MH_dev_164

Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation ( s ) of the low-density lipoprotein receptor ( LDL-R ) , i . e . autosomal dominant hypercholesterolemia type 1 ( P07327 REA ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 REA ) , or gain-of-function mutations of proprotein convertase subtilisin / kexin type 9 ( Q8NBP7 ) ( P00326 REA ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly / secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 MEN , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction / regression , but animal models provide encouraging results .

P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2 + or Ca2 + to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2 + or Ca2 + . TnC and P62158 in the presence of either Ca2 + or Mg2 + bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2 + but , in the presence of Mg2 + , did not bind to TnC and only bound weakly to P62158 . DB00623 MENMAX DB00623 MEN bound to TnC and P62158 only in the presence of Ca2 + . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins . ( ABSTRACT TRUNCATED AT 250 WORDS )

Association analysis between the A118G polymorphism in the P35372 REA gene and treatment response to venlafaxine XR in generalized anxiety disorder . Patients diagnosed with generalized anxiety disorder ( Q99259 REA ) exhibit differential responses to standard antidepressant pharmacotherapy . Mounting evidence demonstrates that genetic differences may be implicated in treatment response in disorders like Q99259 REA . In this study , we examined whether the P35372 REA gene , which has been implicated in antidepressant treatment response in major depressive disorder , also has an effect in Q99259 REA . In our study , 156 patients diagnosed with Q99259 REA received venlafaxine XR treatment as part of an 18 - month relapse prevention study . Genotypes were obtained for the P35372 REA functional variant A118G for the entire sample ( n = 151 ) ; however , only the European American population was considered ( n = 108 ) for pharmacogenetic analysis . We found no significant association between A118G and antidepressant treatment response in our Q99259 REA population . Future studies that include different single nucleotide polymorphisms of the P35372 REA gene as well as larger populations will need to be conducted to further elucidate the pharmacogenetic role of the endogenous opioid system in anxiety disorders .

Phosphodiesterase - 4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 REA ) in P29320 REA - 293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 REA ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta 2AR ( beta 2 - adrenergic receptor ) . In the present paper , we show that challenge of P29320 REA - 293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta 2AR - GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 REA to become phosphorylated by PKA ( DB02527 - dependent protein kinase ) . This action is facilitated when DB02527 - specific DB05876 ( phosphodiesterase - 4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) - mediated knockdown of Q07343 REA and Q08499 REA . DB05876 - selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 REA , phosphorylation of the beta 2AR by P25098 REA , membrane translocation of beta-arrestin and internalization of beta 2ARs . DB05876 - selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 REA in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 REA - 293beta2 cells acts to stimulate PKA phosphorylation of P25098 REA , with consequential effects on P25098 REA membrane recruitment and P25098 REA - mediated phosphorylation of the beta 2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 REA , influencing the rate of P25098 REA phosphorylation of the beta 2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 REA from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 .

Association of polymorphisms in nicotinic acetylcholine receptor alpha 4 subunit gene ( P43681 REA ) , mu-opioid receptor gene ( P35372 REA ) , and ethanol-metabolizing enzyme genes with alcoholism in Korean patients . Findings obtained from several studies indicate that ethanol enhances the activity of alpha 4beta2 neuronal nicotinic acetylcholine receptor and support the possibility that a polymorphism of the nicotinic acetylcholine receptor alpha 4 subunit gene ( P43681 REA ) modulates enhancement of nicotinic receptor function by ethanol . To identify the association between the CfoI polymorphism of the P43681 REA and alcoholism , we examined distribution of genotypes and allele frequencies in Korean patients diagnosed with alcoholism ( n = 127 ) and Korean control subjects without alcoholism ( n = 185 ) with polymerase chain reaction-restriction fragment length polymorphism methods . We were able to detect the association between the CfoI polymorphism of the P43681 REA and alcoholism in Korean patients ( genotype P = . 023 ; allele frequency P = . 047 ) . The genotypes and allele frequencies of known polymorphisms in other alcoholism candidate genes , such as alcohol metabolism-related genes [ alcohol dehydrogenase 2 ( P00325 REA ) , aldehyde dehydrogenase 2 ( P05091 REA ) , alcohol dehydrogenase 3 ( P00326 REA ) , and cytochrome P450 2E1 ( P05181 REA ) ] and mu-opioid receptor gene ( P35372 REA ) , were studied . The polymorphisms of P00325 REA , P05091 REA , and P05181 REA were significantly different in Korean patients with alcoholism and Korean control subjects without alcoholism , but P00326 REA and P35372 REA did not differ between the two groups .

