274 - P00519 and by interleukin ( IL ) -4 . MATERIALS AND METHODS : We used Ba/ P13726 ( Q92817 -tetOFF) cells to study the downregulation of Q92835 by P11274 - P00519 and bone marrow-derived macrophages to study Q92835 's downregulation by P05112 . RESULTS : We show herein that P11274 - P00519 downregulates Q92835 , but not O15357 or P60484 , and this can be blocked with the Src kinase inhibitor Q99463 , which inhibits the tyrosine phosphorylation of Q92835 , or with the proteasomal inhibitor MG-132 . We also show , using anti- Q92835 immunoprecipitates , that c-Cbl and Cbl-b are associated with Q92835 and that P11274 - P00519 induces Q92835 's polyubiquitination . This ubiquitination can be blocked with Q99463 , consistent with the tyrosine phosphorylation of Q92835 acting as a signal for its ubiquitination . In bone marrow-derived macrophages , P05112 also leads to the proteasomal degradation of Q92835 but , unlike in Ba/ P13726 ( Q92817 -tetOFF) cells , O15357 is also proteasomally degraded and the degradation of both inositol phosphatases can be prevented with Q99463 or MG-132 . CONCLUSION : Our results suggest that Q92835 protein levels can be reduced via P11274 - P00519 and/or Src family member-induced tyrosine phosphorylation of Q92835 because this triggers its polyubiquitination and degradation within the proteasome .

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interacts_with DB01296?

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Prolyl-4-hydroxylase domain protein 2 controls NF-κB/p65 transactivation and enhances the catabolic effects of inflammatory cytokines on cells of the nucleus pulposus . Prolyl-4-hydroxylase ( P20941 ) proteins are key in sensing tissue hypoxia . In nucleus pulposus ( NP ) cells , our previous work demonstrated that P20941 isoforms have a differential contribution in controlling hypoxia-inducible factor ( HIF ) -α degradation and activity . Recently we have shown that a regulatory relationship exists between Q9H6Z9 and inflammatory cytokines in NP cells . With respect to Q9GZT9 , the most abundant P20941 isoform in NP cells , very little is known concerning its function and regulation under inflammatory conditions that characterize intervertebral disc degeneration . Here , we show that Q9GZT9 is a potent regulator of the catabolic activities of P01375 -α ; silencing of Q9GZT9 significantly decreased P01375 -α-induced expression of catabolic markers including P31431 , P08254 , P45452 , and Q9UNA0 , as well as several inflammatory cytokines and chemokines , while partially restoring aggrecan and collagen II expression . Use of NF-κB reporters with ShPHD2 , SiHIF-1α , as well as p65(-/-) , Q9GZT9 (-/-) , and Q9H6Z9 (-/-) cells , shows that Q9GZT9 serves as a co-activator of NF-κB/p65 signaling in Q9BYW2 -independent fashion . Immunoprecipitation of endogenous and exogenously expressed tagged proteins , as well as fluorescence microscopy , indicates that following P01375 -α treatment , Q9GZT9 interacts and co-localizes with p65 . Conversely , loss of function experiments using lentivirally delivered Sh-p65 , Sh-IKKβ , and NF-κB inhibitor confirmed that cytokine-dependent Q9GZT9 expression in NP cells requires NF-κB signaling . These findings clearly demonstrate that Q9GZT9 forms a regulatory circuit with P01375 -α via NF-κB and thereby plays an important role in enhancing activity of this cytokine . We propose that during disc degeneration Q9GZT9 may offer a therapeutic target to mitigate the deleterious actions of P01375 -α , a key proinflammatory cytokine . DB02426 effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 and P05141 may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 -/- mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 -independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 -/- brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 -independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 -/- mice . However , in liver , only Ant2 mRNA was found , whereas in brown adipose tissue , Ant1 and Ant2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 isoform mediates fatty-acid-induced uncoupling , whereas the P12235 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria . Inhibition of cyclin-dependent kinases , GSK-3beta and CK1 by hymenialdisine , a marine sponge constituent . BACKGROUND : Over 2000 protein kinases regulate cellular functions . Screening for inhibitors of some of these kinases has already yielded some potent and selective compounds with promising potential for the treatment of human diseases . RESULTS : The marine sponge constituent hymenialdisine is a potent inhibitor of cyclin-dependent kinases , glycogen synthase kinase-3beta and casein kinase 1 . DB02950 competes with DB00171 for binding to these kinases . A P24941 -hymenialdisine complex crystal structure shows that three hydrogen bonds link hymenialdisine to the Glu81 and Leu83 residues of P24941 , as observed with other inhibitors . DB02950 inhibits Q00535 /p35 in vivo as demonstrated by the lack of phosphorylation/down-regulation of Pak1 kinase in E18 rat cortical neurons , and also inhibits GSK-3 in vivo as shown by the inhibition of P46821 phosphorylation . DB02950 also blocks the in vivo phosphorylation of the microtubule-binding protein tau at sites that are hyperphosphorylated by GSK-3 and Q00535 /p35 in Alzheimer 's disease ( cross-reacting with Alzheimer's-specific AT100 antibodies ) . CONCLUSIONS : The natural product hymenialdisine is a new kinase inhibitor with promising potential applications for treating neurodegenerative disorders . P01579 -induced P56817 expression is mediated by activation of O60674 and P27361 /2 signaling pathways and direct binding of P42224 to P56817 promoter in astrocytes . P56817 ( P56817 ) is an essential enzyme for the production of beta amyloid . Since we found that injection of interferon-gamma ( P01579 ) into young mouse brains increased P56817 expression in astrocytes , we investigated molecular mechanisms underlying this process by cloning a putative P56817 promoter . P56817 promoter activity was differentially regulated by P01579 in a region specific manner and down-regulated by an inhibitor of O60674 ( O60674 ) . A dominant negative mutant of signal transducer and activator of transcription 1 ( P42224 ) expression suppressed P56817 promoter activity , and this was rescued by transfecting wild type P42224 . Electrophoretic mobility shift assay and promoter activity assays indicated that P42224 binds directly to the putative P42224 binding sequence of P56817 promoter . Because P01579 treatment induced P42224 phosphorylation , we examined whether the expression of a suppressor of cytokine signaling ( Q9NSE2 ) , negative regulator of O60674 , suppresses P56817 promoter activity . The results show that O15524 or O14543 expression suppressed P56817 promoter by blocking phosphorylation of Tyr701 residue in P42224 . Also , because P01579 treatment specifically potentiated extracellular signal regulated Q96HU1 kinase ( P29323 ) 1/2 activation , pretreatment of mitogen-activated or extracellular signal-regulated protein kinase ( MEK ) inhibitor , PD98059 , significantly attenuated P01579 -induced P56817 promoter activity and protein expression through blocking phosphorylation of Ser727 residue in P42224 , suggesting that P27361 /2 is associated with P01579 -induced P42224 signaling cascade . Taken together , our results suggest that P01579 activates O60674 and P27361 /2 and then phosphorylated P42224 binds to the putative P42224 binding sequences in P56817 promoter region to modulate P56817 protein expression in astrocytes . Optical scanner for immunoassays with up-converting phosphorescent labels . A 2-D optical scanner was developed for the imaging and quantification of up-converting phosphor ( P25874 ) labels in immunoassays . With resolution better than 500 microm , a scan rate of 0.4 mm/s , and a 1-2 % coefficient of variation for repeatability , this scanner achieved a detection limit of fewer than 100 P25874 particles in an 8.8 . x 10(4) microm(2) area and a dynamic range that covered more than three orders of magnitude . Utilizing this scanner , a microfluidic chip immunoassay for the cytokine interferon-gamma ( P01579 ) was developed : concentrations as low as 3 pM ( 50 pg/mL ) were detected from 100 microL samples with a total assay time of under an hour , including the 8 min readout . For this P25874 -based assay , 2-D images of the capture antibody lines were scanned , image processing techniques were employed to extract the P25874 emission signals , a response curve that spanned 3-600 pM P01579 was generated , and a five-parameter logistic mathematical model was fitted to the data for determination of unknown P01579 concentrations . Relative to common single-point or 1-D scanning optical measurements , our results suggest that a simple 2-D imaging system can speed assay development , reduce errors , and improve accuracy by characterizing the spatial distribution and uniformity of surface-captured optical labels as a function of assay conditions and device parameters . Expression of vitamin D receptor ( P11473 ) , cyclooxygenase-2 ( P35354 ) and 15-hydroxyprostaglandin dehydrogenase ( P15428 ) in benign and malignant ovarian tissue and DB00146 ( 25(OH2)D3 ) and prostaglandin E2 ( DB00917 ) serum level in ovarian cancer patients . Ovarian carcinomas are associated with increased inflammation which is based upon an up-regulation of inducible cyclooxygenase-2 ( P35354 ) . Moreover , based on our previous published data , the extra-renal vitamin D metabolism seems to be dysregulated in comparison to healthy tissue . In order to gain further insight into the prostaglandin ( PG ) - and vitamin D-metabolism in ovarian carcinomas , the study aimed to evaluate the expression of the PG metabolising enzymes P35354 and 15-hydroxyprostaglandin dehydrogenase ( P15428 ) compared to the vitamin D receptor ( P11473 ) in benign and malignant ovarian tissues . Additionally , we determined the DB00146 ( 25(OH2)D3 ) serum levels . Expression of P11473 , P35354 and P15428 was determined by Western blot analysis . Serum levels of 25(OH2)D3 and DB00917 were measured by chemiluminescence-based and colorimetric immunoassay . We detected significantly higher expressions of the PG metabolising enzymes P15428 and P35354 in malignant tissue and DB00917 serum levels were 2-fold higher in tumour patients . Furthermore , we found an inverse correlation to the P11473 -expression which was 62.1 % lower in malignant tissues compared to that in benign tissues . Surprisingly , we could not detect any differences between the 25(OH2)D3 serum levels in either group ( n=20 ) . These data suggest a correlation between PG- and vitamin D-metabolism in ovarian carcinomas . Role of chronic inhibition of dopamine-metabolizing enzymes in the regulation of renal sodium and phosphate excretion in the rat remnant kidney . BACKGROUND/AIMS : The present study examined the effects of chronic selective or combined inhibition of type A monoamine oxidase ( MAO ) and catechol-O-methyltransferase ( P21964 ) on daily urinary excretion of dopamine and metabolites and on natriuresis and phosphaturia in 3/4 nephrectomized ( 3/4nx ) and Sham rats . METHODS : The 3/4nx and Sham rats were placed in metabolic cages and received the P21397 -selective inhibitor Ro-411049 ( 7.5 mg x kg(-1) bid ) and/or the P21964 -selective inhibitor DB03336 3-202 ( 30 mg x kg(-1) bid ) orally for 3 days during high sodium diet . RESULTS : Selective P21964 inhibition increased the urinary excretion of the deaminated metabolite ( 3,4-dihydroxyphenylacetic acid , DOPAC ) and decreased the urinary excretion of the methylated ( 3-methoxytyramine , 3-MT ) and deaminated plus methylated metabolite ( homovanillic acid , HVA ) in both groups . Selective P21397 inhibition increased the urinary excretion of 3-MT and reduced the urinary excretion of both DOPAC and HVA in either 3/4nx or Sham rats . Combined inhibition of P21397 and P21964 did not significantly change the urinary excretion of DOPAC and markedly decreased the urinary excretion of 3-MT and HVA in both groups . Selective or combined inhibition of P21397 and P21964 did not alter the daily urinary excretion of dopamine , sodium or phosphate in either 3/4nx or Sham rats . CONCLUSIONS : Chronic selective or combined inhibition of P21397 and P21964 is not of major importance in regulating the dopamine-dependent natriuresis and phosphaturia in either 3/4nx or Sham rats . P17936 sensitizes prostate cancer cells to interferon-gamma-induced apoptosis . OBJECTIVE : P01308 -like growth factor binding protein-3 ( P17936 ) has been shown to exhibit diverse