interacts_with DB00244?
Intracolonic administration of zileuton , a selective P09917 inhibitor , accelerates healing in a rat model of chronic colitis . BACKGROUND : 5-Lipoxygenase products play a part in inflammatory response . AIMS : The effect of intracolonic administration of zileuton ( a P09917 inhibitor ) on colonic damage and eicosanoid local release was assessed in a rat model of colitis . METHODS : Ninety rats with trinitrobenzenesulphonic acid induced colitis were randomised to receive placebo , DB00244 ( 50 mg/kg ) , or zileuton ( 50 mg/kg ) intracolonically for four weeks . Local eicosanoid release was monitored by intracolonic dialysis throughout the study . The colon was removed for macroscopic and histological assessment at weeks 1 , 2 , and 4 after colitis induction in 10 rats of each group . RESULTS : Zileuton significantly reduced macroscopic damage score after four weeks of treatment in comparison with the other two groups ( p = 0.034 ) . In addition , zileuton administration significantly increased the intracolonic release of both thromboxane B2 at week 1 ( p = 0.05 ) and prostaglandin E2 at weeks 2 and 4 ( p < 0.05 ) . Zileuton and DB00244 decreased leukotriene B4 release by 90 % at day 3 . CONCLUSIONS : Intracolonic zileuton , compared with DB00244 and placebo , seems to improve the course of the disease in a model of chronic colitis . This effect may be related to an increased and maintained production of prostaglandin E2 together with inhibition of leukotriene B4 synthesis . 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 . DB00233 inhibits O15111 alpha phosphorylation of P25963 in mouse intestinal epithelial cells . P01375 alpha ( TNFalpha ) -stimulated nuclear factor ( NF ) kappaB activation plays a key role in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Phosphorylation of NFkappaB inhibitory protein ( IkappaB ) leading to its degradation and NFkappaB activation , is regulated by the multimeric O15111 complex , including IKKalpha and IKKbeta . We recently reported that DB00244 ( DB00244 ) inhibits TNFalpha-regulated IkappaB degradation and NFkappaB activation . To determine the mechanism of DB00244 inhibition of IkappaB degradation , we studied young adult mouse colon ( YAMC ) cells by immunodetection and in vitro kinase assays . We show DB00244 inhibits TNFalpha-stimulated phosphorylation of P25963 in intact YAMC cells . Phosphorylation of a glutathione S-transferase- P25963 fusion protein by cellular extracts or immunoprecipitated IKKalpha isolated from cells treated with TNFalpha is inhibited by DB00244 . Recombinant IKKalpha and IKKbeta autophosphorylation and their phosphorylation of glutathione S-transferase- P25963 are inhibited by DB00244 . However , IKKalpha serine phosphorylation by its upstream kinase in either intact cells or cellular extracts is not blocked by DB00244 . Surprisingly , immunodepletion of cellular extracts suggests IKKalpha is predominantly responsible for P25963 phosphorylation in intestinal epithelial cells . In summary , DB00244 inhibits TNFalpha-stimulated IKKalpha kinase activity toward P25963 in intestinal epithelial cells . These findings suggest a novel role for DB00244 in the management of Q9UKU7 by disrupting TNFalpha activation of NFkappaB . 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 . 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 . 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 . 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 . 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 . Large-scale association study for structural soundness and leg locomotion traits in the pig . BACKGROUND : Identification and culling of replacement gilts with poor skeletal conformation and feet and leg ( FL ) unsoundness is an approach used to reduce sow culling and mortality rates in breeding stock . Few candidate genes related to soundness traits have been identified in the pig . METHODS : In this study , 2066 commercial females were scored for 17 traits describing body conformation and FL structure , and were used for association analyses . Genotyping of 121 SNPs derived from 95 genes was implemented using Sequenom 's MassARRAY system . RESULTS : Based on the association results from single trait and principal components using mixed linear model analyses and false discovery rate testing , it was observed that P02649 , P34820 , P30988 , P08123 , P20849 , DKFZ , P35555 and VDBP were very highly significantly ( P < 0.001 ) associated with body conformation traits . The genes P09917 , P34820 , P30988 , O00300 , P30559 and Q9UBV4 were very highly significantly ( P < 0.001 ) associated with FL structures , and P02649 , P30988 , P08123 , P30968 , Q14623 , P42898 and Q9UBV4 were highly significantly ( P < 0.01 ) associated with overall leg action . Strong linkage disequilibrium between P30988 and P08123 on SSC9 was detected , and haplotype -ACGACC- was highly significantly ( P < 0.01 ) associated with overall leg action and several important FL soundness traits . CONCLUSION : The present findings provide a comprehensive list of candidate genes for further use in fine mapping and biological functional analyses . Leukotriene B4 amplifies NF-κB activation in mouse macrophages by reducing O15524 inhibition of MyD88 expression . Activation of NF-κB and P09917 -mediated ( P09917 -mediated ) biosynthesis of the lipid mediator leukotriene B4 ( LTB4 ) are pivotal components of host defense and inflammatory responses . However , the role of LTB4 in mediating innate immune responses elicited by specific TLR ligands and cytokines is unknown . Here we have shown that responses dependent on MyD88 ( an adaptor protein that mediates signaling through all of the known TLRs , except O15455 , as well as IL-1β and Q14116 ) are reduced in mice lacking either P09917 or the LTB4 receptor BTL1 , and that macrophages from these mice are impaired in MyD88-dependent activation of NF-κB . This macrophage defect was associated with lower basal and inducible expression of MyD88 and reflected impaired activation of P42224 and overexpression of the P42224 inhibitor O15524 . Expression of MyD88 and responsiveness to the O00206 ligand LPS were decreased by Stat1 siRNA silencing in WT macrophages and restored by Socs1 siRNA in P09917 -deficient macrophages . These results uncover a pivotal role in macrophages for the GPCR Q15722 in regulating activation of NF-κB through Stat1-dependent expression of MyD88 . 