Modulating micro-opioid receptor phosphorylation switches agonist-dependent signaling as reflected in PKCepsilon activation and dendritic spine stability . A new role of G protein-coupled receptor ( GPCR ) phosphorylation was demonstrated in the current studies by using the μ-opioid receptor ( P35372 REA ) as a model . Morphine induces a low level of receptor phosphorylation and uses the PKCε pathway to induce P29323 REA phosphorylation and receptor desensitization , whereas etorphine , fentanyl , and [ D-Ala 2 , N-Me-Phe 4 , Gly 5 - ol ] - enkephalin ( DAMGO ) induce extensive receptor phosphorylation and use the β-arrestin 2 pathway . Blocking P35372 REA phosphorylation ( by mutating Ser 363 , Thr 370 and Ser 375 to Ala ) enabled etorphine , fentanyl , and DAMGO to use the PKCε pathway . This was not due to the decreased recruitment of β-arrestin 2 to the receptor signaling complex , because these agonists were unable to use the PKCε pathway when β-arrestin 2 was absent . In addition , overexpressing G protein-coupled receptor kinase 2 ( P25098 REA ) decreased the ability of morphine to activate PKCε , whereas overexpressing dominant-negative P25098 REA enabled etorphine , fentanyl , and DAMGO to activate PKCε . Furthermore , by overexpressing wild-type P35372 REA and a phosphorylation-deficient mutant in primary cultures of hippocampal neurons , we demonstrated that receptor phosphorylation contributes to the differential effects of agonists on dendritic spine stability . Phosphorylation blockage made etorphine , fentanyl , and DAMGO function as morphine in the primary cultures . Therefore , agonist-dependent phosphorylation of GPCR regulates the activation of the PKC pathway and the subsequent responses .

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

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

Effects of the total saponins from Rosa laevigata Michx fruit against acetaminophen-induced liver damage in mice via induction of autophagy and suppression of inflammation and apoptosis . The effect of the total saponins from Rosa laevigata Michx fruit ( RLTS ) against acetaminophen ( DB00316 MEN ) - induced liver damage in mice was evaluated in the present paper . The results showed that RLTS markedly improved the levels of liver SOD , CAT , DB00143 , DB00143 - Px , MDA , NO and P35228 REA , and the activities of serum ALT and Q9NRA2 caused by DB00316 MEN . Further research confirmed that RLTS prevented fragmentation of DNA and mitochondrial ultrastructural alterations based on TdT-mediated dUTP nick end labeling ( TUNEL ) and transmission electron microscopy ( TEM ) assays . In addition , RLTS decreased the gene or protein expressions of cytochrome P450 ( P05181 REA ) , pro-inflammatory mediators ( IL - 1β , P05112 REA , P05231 REA , P01375 REA - α , P35228 REA , Bax , HMGB - 1 and P35354 REA ) , pro-inflammatory transcription factors ( NF-κB and AP - 1 ) , pro-apoptotic proteins ( cytochrome C , p53 , caspase - 3 , caspase - 9 , p-JNK , p-p 38 and p - P29323 REA ) , and increased the protein expressions of Bcl - 2 and Bcl-xL . Moreover , the gene expression of P22301 REA , and the proteins including LC3 , Q14457 REA and Atg 5 induced by DB00316 MEN were even more augmented by the extract . These results demonstrate that RLTS has hepatoprotective effects through antioxidative action , induction of autophagy , and suppression of inflammation and apoptosis , and could be developed as a potential candidate to treat DB00316 MEN - induced liver damage in the future .

P35372 REA phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala 2 , MePhe 4 , Glyol 5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin ( 1-31 ) , enkephalins , and dynorphin A ( 1-17 ) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 SUB and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor / agonist / G-protein complexes and / or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies .

Regulation of P14061 REA and P18405 REA in lymphocytes . We previously reported lymphocyte expression of genes encoding enzymes required for steroid metabolism ; however , only 17beta - HSD and 5alpha - reductase showed significant enzyme activity . We now investigate regulation of lymphocyte expression for genes encoding 17beta - HSD and 5alpha - reductase . Cultured human T and B lymphoid cell lines and peripheral blood mononuclear cells were treated with known regulators of steroidogenic gene expression including forskolin , PMA , ionomycin , various steroids , interleukin ( IL ) - 4 , and P05231 REA . Treatment with 10 or 50 microM forskolin resulted in a 20-60 % reduction of expression for P14061 REA ( encoding 17beta - HSD I ) in T and B lymphoid cell lines and peripheral blood mononuclear cells , although such a change was not observed in the expression of P18405 REA ( encoding 5alpha - reductase I ) . No significant changes were found when cells were treated for 24 h with various concentrations of PMA or ionomycin . Incubation with 10 ( - 9 ) to 10 ( - 7 ) M androstenedione or estradiol increased expression of P14061 REA , while testosterone decreased the expression of this gene . P18405 REA expression was increased in the presence of 5alpha - DB02901 MEN although no consistent changes were observed when the cells were treated with testosterone . Other steroids , including dexamethasone , progesterone , and 6 - hydroxypregnanolone , produced no effects on expression of either P14061 REA or P18405 REA . Treatment with 0.1- 10 ng / ml of P05112 REA or P05231 REA also did not effect significant changes in gene expression . These data implicate the involvement of the DB02527 - protein kinase signal transduction pathway in regulating lymphocyte expression of P14061 REA . Furthermore , it appears that lymphocyte P14061 REA and P18405 REA are regulated to some extent by specific steroids .