biological actions , including IGF-independent effects on cell growth and cell death . Here we report that P17936 sensitizes prostate cancer cells to interferon-gamma ( P01579 ) -induced apoptosis and inhibition of cell proliferation . DESIGN : The cell growth or cell death of prostate cells in response to the treatments of IGFBPs and/or P01579 was measured , and the signaling pathways mediating these actions assessed . RESULTS : Cell proliferation was minimally affected when M12 prostate cancer cells were treated with exogenous P17936 ( 1-5 microg/ml ) , P08833 ( 1-5 microg/ml ) or P01579 ( 20 U/ml ) . However , strong inhibition of cell growth and significant apoptosis were observed when M12 cells were co-treated with P17936 and P01579 , but not with P08833 and P01579 . These effects were IGF-independent and appear not to require intracellular localization of P17936 , as similar results were obtained with mutants of P17936 that either could not bind IGF or has impaired ability to be internalized . Further analyses revealed that P17936 , but not P08833 , could significantly enhance the weak tyrosine phosphorylation of P42224 induced by P01579 ( 20 U/ml ) alone . The P17936 -promoted apoptosis in the presence of P01579 could also be abrogated by blockade of the P42345 pathway with its pharmacological inhibitors , LY294002 or rapamycin . CONCLUSIONS : These results demonstrated that in a cancer cell line not responsive to exogenous P17936 alone , P17936 sensitized the cells to the anti-proliferative , proapoptotic actions of P01579 through an IGF-independent , P42224 - and P42345 -dependent mechanism . Effects of Q9H7B4 overexpression on transformation , serum dependence , and apoptosis sensitivity in NIH3T3 cells . The Q9H7B4 ( Q9H7B4 ) gene was found to encode a novel histone methyltransferase involved in human cancer cells . It could specifically methylate histone H3 at lysine 4 and activate the transcription of a set of downstream genes , including of several oncogenes ( e.g. , N-Myc , CrkL , Wnt10b , Q13029 , and hTERT ) and genes involved in the control of cell cycle ( e.g. , P12004 P55008 and P24941 ) and signal transduction ( e.g. , P42224 , Q16584 , and P42338 ) . To determine the effects of Q9H7B4 overexpression on cell transformation , serum dependence and apoptosis sensitivity , we expressed Q9H7B4 in NIH3T3 cells , and these cells showed several transformed phenotypes as demonstrated by foci formation and colony growth in soft agar . Besides , these transfectants also showed increased serum dependence and depression of sensitivity to apoptosis induced by dexamethasone . These findings lend further understanding to the role of Q9H7B4 in the genesis of human cancers and might throw light on the development of novel therapeutic approaches to human cancers . The glucagon-like peptide-1 receptor agonist , liraglutide , attenuates the progression of overt diabetic nephropathy in type 2 diabetic patients . Diabetic nephropathy ( DN ) is the leading cause of end-stage renal disease . Glucagon-like peptide-1 ( P0C6A0 ) is one of the incretins , gut hormones released from the intestine in response to food intake . P43220 ( P43220 ) agonists have been used to treat type 2 diabetes . Here , we studied the effect of the administration of a P43220 agonist , liraglutide , on proteinuria and the progression of overt DN in type 2 diabetic patients . Twenty-three type 2 diabetic patients with overt DN , who had already been treated with blockade of renin-angiotensin system under dietary sodium restriction , were given liraglutide for a period of 12 months . Treatment with liraglutide caused a significant decrease in HbA1c from 7.4 ± 0.2 % to 6.9 ± 0.3 % ( p = 0.04 ) , and in body mass index ( BMI ) from 27.6 ± 0.9 kg/m² to 26.5 ± 0.8 kg/m² after 12 months ( p < 0.001 ) , while systolic blood pressure did not change . The progression of DN was determined as the rate of decline in estimated glomerular filtration rate ( eGFR ) . The 12-month administration of liraglutide caused a significant decrease in proteinuria from 2.53 ± 0.48 g/g creatinine to 1.47 ± 0.28 g/g creatinine ( p = 0.002 ) . The administration of liraglutide also substantially diminished the rate of decline in eGFR from 6.6 ± 1.5 mL/min/1.73 m²/year to 0.3 ± 1.9 mL/min/1.73 m²/year ( p = 0.003 ) . DB06655 can be used not only for reducing HbA1c and BMI , but also for attenuating the progression of nephropathy in type 2 diabetic patients . Chromosomal localization of the genes encoding P37088 , P62324 , P01579 and P21397 on chicken chromosome 1 by fluorescence in-situ hybridization . DB00126 is dispensable for oxygen sensing in vivo . Prolyl-4-hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl-4-hydroxylase domain ( P20941 ) enzymes requires oxygen and 2-oxoglutarate as cosubstrates with iron(II) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo(-/-) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo(-/-) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum- and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF-1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl(2) . A Cys201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione . Suppressive effects of glucagon-like peptide-1 on interferon-gamma-induced nitric oxide production in insulin-producing cells is mediated by inhibition of tumor necrosis factor-alpha production . During the development of Type 1 diabetes , inflammatory cytokines are known to induce the expression of inducible nitric oxide synthase ( P35228 ) in pancreatic islets , and subsequent production of nitric oxide ( NO ) contributes to beta cell destruction . Glucagon-like peptide-1 ( P0C6A0 ) has been shown to reduce cytokine-induced apoptosis of beta cells . In this study , we investigated whether P0C6A0 affects cytokine-induced NO production , resulting in the inhibition of beta-cell apoptosis . We treated MIN6N8a mouse beta cells with interferon ( IFN ) -gamma in the presence or absence of P0C6A0 and found that P01579 treatment induced P35228 mRNA expression and NO production , which was significantly inhibited by treatment with P0C6A0 . Blocking of P43220 signaling via the cyclic AMP and phosphatidylinositol 3-kinase pathway did not directly affect the suppressive effect of P0C6A0 on IFN- gamma-induced P35228 mRNA expression . Further studies revealed that P01579 induced the expression of P01375 mRNA and protein , which synergistically induced NO production , and P0C6A0 treatment inhibited this induction of P01375 . To examine whether the reduction of P01375 by P0C6A0 treatment plays a role in suppressing NO production , we treated MIN6N8a cells with P01579 in the presence of anti- P01375 neutralizing antibody and found that NO production was reduced . In addition , treatment of mouse islets with P0C6A0 inhibited the expression of P35228 and TNFmRNA . These results suggest that P0C6A0 inhibits P01579 -induced NO production by suppression of P01375 production . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . Nonsteroidal anti-inflammatory drugs ( NSAID ) sparing effects of glucosamine hydrochloride through N-glycosylation inhibition ; strategy to rescue stomach from NSAID damage . Gastrointestinal or cardiovascular complications limit nonsteroidal anti-inflammatory drugs ( NSAID ) prescription . DB01296 hydrochloride ( GS-HCl ) alternatively chosen , but debates still exist in its clinical efficiency . P35354 instability through inhibiting P35354 N-glycosylation of GS-HCl raised the possibility of NSAID sparing effect . Study was done to determine whether combination treatment of glucosamine and NSAID contributes to gastric safety through NSAID sparing effect . IEC-6 cells were stimulated with P01375 -α and compared the expressions of inflammatory mediators after indomethacin alone or combination of indomethacin and GS-HCl by Western blotting and RT-PCR . C57BL/6 mice injected with type II collagen to induce arthritis were treated with indomethacin alone or combination of reduced dose of indomethacin and GS-HCl after 3 weeks . P01375 -α increased the expression of P35354 , P35228 and inflammatory cytokines , but GS-HCl significantly attenuated P01375 -α-induced P35354 expression . Decreased P35354 after GS-HCl was caused by N-glycosylation inhibition as much as tunicamycin . Combination of reduced dose of indomethacin and GS-HCl significantly reduced the expressions of P05362 , P19320 , P10145 , IL-1β , P08253 , P09237 , P14780 , and P24347 mRNA as well as NF-κB activation better than high dose indomethacin alone . These NSAID sparing effect of GS-HCl was further proven in collagen-induced arthritis model . Combination of GS-HCl and 2.5 mg/kg indomethacin showed significant protection from gastric damages as well as efficacious anti-arthritic effect . Taken together , P35354 N-glycosylation inhibition by GS-HCl led to indomethacin sparing effects , based on which combination of GS-HCl and reduced dose of NSAID can provide the strategy to secure stomach from NSAID-induced gastric damage as well as excellent anti-arthritic effects . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . DB04901 signals B- Q9NZ71 cells and inhibits the activities of NF-κB-induced P25490 - and snail-resistant factors : mechanism of sensitization to apoptosis by chemoimmunotherapeutic drugs . DB04901 ( anti- P33681 monoclonal antibody ) is a primatized ( human IgG1 constant regions and cynomologus macaque variable regions ) monoclonal antibody that is currently in clinical trials . DB04901 inhibits tumor cell proliferation through possibly cell signaling-mediated effects . Thus , we hypothesized that galiximab may signal the tumor cells and modify intracellular survival/antiapoptotic pathways such as the NF-κB pathway . This hypothesis was tested using various P33681 (+) Burkitt B- Q9NZ71 ( non-Hodgkin lymphomas ) cell lines as models . Treatment of B- Q9NZ71 cells with galiximab ( 25-100 μg/mL ) resulted in significant inhibition of NF-κB activity and its target resistant factors such as P25490 , Snail , and Bcl-2/Bcl-XL . Treatment of B- Q9NZ71 cells with galiximab sensitized the tumor cells to both DB00515 ( DB00515 ) - and P50591 -induced apoptosis . The important roles of P25490 - and Snail-induced inhibition by galiximab in the sensitization to CCDP and P50591 were corroborated following transfection of Raji cells with P25490 or Snail short interfering RNA . The transfected cells were shown to become sensitive to both CCDP- and P50591 -induced apoptosis in the absence of galiximab . Furthermore , knockdown of P25490 or Snail inhibited Bcl-XL . The involvement of Bcl-XL inhibition in sensitization was corroborated by the use of the pan-Bcl-2 inhibitor 2MAM-3 whereby the treated cells were sensitive to both DB00515 - and P50591 -induced apoptosis . These findings show that galiximab inhibits the NF-κB/Snail/ P25490 /Bcl-XL circuit that regulates drug resistance in B- Q9NZ71 and in combination with cytotoxic drugs results in apoptosis . The findings also support the therapeutic application of the combination of galiximab and cytotoxic drugs in the treatment of drug-resistant P33681 -positive B-cell malignancies . Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . Retinoids induce P22736 -dependent apoptosis in mouse thymocytes . P22736 is a transcription factor , which plays a determinant role in mediating T cell receptor-induced cell death of thymocytes . In addition to regulation of transcription , P22736 contributes to apoptosis induction by targeting mitochondria , where it can convert Bcl-2 , an anti-apoptotic protein into a proapoptotic molecule . Previous studies have demonstrated that retinoids are actively produced in the mouse thymus and can induce a transcription-dependent apoptosis in mouse thymocytes . Here we show that retinoic acids induce the expression of P22736 , and retinoid-induced apoptosis is completely dependent on P22736 , as retinoids were unable to induce apoptosis in P22736 null thymocytes . In wild-type thymocytes retinoids induced enhanced expression of the apoptosis-related genes P48023 , P50591 , NDG-1 , Gpr65 and Bid , all of them in a P22736 -dependent manner . The combined action of these proteins led to Caspase 8-dependent Bid cleavage in the mitochondria . In addition , we could demonstrate the P22736 -dependent induction of P42224 leading to enhanced Bim expression , and the mitochondrial translocation of P22736 leading to the exposure of the Bcl-2/BH3 domain . The retinoid-induced apoptosis was dependent on both Caspase 8 and P42224 . Our data together indicate that retinoids induce a P22736 -dependent cell death program in thymocytes activating the mitochondrial pathway of apoptosis .