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 . 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 . Pro12Ala sequence variant of the P37231 gene is associated with postprandial hypertriglyceridemia in non-E3/E3 patients with the metabolic syndrome . BACKGROUND : Postprandial hypertriglyceridemia , a component of the metabolic syndrome , has varied etiology and involves many genes related to triglyceride metabolism . Variations in these genes may affect postprandial hypertriglyceridemia in the context of the metabolic syndrome . METHODS : We orally administered 60 g of fat overload to 74 patients with the metabolic syndrome . We then measured baseline concentrations of cholesterol , triglycerides , HDL cholesterol , apolipoprotein AI , apolipoprotein B , uric acid , and uric acid excretion ; we also performed homeostasis model assessments of insulin resistance and insulin sensitivity . At 3 h , we measured triglycerides , cholesterol , apolipoprotein AI , and apolipoprotein B . Patients were considered to have postprandial hypertriglyceridemia if the difference in plasma triglycerides between baseline and 3 h after the test was 1.71 mmol/L or more . We also measured anthropometrical variables and classified the patients according to their peroxisome proliferative activated receptor , gamma ( P37231 ) gene and apolipoprotein E ( P02649 ) genotype . RESULTS : Postprandial hypertriglyceridemia occurred in 64.7 % of patients with the Ala12 allele vs 19.9 % of the Pro12Pro patients , ( P = 0.00032 ; odds ratio , 7.6 ) , and in 87.5 % of the patients with both the Ala12 allele and the non-E3/E3 P02649 genotype ( odds ratio , 23.8 ) . Logistic regression analysis showed that P37231 and P02649 sequence variants were associated with the presence of postprandial hypertriglyceridemia . CONCLUSION : The Pro12Ala P37231 sequence variant together with a non-E3/E3 P02649 genotype is associated with a high risk for postprandial hypertriglyceridemia in patients with the metabolic syndrome , indicating a close association between these genes and the regulation of lipoproteinase clearance . 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 . 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 . 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 . Nonsteroidal anti-inflammatory drugs and inflammatory bowel disease : current perspectives . Mechanisms underlying the gastric toxicity of nonsteroidal anti-inflammatory drugs ( NSAIDs ) have been extensively investigated , whereas those leading to intestinal damage are not completely understood . Several hypotheses have been put forward on the pathophysiology of intestinal damage by NSAIDs : enhanced intestinal permeability , inhibition of cyclooxygenase ( P36551 ) , enterohepatic recirculation , and formation of adducts . The effects of P35354 selective inhibitors , which appear to have better gastric tolerability when compared to nonselective NSAIDs , on normal and inflamed intestinal mucosa ( as in Crohn 's disease or ulcerative colitis ) are still largely unexplored . If P35354 inhibition plays a key role in suppressing the inflammatory process , recent evidence suggests that P35354 products are involved in maintaining the integrity of intestinal mucosa , in the healing of gastrointestinal ulcers and in the modulation of inflammatory bowel disease ( Q9UKU7 ) . Animal models of intestinal inflammation have so far yielded conflicting results on the effects of P35354 selective inhibitors on the intestinal mucosa . It is now clear that NSAIDs do not act through cyclooxygenase inhibition , but also have different targets such as nuclear factor-kappaB and/or peroxisome proliferator-activated receptors gamma . The peculiar pharmacological profile of each compound may help to explain the different impact of each NSAID on the inflammatory process and on Q9UKU7 . Notably , the salicylic acid derivative DB00244 is widely used in the treatment of Q9UKU7 and is believed to act through nuclear factor-kappaB inhibition . Although the use of P35354 selective inhibitors remains contraindicated in patients with Q9UKU7 , studying their effects on intestinal mucosa may offer new insights into their subcellulars mechanisms of action and open new avenues for the development of novel therapies for Q9UKU7 . Genome-wide microarray-based analysis of miRNAs expression in patients with acute-on-chronic liver failure . BACKGROUND : Acute-on-chronic liver failure ( ACLF ) is a severe clinical syndrome that may cause a high mortality . However , the mechanism is still not clear . Characterization of the microRNA ( miRNA ) profiles in ACLF patients may provide new clues to the pathogenesis and management of this syndrome . METHODS : Genome-wide microarray was performed to compare the different miRNA expression profiles in peripheral blood mononuclear cells of a pair of monozygotic twins , an ACLF patient and an HBV asymptomatic carrier ( AsC ) . The case-control miRNA profiles were compared and confirmed by quantitative reverse transcription-polymerase chain reaction in 104 ACLF patients and 96 AsCs . A combined computational prediction algorithm was used to predict the potential target genes . RESULTS : Forty-five miRNAs were increased and eight miRNAs were decreased in the ACLF group . The expressions of hsa-let-7a and hsa-miR-16 were increased by 8.58- and 8.63-fold in ACLF patients compared with that in AsCs , respectively ( P < 0.001 ) . Q9Y2G2 , P10415 , Q9NZN1 , Q06643 , Q9ULW2 and Q92838 were identified as the target genes of hsa-miR-16 ; Q8IVH8 , P35372 , Q9NZI8 and Q5T4B2 were verified as the target genes of hsa-let-7a . CONCLUSIONS : Our results showed that there is a close relationship between specific miRNAs of peripheral blood mononuclear cells and ACLF. hsa-miR-16 and hsa-let-7a may contribute to the development of ACLF . Sulfasalazine inhibits lyso- Q15004 : acetyl-COA acetyltransferase . Sulfasalazine produced a dose-dependent inhibition of the enzymatic synthesis of platelet-activating factor ( Q15004 ) in lysates of rat pleural neutrophils , with an IC50 of 50 microM . Major metabolites of sulfasalazine , DB00244 and sulfapyridine , inhibited this enzymatic synthesis at much higher concentrations . Inhibition of arachidonate P09917 by sulfasalazine and its major metabolites was also observed at higher concentrations ( 2-3 mM ) . Because Q15004 is a potent mediator of inflammatory responses , an inhibition of Q15004 synthesis by sulfasalazine may contribute to its therapeutic actions in conditions such as ulcerative colitis and rheumatic illnesses . