interacts_with DB00887?
DB00207 fails to reduce inflammation in cystic fibrosis airway epithelial cells . Cystic fibrosis is a hereditary disease caused by a mutation in the Cystic Fibrosis Transmembrane conductance Regulator ( P13569 ) gene that encodes a chloride ( Cl(-) ) channel . Cystic fibrosis pulmonary pathophysiology is characterised by chronic inflammation and bacterial infections . DB00207 , a macrolide antibiotic , has shown promising anti-inflammatory properties in some inflammatory pulmonary diseases . Moreover , all clinical studies have presented an improvement of the respiratory condition of cystic fibrosis patients , but the molecular and cellular mechanisms remain unknown . The aim of this study was to investigate , in bronchial epithelial cells , the effects of azithromycin on inflammatory pathways involved in cystic fibrosis . We have analysed the effects of azithromycin on cystic fibrosis and non-cystic fibrosis bronchial epithelial cell lines but also in non-immortalized non-cystic fibrosis human glandular cells . To create an inflammatory context , cells were treated with Tumor Necrosis Factor ( P01375 ) -α or Interleukin (IL)1-β . Activation of the NF-κB pathway was investigated by luciferase assay , western blotting , and by Förster Resonance Energy Transfer imaging , allowing the detection of the interaction between the transcription factor and its inhibitor in live cells . In all conditions tested , azithromycin did not have an anti-inflammatory effect on the cystic fibrosis human bronchial epithelial cells and on P13569 -inhibited primary human bronchial glandular cells . More , our data showed no effect of azithromycin on IL-1β- or P01375 -α-induced P10145 secretion and NF-κB pathway activation . Taken together , these data show that azithromycin is unable to decrease in vitro inflammation in cystic fibrosis cells from airways . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . DCEBIO stimulates Cl- secretion in the mouse jejunum . We investigated the effects of 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one(DCEBIO) on the Cl- secretory response of the mouse jejunum using the Ussing short-circuit current ( Isc ) technique . DCEBIO stimulated a concentration-dependent , sustained increase in Isc ( EC50 41 +/- 1 microM ) . Pretreating tissues with 0.25 microM forskolin reduced the concentration-dependent increase in Isc by DCEBIO and increased the EC50 ( 53 +/- 5 microM ) . DB00887 blocked ( 82 +/- 5 % ) the DCEBIO-stimulated Isc consistent with Cl- secretion . DCEBIO was a more potent stimulator of Cl- secretion than its parent molecule , 1-ethyl-2-benzimidazolinone . DB01016 or P16860 reduced the DCEBIO-stimulated Isc by > 80 % indicating the participation of P13569 in the DCEBIO-stimulated Isc response . Clotrimazole reduced DCEBIO-stimulated Isc by 67 +/- 15 % , suggesting the participation of the intermediate conductance Ca2+-activated K+ channel ( IKCa ) in the DCEBIO-activated Isc response . In the presence of maximum forskolin ( 10 microM ) , the DCEBIO response was reduced and biphasic , reaching a peak response of the change in Isc of 43 +/- 5 microA/cm2 and then falling to a steady-state response of 17 +/- 10 microA/cm2 compared with DCEBIO control tissues ( 61 +/- 6 microA/cm2 ) . The forskolin-stimulated Isc in the presence of DCEBIO was reduced compared with forskolin control tissues . Similar results were observed with DCEBIO and 8-BrcAMP where adenylate cyclase was bypassed . H89 , a PKA inhibitor , reduced the DCEBIO-activated Isc , providing evidence that DCEBIO increased Cl- secretion via a DB02527 /PKA-dependent manner . These data suggest that DCEBIO stimulates Cl- secretion of the mouse jejunum and that DCEBIO targets components of the Cl- secretory mechanism . Refined localization and yeast artificial chromosome ( YAC ) contig -- mapping of genes and DNA segments in the 7q21-q32 region . The chromosome