Synergistic proapoptotic effects of the two tyrosine kinase inhibitors pazopanib and lapatinib on multiple carcinoma cell lines . DB06589 MEN and lapatinib are two tyrosine kinase inhibitors that have been designed to inhibit the P15692 REA tyrosine kinase receptors 1 , 2 and 3 ( pazopanib ) , and the P00533 REA and P04626 REA receptors in a dual manner ( lapatinib ) . DB06589 MEN has also been reported to mediate inhibitory effect on a selected panel of additional tyrosine kinases such as P09619 REA and c-kit . Here , we report that pazopanib and lapatinib act synergistically to induce apoptosis of A549 non-small-cell lung cancer cells . Systematic assessment of the kinome revealed that both pazopanib and lapatinib inhibited dozens of different tyrosine kinases and that their combination could suppress the activity of some tyrosine kinases ( such as c - DB00134 ) that were not or only partially affected by either of the two agents alone . We also found that pazopanib and lapatinib induced selective changes in the transcriptome of A549 cells , some of which were specific for the combination of both agents . Analysis of a panel of unrelated human carcinoma cell lines revealed a signature of 52 genes whose up - or downregulation reflected the combined action of pazopanib and lapatinib . Indeed , pazopanib and lapatinib exerted synergistic cytotoxic effects on several distinct non-small-cell lung cancer cells as well as on unrelated carcinomas . Altogether , these results support the contention that combinations of tyrosine kinase inhibitors should be evaluated for synergistic antitumor effects . Such combinations may lead to a ' collapse ' of pro-survival signal transduction pathways that leads to apoptotic cell death .

Molecular and biochemical analysis of calmodulin interactions with the calmodulin-binding domain of plant glutamate decarboxylase . We previously provided what to our knowledge is the first evidence that plant glutamate decarboxylase ( Q99259 REA ) is a calmodulin ( P62158 ) - binding protein . Here , we studied the Q99259 REA P62158 - binding domain in detail . A synthetic peptide of 26 amino acids corresponding to this domain forms a stable complex with Ca2 + / P62158 with a 1:1 stoichiometry , and amino acid substitutions suggest that tryptophan - 485 has an indispensable role in P62158 binding . Chemical cross-linking revealed specific P62158 / Q99259 REA interactions even in the absence of Ca2 + . However , increasing KCI concentrations or deletion of two carboxy-terminal lysines abolished these interactions but had a mild effect on P62158 / Q99259 REA interactions in the presence of Ca2 + . We conclude that in the presence of Ca ( 2 + ) - hydrophobic interactions involving tryptophan - 485 and electrostatic interactions involving the carboxy-terminal lysines mediate P62158 / Q99259 REA complex formation . By contrast , in the absence of Ca2 + , P62158 / Q99259 REA interactions are essentially electrostatic and involve the carboxy-terminal lysines . In addition , a tryptophan residue and carboxy-terminal lysines are present in the P62158 - binding domain of an Arabidopsis Q99259 REA . Finally , we demonstrate that petunia Q99259 REA activity is stimulated in vitro by Ca2 + / P62158 . Our study provides a molecular basis for Ca ( 2 + ) - dependent P62158 / Q99259 REA interactions and suggests the possible occurrence of Ca ( 2 + ) - independent P62158 / Q99259 REA interactions .

P35372 REA and P00533 REA contribute to skin pigmentation differences between Indigenous Americans and Europeans . Contemporary variation in skin pigmentation is the result of hundreds of thousands years of human evolution in new and changing environments . Previous studies have identified several genes involved in skin pigmentation differences among African , Asian , and European populations . However , none have examined skin pigmentation variation among Indigenous American populations , creating a critical gap in our understanding of skin pigmentation variation . This study investigates signatures of selection at 76 pigmentation candidate genes that may contribute to skin pigmentation differences between Indigenous Americans and Europeans . Analysis was performed on two samples of Indigenous Americans genotyped on genome-wide SNP arrays . Using four tests for natural selection - - locus-specific branch length ( LSBL ) , ratio of heterozygosities ( lnRH ) , Tajima ' s D difference , and extended haplotype homozygosity ( EHH ) - - we identified 14 selection-nominated candidate genes ( SNCGs ) . SNPs in each of the SNCGs were tested for association with skin pigmentation in 515 admixed Indigenous American and European individuals from regions of the Americas with high ground-level ultraviolet radiation . In addition to Q71RS6 and Q9UMX9 , genes previously associated with European / non-European differences in skin pigmentation , P35372 REA and P00533 REA were associated with variation in skin pigmentation in New World populations for the first time .

Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 REA by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 REA ) plays a key role in regulating inflammation . DB01656 MEN , a phosphodiesterase ( PDE ) 4 - selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 REA ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 REA up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 REA is up-regulated in the context of the complex pathogenesis and medications of P48444 REA may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 REA exacerbation , to up-regulate PDE 4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE 4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE 4B2 . PKA-Cβ phosphorylates p65 in a DB02527 - dependent manner . Moreover , Ser 276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE 4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE 4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor .

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

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

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