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . 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 . 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 . 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 . P09917 deficiency reduces acetaminophen-induced hepatotoxicity and lethality . 5-Lipoxygenase ( P09917 ) converts arachidonic acid into leukotrienes ( LTs ) and is involved in inflammation . At present , the participation of P09917 in acetaminophen ( DB00316 ) -induced hepatotoxicity and liver damage has not been addressed . P09917 deficient ( 5-LO⁻/⁻ ) mice and background wild type mice were challenged with DB00316 ( 0.3-6 g/kg ) or saline . The lethality , liver damage , neutrophil and macrophage recruitment , LTB₄ , cytokine production , and oxidative stress were assessed . DB00316 induced a dose-dependent mortality , and the dose of 3 g/kg was selected for next experiments . DB00316 induced LTB4 production in the liver , the primary target organ in DB00316 toxicity . Histopathological analysis revealed that 5-LO⁻/⁻ mice presented reduced DB00316 -induced liver necrosis and inflammation compared with WT mice . DB00316 -induced lethality , increase of plasma levels of aspartate aminotransferase and alanine aminotransferase , liver cytokine ( IL-1β , P01375 -α , IFN- γ , and P22301 ) , superoxide anion , and thiobarbituric acid reactive substances production , myeloperoxidase and N-acetyl-β-D-glucosaminidase activity , Nrf2 and gp91(phox) mRNA expression , and decrease of reduced glutathione and antioxidant capacity measured by 2,2'-azinobis ( 3-ethylbenzothiazoline 6-sulfonate ) assay were prevented in 5-LO⁻/⁻ mice compared to WT mice . Therefore , P09917 deficiency resulted in reduced mortality due to reduced liver inflammatory and oxidative damage , suggesting P09917 is a promising target to reduce DB00316 -induced lethality and liver inflammatory/oxidative damage . 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 . DB00945 -mediated P35354 transcript stabilization via sustained p38 activation in human intestinal myofibroblasts . Acetylsalicylic acid ( aspirin ) is a cyclooxygenase ( P36551 ) inhibitor , yet some of its therapeutic effects are thought to derive from mechanisms unrelated to prostaglandin synthesis inhibition . In human intestinal myofibroblasts , aspirin , at therapeutic doses , had the unexpected effect of inducing prolonged P35354 expression . This induction was especially pronounced when cells were treated with interleukin-1alpha ( IL-1 ) plus aspirin for 24 h . Sodium salicylate , a poor P36551 inhibitor , likewise enhanced IL-1-mediated P35354 gene expression whereas DB00244 ( DB00244 ) or indomethacin had no effect . The P35354 transcriptional rate , measured by nuclear runoff analysis and heterogeneous nuclear RNA reverse transcription-polymerase chain reaction , was only modestly elevated by aspirin treatment . In contrast , aspirin treatment dramatically stabilized the P35354 message . The P35354 mRNA half-life in IL-1 treated cells was 1 h and was increased in excess of 5 h in IL-1 + aspirin-treated cells . Phosphorylation of p38 MAPK was enhanced in aspirin-treated cells ( but not in cells treated with DB00244 or indomethacin ) for up to 24 h after treatment . Inhibition of p38 activity negated aspirin-mediated P35354 mRNA stabilization and the resultant increase in P35354 mRNA and protein levels . The modest transcriptional response seen in aspirin treated cells was also abolished by p38 inhibition . We conclude that aspirin enhances P35354 expression via sustained activation of p38 , which results in prolonged stabilization of the P35354 message and a slightly elevated transcription rate . DB00945 also enhanced steady-state mRNA levels of other IL-1 modulated genes ( IL-1beta , P05231 , groalpha , and TNFalpha ) that are likewise regulated at the level of message stability via p38 activation . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . 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 . Systems pharmacology dissection of the anti-inflammatory mechanism for the medicinal herb Folium eriobotryae . Inflammation is a hallmark of many diseases like diabetes , cancers , atherosclerosis and arthritis . Thus , lots of concerns have been raised toward developing novel anti-inflammatory agents . Many alternative herbal medicines possess excellent anti-inflammatory properties , yet their precise mechanisms of action are yet to be elucidated . Here , a novel systems pharmacology approach based on a large number of chemical , biological and pharmacological data was developed and exemplified by a probe herb Folium Eriobotryae , a widely used clinical anti-inflammatory botanic drug . The results show that 11 ingredients of this herb with favorable pharmacokinetic properties are predicted as active compounds for anti-inflammatory treatment . In addition , via systematic network analyses , their targets are identified to be 43 inflammation-associated proteins including especially P35354 , P09917 , P37231 , P01375 and Q04206 that are mainly involved in the mitogen-activated protein kinase ( MAPK ) signaling pathway , the rheumatoid arthritis pathway and NF-κB signaling pathway . All these demonstrate that the integrated systems pharmacology method provides not only an effective tool to illustrate the anti-inflammatory mechanisms of herbs , but also a new systems-based approach for drug discovery from , but not limited to , herbs , especially when combined with further experimental validations . 