localizations for 159 gene and DNA segments have been refined to one of five intervals in the 7q21-132 region through hybridization analysis with a panel of somatic cell hybrid lines . Seventy-two of these chromosome 7 markers are also mapped on common or overlapping yeast artificial chromosome ( YAC ) clones . In addition , the breakpoints of chromosome rearrangement contained in five of the somatic cell hybrid lines have been defined by flanking probes within YAC contigs . To provide a framework for further mapping of the 7q21-q32 region , we have established the physical order of a set of reference markers : cen-( P08123 -D7S15- P08684 - P27169 )-D7S456- ( brea kpoint contained in cell hybrid 1EF2/3/K017 ) - P08236 -D7S186-ASL- ( P08183 - P21439 - P62879 -EPO- P22303 ) -D7S238- ( proximal breakpoint in GM1059-Rag5 ) -D7S240-(CUTL1- P05121 )- ( breakp oints in 1CF2/5/K016 and 2068Rag22-2 ) -( P31323 -D7S13)- P07942 - ( breakpoint in JSR-17S ) -DLD-D7S16-MET- P09544 - P13569 -D7S8-tel . Regulation of channels by the serum and glucocorticoid-inducible kinase - implications for transport , excitability and cell proliferation . The serum and glucocorticoid-inducible kinase O00141 stimulates the Na+ channels ENaC and Q14524 , the K+ channels P48048 , Kv1.3 , and P15382 / P51787 , the cation conductance induced by 4F2/ Q01650 and the chloride conductance induced by P13569 . The isoforms Q9HBY8 and Q96BR1 have similarly been shown to regulate ENaC , Q14524 , Kv1.3 and P15382 / P51787 . The kinases regulate channel abundance in the plasma membrane in part by inhibition of the ubiquitin ligase Q96PU5 and in part by interaction with trafficking molecules such as the Na+/H+ exchanger regulating factor Q15599 . An in vivo role of O00141 mediated ENaC channel regulation in renal salt excretion and blood pressure control is documented by the impaired ability of O00141 knockout mice to adequately reduce renal Na+ output and maintain blood pressure during dietary salt restriction and by enhanced blood pressure in individuals carrying certain polymorphisms in the O00141 gene . The in vivo physiological significance of O00141 dependent regulation of the other channels remains to be shown even though circumstantial evidence points to involvement in the regulation of epithelial transport , cell volume , cell proliferation , cardiac action potential and neuroexcitability . There is little doubt that further channels will be identified which are modulated by the SGKs and that further in vivo physiological functions will be defined where channel regulation by the SGKs plays a critical role . 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 . DB01645 stimulates electrogenic Cl(-) secretion in mouse jejunum . We used the short-circuit current ( I(sc) ) technique to investigate the effects of the isoflavone genistein on the electrogenic Cl(-) secretion of the mouse jejunum . DB01645 stimulated a sustained increase in I(sc) that was dose dependent . DB00887 inhibited 76 +/- 5 % of the genistein-stimulated I(sc) consistent with activation of Cl(-) secretion . DB01645 failed to stimulate I(sc) following maximal activation of the DB02527 pathway by forskolin . In addition , forskolin had a reduced effect on I(sc) of the mouse jejunum in the presence of genistein . DB01016 , a blocker of P13569 , eliminated the genistein-stimulated increase of I(sc) and reduced the forskolin-activated I(sc) . Clotrimazole , a Ca(2+)-activated K(+) channel blocker , failed to reduce the genistein-stimulated I(sc) . Vanadate , a blocker of tyrosine-dependent phosphatases , reduced the genistein-activated I(sc) . Tyrphostin A23 , a tyrosine kinase inhibitor , reduced basal I(sc) , after which genistein failed to stimulate I(sc) . These data suggest that genistein activated a sustained Cl(-) secretory response of the mouse jejunum and that the effect of genistein was via a tyrosine-dependent phosphorylation pathway . 