MH_train_1296

MH_train_1296

MH_train_1296

interacts_with DB00317?

multiple_choice

Beneficial behavioural and neurogenic effects of agomelatine in a model of depression/anxiety . DB06594 ( S20098 ) is a novel antidepressant drug with melatonergic agonist and P28335 receptor antagonist properties , displaying antidepressant/anxiolytic-like properties in animal models and in humans . In a depression/anxiety-like mouse model in which the response of the Q9Y251 axis is blunted , we investigated whether agomelatine could reverse behavioural deficits related to depression/anxiety compared to the classical selective serotonin reuptake inhibitor , fluoxetine . Adult mice were treated for 8 wk with either vehicle or corticosterone ( 35 μg/ml.d ) via drinking water . During the final 4 wk , animals were treated with vehicle , agomelatine ( 10 or 40 mg/kg i.p. ) or fluoxetine ( 18 mg/kg i.p. ) and tested in several behavioural paradigms and also evaluated for home-cage activity . Our results showed that the depressive/anxiety-like phenotype induced by corticosterone treatment is reversed by either chronic agomelatine or fluoxetine treatment . Moreover , agomelatine increased the dark/light ratio of home-cage activity in vehicle-treated mice and reversed the alterations in this ratio induced by chronic corticosterone , suggesting a normalization of disturbed circadian rhythms . Finally , we investigated the effects of this new antidepressant on neurogenesis . DB06594 reversed the decreased cell proliferation in the whole hippocampus in corticosterone-treated mice and increased maturation of newborn neurons in both vehicle- and corticosterone-treated mice . Overall , the present study suggests that agomelatine , with its distinct mechanism of action based on the synergy between the melatonergic agonist and P28335 antagonist properties , provides a distinct antidepressant/anxiolytic spectrum including circadian rhythm normalization . Identification of N-(4-piperidinyl)-4-(2,6-dichlorobenzoylamino)-1H-pyrazole-3-carboxamide ( DB05037 ) , a novel cyclin dependent kinase inhibitor using fragment-based X-ray crystallography and structure based drug design . The application of fragment-based screening techniques to cyclin dependent kinase 2 ( P24941 ) identified multiple ( > 30 ) efficient , synthetically tractable small molecule hits for further optimization . Structure-based design approaches led to the identification of multiple lead series , which retained the key interactions of the initial binding fragments and additionally explored other areas of the DB00171 binding site . The majority of this paper details the structure-guided optimization of indazole ( 6 ) using information gained from multiple ligand- P24941 cocrystal structures . Identification of key binding features for this class of compounds resulted in a series of molecules with low nM affinity for P24941 . Optimisation of cellular activity and characterization of pharmacokinetic properties led to the identification of 33 ( DB05037 ) , which is currently being evaluated in clinical trials for the treatment of human cancers . P06401 membrane component 1 is a functional part of the glucagon-like peptide-1 ( P0C6A0 ) receptor complex in pancreatic β cells . Glucagon-like peptide-1 ( P0C6A0 ) is an incretin hormone that regulates glucose homeostasis . Because of their direct stimulation of insulin secretion from pancreatic β cells , P43220 ( P43220 ) agonists are now important therapeutic options for the treatment of type 2 diabetes . To better understand the mechanisms that control the insulinotropic actions of P0C6A0 , affinity purification and mass spectrometry ( AP-MS ) were employed to uncover potential proteins that functionally interact with the P43220 . AP-MS performed on Chinese hamster ovary cells or MIN6 β cells , both expressing the human P43220 , revealed 99 proteins potentially associated with the P43220 . Three novel P43220 interactors ( O00264 , Rab5b , and Rab5c ) were further validated through co-immunoprecipitation/immunoblotting , fluorescence resonance energy transfer , and immunofluorescence . Functional studies revealed that overexpression of O00264 , a novel cell surface receptor that associated with liganded P43220 , enhanced P0C6A0 -induced insulin secretion ( GIIS ) with the most robust effect . Knockdown of O00264 in β cells decreased GIIS , indicative of positive interaction with P43220 . To gain insight mechanistically , we demonstrated that the cell surface O00264 ligand P4-BSA increased GIIS , whereas its antagonist AG-205 decreased GIIS . It was then found that O00264 increased P0C6A0 -induced DB02527 accumulation . O00264 activation and GIIS induced by P4-BSA could be blocked by inhibition of adenylyl cyclase/ O95398 signaling or the P01133 receptor-PI3K signal transduction pathway . These data reveal a dual mechanism for O00264 -increased GIIS mediated through DB02527 and P01133 receptor signaling . In conclusion , we identified several novel P43220 interacting proteins . O00264 expressed on the cell surface of β cells was shown to interact with the activated P43220 to enhance the insulinotropic actions of P0C6A0 . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . DB09045 for the treatment of type 2 diabetes . DB09045 is a novel glucagon-like peptide 1 ( P0C6A0 ) receptor agonist with a unique structure that supports once-weekly dosing in patients with type 2 diabetes ( T2DM ) , most of whom have a big pill burden . It appears to be efficacious in reducing hemoglobin A1c ( HbA1c ) up to 1.59 % and promotes modest weight loss up to 3 kg with a low incidence of hypoglycemia and mild to moderate gastrointestinal adverse events . Convenient weekly dosing could improve compliance and help attain sustained glycemic goals . Addressing obesity is an integral part of T2DM management and weight loss may contribute to better glycemic and cardiovascular benefits . Results of ongoing clinical trials on cardiovascular safety are important to determine the risk-to-benefit ratio . As with any drug , patient selection and ongoing monitoring will be important . If approved , dulaglutide will be one of the first weekly P43220 agonist to be available in a ready-to-use pen device with an automatic injector . Critical role of both p27KIP1 and p21CIP1/ P38936 in the antiproliferative effect of ZD1839 ( ' DB00317 ' ) , an epidermal growth factor receptor tyrosine kinase inhibitor , in head and neck squamous carcinoma cells . High expression of the epidermal growth factor receptor ( P00533 ) has been implicated in the development of squamous-cell carcinomas of head and neck ( SCCHN ) . ZD1839 ( ' DB00317 ' ) is an orally active , selective P00533 -TKI ( P00533 -tyrosine kinase inhibitor ) that blocks signal transduction pathways implicated in proliferation and survival of cancer cells , and other host-dependent processes promoting cancer growth . We have demonstrated that ZD1839 induces growth arrest in SCCHN cell lines by inhibiting P00533 -mediated signaling . Cell cycle kinetic analysis demonstrated that ZD1839 induces a delay in cell cycle progression and a P55008 arrest together with a partial G2/M block ; this was associated with increased expression of both p27( P46527 ) and P38936 (CIP1/ P38936 ) cyclin-dependent kinase ( CDK ) inhibitors . The activity of P24941 , the main target of CIP/ Q99828 CDK inhibitors , was reduced in a dose-dependent fashion after 24 h of ZD1839 treatment and this effect correlated to the increased amount of p27( P46527 ) and P38936 (CIP1/ P38936 ) proteins associated with P24941 -cyclin-E and P24941 -cyclin-A complexes . In addition , ZD1839-induced growth inhibition was significantly reduced in cell transfectants expressing p27( P46527 ) or P38936 (CIP1/ P38936 ) antisense constructs . Overall , these results as well as the timing of the effect of ZD1839 on P55008 arrest and p27( P46527 ) and P38936 (CIP1/ P38936 ) upregulation , suggest a mechanistic connection between these events . Effects of phenacetin and its metabolite p-phenetidine on P23219 and P35354 activities and expression in vitro . The present study was aimed to test the possible cyclooxygenase ( P36551 ) -1/ P35354 selectivity of the old analgesic drug phenacetin and its metabolite p-phenetidine , which exhibits high renal toxicity . DB00316 ( acetaminophen ) , the main metabolite of phenacetin with low renal toxicity , and indomethacin were selected as reference compounds . Collagen-stimulated platelet thromboxane B2 ( TxB2 ) production and phorbol 12-myristate-13-acetate ( PMA ) -induced neutrophil prostaglandin E2 ( DB00917 ) synthesis were used as indicators for P23219 and P35354 activity , respectively . DB03783 was even less potent than paracetamol to reduce the production of both TxB2 and DB00917 , and no clear preference for either of the P36551 -enzymes was seen . P-phenetidine was a more potent inhibitor , already at nanomolar level , of the synthesis of these prostanoids than indomethacin and showed some preference to P35354 inhibition . Somewhat higher , micromolar , concentrations of p-phenetidine also reduced P35354 expression in neutrophils . We suggest that the very potent inhibitory activity of p-phenetidine on DB00917 synthesis combined with the reduction of P35354 expression could explain the renal papillary necrosis in phenacetin kidney . Disabling the mitotic spindle and tumor growth by targeting a cavity-induced allosteric site of survivin . Survivin is a member of the inhibitor of apoptosis protein family and has an essential role in mitosis . Survivin is overexpressed in a large variety of human cancers and represents an attractive target for cancer therapy . P00533 and Her/neu-transformed human tumors in particular exhibit high levels of survivin . The survivin protein forms dimers through a conserved region that is critical for subcellular localization and biological functions of the protein . We identified small molecules that target a specific cavity adjacent to the survivin dimerization surfaces . P28222 , a lead compound identified in the screen , can bind to the survivin protein at the intended target site . Moreover , P28222 alters spindle formation , causing mitotic arrest and cell death , and inhibits tumor growth in vitro and in vivo . Cell death occurs in premetaphase stage following mitotic arrest and is not a consequence of general toxicity . Thus , the study validates a novel therapeutic target site in the survivin protein and provides a promising strategy to develop a new class of therapeutic small molecules for the treatment of human cancers . P00533 changes during breast cancer metastasis . BACKGROUND : Since the anti- P04626 monoclonal antibody trastuzumab made its triumphant advance into breast cancer therapy , new selective agents , including pan-HER inhibitors are entering clinical practice . MATERIALS AND METHODS : This study investigates the expression of the four HER-family members , P00533 -4 , in 48 primary breast carcinomas ( P10515 ) and corresponding distant metastases ( Q05682 ) by immunohistochemistry and fluorescence in situ hybridisation . RESULTS : Concordance rate between P10515 and Q05682 was 79 % for HERI and P04626 , 67 % for P21860 and 56 % or Q15303 . Expression of P00533 -3 was associated with poor prognosis compared to HER-negative disease ( p = 0.036 ) . Q15303 overexpression was associated with a better outcome ( p = 0.003 ) . CONCLUSION : Though most tumours demonstrate a stable HER expression pattern , both loss and acquisition of HER receptor overexpression can occur during metastasis . Q15303 overexpression predicts prolonged survival compared to receptor negative disease , while the opposite is true for P00533 -3 . Consequences for modem antibody therapy are discussed . Isolation and characterization of cultured human conjunctival goblet cells . PURPOSE : To isolate and characterize goblet cells from normal human conjunctival tissue to determine whether epidermal growth factor ( P01133 ) receptors are present and whether P01133 can influence goblet cell proliferation . METHODS : Goblet cells were isolated from explant cultures established from normal conjunctival tissue harvested from patients during periocular surgery . The cells were grown in RPMI culture medium supplemented with 10 % fetal bovine serum and characterized using morphology , histochemistry , indirect immunofluorescence microscopy , molecular biology , and biochemistry . Proliferation was determined with a MTT proliferation assay after exposing goblet cells , which had been serum deprived for 48 hours , to increasing concentrations of epidermal growth factor ( P01133 ; 0-80 ng/mL ) for 24 hours . RESULTS : Goblet cells were isolated from conjunctival explants by scraping nongoblet cells from the culture dish . Human goblet cells exhibited positive reactivity with alcian blue-periodic acid Schiff ( DB00233 ) reagent , goblet cell-specific cytokeratin-7 , Q9Y251 lectin , and P98088 , but negative reactivity to the stratified squamous epithelial cell marker , cytokeratin-4 . The mRNA for P98088 was detected using RT-PCR . The presence of the P01133 receptors P00533 , ErbB2 , and ErbB3 was confirmed through Western blot analysis of cell lysates . P01133 elicited a concentration-dependent increase in goblet cell proliferation of 160 % +/- 0.5 % , 188 % +/- 0.45 % , 293 % +/- 1.3 % , and 220 % +/- 0.5 % of control values with 10 , 20 , 40 , and 80 ng/mL P01133 , respectively . CONCLUSIONS : Human goblet cells that retain characteristics of goblet cells in vivo can be cultured . P01133 receptors are present in human goblet cells , and P01133 stimulates their proliferation . The role of celecoxib in Rad51 expression and cell survival affected by gefitinib in human non-small cell lung cancer cells . Celecoxib ( DB00482 ) is a cyclooxygenase-2 ( P35354 ) selective inhibitor and gefitinib ( DB00317 (R) , ZD1839 ) is a selective epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor for human non-small cell lung cancer ( NSCLC ) . The addition of celecoxib to gefitinib to prolong the survival of patients with NSCLC still remains controversial and needs to be investigated . The Rad51 protein is essential for homologous recombination repair , and is overexpressed in chemo- or radioresistant carcinomas . In this study , we characterize the role of celecoxib in the cytotoxicity , P27361 /2 activation and Rad51 expression affected by gefitinib in NSCLC cells . We show that celecoxib can enhance the cytotoxicity induced by gefitinib in NSCLC cells . Treatment with celecoxib alone has no effect on the P27361 /2 activation , Rad51 mRNA and protein levels , however , combined treatment with gefitinib results in a significant reduction of phospho- P27361 /2 and Rad51 protein levels , and triggers the degradation of Rad51 via a 26S proteasome-dependent pathway . Expression of constitutively active Q02750 /2 vectors ( Q02750 /2-CA ) significantly rescues the decreased P27361 /2 activity , and restores Rad51 protein levels and cell survival under co-treatment with gefitinib and celecoxib . Furthermore , blocking P27361 /2 activation by U0126 ( Q02750 /2 inhibitor ) and knocking down Rad51 expression by transfection with small interfering RNA of Rad51 can enhance the cytotoxicity of celecoxib . [ DB05812 acetate : a novel therapeutic option in hormone-refractory prostate cancer ] . Until recently , only therapy with docetaxel and prednisone has been shown to prolong survival in men with hormonorefractory metastatic prostate cancer . With approvals of sipuleucel-T , cabazitaxel , and abiraterone acetate , all based on improvement in overall survival , the scenary for management of men with metastatic prostate cancer has dramatically changed . DB05812 acetate was developed to specifically inhibit cytochrome P450 (CYP)17A1 , which is an essential enzyme in the biosynthesis of testosterone . In the phase III , the trial treatment with abiraterone acetate plus prednisone prolongs overall survival relative to prednisone alone in patients with metastatic castration-resistant prostate cancer who have disease progression after treatment with docetaxel and associated with an acceptable tolerability profile , which was generally similar to that of the placebo plus prednisone group . However , adverse events resulting from elevated mineralocorticoid levels because of P05093 inhibition , fluid retention and oedema , hypokalaemia , hypertension occurred in significantly more in abiraterone acetate plus prednisone than in placebo plus prednisone . Drugs targeting nitric oxide synthase for migraine treatment . INTRODUCTION : Ample evidence that nitric oxide ( NO ) is a causative molecule in migraine has encouraged research to develop drugs that target the NO-cGMP cascade for migraine treatment . NO synthase ( NOS ) inhibition is an innovative therapeutic principle . AREAS COVERED : This paper reviews the rationale underlying NOS inhibition in migraine treatment . It also provides a review on the efficacy and safety data for NOS inhibitors ( nonselective NOS inhibitor L-N(G)-methyl-arginine hydrochloride [ L-NMMA ] , selective inducible NOS [ P35228 ] inhibitors GW273629 and GW274150 , combined neuronal NOS [ P29475 ] inhibitor and P28222 /1D receptor agonist DB06096 ) in acute or preventive migraine treatment . EXPERT OPINION : The data highlighted herein , from four placebo-controlled trials and 1 open-labeled clinical trial using 4 different NOS inhibitors on a total of 705 patients , provide convincing efficacy data only for the nonselective NOS inhibitor L-NMMA . Unfortunately , this NOS inhibitor raises cardiovascular safety concerns and has an unfavorable pharmacokinetic profile . As experimental studies predicted , P35228 inhibitors are ineffective in migraine . Still , upcoming selective P29475 inhibitors are a hope for migraine treatment , with the P29475 isoform being most clearly involved in trigeminovascular transmission and central sensitization . Future studies should help to clarify whether NOS inhibition is equally fruitful in acute and preventive treatment . It should also clarify if P29475 inhibition holds promise as a therapeutic tool for the treatment of chronic migraine and other forms of headache . Structural basis for P01133 receptor inhibition by the therapeutic antibody DB05774 . Therapeutic anticancer strategies that target and inactivate the epidermal growth factor receptor ( P00533 ) are under intense study in the clinic . Here we describe the mechanism of P00533 inhibition by an antibody drug DB05774 . DB05774 is a fully human antibody that has similar antitumor potency as the chimeric cetuximab/Erbitux and might represent a safer therapeutic alternative . We report the X-ray crystal structure of the Fab fragment of DB05774 ( Fab11F8 ) in complex with the entire extracellular region and with isolated domain III of P00533 . We compare this to our previous study of the cetuximab/ P00533 interaction . Fab11F8 interacts with a remarkably similar epitope , but through a completely different set of interactions . Both the similarities and differences in binding of these two antibodies have important implications for the development of inhibitors that could exploit this same mechanism of P00533 inhibition . Simvastatin in primary biliary cirrhosis : effects on serum lipids and distinct disease markers . BACKGROUND/AIMS : Primary biliary cirrhosis ( P10515 ) is an autoimmune disease of the liver with inflammation of small and middle-sized bile ducts . Serum lipids are frequently elevated , but the use of a lipid lowering drug therapy in P10515 is still a matter of debate . Application of an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase in hypercholesterolemic P10515 patients was therefore the subject of the present study . METHODS : Six female patients ( aged 46.5 ( 32-61 ) years ; median ( range ) ) were treated with the P04035 inhibitor simvastatin ( 5 or 20 mg/day ) . Levels of serum total cholesterol , low-density lipoprotein ( LDL ) cholesterol and high-density lipoprotein ( HDL ) cholesterol were determined prior to and after 2 months of treatment . Concentrations of serum markers of cholestasis , antimitochondrial antibodies ( AMA ) , and immunoglobulins A , G and M were also assessed . RESULTS : Simvastatin significantly ( P < 0.05 ) reduced serum levels of total cholesterol , LDL cholesterol , alkaline phosphatase , -glutamyltransferase , and immunoglobulin M ( by 19 , 26 , 12 , 37 and 14 % , respectively ) . CONCLUSIONS : The lipid lowering potency of the P04035 inhibitor simvastatin was confirmed in hypercholesterolemic patients with P10515 . The drug might also prove useful as modulator of cholestasis and of immune response in this disease . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Targeting of P00533 tyrosine kinase by ZD1839 ( " DB00317 " ) in androgen-responsive prostate cancer in vitro . P00533 , highly expressed in a variety of human malignancies , is correlated with poor tumour differentiation , high tumour growth and metastatic rate . P01133 and several other ligands , such as transforming growth factor-alpha , amphiregulin , heparin-binding P01133 , and betacellulin , activate Ras/Raf mitogen-activated protein kinases ( MAPKs ) and phosphatidyl inositol 3'-kinase ( PI3K ) /Akt signalling pathways . Therefore , P00533 can regulate multiple processes , i.e. , gene expression , cellular proliferation , angiogenesis , and inhibition of apoptosis , which contribute to the development of malignancy . In this review , we discuss the inhibition of P00533 by the specific tyrosine kinase inhibitor DB00317 ( ZD1839 ) focusing on its effects in prostate cancer . Vascular endothelial growth factor gene polymorphism is associated with calcium oxalate stone disease . Growth factor-related genes regulate cell growth , differentiation and apoptosis in the kidney in response to cellular injury . One of the theories of stone formation is that cellular injury , and thus growth factors , play a role . We therefore investigated the association between growth factor genes and calcium oxalate stone disease . The most frequently seen polymorphism of the vascular endothelial growth factor ( P15692 ) gene is Bst U I C/T , which is located upstream at the -460th nucleotide . Other growth factor-related gene polymorphisms include the cytochrome P450c17alpha enzyme ( P05093 ) gene MspA I C/T polymorphism at the 5'-UTR promoter region , the epidermal growth factor receptor ( P00533 ) gene Bsr I polymorphism ( A to T ) at position 2,073 , and the insulin-like growth factor-2 ( IGF-2 ) gene Apa I A/G at exon 9 . All four polymorphisms were used as genetic markers in this study in the search for an association between stone disease and growth factor related genes . A normal control group of 230 healthy people , and 230 patients with calcium oxalate stone , were examined . The polymorphism was seen following polymerase chain reaction based restriction analysis . The result revealed a significant difference between normal individuals and stone patients ( P=0.0003 , Fisher 's exact test ) in the distribution of the P15692 gene polymorphism as well as an odds ratio of 1.30 ( 95 % confidence interval=0.993-1.715 ) per copy of the " T " allele . Whereas , the IGF-2 , P00533 and P05093 gene polymorphisms did not reveal a significant association with stone disease . We conclude that the P15692 gene Bst U I polymorphism is a suitable genetic marker of urolithiasis . Adenoid cystic carcinomas of the breast have low Topo IIα expression but frequently overexpress P00533 protein without P00533 gene amplification . Adenoid cystic carcinoma of the breast is a rare subtype of breast cancer with basal-like features . Published studies on breast adenoid cystic carcinoma are limited , resulting in relatively scarce information on the value of predictive tumor markers . We studied 20 primary cases of adenoid cystic carcinoma of the breast for expression of estrogen receptor , progesterone receptor , androgen receptor , epidermal growth factor receptor , HER-2/neu , and topoisomerase IIα using immunohistochemistry and fluorescent in situ hybridization methods . Estrogen and progesterone receptor expression were detected in 1 case each . All tumors were uniformly negative for Her-2/neu expression . P10275 and topoisomerase IIα expression were weakly positive in three cases and 7 cases , respectively . P00533 overexpression was detected in 13 cases ( 65 % of all cases ) . Amplification of P11388 or HER-2/neu gene was not detected in any of the cases . Our study shows that the majority of adenoid cystic carcinomas of the breast do not overexpress Her-2/neu , topoisomerase IIα , or estrogen receptor , and thus , they are unlikely to respond to therapies targeting these proteins . However , these tumors frequently over-express epidermal growth factor receptor , indicating a potential benefit from anti-epidermal growth factor receptor therapy for patients with advanced adenoid cystic carcinomas of the breast .