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 . An anti-inflammatory mechanism of taurine conjugated DB00244 against experimental colitis : taurine chloramine potentiates inhibitory effect of DB00244 on IL-1beta-mediated NFkappaB activation . Previously , we reported that oral administration of taurine conjugated DB00244 , a colon-specific prodrug of DB00244 ( DB00244 ) , is effective in ameliorating experimental colitis and taurine elicits an additive anti-inflammatory effect upon cotreatment with DB00244 . To explore a molecular mechanism for the anti-inflammatory property of the prodrug , we investigated the effect of the conjugate on IL-1beta-mediated NFkappaB activation . In human colon carcinoma Caco-2 and HCT116 cells , NFkappaB activity was accessed by a luciferase reporter assay and P05231 secretion . Protein levels were determined by Western blotting . P05231 levels were monitored by an Elisa kit . Treatment with either DB00244 or taurine chloramine ( TauCl ) inhibited IL-1beta-mediated NFkappaB dependent luciferase expression and P05231 secretion . In HCT116 cells , the inhibitory effect by TauCl or DB00244 was through preventing IL-1beta-induced O15111 activation and subsequently interfering with P25963 degradation and p65 nuclear accumulation . Furthermore , combined TauCl/ DB00244 treatment interfered additively with the activation process , leading to additive inhibitory effect on IL-1beta-mediated NFkappaB activation . Our results suggest that the anti-inflammatory effect of the prodrug on experimental colitis is attributed to the inhibition of the IL-1beta-mediated NFkappaB activation and the taurine effect is through TauCl potentiating the ability of DB00244 to inhibit IL-1beta dependent NFkappaB activation . 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 . 4- DB00233 , in contrast to DB00244 , has no effect on arachidonic acid metabolism in human neutrophils , or on the free radical 1,1-diphenyl-2-picrylhydrazyl . 5- DB00233 seems to be the active moiety of sulfasalazine in the treatment of chronic inflammatory bowel disease . Even if the precise mode of action is obscure , it is assumed that two of the main mechanisms are inhibitory effects on the lipoxygenation of arachidonic acid and interaction with free radicals . As DB00233 has been claimed to be beneficial in the topical treatment of ulcerative colitis , it was tested whether this drug possesses any influence on the P09917 activity in human neutrophils in vitro or whether it acts as a radical scavenger . The change of the amino residue from carbon-5 to carbon-4 abolished the effect in the two systems tested . The reported clinical observations on DB00233 in the treatment of chronic inflammatory bowel disease remain to be confirmed and can not be explained by interference with arachidonic acid metabolism or free oxygen radicals . 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 . Intestinal antiinflammatory effect of DB00244 is dependent on peroxisome proliferator-activated receptor-gamma . DB00244 ( DB00244 ) is an antiinflammatory drug widely used in the treatment of inflammatory bowel diseases . It is known to inhibit the production of cytokines and inflammatory mediators , but the mechanism underlying the intestinal effects of DB00244 remains unknown . Based on the common activities of peroxisome proliferator-activated receptor-gamma ( P37231 ) ligands and DB00244 , we hypothesized that this nuclear receptor mediates DB00244 therapeutic action . To test this possibility , colitis was induced in heterozygous P37231 (+/-) mice and their wild-type littermates , which were then treated with DB00244 . DB00244 treatment had a beneficial effect on colitis only in wild-type and not in heterozygous mice . In epithelial cells , DB00244 increased P37231 expression , promoted its translocation from the cytoplasm to the nucleus , and induced a modification of its conformation permitting the recruitment of coactivators and the activation of a peroxisome-proliferator response element-driven gene . Validation of these results was obtained with organ cultures of human colonic biopsies . These data identify P37231 as a target of DB00244 underlying antiinflammatory effects in the colon . 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 . 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 . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Possible mode of action of DB00244 . Despite the extensive use of sulfasalazine ( SAS ) and/or DB00244 ( DB00244 ) in patients with inflammatory bowel disease and , more recently , rheumatoid arthritis , their mode of action has not been elucidated so far . None of the numerous pharmacological and biochemical effects described , including immunosuppressive , antifolate , and modulatory actions on lymphocyte and leukocyte functions , could be defined unequivocally as mediating their beneficial activity . Recently , interest has focused on actions of SAS and DB00244 on the various enzymes of the arachidonic acid cascade . Mucosa of patients with inflammatory bowel disease generates excessive amounts of cyclooxygenase products such as prostaglandins ( PG ) as well as P09917 products such as leukotriene ( LT ) B4 and sulfidopeptide-LT . Both PG and LT exert proinflammatory actions and are potentially important mediators of mucosal inflammation . SAS and DB00244 , however , have been found to inhibit PG synthesis under certain experimental conditions only , while increasing PG formation under other conditions . While SAS was found to inhibit colonic LTB4 synthesis , DB00244 was reported to selectively affect the cyclooxygenase pathway of arachidonate metabolism in this tissue . Our results demonstrate that , like the parent compound , the metabolite DB00244 in a dose-dependent manner inhibits release of LTB4 and sulfidopeptide-LT from normal human colonic mucosa ( IC50 3.5 and 3.7 mmol/liter , respectively ) . Indomethacin , which has no efficacy in the treatment of patients with inflammatory bowel disease , on the other hand , selectively inhibited DB00917 formation in normal and inflamed colonic mucosa ( IC50 1.7 and 1.0 mmol/liter , respectively ) without reducing synthesis of LTB4 or sulfidopeptide-LT. ( ABSTRACT TRUNCATED AT 250 WORDS ) Cyanidin-3-glucoside suppresses cytokine-induced inflammatory response in human intestinal cells : comparison with DB00244 . The potential use of polyphenols in the prevention and treatment of chronic inflammatory diseases has been extensively investigated although the mechanisms involved in cellular signaling need to be further elucidated . Cyanidin-3-glucoside is a typical anthocyanin of many pigmented fruits and vegetables widespread in the human diet . In the present study , the protection afforded by cyanidin-3-glucoside against cytokine-triggered inflammatory response was evaluated in the human intestinal HT-29 cell line , in comparison with DB00244 , a well-established anti-inflammatory drug , used in inflammatory bowel disease . For this purpose , some key inflammatory mediators and inflammatory enzymes were examined . Our data showed that cyanidin-3-glucoside reduced cytokine-induced inflammation in intestinal cells , in terms of NO , DB00917 and P10145 production and of P35228 and P35354 expressions , at a much lower concentration than DB00244 , suggesting a higher anti-inflammatory efficiency . Interestingly , cyanidin-3-glucoside and DB00244 neither prevented IkB-α degradation nor the activation of NF-kB , but significantly reduced cytokine-induced levels of activated P42224 accumulated in the cell nucleus . In addition , we established that phosphorylated p38 MAPK was not involved in the protective effect of cyanidin-3-glucoside or DB00244 . Taking into account the high concentrations of dietary anthocyanins potentially reached in the gastrointestinal tract , cyanidin-3-glucoside may be envisaged as a promising nutraceutical giving complementary benefits in the context of inflammatory bowel disease . 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 . In vitro effects of E3040 , a dual inhibitor of P09917 and thromboxane A(2) synthetase , on eicosanoid production . In vitro pharmacological profiles of E3040 , 6-hydroxy-5 , 7-dimethyl-2-(methylamino)-4-(3-pyridylmethyl) benzothiazole were investigated . Against the P09917 activity of rat basophilic leukemia cells , E3040 and zileuton ( a P09917 inhibitor ) had an IC(50) of 0.23 and 0.93 microM , respectively . Against the thromboxane A(2) synthetase activity of human platelets , E3040 had an IC(50) of 0.01 microM , which was comparable to that of OKY-1581 ( sodium ( E ) -3- [ 4-(3-pyridylmethyl) phenyl ] -2-methylacrylate , a thromboxane A(2) synthetase inhibitor ) . Against cyclooxygenase activity of sheep seminal vesicles , E3040 showed no inhibition ( IC(50) , > 300 microM ) . Sulfasalazine and DB00244 , therapeutic drugs for inflammatory bowel disease , inhibited P09917 activity with an IC(50) of 293 and 970 microM , respectively . Sulfasalazine inhibited thromboxane A(2) synthetase activity with an IC(50) of 20 microM . In rat peritoneal leukocytes , E3040 inhibited leukotriene B(4) and thromboxane B(2) production with an IC(50) of 0.17 and 0.24 microM , respectively . E3040 inhibited leukotriene B(4) production in human neutrophils and thromboxane B(2) production in human platelets ( IC(50) of 0.21 and 0.09 microM , respectively ) . These results indicated that E3040 potently inhibited P09917 and thromboxane A(2) synthetase and blocked leukotriene B(4) and thromboxane B(2) production in rat peritoneal and human blood cells . Inhibition of P09917 pathway of arachidonic acid metabolism in human neutrophils by sulfasalazine and DB00244 . The possible effect of sulfasalazine , DB00244 , and acetyl- DB00244 on endogenous arachidonic acid release and metabolism was studied in human polymorphonuclear leukocytes ( PMNs ) . A new in vitro assay was used by which [1-14C]arachidonic acid is incorporated by purified peripheral PMNs until steady state was obtained ( 5 hr ) . After preincubation with the test drugs prior to activation with calcium ionophore A23187 , the released eicosanoids were isolated by extraction and thin-layer chromatography ( TLC ) and quantitated by autoradiography and laser densitometry . Median drug concentrations needed for 50 % inhibition of leukotriene B4 and 5-hydroxyeicosatetraenoic acid ( 5-HETE ) release was 4-5 mM ( range 1-9 mM ) for both sulfasalazine and DB00244 . The acetylated derivative of DB00244 was ineffective . The present data suggest that inhibition of arachidonic acid lipoxygenation may be an essential action of sulfasalazine and its active metabolite , DB00244 . Interference with lipoxygenase enzymes , rather than a steroid-like inhibition of arachidonic acid release from intracellular phospholipids , seems to be the mode of action . 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 . Fibroblast growth factors mobilize peritoneal macrophage intracellular calcium . Macrophages have been implicated in the propagation of inflammatory disease . The evidence linking macrophages to inflammation stems from their elicited responses to various extracellular ligands eventually culminating in the elaboration of a variety of inflammatory mediators . As part of an investigation of fibroblast growth factors role in promoting inflammation , we examined one aspect of transmembrane signal transduction , intracellular calcium mobilization following culture of murine peritoneal macrophages with acidic and basic fibroblast growth factor . Peritoneal macrophages displayed a rapid rise in cytosolic calcium from a basal level of 147.6 +/- 25.4 nM to 261.9 +/- 49.9 nM at 3.5 minutes following culture with acidic fibroblast growth . A similar rise in calcium was noted with basic fibroblast growth factor . Titration revealed the maximal effective dose of P05230 and P09038 with respect to calcium response to be 10 ng/ml . Using blockers of both voltage and non-voltage gated channels , the FGF induced rise in cytosolic calcium was specifically abolished . Similarly , using specific P09917 ( A69412 ) or cyclooxygenase ( Indomethacin ) blockers , the P05230 induced rise in maximal calcium response was reduced by 41 % and 96 % respectively . On the basis of these data , we speculate on some possible roles that FGF may play in the inflammatory response . Arachidonic acid metabolism in Q9HBL0 -induced chronic and immunologic enteritis in rats , and the effect of DB00244 . Inflammation of the rat distal intestine was induced by intradermal sensitization and subsequent multiple intrajejunal challenge with the hapten 2,4,6-trinitrobenzenesulphonic acid ( TNBS ) via an implanted catheter . The time course of the inflammatory reaction was followed by determination of the enteritis score and measurement of in