MH_train_1297

MH_train_1297

MH_train_1297

interacts_with DB00104?

multiple_choice

Distribution of polyamines and their biosynthetic enzymes in intestinal adaptation . P11926 ( ODC ) and the polyamines have been shown to be important for growth processes in the intestinal mucosa . The highest activity of ODC is found in the differentiated , nonproliferating villus-tip cells rather than in the rapidly proliferating undifferentiated crypt cells . During poststarvation refeeding and lactation , we now show that increases in ODC activity paralleled the time course of mucosal hyperplasia and thymidine incorporation . Increases in ODC ( threefold ) were similar in villus and crypt cells , and the villus-crypt gradient of decreasing ODC activity ( 40:1 ) was maintained . The activity of the other polyamine biosynthetic enzyme , S-adenosylmethionine decarboxylase ( P18827 ) , was highest in the crypt cells in the basal state and increased throughout the entire villus-crypt axis during refeeding and lactation , preserving a villus-crypt gradient opposite to that of ODC . During hyperplasia , all three polyamines increased . DB01917 was highest in the villus-tip cells , paralleling ODC activity , whereas spermidine and spermine were highest in the crypt cells and paralleled the distribution of P18827 activity . Thus P18827 activity and spermidine and spermine content may play a more important role than ODC and putrescine in regulation of intestinal mucosal proliferation . It is also possible that the threefold increases in the low levels of ODC in the crypt cells are adequate to trigger cell proliferation , whereas the higher ODC levels in villus cells may represent an association with the differentiation of the enterocytes . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . Potentiation of P01138 -induced neurite outgrowth in PC12 cells by papaverine : role played by P98160 -γ , IP3 receptors . DB01113 , an inhibitor of phosphodiesterase ( PDE ) 10A , is gaining attention for its potential in the treatment of neuropsychiatric diseases such as schizophrenia . However , the precise mechanisms underlying the putative neuroprotective/neurotrophic actions of papaverine remain unclear . Thus , we investigated the effects of papaverine on nerve growth factor ( P01138 ) -induced neurite outgrowth in PC12 cells . DB01113 potentiated P01138 -induced neurite outgrowth in PC12 cells in a concentration-dependent manner . In contrast , the selective Q9Y233 inhibitor MP-10 had no effect on P01138 -induced neurite outgrowth . The potentiation of P01138 -induced neurite outgrowth by papaverine was blocked by the P98160 -γ inhibitor U73122 . Furthermore , papaverine 's potentiation of P01138 -induced neurite outgrowth was also blocked by the co-administration of inositol 1,4,5-trisphosphate ( IP(3) ) receptor antagonists ( xestospongin C and 2-aminoethoxydiphenyl borate ( 2- Q9H4A4 ) ) and by reduced expression of IP(3) receptor gene ( i.e. , itpr1 and itpr3 ) by siRNA . Our findings suggest that papaverine could potentiate P01138 -induced neurite outgrowth , and that activation of P98160 -γ and IP(3) receptors might be involved in the mechanism underlying papaverine 's potentiation of neurite outgrowth in PC12 cells . DB05229 sodium , a stable prostacyclin analogue , elicits dilation of isolated porcine retinal arterioles : roles of P29474 and potassium channels . PURPOSE : DB01240 ( DB01240 ) is usually described as an endoEDRFsthelium-derived relaxing factor , but the vasoreactivity to DB01240 in the retinal arterioles and the underlying mechanisms are not fully understood . We examined the effects of DB01240 on the retinal microcirculation using beraprost sodium ( BPS ) , a stable DB01240 analogue , and the signaling mechanisms involved in this vasomotor activity . METHODS : Porcine retinal arterioles were isolated , cannulated , and pressurized without flow in vitro . Video microscopic techniques recorded the diametric responses to BPS . RESULTS : DB05229 sodium elicited dose-dependent ( 0.1 pM-0.1 μM ) vasodilation of the retinal arterioles that was abolished by the P43119 ( IP ) antagonist CAY10441 . DB05229 sodium-induced vasodilation decreased by 50 % after the endothelium was removed and was inhibited by the nitric oxide ( NO ) synthase inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) comparable with denudation . Inhibition of soluble guanylyl cyclase by 1H-1,2,4-oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) and blockage of protein kinase A ( PKA ) by Rp-8-Br-cAMPS were comparable to L-NAME . DB05229 sodium-induced vasodilation was also inhibited by the nonselective potassium channel inhibitor , tetraethylammonium , and the adenosine triphosphate-sensitive potassium ( KATP ) channel blocker , glibenclamide . Residual vasodilation in the presence of glibenclamide decreased further with subsequent application of ODQ . CONCLUSIONS : DB05229 sodium , a stable DB01240 analogue , causes vasodilation of the retinal arterioles mediated via the IP receptor . The current findings suggest that BPS elicits endothelium-dependent and -independent dilation of the retinal arterioles mediated by NO induced by activation of PKA in the endothelium and the KATP channel activation in the vascular smooth muscle , respectively . CP-101,606 , a potent neuroprotectant selective for forebrain neurons . The neuroprotective activity of ( 1S,2S ) -1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol ( CP-101,606 ) , an N-methyl-D-aspartate ( DB01221 ) receptor antagonist structurally similar to ( (+/-)-(R*,S*)-alpha-(4-hydroxyphenyl)-beta-methyl-4-(phenylmethyl)-1-++ +piperidineethanol ( ifenprodil ) , was investigated in neurons in primary culture . CP-101,606 potently and efficaciously protected hippocampal neurons from glutamate toxicity but was > 900-fold less effective for cerebellar granule neurons . The neuroprotective activity in the hippocampal neurons is mediated through a high affinity binding site distinct from the agonist and thienylcyclohexylpiperidine ( DB01520 ) binding sites of the DB01221 receptor . Autoradiography indicates the CP-101,606 binding site is localized in forebrain , most notably in hippocampus and the outer layers of cortex . The functional selectivity for hippocampal neurons , forebrain localization of binding sites , and structural relation to ifenprodil suggest that CP-101,606 is an DB01221 antagonist highly selective for Q13224 subunit containing receptors . Expression of hypothalamic peptides in mice lacking neuronal nitric oxide synthase : reduced beta- P17813 immunoreactivity in the arcuate nucleus . The gas nitric oxide ( NO ) is an important messenger in brain signaling . Along with many other functions , NO is thought to influence the expression and/or release of various hypothalamic hormones ( corticotropin-releasing hormone ( P06850 ) , gonadotropin-releasing hormone ( DB00644 ) and vasopressin ) . To learn more about the role of NO in neuroendocrine mechanisms , we studied in mutant mice lacking neuronal isoform of NO synthase ( P29475 ) the cellular expression of P06850 , neurophysin ( the carrier protein of vasopressin/oxytocin ) and pro-opiomelanocortin ( P01189 ) , as well as of the P01189 -derived peptides beta-endorphin ( beta- P17813 ) , alpha-melanocyte-stimulating hormone ( alpha-MSH ) and corticotropin ( DB01285 ) by use of immunohistochemistry and in situ hybridization . Additionally , the remaining NO-generating capacities of the P29475 minus mice were investigated by NADPH-diaphorase histochemistry and citrulline immunohistochemistry as well as by immunohistochemical localization and Western blot analysis of endothelial NOS ( P29474 ) and P29475 isoforms . Amongst all hypothalamic peptides under investigation , only beta- P17813 was found to be altered in mutant mice . A morphometric analysis of beta- P17813 producing neurons of the arcuate nucleus revealed that significantly less cells were immunoreactive in mutant mice , whereas the expression of the precursor P01189 as well as of other P01189 -derived peptides was found to be unchanged . In addition to that , fewer beta- P17813 -immunoreactive fibers were found in the paraventricular nucleus of P29475 minus mice in comparison to wild-type animals . Hence , the reduction of hypothalamic beta- P17813 is probably a posttranslational event that might reflect a disturbed endorphinergic innervation of those hypothalamic neurons which normally express P29475 . Aggressive inherited and sporadic medullary thyroid carcinomas display similar oncogenic pathways . P07949 oncogene mutations are found in familial medullary thyroid carcinomas ( P04629 ) and in one-third of sporadic cases . Oncogenic mechanisms involved in non- P07949 mutated sporadic P04629 remain unclear . To study alterations associated with the development of both inherited and sporadic P04629 , pangenomic DNA microarrays were used to analyze the transcriptome of 13 MTCs ( four familial and nine sporadic ) . By using an Q9UNW9 test , a list of 173 gene sequences with at least a twofold change expression was obtained . A subset of differentially expressed genes was controlled by real-time quantitative PCR and immunohistochemistry on a larger collection of MTCs . The expression pattern of those genes allowed us to distinguish two groups of sporadic tumors . The first group displays an expression profile similar to that expressed by inherited RET634 tumors . The second presents an expression profile close to that displayed by inherited RET918 tumors and includes tumors from patients with distant metastases . It is characterized by the overexpression of genes involved in proliferation and invasion ( P21246 , Q9NQ30 , and P40199 ) or matrix remodeling ( P02452 , P08123 , and FAP ) . Interestingly , RET918 tumors showed overexpression of the P21246 gene , encoding pleiotrophin , a protein associated with metastasis . Using a P04629 cell line , silencing of P07949 induced the inhibition of P21246 gene expression . Overall , our results suggest that familial P04629 and sporadic P04629 could activate similar oncogenic pathways . Carcinoma ex pleomorphic adenoma of the salivary gland : an immunohistochemical study . The proliferative activity of the tumor cells and the expression of tumor-associated genes and sex steroid hormone receptors were investigated immunohistochemically in ten cases of carcinoma ex pleomorphic adenoma ( Ca-ex-PA ) of the salivary glands . These were analyzed in benign and malignant components separately , and then were compared with ten cases of the other malignant tumors [ adenocarcinomas , not otherwise specified ( ACN ) and salivary duct carcinomas ( P18827 ) ] and ten cases of pleomorphic adenomas ( PA ) . The results obtained in this study were as follows : ( 1 ) malignant component of Ca-ex-PA showed a higher incidence of P12004 and Ki67 than benign component of Ca-ex-PA . A significant difference between benign component of Ca-ex-PA and PA was not observed . ( 2 ) A significant difference in the incidence of p53 , c-erbB-2 , P00533 overexpression was observed only between malignant component of Ca-ex-PA and benign component of Ca-ex-PA . ( 3 ) The incidence of P12004 , Ki67 , p53 , c-erbB-2 overexpression in malignant component of Ca-ex-PA showed the highest data among the four groups . These results suggest that Ca-ex-PA acquired the particular biological behavior in contrast to the other salivary neoplasms in the long-standing process while PA undergoes malignant transformation . Frequency of mutations in the genes associated with hereditary sensory and autonomic neuropathy in a UK cohort . The hereditary sensory and autonomic neuropathies ( HSAN , also known as the hereditary sensory neuropathies ) are a clinically and genetically heterogeneous group of disorders , characterised by a progressive sensory neuropathy often complicated by ulcers and amputations , with variable motor and autonomic involvement . To date , mutations in twelve genes have been identified as causing HSAN . To study the frequency of mutations in these genes and the associated phenotypes , we screened 140 index patients in our inherited neuropathy cohort with a clinical diagnosis of HSAN for mutations in the coding regions of O15269 , P51149 , Q9H4A3 / Q9H4A3 , Q9H6L5 , P04629 ( P04629 ) and P01138 . We identified 25 index patients with mutations in six genes associated with HSAN ( O15269 , P51149 , Q9H4A3 / Q9H4A3 , Q9H6L5 , P04629 and P01138 ) ; 20 of which appear to be pathogenic giving an overall mutation frequency of 14.3 % . Mutations in the known genes for HSAN are rare suggesting that further HSAN genes are yet to be identified . The p.Cys133Trp mutation in O15269 is the most common cause of HSAN in the UK population and should be screened first in all patients with sporadic or autosomal dominant HSAN . p38 Q96HU1 kinase regulates stem cell apoptosis in human hematopoietic failure . Myelodysplastic syndromes ( P43034 ) are clonal stem cell disorders that lead to ineffective hematopoiesis and are common causes of low blood counts in the elderly . The exact molecular mechanisms regulating increased stem apoptosis in these disorders are not well defined . p38 MAPK activation is important in regulating the growth inhibitory signals of P01375 , TGF-beta and Interferons on human hematopoiesis . Our findings show that p38 MAPK is overactivated in myelodysplasia bone marrows and regulates hematopoietic stem cell apoptosis . Inhibition of p38 MAPK by genetic or pharmacologic means decreases apoptosis and stimulates in vitro hematopoiesis from primary P43034 hematopoietic progenitors . These studies point to the potential efficacy of selective p38alpha inhibitor , DB05412 , in human bone marrow failure . Reduction of burn scar formation by halofuginone-eluting silicone gel sheets : a controlled study on nude mice . Burn scar formations can cause disfiguration and loss of dermal function . The purpose of this study was to examine whether application of modified silicone gel sheets with an antifibrotic drug halofuginone-eluting hybrid surface produce an effect on scar development . There were a total of 2 animal groups . The athymic nude mice ( nu/nu ) of both groups underwent transplantation of full-thickness human skin grafts onto their backs and setting of partial thickness burn injury . The status of local scar development was observed over a period of 3 months after the application of silicone gel sheets and also after application of surface-modified halofuginone-eluting silicone gel sheets . Subsequently , via real-time polymerase chain reaction , the cDNA levels from key mediators of scar formation ( transforming growth factor beta , P02452 , connective tissue growth factor , fibroblast growth factor 2 , matrix metalloproteinase 2 , matrix metalloproteinase 9 ) were established and statistically evaluated . In comparison with uncoated silicone gel sheets , the application of halofuginone-eluting silicone gel sheets lead to a significant difference in gene expression activity in scar tissue . DB04866 -eluting hybrid surface silicone gel sheets significantly increase the antiscarring effect of adhesive silicone gel sheets by deceleration and downregulation of scar development by normalization of the expression activity . Sexually dimorphic stress and pro-inflammatory cytokine responses to an intravenous corticotropin-releasing hormone challenge of Brahman cattle following transportation . This study was designed to characterize potential sexually dimorphic stress and immunological responses following a corticotropin-releasing hormone ( P06850 ) challenge in beef cattle . Six female ( heifers ) and six male ( bulls ) Brahman calves ( 264 ± 12 d of age ) were administered P06850 intravenously ( 0.5 µg of P06850 /kg body mass ) after which serum concentrations of cortisol increased from 0.5 h to 4 h . From 1 h to 4 h after P06850 administration , serum cortisol concentrations were greater in heifers than in bulls . In all cattle , increased serum concentrations of P01375 -α , P05231 and IFN-γ were observed from 2.5 h to 3 h after P06850 , with greater concentrations of IFN-γ and P05231 in heifers than bulls . Heifer total leukocyte counts decreased 1 h after P06850 administration , while bull leukocyte counts and percent neutrophils decreased 2 h after P06850 administration . Heifers had greater rectal temperatures than bulls , yet rectal temperatures did not change following administration of P06850 . There was no effect of P06850 administration on heart rate . However , bulls tended to have increased heart rate 2 h after P06850 administration than before P06850 . Heifer heart rate was greater than bulls throughout the study . These data demonstrate that acute P06850 administration can elicit a pro-inflammatory response , and cattle exhibit a sexually dimorphic pro-inflammatory cytokine and cortisol response to acute P06850 administration . Inhibitory effects of a luteinizing hormone-releasing hormone agonist on basal and epidermal growth factor-induced cell proliferation and metastasis-associated properties in human epidermoid carcinoma A431 cells . The purpose of this study was to investigate the effects of a potent P01148 agonist , [D- DB00150 (6)] P01148 on the basal and P01133 -induced cell proliferation and the metastasis-associated properties in A431 human epidermoid carcinoma . [D- DB00150 (6)] P01148 time-dependently inhibited the basal and P01133 -stimulated growth of A431 cancer cells . It is assumed that phosphorylation/dephosphorylation of cellular proteins is highly related to cell growth . This study demonstrates that [D- DB00150 (6)] P01148 decreased the basal and P01133 -induced total cellular kinase activity , particularly the tyrosine phosphorylation of several cellular proteins including the P00533 . In contrast , [D- DB00150 (6)] P01148 did not cause detectable changes in basal and P01133 -stimulated serine/threonine phosphorylation of A431 cellular proteins . The inhibitory effect of [D- DB00150 (6)] P01148 on A431 cell proliferation was associated with apoptosis as evidenced by the cell morphology and DNA integrity ( ladder pattern ) , the expression of interleukin 1beta-converting enzyme ( ICE ) and activation of caspase . Furthermore , P01133 could rescue the remaining attached A431 cells following [D- DB00150 (6)] P01148 treatment for 48 hr , which suggests that limited exposure to [D- DB00150 (6)] P01148 did not channel all cells to irreversible apoptotic process . We also determined the effects of [D- DB00150 (6)] P01148 on metastasis-associated properties in A431 cells . [D- DB00150 (6)] P01148 reduced both basal and P01133 -stimulated secretion of P14780 and P08253 . In addition , [D- DB00150 (6)] P01148 suppressed the basal and P01133 -induced invasive activity of A431 cells based on an in vitro invasion assay . In conclusion , this study indicates that [D- DB00150 (6)] P01148 may act partly through activating tyrosine phosphatase activity to inhibit cell proliferation and the metastasis-associated properties of A431 cancer cells . Our work suggests that [D- DB00150 (6)] P01148 may be therapeutically useful in limiting the tumor growth and metastasis of some neoplasms . Suppression of cyclooxygenase-2 overexpression by 15S-hydroxyeicosatrienoic acid in androgen-dependent prostatic adenocarcinoma cells . Emerging reports now implicate alterations of arachidonic acid ( AA ) metabolism with prostate carcinogenesis . To test this hypothesis , androgen-primed benign hyperplastic ( BHC ) and malignant tumorigenic ( P04629 ) cells derived from the Lobund-Wistar rat model of autochthonous prostate adenocarcinoma were incubated with (14)C-AA . Our data using MTCs revealed enhanced dual metabolism of (14)C-AA via P36551 to generate increased PGE(2) and via P09917 ( P28300 ) to generate increased 5S-HETE in tumorigenic cells . Western blot of MTCs revealed upregulation of P35354 expression . This paralleled the increased biosynthesis of PGE(2) . Since some polyunsaturated fatty acids have been reported to modulate AA metabolism and tumorigenesis , we primed the cells with either gamma-linolenic acid ( GLA ) or its in vivo metabolite , 15S-HETrE , prior to incubation with AA . Our data revealed suppression of P35354 expression/PGE(2) biosynthesis . In parallel , priming cells with 15S-HETrE resulted in greater suppression of P35354 expression/PGE(2) biosynthesis . These findings suggest that 15S-HETrE could function in vivo after dietary intake of GLA to suppress DB02901 -enhanced prostatic P35354 expression/PGE(2) biosynthesis and , thus , alleviate tumor growth and progression . A water-soluble homodimeric serine palmitoyltransferase from Sphingomonas paucimobilis EY2395T strain . Purification , characterization , cloning , and overproduction . DB00133 palmitoyltransferase ( P21549 , EC ) is a key enzyme in sphingolipid biosynthesis and catalyzes the decarboxylative condensation of l-serine and palmitoyl-coenzyme A to 3-ketodihydrosphingosine . We found that the Gram-negative obligatory aerobic bacteria Sphingomonas paucimobilis EY2395(T) have significant P21549 activity and purified P21549 to homogeneity . This enzyme is a water-soluble homodimeric protein unlike eukaryotic enzymes , known as heterodimers composed of tightly membrane-bound subunits , named O15269 and O15270 . The purified P21549 shows an absorption spectrum characteristic of a pyridoxal 5'-phosphate-dependent enzyme . The substrate specificity of the Sphingomonas P21549 is less strict than the P21549 complex from Chinese hamster ovary cells . We isolated the P21549 gene encoding 420 amino acid residues ( M(r) 45,041 ) and succeeded in overproducing the P21549 protein in Escherichia coli , in which the product amounted to about 10-20 % of the total protein of the cell extract . Sphingomonas P21549 shows about 30 % homology with the enzymes of the alpha-oxamine synthase family , and amino acid residues supposed to be involved in catalysis are conserved . The recombinant P21549 was catalytically and spectrophotometrically indistinguishable from the native enzyme . This is the first successful overproduction of an active enzyme in the sphingolipid biosynthetic pathway . Sphingomonas P21549 is a prototype of the eukaryotic enzyme and would be a useful model to elucidate the reaction mechanism of P21549 . Pharmacological characterization of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel selective P09917 -activating protein inhibitor that reduces acute and chronic inflammation . Leukotrienes ( LTs ) are proinflammatory lipid mediators synthesized by the conversion of arachidonic acid ( AA ) to P01374 (4) by the enzyme P09917 ( P09917 ) in the presence of P09917 -activating protein ( P20292 ) . 3-[3-tert-Butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) is a novel selective P20292 inhibitor in development for the treatment of respiratory conditions such as asthma . In a rat ex vivo whole-blood calcium ionophore-induced Q06643 (4) assay , DB05225 ( administered orally at 1 mg/kg ) displayed > 50 % inhibition for up to 6 h with a calculated EC(50) of approximately 60 nM . When rat lung was challenged in vivo with calcium ionophore , DB05225 inhibited Q06643 (4) and cysteinyl leukotriene ( CysLT ) production with ED(50) values of 0.8 and 1 mg/kg , respectively . In this model , the EC(50) derived from plasma DB05225 was approximately 330 nM for inhibition of both Q06643 (4) and CysLT . In an acute inflammation setting , DB05225 displayed dose-dependent inhibition of Q06643 (4) , CysLT , and plasma protein extravasation induced by peritoneal zymosan injection . In a model of chronic lung inflammation using ovalbumin-primed and challenged BALB/c mice , DB05225 reduced the concentrations of eosinophil peroxidase , CysLTs , and interleukin-5 in the bronchoalveolar lavage fluid . Finally , DB05225 increased survival time in mice exposed to a lethal intravenous injection of platelet-activating factor . In summary , DB05225 is a novel , potent and selective P20292 inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute and chronic inflammation and in a model of lethal shock . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . P06850 ( P06850 ) depresses n-methyl-D-aspartate receptor-mediated current in cultured rat hippocampal neurons via P06850 receptor type 1 . P06850 , the primary regulator of the neuroendocrine responses to stress , has been shown to modulate synaptic efficacy and the process of learning and memory in hippocampus . However , effects of P06850 on N-methyl-d-aspartate ( DB01221 ) receptor , the key receptor for synaptic plasticity , remain unclear . In primary cultured hippocampal neurons , using the technique of whole-cell patch-clamp recordings , we found that P06850 ( 1 pmol/liter to 10 nmol/liter ) inhibited DB01221 -induced currents in a dose-dependent manner . This effect was reversed by the P06850 receptor type 1 ( P34998 ) antagonist antalarmin but not by the Q13324 antagonist astressin-2B , suggesting that P34998 mediated the inhibitory effect of P06850 . Investigations on the signaling pathways of P06850 showed that P06850 dose-dependently induced phosphorylated phospholipase C ( P98160 ) -beta3 expression and increased intracellular DB02527 content in these cells . Blocking P98160 activity with U73122 prevented P06850 -induced depression of DB01221 current , whereas blocking protein kinase A ( H89 ) and adenylate cyclase ( SQ22536 ) failed to affect the P06850 -induced depression of DB01221 current . Application of inositol-1,4,5-triphosphate receptor ( IP(3)R ) antagonist , Ca(2+) chelators or protein kinase C ( PKC ) inhibitors also mainly blocked P06850 -induced depression of DB01221 currents , suggesting involvement of P98160 /IP(3)R/Ca(2+)and P98160 /PKC signaling pathways in P06850 down-regulation of DB01221 receptors . Our results suggest that P06850 may exert neuromodulatory actions on hippocampus through regulating DB01221 receptor function . DB00104 is the favorable alternative for cisplatin resistance reversal of ovarian cancer in vitro and in nude mice in vivo . This study aimed to observe the effects of octreotide ( O75051 ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo . MTT method and flow cytometry were used to investigate the effect of cisplatin , O75051 or the combination of these two compounds on the proliferation and apoptosis of SKOV3- O60220 cells . The size and weight of xenograft tumors from the nude mice model were measured . Real-time PCR was used to detect the mRNA expression of P30874 , P08183 , Q92887 , Q86UG4 -pi and P00533 in SKOV3/ O60220 cells following the different treatment . At the concentration of 2.5-20 g/ml , O75051 significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3- O60220 cells ' response to cisplatin . Unchanged expression was found in P30874 on the SKOV3/ O60220 cell in vitro after O75051 treatment , but increased expression in vivo ( p < 0.05 ) . O75051 increased Q86UG4 -pi expression ( p < 0.05 ) and reduced Q92887 and P00533 expression ( p < 0.05 ) in a dose-dependent manner . The similar results were obtained in mice in vivo experiment , except the reduced expression of Q86UG4 -pi . It is suggested that O75051 could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface P30874 , and reverse cisplatin resistance through inhibition of Q92887 , P00533 , and even Q86UG4 -pi expressions . DB00104 -targeted liposomes loaded with CPT-11 enhanced cytotoxicity for the treatment of medullary thyroid carcinoma . Medullary thyroid carcinoma ( P04629 ) is a rare endocrine tumor that frequently metastasizes , but treatment with irinotecan ( CPT-11 ) is limited because of side effects . P04629 is known to overexpress the somatostatin receptor subtype 2 ( P30874 ) . DB00104 ( Oct ) is a somatostatin analogue that has a high binding affinity for SSTR and can be used as a tumor-targeting ligand . We prepared Oct-targeted liposomes loaded with CPT-11 using Oct-poly ( ethylene glycol ) ( PEG ) -lipid and evaluated Oct-mediated association and cytotoxicity of the liposomes with an P04629 cell line TT . The association of higher concentrations of modified Oct-targeted liposomes with TT cells was significantly higher than PEGylated liposomes and was significantly inhibited by empty Oct-targeted liposomes but not by free Oct . With exposure for 96 h , the cytotoxicity of Oct-targeted liposomal CPT-11 ( IC50 : 1.05 ± 0.47 μM ) was higher than free CPT-11 ( IC50 : 3.76 ± 0.61 μM ) or PEGylated liposomal CPT-11 ( IC50 : 3.05 ± 0.28 μM ) . In addition , empty Oct-targeted liposomes showed significantly higher cytotoxicity than empty nontargeted liposomes at a concentration where free Oct did not show cytotoxicity , suggesting that Oct as a ligand showed cytotoxicity . Moreover , Oct-targeted liposomal CPT-11 led to significantly higher antitumor activity and prolonged the survival time compared with nontargeted liposomal and free CPT-11 at a one-third dose and lower administration times with free CPT-11 . These findings indicated that Oct-targeted liposomes loaded with CPT-11 may offer considerable potential for P04629 chemotherapy because cytotoxicity of both CPT-11 and Oct was enhanced by effective cellular uptake via P30874 . Organization and regulation of paraventricular nucleus glutamate signaling systems : N-methyl-D-aspartate receptors . Stress activation of the hypothalamo-pituitary-adrenocortical ( Q9Y251 ) axis is mediated in part by glutamatergic neurotransmission . The precise nature of glutamate effects on stress-integrative hypothalamic paraventricular nucleus ( PVN ) neurons remains to be determined . Therefore , the current study was designed to delineate the organization of glutamate/ DB01221 receptor systems in the PVN and to assess regulation of PVN glutamate receptor subunit expression by chronic intermittent stress and glucocorticoids . Immunohistochemical studies verified that N-methyl-D-aspartate ( DB01221 ) receptor subunit proteins Q9UHB4 and Q12879 /2B are expressed in the medial parvocellular PVN , indicating the potential for DB01221 receptor regulation of corticotropin-releasing hormone ( P06850 ) release . Dual-label confocal analysis revealed that P06850 neurons are apposed by vesicular glutamate transporter 2 ( Q9P2U8 ) -containing terminals , consistent with glutamatergic innervation from hypothalamus and/or brainstem . In situ hybridization analysis revealed a significant and selective stress-induced decrease ( 37 % ) in Q13224 subunit mRNA expression in the P06850 -containing region of the PVN . No changes were observed for Q9UHB4 or Q12879 mRNAs . In contrast , none of the subunits investigated showed altered expression following adrenalectomy with or without low/high-dose corticosterone replacement . Thus , the observed stress regulation is likely mediated by neurogenic mechanisms in the PVN and upstream stress-transducing neurocircuitry . Because a loss of Q13224 subunit inclusion in NR receptors would likely confer increased Ca(++) conductance and faster deactivation kinetics , the stress-induced decrease in Q13224 mRNA is consistent with enhanced glutamate signaling in the PVN following chronic stress and , perhaps , increased basal Q9Y251 activity and more rapid and/or more robust Q9Y251 responses to stress .