vitro eicosanoid formation of homogenates of the gut after 0 , 1 , 2 , 4 , 7 , 14 and 21 days of local daily challenge with 0.08 % TNBS . There was a small initial increase of eicosanoid formation , reached at days 1 and 2 , followed by a significant increase in metabolism of arachidonic acid on day 21 . Although at day 1 a four-fold increase in inflammation score was reached , no further significant changes were observed during the following 3 weeks . The greatest increase in metabolite formation was observed in prostanoids TxB(2) , PGE(2). and P52209 (2) and the P09917 product LTC(4) , whereas minor changes were found for Q06643 (4) and other lipoxygenase products such as 12- and 15-HETE . The formation of these metabolites was already inhibited by low-dose DB00244 ( DB00244 ) , given orally twice daily during the 3 weeks challenge period , while the enteritis score was affected dosedependently . Nonsteroidal anti-inflammatory medications are cytostatic against human vestibular schwannomas . Vestibular schwannomas ( VSs ) are the most common tumors of the cerebellopontine angle . Significant clinical need exists for pharmacotherapies against VSs . Motivated by previous findings that immunohistochemical expression of cyclooxygenase 2 ( P35354 ) correlates with VS growth rate , we investigated the role of P35354 in VSs and tested P35354 inhibiting salicylates against VSs . P35354 was found to be aberrantly expressed in human VS and primary human VS cells in comparison with control human nerve specimens and primary Schwann cells ( SCs ) , respectively . Furthermore , levels of prostaglandin E2 , the downstream enzymatic product of P35354 , were correlated with primary VS culture proliferation rate . Because P35354 inhibiting salicylates such as aspirin are well tolerated and frequently clinically used , we assessed their repurposing for VS . Changes in proliferation , cell death , and cell viability were analyzed in primary VS cultures treated with aspirin , sodium salicylate , or DB00244 . These drugs neither increased VS cell death nor affected healthy SCs . The cytostatic effect of aspirin in vitro was in concurrence with our previous clinical finding that patients with VS taking aspirin demonstrate reduced tumor growth . Overall , this work suggests that P35354 is a key modulator in VS cell proliferation and survival and highlights salicylates as promising pharmacotherapies against VS . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . P00747 activator inhibitor-1 supports P10145 -mediated neutrophil transendothelial migration by inhibition of the constitutive shedding of endothelial P10145 /heparan sulfate/syndecan-1 complexes . The endothelium is the primary barrier to leukocyte recruitment at sites of inflammation . Neutrophil recruitment is directed by transendothelial gradients of P10145 that , in vivo , are bound to the endothelial cell surface . We have investigated the identity and function of the binding site(s) in an in vitro model of neutrophil transendothelial migration . In endothelial culture supernatants , P10145 was detected in a trimolecular complex with heparan sulfate and syndecan-1 . Constitutive shedding of P10145 in this form was increased in the presence of a neutralizing Ab to plasminogen activator inhibitor-1 ( P05121 ) , indicating a role for endothelial plasminogen activator in the shedding of P10145 . Increased shedding of P10145 /heparan sulfate/syndecan-1 complexes was accompanied by inhibition of neutrophil transendothelial migration , and aprotinin , a potent plasmin inhibitor , reversed this inhibition . Platelets , added as an exogenous source of P05121 , had no effect on shedding of the complexes or neutrophil migration . Our results indicate that P10145 is immobilized on the endothelial cell surface through binding to syndecan-1 ectodomains , and that plasmin , generated by endothelial plasminogen activator , induces the shedding of this form of P10145 . P05121 appears to stabilize the chemoattractant form of P10145 at the cell surface and may represent a therapeutic target for novel anti-inflammatory strategies . 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 . 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 . Predictive value of circulating interleukin-6 and heart-type fatty acid binding protein for three months clinical outcome in acute cerebral infarction : multiple blood markers profiling study . INTRODUCTION : There is no single blood marker for predicting the prognosis in ischemic stroke . A combination of multiple blood markers may enhance the ability to predict long-term outcome following ischemic stroke . METHODS : Blood concentrations of neuronal markers ( neuron-specific enolase , visinin-like protein 1 , heart type fatty acid binding protein ( hFABP ) and neuroglobin ) , astroglial markers ( P04271 and glial fibrillary acidic protein ) , inflammatory markers ( P05231 , P01375 -α , and P02741 ) , blood-brain barrier marker ( matrix metalloproteinase 9 ) , and haemostatic markers ( D-dimer and P05121 ) were measured within 24 hours after stroke onset . The discrimination and reclassification for favorable and poor outcome were compared after adding individual or a combination of blood markers to the clinical model of stroke outcome . RESULTS : In multivariate analysis , natural log-transformed ( log ) P05231 ( odds ratio ( OR ) : 1.75 , 95 % CI : 1.25 to 2.25 , P=0.001 ) and loghFABP ( OR : 3.23 , 95 % CI : 1.44 to 7.27 , P=0.005 ) were independently associated with poor outcome . The addition of a single blood marker to the clinical model did not improve the discriminating ability of the clinical model of stroke outcome . However , the addition of the combination of logIL-6 and loghFABP to the clinical model showed improved discrimination ( area under receiver operating characteristic ( AUROC ) curve : 0.939 versus 0.910 , P=0.03 ) and reclassification performance ( net reclassification improvement index : 0.18 , P=0.005 ) . CONCLUSIONS : A combination of circulating P05231 and hFABP level has an additive clinical value for the prediction of stroke outcome . Effects of sulfasalazine and a sulfasalazine analogue on the formation of lipoxygenase and cyclooxygenase products . A sulfasalazine analogue , 