MH_train_1298

MH_train_1298

MH_train_1298

interacts_with DB00912?

multiple_choice

DB04540 attenuates linoleic acid-induced endothelial cell activation . Vascular endothelial cell activation and dysfunction are critical early events in atherosclerosis . Even though very low or high levels of cholesterol can compromise cellular functions , cholesterol is a critical membrane component and may protect the vascular endothelium from oxidative stress and polyunsaturated fatty acid-mediated inflammatory responses . We have previously shown that the parent omega-6 fatty acid linoleic acid can markedly activate vascular endothelial cells . We now propose that membrane cholesterol can modify and inhibit linoleic acid-mediated endothelial cell dysfunction . To test this hypothesis , pulmonary artery endothelial cells were incubated with cholesterol ( 0 to 100 micromol/L ) for 24 hours and then treated with 90 micromol/L of linoleic acid ( 18:2n-6 ) for 6 to 24 hours . In control cells , treatment with linoleic acid reduced intracellular glutathione levels and induced the DNA binding activity of nuclear factor-kappaB ( NF-kappaB ) leading to the upregulation of interleukin-6 ( P05231 ) . In addition , the expression of endothelial nitric oxide synthase ( P29474 ) was altered , with linoleic acid increasing P29474 activity . In contrast , enrichment with cholesterol enhanced glutathione levels and reduced the linoleic acid-induced activation of NF-kappaBand the production of P05231 . Prior exposure to 50 micromol/L cholesterol also prevented the fatty acid-induced increase in P29474 activation . DB04540 loading activated peroxisome proliferator-activated receptor-gamma ( P37231 ) , a nuclear receptor that can decrease inflammatory responses . Furthermore , the P37231 agonist thiazolidinedione markedly downregulated the NF-kappaB activation mediated by linoleic acid . Our data suggest that signaling pathways linked to endothelial cell activation by prooxidant and proinflammatory insults may be influenced by cellular cholesterol levels . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Permanent neonatal diabetes mellitus in China . BACKGROUND : Permanent neonatal diabetes mellitus ( PNDM ) is a rare disease , which is defined as the onset of diabetes before the age of 6 months with persistence through life . Infants with Q14654 or Q09428 genetic mutations may respond to oral sulfonylurea therapy . Currently , there are limited studies about the genetic analysis and long-term follow-up of PNDM . CASE PRESENTATION : We report four cases of PNDM . None of the infants or their parents had P01308 , Q14654 , or Q09428 genetic mutations . One infant underwent continuous subcutaneous insulin infusion ( CSII ) and the other infants underwent multiple injections of insulin ( MII ) . In these infants , PNDM persisted from 35 months to 60 months of follow-up . Three infants maintained fairly stable blood sugar levels , and one infant had poor sugar control . CONCLUSIONS : We suggest that all of the infants with PNDM should undergo genetic evaluation . For infants without Q14654 and Q09428 genetic mutations , oral sulfonylurea should not be considered as treatment . CSII is a useful method for overcoming the difficulties of diabetes , and it may also improve the quality of life of both infants and their parents . Peroxisome proliferator-activated receptor-gamma and its ligands attenuate biologic functions of human natural killer cells . Interferon-gamma ( P01579 ) production and cytolytic activity are 2 major biologic functions of natural killer ( NK ) cells that are important for innate immunity . We demonstrate here that these functions are compromised in human NK cells treated with peroxisome proliferator-activated-gamma ( P37231 ) ligands via both P37231 -dependent and -independent pathways due to variation in P37231 expression . In P37231 -null NK cells , 15-deoxy-Delta(12,14) prostaglandin J(2) ( 15d-PGJ(2) ) , a natural P37231 ligand , reduces P01579 production that can be reversed by MG132 and/or chloroquine , and it inhibits cytolytic activity of NK cells through reduction of both conjugate formation and Q07108 expression . In PPARgamma-positive NK cells , P37231 activation by 15d-PGJ(2) and ciglitazone ( a synthetic ligand ) leads to reduction in both mRNA and protein levels of P01579 . Overexpression of P37231 in P37231 -null NK cells reduces P01579 gene expression . However , P37231 expression and activation has no effect on NK cell cytolytic activity . In addition , 15d-PGJ(2) but not ciglitazone reduces expression of Q07108 in human NK cells , whereas P16070 expression is not affected . These results reveal novel pathways regulating NK cell biologic functions and provide a basis for the design of therapeutic agents that can regulate the function of NK cells within the innate immune response . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . Effect of cholinesterase inhibitors on acetylcholine and insulin induced glucose uptake and certain hepatic enzymes in pigeon liver : an in vitro study . P01308 and acetylcholine ( ACh ) are both known to promote glucose uptake by liver of birds . DB03128 induced glucose uptake can be predictably potentiated by inhibiting acetylcholinesterase activity . Monocrotophos , acothione ( organophosphorus compound ) and prostigmine are known inhibitors of acetylcholinesterase ( P22303 ) . In the present study the action of these three inhibitors of P22303 alone as well as in combination with insulin and acetylcholine on in vitro glucose uptake by pigeon liver slices was investigated . Both organophorus compounds potentiated the action of insulin as well as acetycholine mediated glucose uptake by liver slices while prostigmine had inhibitory influence . The three compounds also induced alterations in enzyme activities in the liver slices . These results are discussed in detail in the text . Andrographis paniculata extract ( DB05767 ) for active ulcerative colitis . OBJECTIVES : Andrographis paniculata has in vitro inhibitory activity against P01375 -α , IL-1β and NF-κB . A pilot study of A. paniculata extract ( DB05767 ) suggested similar efficacy to mesalamine for ulcerative colitis . METHODS : A randomized , double-blind , placebo-controlled trial evaluated the efficacy of A. paniculata extract ( DB05767 ) in 224 adults with mild-to-moderate ulcerative colitis . Patients were randomized to A. paniculata extract ( DB05767 ) 1,200 mg or 1,800 mg daily or placebo for 8 weeks . RESULTS : In total , 45 and 60 % of patients receiving A. paniculata 1,200 mg and 1,800 mg daily , respectively , were in clinical response at week 8 , compared with 40 % of those who received placebo ( P=0.5924 for 1,200 mg vs. placebo and P=0.0183 for 1,800 mg vs. placebo ) . In all , 34 and 38 % of patients receiving A. paniculata 1,200 mg and 1,800 mg daily , respectively , were in clinical remission at week 8 , compared with 25 % of those who received placebo ( P=0.2582 for 1,200 mg vs. placebo and P=0.1011 for 1,800 mg vs. placebo ) . Adverse events developed in 60 and 53 % of patients in the A. paniculata 1,200 mg and 1,800 mg daily groups , respectively , and 60 % in the placebo group . CONCLUSIONS : Patients with mildly to moderately active ulcerative colitis treated with A. paniculata extract ( DB05767 ) at a dose of 1,800 mg daily were more likely to achieve clinical response than those receiving placebo . Mechanisms of regulation of P61073 / P48061 ( P48061 ) -dependent migration and homing in multiple myeloma . The mechanisms by which multiple myeloma ( MM ) cells migrate and home to the bone marrow are not well understood . In this study , we sought to determine the effect of the chemokine P48061 ( P48061 ) and its receptor P61073 on the migration and homing of MM cells . We demonstrated that P61073 is differentially expressed at high levels in the peripheral blood and is down-regulated in the bone marrow in response to high levels of P48061 . P48061 induced motility , internalization , and cytoskeletal rearrangement in MM cells evidenced by confocal microscopy . The specific P61073 inhibitor DB06809 and the anti- P61073 antibody MAB171 inhibited the migration of MM cells in vitro . P61073 knockdown experiments demonstrated that P48061 -dependent migration was regulated by the P13K and P29323 / MAPK pathways but not by p38 MAPK . In addition , we demonstrated that DB06809 inhibited the homing of MM cells to the bone marrow niches using in vivo flow cytometry , in vivo confocal microscopy , and whole body bioluminescence imaging . This study , therefore , demonstrates that P48061 / P61073 is a critical regulator of MM homing and that it provides the framework for inhibitors of this pathway to be used in future clinical trials to abrogate MM trafficking . DB04941 , an antisecretory antidiarrheal proanthocyanidin oligomer extracted from Croton lechleri , targets two distinct intestinal chloride channels . DB04941 , a purified proanthocyanidin oligomer extracted from the bark latex of Croton lechleri , is in clinical trials for secretory diarrheas of various etiologies . We investigated the antisecretory mechanism of crofelemer by determining its effect on the major apical membrane transport and signaling processes involved in intestinal fluid transport . Using cell lines and measurement procedures to isolate the effects on individual membrane transport proteins , crofelemer at 50 microM had little or no effect on the activity of epithelial Na(+) or K(+) channels or on DB02527 or calcium signaling . DB04941 inhibited the cystic fibrosis transmembrane regulator ( P13569 ) Cl(-) channel with maximum inhibition of approximately 60 % and an IC(50) approximately 7 microM . DB04941 action at an extracellular site on P13569 produced voltage-independent block with stabilization of the channel closed state . DB04941 did not affect the potency of glycine hydrazide or thiazolidinone P13569 inhibitors . DB04941 action resisted washout , with < 50 % reversal of P13569 inhibition after 4 h . DB04941 was also found to strongly inhibit the intestinal calcium-activated Cl(-) channel Q5XXA6 by a voltage-independent inhibition mechanism with maximum inhibition > 90 % and IC(50) approximately 6.5 microM . The dual inhibitory action of crofelemer on two structurally unrelated prosecretory intestinal Cl(-) channels may account for its intestinal antisecretory activity . A randomized , placebo-controlled study of the effects of the p38 MAPK inhibitor SB- DB05250 on blood biomarkers of inflammation in P48444 patients . The p38 mitogen-activated protein kinase ( MAPK ) signaling upregulates inflammation and is known to be increased in chronic obstructive pulmonary disease ( P48444 ) . The authors assessed the pharmacology of the novel p38 MAPK inhibitor SB- DB05250 using blood biomarkers in P48444 . Seventeen P48444 patients ( forced expiratory volume in 1 second 50 % -80 % predicted ) using short-acting bronchodilators participated in a double-blind , double-dummy , randomized , crossover study . Patients received single oral doses of SB- DB05250 7.5 mg and 25 mg , prednisolone 10 mg and 30 mg , and placebo . Blood was obtained predose and at 1 , 2 , 6 , and 24 hours postdose . Whole-blood sorbitol-induced phosphorylated ( p ) heat shock protein ( HSP ) 27 levels as a marker of p38 pathway activation and lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -alpha production were assessed . Both doses of SB- DB05250 , but not prednisolone , significantly ( P < .0001 ) reduced weighted mean ( WM ) pHSP27 ( 0-6 hours ) by 58 % compared with placebo . WM P01375 production ( 0-24 hours ) was significantly reduced compared with placebo by SB- DB05250 25 mg ( 40 % , P = .005 ) and 7.5 mg ( 33.4 % , P = .02 ) , while prednisolone 30 mg and 10 mg caused 81.5 % and 58.2 % suppression , respectively ( both P < .0001 ) . SB- DB05250 inhibited the p38 MAPK pathway to a greater degree than prednisolone did . SB- DB05250 inhibited P01375 production . SB- DB05250 is a potent p38 MAPK inhibitor that potentially suppresses inflammation in P48444 . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Modulation of Q8IVT5 activity in Caenorhabditis elegans by Zn ions , P25116 kinase and PP2A phosphatase . Vulval differentiation in Caenorhabditis elegans is controlled by a conserved signal transduction pathway mediated by Ras and a kinase cascade that includes Raf , Mek and MAPK . Activation of this cascade is positively regulated by a number of proteins such as Q8IVT5 ( kinase suppressor of Ras ) , Q09428 -8/ Q5T124 -2 , Q09428 -6/PP2A-B and P05231 -1 . We describe the functional characterization of sur-7 and several genes that regulate signaling downstream of ras . We identified sur-7 by isolating a mutation that suppresses an activated ras allele , and showed that Q09428 -7 is a divergent member of the cation diffusion facilitator family of heavy metal ion transporters that is probably localized to the endoplosmic recticulum membrane and regulates cellular Zn(2+) concentrations . Genetic double mutant analyses suggest that the Q09428 -7-mediated effect is not a general toxic response . Instead , Zn(2+) ions target a specific step of the pathway , probably regulation of the scaffolding protein Q8IVT5 . Biochemical analysis in mammalian cells indicates that high Zn(2+) concentration causes a dramatic increase of Q8IVT5 phosphorylation . Genetic analysis also indicates that PP2A phosphatase and P25116 kinase act downstream of Raf to positively and negatively regulate Q8IVT5 activity , respectively . High-fat diet exacerbates renal dysfunction in SHR : reversal by induction of P09601 -adiponectin axis . High-dietary fat intake is a major risk factor for development of metabolic and cardiovascular-renal dysfunction including obesity , coronary artery disease , hypertension , and chronic renal failure . We examined the effect of a high-fat diet on renal function and morphology in spontaneously hypertensive rats ( SHR ) , a phenotype designed to mimic metabolic syndrome . High-fat diet induced increase ( P < 0.05 ) in blood pressure , body weight , and renal lipid deposition in these rats . This increase in body weight was accompanied by elevations ( P < 0.05 ) of blood glucose and low-density lipoprotein ( LDL ) levels , a decrease ( P < 0.05 ) in adiponectin and increases ( P < 0.05 ) in plasma monocyte chemotactic protein-1 ( P13500 ) along with renal macrophage infiltration . These pathophysiological perturbations were attenuated ( P < 0.05 ) by heme oxygenase-1 ( P09601 ) induction by treatment with cobalt protoporphyrin ( CoPP ) . Further effects of CoPP included increased ( P < 0.05 ) renal expression of adiponectin along with enhancement ( P < 0.05 ) of pAKT , pAMPK , and p- P29474 in SHRs fed a high-fat diet . Prevention of such beneficial effects of CoPP by the concurrent administration of the heme-HO inhibitor stannous mesoporphyrin ( DB04912 ) corroborates the role of HO system in mediating such effects . Taken together , our results demonstrate that high-fat diet induces a metabolic syndrome-like phenotype in hypertensive rats , which is amenable to rescue by increases in P09601 - and adiponectin-dependent pathways . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . P16070 -targeted docetaxel conjugate for cancer cells and cancer stem-like cells : a novel hyaluronic acid-based drug delivery system . A P16070 -targeted macromolecular conjugate of docetaxel was prepared via a pH-sensitive linkage to hyaluronic acid and was characterized using NMR , gel permeation chromatography , and differential scanning calorimetry . The conjugated species were further evaluated in terms of drug release , cytotoxicity , cellular uptake , cell cycle inhibition , and subacute toxicity in mice . Cellular microscopic studies revealed that P16070 -expressing cells including MCF-7 cancer stem cells and MDA-MB-231 metastatic breast cancer cells had internalized the conjugates via a selective receptor-mediated mechanism , leading to cell cycle arrest in the G2/M phase . DB08818 -docetaxel conjugates showed specific toxicity only in P16070 -expressing cells in vitro , along with a decreased risk of neutropenia and dose-dependent mortality in vivo . DB08818 -drug conjugates represent a promising and efficient platform for solubilization of sparingly soluble molecules as well as active and selective targeted delivery to cancer cells and cancer stem cells . DB01819 cycling via mitochondrial phosphoenolpyruvate carboxykinase links anaplerosis and mitochondrial GTP with insulin secretion . Pancreatic beta-cells couple the oxidation of glucose to the secretion of insulin . Apart from the canonical K( DB00171 )-dependent glucose-stimulated insulin secretion ( GSIS ) , there are important K( DB00171 )-independent mechanisms involving both anaplerosis and mitochondrial GTP ( mtGTP ) . How mtGTP that is trapped within the mitochondrial matrix regulates the cytosolic calcium increases that drive GSIS remains a mystery . Here we have investigated whether the mitochondrial isoform of phosphoenolpyruvate carboxykinase ( Q16822 ) is the GTPase linking hydrolysis of mtGTP made by succinyl- DB01992 synthetase ( SCS-GTP ) to an anaplerotic pathway producing phosphoenolpyruvate ( PEP ) . Although cytosolic PEPCK ( P35558 ) is absent , Q16822 message and protein were detected in P01308 -1 832/13 cells , rat islets , and mouse islets . PEPCK enzymatic activity is half that of primary hepatocytes and is localized exclusively to the mitochondria . Novel (13)C-labeling strategies in P01308 -1 832/13 cells and islets measured substantial contribution of Q16822 to the synthesis of PEP . As high as 30 % of PEP in P01308 -1 832/13 cells and 41 % of PEP in rat islets came from Q16822 . The contribution of Q16822 to overall PEP synthesis more than tripled with glucose stimulation . Silencing the Q16822 gene completely inhibited GSIS underscoring its central role in mitochondrial metabolism-mediated insulin secretion . Given that mtGTP synthesized by SCS-GTP is an indicator of TCA flux that is crucial for GSIS , Q16822 is a strong candidate to link mtGTP synthesis with insulin release through anaplerotic PEP cycling .

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interacts_with DB00203?

multiple_choice

DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . Hyperhomocysteinemia and P42898 C677T and A1298C polymorphisms are associated with chronic allograft nephropathy in renal transplant recipients . Hyperhomocysteine has been reported to be an important risk factor for the development of atherosclerosis . Identification of risk factors , such as hyperhomocysteinemia , is crucial for a better understanding of the events that lead to degenerative processes in the vascular system and for a correct understanding of the potential role of methylene- DB00116 reductase enzymes ( P42898 ) to help in the treatment of vascular disease observed in chronic allograft nephropathy ( P35658 ) . In this study we analyzed the plasma homocysteine concentrations and P42898 C677T and A1298C polymorphism frequencies among 110 renal transplant recipients ( 53 with P35658 and 57 with normal renal function ) . All recipients had undergone renal transplantation at least 12 months prior to this investigation to establish a possible correlation with the posttransplant outcome . Plasma homocysteine concentrations were measured by liquid chromatography-tandem mass spectrometry and P42898 polymorphisms were investigated by the PCR-RFLP technique . The results demonstrated that in renal transplant recipients , hyperhomocysteinemia in addition to the presence of the allelic variants for both P42898 polymorphisms ( 677T/1298C ) might play a role as an additional risk factor for P35658 . We understand that analysis of these polymorphisms might have a role in the P35658 process . Therefore , studies to evaluate their presence in renal transplant patients may be extremely useful to individualize immunosuppressive protocols to inhibit or retard the progression of P35658 . OSU-03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier : Implications for Anti-Cancer Therapies . We examined the interaction between OSU-03012 ( also called AR-12 ) with phosphodiesterase 5 ( O76074 ) inhibitors to determine the role of the chaperone glucose-regulated protein ( P11021 ) / P11021 / P11021 in the cellular response . DB00203 ( Viagra ) interacted in a greater than additive fashion with OSU-03012 to kill stem-like GBM cells . Treatment of cells with OSU-03012/sildenafil : abolished the expression of multiple oncogenic growth factor receptors and plasma membrane drug efflux pumps and caused a rapid degradation of P11021 and other HSP70 and HSP90 family chaperone proteins . Decreased expression of plasma membrane receptors and drug efflux pumps was dependent upon enhanced Q9NZJ5 -eIF2α- P18848 - P35638 signaling and was blocked by P11021 over-expression . In vivo OSU-03012/sildenafil was more efficacious than treatment with celecoxib and sildenafil at killing tumor cells without damaging normal tissues and in parallel reduced expression of P08183 and Q9UNQ0 in the normal brain . The combination of OSU-03012/sildenafil synergized with low concentrations of sorafenib to kill tumor cells , and with lapatinib to kill P00533 over-expressing tumor cells . In multiplex assays on plasma and human tumor tissue from an OSU-03012/sildenafil treated mouse , we noted a profound reduction in uPA signaling and identified FGF and P23458 /2 as response biomarkers for potentially suppressing the killing response . Inhibition of FGFR signaling and to a lesser extent P23458 /2 signaling profoundly enhanced OSU-03012/sildenafil lethality . P15692 -dependent and PDGF-dependent dynamic neurovascular reconstruction in the neurohypophysis of adult mice . Hypothalamo-neurohypophysial system ( HNS ) releases arginine vasopressin ( AVP ) and oxytocin ( P01178 ) from axonal terminals of the neurohypophysis ( NH ) into blood circulation for controlling body fluid homeostasis and lactation . Chronic osmotic and suckling stimulations have been shown to cause neurovascular and neuroglial reconstruction in the NH of adult mammals and no study has been reported for vascular dynamics . The aim of this study was to elucidate the occurrence of continuous angiogenesis and growth factor-dependent neurovascular reconstruction in the NH of adult mice . Active proliferation of endothelial cells and oligodendrocyte progenitor cells ( OPCs ) was observed using the immunohistochemistry of bromodeoxyuridine and Ki-67 . P15692 ( P15692 ) and P15692 receptor 2 ( P35968 ( P35968 ) ) were highly expressed at pituicytes and endothelial cells respectively . Moreover , prominent expression of platelet-derived growth factor B ( PDGFB ) and PDGF receptor beta was observed at P01178 -containing axonal terminals and pericytes respectively . Administration of the selective tyrosine kinase inhibitor DB04849 for VEGFRs and STI571 for PDGFRs significantly decreased proliferation of endothelial cells and OPCs . Moreover , DB04849 treatment decreased vascular density by facilitating apoptosis of endothelial cells and the withdrawal of its treatment led to remarkable rebound proliferation of endothelial cells , so that vascular density rapidly returned to normal levels . DB04849 decreased the density of both AVP- and P01178 -containing axonal terminals , whereas STI571 selectively decreased the density of AVP-containing ones . Thus , this study demonstrates that the signaling pathways of P15692 and PDGF are crucial mediators for determining proliferation of endothelial cells and OPCs and the density of AVP- and P01178 -containing axonal terminals in the HNS . Leukotriene A4 hydrolase . Inhibition by bestatin and intrinsic aminopeptidase activity establish its functional resemblance to metallohydrolase enzymes . DB03424 , an inhibitor of aminopeptidases , was also a potent inhibitor of leukotriene ( LT ) A4 hydrolase . On isolated enzyme its effects were immediate and reversible with a Ki = 201 +/- 95 mM . With erythrocytes it inhibited LTB4 formation greater than 90 % within 10 min ; with neutrophils it inhibited LTB4 formation by only 10 % during the same period , increasing to 40 % in 2 h . DB03424 inhibited P09960 hydrolase selectively ; neither P09917 nor 15-lipoxygenase activity in neutrophil lysates was affected . Purified P09960 hydrolase exhibited an intrinsic aminopeptidase activity , hydrolyzing L-lysine-p-nitroanilide and L-leucine-beta-naphthylamide with apparent Km = 156 microM and 70 microM and Vmax = 50 and 215 nmol/min/mg , respectively . Both P09960 and bestatin suppressed the intrinsic aminopeptidase activity of P09960 hydrolase with apparent Ki values of 5.3 microM and 172 nM , respectively . Other metallohydrolase inhibitors tested did not reduce P09960 hydrolase/aminopeptidase activity , with one exception ; captopril , an inhibitor of angiotensin-converting enzyme , was as effective as bestatin . The results demonstrate a functional resemblance between P09960 hydrolase and certain metallohydrolases , consistent with a molecular resemblance at their putative Zn2(+)-binding sites . The availability of a reversible , chemically stable inhibitor of P09960 hydrolase may facilitate investigations on the role of LTB4 in inflammation , particularly the process termed transcellular biosynthesis . A multicenter , open-label , prospective , randomized , dose-ranging pharmacokinetic study of the anti- P01375 antibody afelimomab in patients with sepsis syndrome . OBJECTIVE : To investigate the pharmacokinetics and safety of afelimomab , a murine antibody fragment against human tumor necrosis factor ( P01375 ) -alpha in patients with sepsis . DESIGN : Multicenter , randomized , open-label , placebo-controlled phase I/II clinical trial . SETTING : Intensive care units of six academic medical centers in the United States . PATIENTS : Forty-eight patients with a clinical diagnosis of sepsis who received standard supportive care and antimicrobial therapy . INTERVENTIONS : Patients received 0.3 , 1.0 , or 3.0 mg/kg afelimomab or placebo intravenously over 20 min . Three patients in each dose group received single doses ; the remaining nine patients in each group received multiple ( nine ) doses at 8-h intervals over 72 h . MEASUREMENTS AND MAIN RESULTS : DB04956 appeared safe and well tolerated . Single- and multiple-dose kinetics were predictable and dose related . The elimination half-life was 44.7 h . DB04956 treatment resulted in increased serum concentrations of P01375 ( includes P01375 -antibody complexes ) and decreased serum interleukin-6 concentrations , whereas no discernible trends were observed in placebo-treated patients . There was no significant treatment effect on 28-day mortality as was expected given the small number of patients . However , overall mortality was significantly ( p = 0.001 ) associated with baseline interleukin-6 concentration . All patients experienced adverse events , but the vast majority were considered unrelated to the study drug and demonstrated no apparent relationship to afelimomab dose . Although 41 % of patients developed human anti-murine antibodies , there were no clinical sequelae . CONCLUSIONS : Multidose therapy with afelimomab was safe , well tolerated , and had predictable linear kinetics . A large randomized trial comparing afelimomab to placebo in patients with well defined sepsis has recently been completed . Sequential treatment of HPV E6 and E7-expressing TC-1 cells with bortezomib and celecoxib promotes apoptosis through p-p38 MAPK-mediated downregulation of cyclin D1 and P24941 . Interruption of the cell cycle is accompanied by changes in several related molecules that result in the activation of apoptosis . The present study was performed to verify the apoptotic effects of sequential treatment with bortezomib and celecoxib in TC-1 cells expressing the human papillomavirus ( HPV ) E6 and E7 proteins . In TC-1 cells sequentially treated with bortezomib and celecoxib , apoptosis was induced through decreased expression of signal transducer and activator of transcription-3 ( P40763 ) , cyclin D1 and cyclin-dependent kinase ( CDK ) 2 , which are major regulators of the G0/ P55008 cell cycle checkpoint . In addition , increased levels of P38936 , P35638 , P11021 and p-p38 MAPK were identified in these cells . The treatment-induced apoptosis was effectively inhibited by treatment with SB203580 , an inhibitor of p-p38 . Moreover , the growth of tumors sequentially treated with bortezomib and celecoxib was retarded compared to the growth of tumors exposed to a single treatment with either bortezomib or celecoxib in vivo . We demonstrated that sequential treatment with bortezomib and celecoxib induced apoptosis via p-p38-mediated G0/ P55008 cell cycle arrest and endoplasmic reticulum ( ER ) stress . Sequential treatment with these two drugs could therefore be a useful therapy for cervical cancer . P03372 ligands : a patent review update . INTRODUCTION : The role of estrogens is mostly mediated by two nuclear receptors ( ERα and ERβ ) and a membrane-associated G-protein ( Q99527 or Q99527 ) , and it is not limited to reproduction , but it extends to the skeletal , cardiovascular and central nervous systems . Various pathologies such as cancer , inflammatory , neurodegenerative and metabolic diseases are often associated with dysfunctions of the estrogenic system . Therapeutic interventions by agents that affect the estrogenic signaling pathway might be useful in the treatment of many dissimilar diseases . AREAS COVERED : The massive chemodiversity of ER ligands , limited to patented small molecules , is herein reviewed . The reported compounds are classified on the basis of their chemical structures . Non-steroidal derivatives , which mostly consist of diphenolic compounds , are further segregated into chemical classes based on their central scaffold . EXPERT OPINION : DB00286 have been used for almost a century and their earlier applications have concerned interventions in the female reproductive functions , as well as the treatment of some estrogen-dependent cancers and osteoporosis . Since the discovery of ERβ in 1996 , the patent literature has started to pay a progressively increasing attention to this newer receptor subtype , which holds promise as a target for new indications , most of which still need to be clinically validated . The PEPvIII-KLH ( DB05374 ) vaccine in glioblastoma multiforme patients . Conventional therapies for glioblastoma multiforme ( GBM ) fail to target tumor cells exclusively , resulting in non-specific toxicity . Immune targeting of tumor-specific mutations may allow for more precise eradication of neoplastic cells . P00533 variant III ( EGFRvIII ) is a tumor-specific mutation that is widely expressed in GBM and other neoplasms and its expression enhances tumorigenicity . This in-frame deletion mutation splits a codon , resulting in a novel glycine at the fusion junction producing a tumor-specific epitope target for cellular or humoral immunotherapy . We have previously shown that vaccination with a peptide that spans the EGFRvIII fusion junction ( PEPvIII-KLH/ DB05374 ) is an efficacious immunotherapy in syngeneic murine models . In this review , we summarize our results in GBM patients targeting this mutation in multiple , multi-institutional Phase II immunotherapy trials . These trials demonstrated that a selected population of GBM patients who received vaccines targeting EGFRvIII had an unexpectedly long survival time . Further therapeutic strategies and potential pitfalls of using this approach are discussed . Proliferative responses to a nonspecific factor produced by irradiated stimulating cells can simulate antigen-specific secondary responses in the primed lymphocyte test . The primed lymphocyte test ( Q02083 ) was utilized to investigate the biologic relationship between alloantigens and soluble environmental antigens . Human lymphocytes primed in vitro to the soluble antigens Candida ( P35658 ) , Tetanus ( TET ) , or Purified Protein Derivative of Tuberculin ( PPD ) gave a strong proliferative response when restimulated with the initial soluble antigen in the presence of irradiated peripheral blood lymphocytes ( PBL ) acting as antigen-presenting cells ( P25054 ) . Surprisingly , the soluble antigen-primed cells as well as alloantigen-primed cells responded to other soluble antigens in the