5'-(2,4-dichlorobenzoyl)2'-hydroxyphenylacetic acid ( CL 42A ) , potently inhibited the formation of P09917 products ( leukotrienes B4 and C4 and 5-hydroxyeicosatetraenoic acid ) by human leukocytes . Half-maximal inhibition of leukotriene production was obtained with 5 and 10 microM CL 42A after stimulation with serum-treated zymosan or ionophore A23187 , respectively . CL 42A was equipotent to nordihydroguaiaretic acid and about 50 times more potent than sulfasalazine and benoxaprofen in studies on the inhibition of LTB4 formation in leukocyte suspensions stimulated with serum-treated zymosan . Furthermore , CL 42A had no inhibitory effect on the production of 15-hydroxyeicosatetraenoic acid after incubation of human leukocytes with ionophore A23187 in the presence of exogenous arachidonic acid . Sulfasalazine inhibited the synthesis of P09917 products ( 5-hydroxyeicosatetraenoic acid and leukotriene B4 : IC50 250 microM , leukotriene C4 : IC50 100 microM ) in a concentration-dependent manner but had no effect on 15-hydroxyeicosatetraenoic acid formation . The metabolites of sulfasalazine , sulfapyridine and DB00244 , and the isomer , DB00233 , were all less potent than sulfasalazine as inhibitors of leukotriene formation . Both CL 42A ( IC50 20 microM ) and sulfasalazine ( IC50 500 microM ) inhibited the synthesis of thromboxane B2 and hydroxyheptadecatrienoic acid in human platelet suspensions after arachidonic acid stimulation . However , while CL 42A inhibited cyclooxygenase , the inhibitory effect of sulfasalazine was exerted mainly on thromboxane synthase . The platelet formation of 12-hydroxyeicosatetraenoic acid was not inhibited by CL 42A whereas sulfasalazine had a weak inhibitory effect . P35354 -dependent and -independent inhibition of proliferation of colon cancer cells by DB00244 . The cyclooxygenase-2 ( P35354 ) /prostaglandin E2 ( DB00917 ) pathway may have a pathogenic role in colorectal cancer ( CRC ) . Recent studies suggest that DB00244 ( DB00244 ) reduces the risk of inflammatory bowel disease-related CRC , but the mechanism by which DB00244 interferes with CRC cell growth remains unknown . In this study , we have examined whether the negative effect of DB00244 on CRC cells is dependent on P35354 / DB00917 axis inhibition . We show that DB00244 down-regulates both constitutive and P01375 or IL-1beta-induced P35354 in HT-115 and HT-29 cells . Inhibition of P35354 by DB00244 occurs at the RNA and protein level , and is associated with a significant decrease in DB00917 synthesis , arrest of growth and enhanced death of CRC cells . However , exogenous DB00917 does not revert the DB00244 -mediated CRC cell proliferation block . DB00244 also inhibits the growth of DLD-1 , a P36551 -deficient CRC cell line , thus suggesting that the anti-proliferative effect of DB00244 on CRC cells is not strictly dependent on the inhibition of P35354 / DB00917 . Taken together our data indicate that DB00244 causes both a P35354 -dependent and -independent inhibition of CRC cell growth . 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 . Review article : new drug formulations , chemical entities and therapeutic approaches for the management of ulcerative colitis . BACKGROUND : Treatment options for ulcerative colitis ( UC ) are expanding with the development of novel drug formulations and dosing regimens and new chemical entities . Although the goals of medical therapy for UC remain unchanged , that is to induce and to maintain remission , focus has also centred on improving patient compliance , modifying the natural course of disease and healing the mucosa . AIM : To examine novel formulations , new chemical entities and novel therapeutic approaches to the management of UC . METHODS : Searches for all studies related to UC treatment in Medline and abstracts from major national and international meetings published in the last 10 years . RESULTS : 5-Aminosalicylic acids ( DB00244 ) remain the standard first-line treatment for patients with mild to moderately active UC . New formulations with altered delivery , and new dosing regimens have demonstrated possible improvements in efficacy compared with historically available preparations and dosing patterns . Once-daily dosing , micropellet formulations,and high-dose tablets offer enhanced efficacy and improved compliance . DB00244 is now recognized as a ligand for peroxisome proliferator activated receptor-gamma ( P37231 ) and it has a role as a chemo-preventive agent in long-standing UC . New colonic release corticosteroid formulations help to limit systemic toxicity ; turmeric , tacrolimus and infliximab have shown promising results . New anti-inflammatory targeted therapies include an anti-CD3 antibody , selective integrin blockers , anti- P60568 antibody and P37231 agonists . CONCLUSION : The evolution of novel oral DB00244 formulations and dosage regimens,and recent development of new molecules have expanded the therapeutic armamentarium of UC . 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 . An open-label trial of the P37231 ligand rosiglitazone for active ulcerative colitis . OBJECTIVES : Previous research has demonstrated that ligands for the gamma subtype of peroxisome proliferator-activated receptors ( PPARs ) reduce inflammation in two different murine models of colitis . This study was designed to examine the potential efficacy of rosiglitazone , a ligand for the gamma subtype of PPARs , as a therapy for active ulcerative colitis . METHODS : Fifteen patients with mild to moderately active ulcerative colitis despite therapy with DB00244 compounds were enrolled in an open-label study of rosiglitazone ( 4 mg b.i.d. p.o. ) for 12 wk . Thirteen of 15 patients were receiving concomitant therapy with corticosteroids and/or immunomodulator medications . Disease activity was measured with the Disease Activity Index . RESULTS : After 12 wk of therapy , four patients ( 27 % ) had achieved clinical remission , of whom three ( 20 % ) also had an endoscopic remission . Four additional patients ( 27 % ) had a clinical response without achieving remission . Two patients were hospitalized with worsened disease activity , and one patient was withdrawn for nephrotic syndrome . CONCLUSIONS : These data suggest that ligands for the gamma subtype of PPARs may