presence of P25054 . By testing primed cells with supernatants derived from irradiated lymphocytes plus soluble antigen , it was apparent that the responses observed were in part due to stimulation by a soluble factor produced by the irradiated " P25054 " in response to the soluble antigen rather than to recognition of widespread cross-reactivity by the primed cells . This " nonspecific " factor production could be diminished by increasing the dose of irradiation to the P25054 or by using a T lymphocyte depleted- ( SRBC-E- ) P25054 population . In addition , certain antigen-reactive T cell clones did not respond to the nonspecific factor to the same degree as the primed bulk cultures . Nevertheless , the recognition of nonspecific stimulation induced by factors produced by irradiated lymphocytes is critical in the interpretation of primed lymphocyte responses to alloantigens or soluble antigens . Pharmacologic blockade of P09917 improves the amyloidotic phenotype of an Alzheimer 's disease transgenic mouse model involvement of γ-secretase . The P09917 ( P09917 ) enzyme is widely distributed within the central nervous system . Previous works showed that this protein is up-regulated in Alzheimer 's disease ( AD ) and that its genetic absence results in a reduction of amyloid β ( Aβ ) levels in Tg2576 mice . In the present study , we examined the effect of P09917 pharmacological inhibition on the amyloidotic phenotype of these mice . Aβ deposition in the brains of mice receiving zileuton , a selective and specific P09917 inhibitor , was significantly reduced when compared with control Tg2576 mice receiving vehicle . This reduction was associated with a similar decrease in brain Aβ peptides levels . Zileuton treatment did not induce any change in the steady state levels of amyloid-β precursor protein ( P05067 ) , P56817 or O14672 . By contrast , it resulted in a significant reduction of presenilin 1 ( P49768 , alias P49768 ) , nicastrin ( Q92542 ) , presenilin enhancer 2 homolog ( PSNEN , alias , Pen-2 ) , and anterior pharynx defective 1 ( P13798 -1 ) , the four components of the γ-secretase complex-at the protein and message level . Furthermore , in vitro studies confirmed that zileuton prevents Aβ formation by modulating γ-secretase complex levels without affecting Notch signaling . These data establish a functional role for P09917 in the pathogenesis of AD-like amyloidosis , whereby it modulates the γ-secretase pathway . They suggest that pharmacological inhibition of P09917 could provide a novel therapeutic opportunity for AD . Inhibition of Q16552 as a pharmacological approach for Q9UKU7 . Several experimental approaches have been utilized , in order to critically examine the roles of Q16552 family members in intestinal inflammation . These approaches have included : ( 1 ) the use of Q16552 and Q96PD4 -deficient mice , ( 2 ) specific antibodies directed against Q16552 , ( 3 ) an Q16552 vaccine , ( 4 ) methods to block the Q16552 receptor and ( 5 ) small-molecule inhibitors of Q16552 . Previous studies found somewhat conflicting results in preclinical models of Inflammatory Bowel Disease ( Q9UKU7 ) , using specific strains of Q16552 -deficient mice . This paper will review the preclinical results using various pharmacological approaches [ specific Q16552 antibodies , an Q16552 receptor fusion protein , IL-12/IL-23 p40 subunit and Q16552 vaccine approaches , as well as a small molecule inhibitor ( Vidofludimus ) ] to inhibit Q16552 in animal models of Q9UKU7 . Recent clinical results in patients with Q9UKU7 will also be discussed for DB09029 ( an Q16552 antibody ) , Brodalumab ( an Q16552 receptor antibody ) and two small-molecule drugs ( Vidofludimus and DB08895 ) , which inhibit Q16552 as part of their overall pharmacological profiles . This review paper will also discuss some pharmacological lessons learned from the preclinical and clinical studies with anti- Q16552 drugs , as related to drug pharmacodynamics , Q16552 receptor subtypes and other pertinent factors . Finally , future pharmacological approaches of interest will be discussed , such as : ( 1 ) Retinoic acid receptor-related orphan nuclear receptor gamma t ( Rorγt ) antagonists , ( 2 ) P10276 ( RARα ) antagonists , ( 3 ) Pim-1 kinase inhibitors and ( 4 ) Dual small-molecule inhibitors of NF-κB and P40763 , like synthetic triterpenoids . Advances in genomic and proteomic studies of non-small-cell lung cancer : clinical and translational research perspective . Recent years have brought tremendous progress in the development of genomic and proteomic platforms to study cancer biology . Tests based on these platforms are helpful in early diagnosis , prognosis , and prediction of treatment benefit . Molecular studies performed on minimally invasive material ( plasma , sputum ) from individuals participating in longitudinal or case-control studies have approximately 70 % -90 % sensitivity and specificity to detect lung cancer . In operable non-small-cell lung cancer , genomic and proteomic studies yield better prognostic information than pathologic staging . There are several examples of successful identification of predictive assays for benefit from chemotherapy ( P07992 , P23921 , P46527 , and p53 expression ) or targeted therapies ( epidermal growth factor receptor [ P00533 ] gene copy number , P00533 activating mutations , P00533 protein expression , serum proteomic profile ) . These markers should be prospectively tested in clinical studies before they can be routinely used in the clinic . Blockade of interleukin-6 signalling with siltuximab enhances melphalan cytotoxicity in preclinical models of multiple myeloma . Signalling through the interleukin ( IL ) -6 pathway induces proliferation and drug resistance of multiple myeloma cells . We therefore sought to determine whether the P05231 -neutralizing monoclonal antibody siltuximab , formerly CNTO 328 , could enhance the activity of melphalan , and to examine some of the mechanisms underlying this interaction . DB09036 increased the cytotoxicity of melphalan in KAS-6/1 , Q16352 -6 , ANBL-6 , and RPMI 8226 human myeloma cell lines ( HMCLs ) in an additive-to-synergistic manner , and sensitized resistant RPMI 8226.LR5 cells to melphalan . These anti-proliferative effects were accompanied by enhanced activation of drug-specific apoptosis in HMCLs grown in suspension , and in HMCLs co-cultured with a human-derived stromal cell line . DB09036 with melphalan enhanced activation of caspase-8 , caspase-9 , and the downstream effector caspase-3 compared with either of the single agents . This increased induction of cell death occurred in association with enhanced Bak activation . Neutralization of P05231 also suppressed signalling through the phosphoinositide 3-kinase/Akt pathway , as evidenced by decreased phosphorylation of Akt , P08133 S6 kinase and Q13541 . Importantly , the siltuximab/melphalan regimen demonstrated enhanced anti-proliferative effects against primary plasma cells derived from patients with myeloma , monoclonal gammopathy of undetermined significance , and amyloidosis . These studies provide a rationale for translation of siltuximab into the clinic in combination with melphalan-based therapies . Q99527 engagement activates endothelial nitric oxide synthase via the P19957 -kinase-Akt pathway in human endothelial cells . 17beta-Estradiol ( E(2) ) is a rapid activator of endothelial nitric oxide synthase ( P29474 ) . The product of this activation event , NO , is a fundamental determinant of cardiovascular homeostasis . We previously demonstrated that E(2)-stimulated endothelial NO release can occur without an increase in cytosolic Ca(2+) . Here we demonstrate for the first time , to our knowledge , that E(2) rapidly induces phosphorylation and activation of P29474 through the phosphatidylinositol 3 ( P19957 ) -kinase-Akt pathway . E(2) treatment ( 10 ng/mL ) of the human endothelial cell line , EA.hy926 , resulted in increased NO production , which was abrogated by the P19957 -kinase inhibitor , LY294002 , and the estrogen receptor antagonist ICI 182 , 780 . E(2) stimulated rapid Akt phosphorylation on serine 473 . As has been shown for vascular endothelial growth factor , P29474 is an E(2)-activated Akt substrate , demonstrated by rapid P29474 phosphorylation on serine 1177 , a critical residue for P29474 activation and enhanced sensitivity to resting cellular Ca(2+) levels . Adenoviral-mediated EA.hy926 transduction confirmed functional involvement of Akt , because a kinase-deficient , dominant-negative Akt abolished E(2)-stimulated NO release . The membrane-impermeant E(2)BSA conjugate , shown to bind endothelial cell membrane sites , also induced rapid Akt and consequent P29474 phosphorylation . Thus , engagement of membrane estrogen receptors results in rapid endothelial NO release through a P19957 -kinase-Akt-dependent pathway . This explains , in part , the reduced requirement for cytosolic Ca(2+) fluxes and describes an important pathway relevant to cardiovascular pathophysiology . DB00203 enhances neurogenesis and oligodendrogenesis in ischemic brain of middle-aged mouse . Adult neural stem cells give rise to neurons , oligodendrocytes and astrocytes . Aging reduces neural stem cells . Using an inducible nestin-CreER( P24752 )/R26R-yellow fluorescent protein ( YFP ) mouse , we investigated the effect of DB00203 , a phosphodiesterase type 5 ( O76074 ) inhibitor , on nestin lineage neural stem cells and their progeny in the ischemic brain of the middle-aged mouse . We showed that focal cerebral ischemia induced nestin lineage neural stem cells in the subventricular zone ( SVZ ) of the lateral ventricles and nestin expressing NeuN positive neurons and adenomatous polyposis coli ( P25054 ) positive mature oligodendrocytes in the ischemic striatum and corpus callosum in the aged mouse . Treatment of the ischemic middle-aged mouse with DB00203 increased nestin expressing neural stem cells , mature neurons , and oligodendrocytes by 33 , 75 , and 30 % , respectively , in the ischemic brain . These data indicate that DB00203 amplifies nestin expressing neural stem cells and their neuronal and oligodendrocyte progeny in the ischemic brain of the middle-aged mouse . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . Expression of the human concentrative nucleotide transporter 1 ( O00337 ) gene correlates with clinical response in patients affected by Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) undergoing a combination treatment with 2-chloro-2'-deoxyadenosine ( DB00242 ) and DB00073 . PURPOSE : Resistance to nucleoside analogues agents is likely to be multifactorial and could involve a number of mechanisms affecting drug penetration , metabolism and targeting . In vitro studies of resistant human cell lines have confirmed that human concentrative nucleoside transporter 1 ( O00337 ) -deficient cells display resistance . EXPERIMENTAL DESIGN : We applied real-time PCR method to assess the mRNA expression of equilibrative and concentrative nucleoside transporter ( hENT1 , O00337 ) , deoxycytidine and deoxyguanosine kinase ( P27707 , Q16854 ) , 5'-nucleotidase ( 5'-NT ) , ribonucleotide reductase catalytic and regulatory ( P23921 , P31350 ) subunits in bone marrow cells from 32 patients with Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) who received 2CdA-based chemotherapy . Responses to chemotherapy , were then correlated to the expression of these markers . RESULTS : All 32 patients enrolled expressed lower levels of O00337 as compared to healthy donors . In univariate analysis , lower expression level of O00337 ( p=0.0021 ) and P31350 ( p=0.02 ) correlated with response to chemotherapy . In particular , patients with low levels of O00337 achieved inferior clinical response . No significant correlation between these genes expression and age , stage of disease was found . This study suggests that nucleotidase expression levels can be used to identify subgroups of WM and SLL patients who will likely respond differently to a 2CdA-based therapy . DB00203 induces angiogenic response in human coronary arteriolar endothelial cells through the expression of thioredoxin , hemeoxygenase and vascular endothelial growth factor . This study was undertaken to investigate the effect of phosphodiesterase-5 ( O76074 ) inhibitor , sildenafil , on angiogenic response in human coronary arteriolar endothelial cells ( HCAEC ) . The cells exposed to sildenafil ( 1-20 microM ) demonstrated significantly accelerated tubular morphogenesis with the induction of thioredoxin-1 ( P10599 -1 ) , hemeoxygenase-1 ( P09601 ) and P15692 . DB00203 induced P15692 and angiopoietin specific receptors such as P35968 , Tie-1 and Tie-2 . This angiogenic response was repressed by tinprotoporphyrin IX ( SnPP ) , an inhibitor of P09601 enzyme activity . DB00203 below 1 muM has no angiogenic effect as evidenced by reduced tuborogenesis . DB00203 along with SnPP inhibited both P15692 and Q15389 ( Ang-1 ) protein expression . Therefore our results demonstrated for the first time that sildenafil is a very potent pro-angiogenic factor . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . Effect of chronic DB00203 treatment on the prostate of C57Bl/6 mice . DB00203 is a potent and selective inhibitor of phosphodiesterase-5 ( O76074 ) and is considered first-line therapy for erectile dysfunction . Nowadays , DB00203 is used extensively throughout the world on patients with pulmonary hypertension . However , few studies have evaluated the possible side effects of chronic DB00203 treatment on the male reproductive system , specifically in the prostate . In the present study , it was demonstrated via morphological and ultrastructural analysis that chronic treatment with DB00203 induced an enhancement of the glandular activity of the prostate . In addition , mice treated with DB00203 showed a significant increase in testosterone serum levels . However , no statistically significant differences were observed in nitric oxide serum levels , or in sGC , P29474 , PSA and TGF-β prostatic expression . In conclusion , the present study suggests that chronic use of DB00203 does not cause evident prostatic damage , and therefore , can be used pharmacologically to treat a variety of disorders .