represent a novel therapy for ulcerative colitis . A double blind , placebo-controlled , randomized trial is warranted . Effects of ropivacaine on eicosanoid release from human granulocytes and endothelial cells in vitro . OBJECTIVE : To examine the effects of ropivacaine , currently being investigated for treatment of ulcerative colitis , on the release of arachidonic acid metabolites . MATERIAL : Human granulocytes and endothelial cells . TREATMENT : Ropivacaine , lidocaine , hydrocortisone , DB00244 or acetylsalicylic acid ( 10-1000 microM ) . METHODS : Leukotriene B4 , 5-hydroxyeicosatetraenoic acid , 6-keto PGF1 alpha and 15-hydroxyeicosatetraenoic acid were measured using immuno assays . Wilcoxon signed rank test was used for statistical calculations . RESULTS : Ropivacaine dose-dependently inhibited zymosan-induced release of leukotriene B4 and 5-hydroxyeicosatetraenoic acid whereas the release after ionophore stimulation was not affected . Ropivacaine was more potent than DB00244 but less potent compared to hydrocortisone . Ropivacaine had only a weak inhibitory effect on the release of 15-hydroxyeicosatetraenoic acid from zymosan- or ionophore-stimulated cells . In contrast to hydrocortisone and DB00244 , ropivacaine only weakly affected the release of 6-keto PGF1 alpha after stimulation with thrombin . CONCLUSIONS : The inhibited release of P09917 products may account for some of the anti-inflammatory effects of ropivacaine seen in the treatment of ulcerative colitis . DB00244 mediates expression of cyclooxygenase-2 and 15-hydroxyprostaglandin dehydrogenase to suppress colorectal tumorigenesis . BACKGROUND : P35354 ( P35354 ) is a key enzyme that produces prostaglandin E2 ( DB00917 ) and plays an important role in colorectal tumor growth . In addition , recent researches focused on 15-hydroxyprostaglandin dehydrogenase ( P15428 ) , which degrades DB00917 . Here we determined the effect of DB00244 ( DB00244 ) on P35354 and P15428 expression and investigated its preventive effect for colorectal cancer ( CRC ) . MATERIALS AND METHODS : HT-29 cells were used in the in vitro experiments . c-Ha-ras transgenic mice were employed in order to explore the chemopreventive effects . Western blotting analysis was performed and the protein expression of P35354 and P15428 was quantified . RESULTS : DB00244 significantly suppressed P35354 expression and induced P15428 expression in HT-29 cells . In the transgenic mice , oral DB00244 intake reduced the incidence of colorectal tumor formation and the tumor size . Furthermore , we observed a down-regulation of P35354 and an up-regulation of P15428 in the tissue from colons of these mice . CONCLUSION : DB00244 exerts a preventive effect against colorectal tumor development through mediation of P35354 and P15428 expression . Selective inhibitors of Q02750 /ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells . The nonsteroidal anti-inflammatory drug ( NSAID ) sulindac prevents experimental colon cancer and can regress precancerous polyps in humans . Sulindac sulfide inhibits cyclooxygenase ( P36551 ) -mediated prostaglandin synthesis and retards the growth of cultured colon cell lines primarily by inducing apoptosis . Given the known role of mitogen-activated protein kinase ( MAPK ) in signal transduction and the regulation of cell survival and death , we determined the effect of sulindac sulfide on MAPK activation , P35354 expression , and apoptosis induction in HCA-7 human colon cancer cells . Sulindac sulfide treatment was associated with activation of ERKp44/42 and p38 MAPK in a dosage- and time-dependent manner , and also activated upstream MEK . Similar results were seen in HCT-15 cells and also with the selective P35354 inhibitor NS398 . ERKp44/42 and p38 activation were accompanied by an induction of P35354 protein expression . Selective inhibitors of sulindac sulfide-induced ERKp44/42 ( PD98059 ) and p38 MAPK ( SB203580 ) activation also suppressed the induction of P35354 by this NSAID . Furthermore , both MAPK inhibitors significantly augmented sulindac sulfide-induced apoptosis , as did suppression of constitutive P35354 using antisense oligonucleotides . In conclusion , MEK/ P29323 and p38 MAPK activation mediate P35354 induction by sulindac sulfide . Selective inhibitors of these MAPKs potentiate apoptosis induction by this NSAID , suggesting a novel strategy for the prevention or treatment of colorectal cancer . Activation of serotonin receptor-2B rescues small-for-size liver graft failure in mice . The implantation of grafts below 30 % of the normal liver volume is associated with a high risk of failure known as small-for-size ( SFS ) syndrome . Strategies to rescue small grafts may have a dramatic impact on organ shortage . Serotonin is a potent growth factor for the liver . The goal of this study was to determine whether enhanced serotonin signaling could prevent the deleterious effects of SFS syndrome . We performed 30 % normal liver volume transplantations in wild-type C57/BL6 and interleukin-6 ( P05231 ) (-/-) mice . Some animals received α-methyl-5-HT ( DOI ) , an agonist of serotonin receptor-2 ( P41595 ) . Endpoints included long-term survival , serum and hepatic markers of liver injury and regeneration , assessment of hepatic microcirculation by intravital fluorescence microscopy and scanning electron microscopy , and transcript levels of a variety of serotonin receptors , tumor necrosis factor α , and P05231 . All recipients of small grafts ( controls ) died within 2-4 days of transplantation , whereas half of those receiving DOI survived permanently . Control animals disclosed major liver injury , including diffuse microvesicular steatosis in hepatocytes , impairment of microcirculation , and a failure of regeneration , whereas these parameters were dramatically improved in animals subjected to DOI . Blockage of P41595 blunted the protective effects of DOI . Whereas P05231 levels were higher in DOI-treated animals , P05231 (-/-) mice were still protected by DOI , suggesting a protective pathway independent of P05231 . CONCLUSION : Serotonin through its action on receptor-2B protects SFS liver grafts from injury and prevents microcirculation and regeneration . The mechanism of hepato-protection is independent of P05231 .