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 . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . DB02527 inhibition of murine intestinal Na/H exchange requires P13569 -mediated cell shrinkage of villus epithelium . BACKGROUND AND AIMS : Unlike the intestine of normal subjects , small-intestinal epithelia of cystic fibrosis patients and cystic fibrosis transmembrane conductance regulator protein-null ( P13569 (-) ) mice do not respond to stimulation of intracellular cyclic adenosine monophosphate with inhibition of electroneutral NaCl absorption . Because P13569 -mediated anion secretion has been associated with changes in crypt cell volume , we hypothesized that P13569 -mediated cell volume reduction in villus epithelium is required for intracellular cyclic adenosine monophosphate inhibition of Na(+)/H(+) exchanger ( primarily P48764 ) activity in the proximal small intestine . METHODS : Transepithelial (22)Na flux across the jejuna of P13569 (+) , P13569 (-) , the basolateral membrane Na(+)/K(+)/2Cl(-) co-transporter protein P55011 (+) , and P55011 (-) mice were correlated with changes in epithelial cell volume of the midvillus region . RESULTS : Stimulation of intracellular cyclic adenosine monophosphate resulted in cessation of Na(+)/H(+) exchanger-mediated Na(+) absorption ( J(ms)(NHE) ) in P13569 (+) jejunum but had no effect on J(ms)(NHE) across P13569 (-) jejunum . Cell volume indices indicated an approximately 30 % volume reduction of villus epithelial cells in P13569 (+) jejunum but no changes in P13569 (-) epithelium after intracellular cyclic adenosine monophosphate stimulation . In contrast , cell shrinkage induced by hypertonic medium inhibited J(ms)(NHE) in both P13569 (+) and P13569 (-) mice . DB00887 treatment to inhibit Cl(-) secretion by blockade of the Na(+)/K(+)/2Cl(-) co-transporter , P55011 , of stimulated P13569 (+) jejunum prevented maximal volume reduction of villus epithelium and recovered approximately 40 % of J(ms)(NHE) . Likewise , J(ms)(NHE) and cell volume were unaffected by intracellular cyclic adenosine monophosphate stimulation in P55011 (-) jejuna . CONCLUSIONS : These findings show a previously unrecognized role of functional P13569 expressed in villus epithelium : regulation of P48764 -mediated Na(+) absorption by alteration of epithelial cell volume . DB00887 blocks P13569 GCl in the native sweat duct . DB00887 is well known for its ability to inhibit the nonconductive Na+-K+-2Cl- cotransporter . We were surprised in preliminary studies to find that bumetanide in the contraluminal bath also inhibited NaCl absorption in the human sweat duct , which is apparently poor in cotransporter activity . Inhibition was accompanied by a marked decrease in the transepithelial electrical conductance . Because the cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channel is richly expressed in the sweat duct , we asked whether bumetanide acts by blocking this anion channel . We found that bumetanide 1 ) significantly increased whole cell input impedance , 2 ) hyperpolarized transepithelial and basolateral membrane potentials , 3 ) depolarized apical membrane potential , 4 ) increased the ratio of apical-to-basolateral membrane resistance , and 5 ) decreased transepithelial Cl- conductance ( GCl ) . These results indicate that bumetanide inhibits P13569 GCl in both cell membranes of this epithelium . We excluded bumetanide interference with the protein kinase A phosphorylation activation process by " irreversibly " phosphorylating P13569 [ by using adenosine 5'-O-(3-thiotriphosphate) in the presence of a phosphatase inhibition cocktail ] before bumetanide application . We then activated P13569 GCl by adding 5 mM DB00171 . DB00887 in the cytoplasmic bath ( 10(-3) M ) inhibited approximately 71 % of this DB00171 -activated P13569 GCl , indicating possible direct inhibition of P13569 GCl . We conclude that bumetanide inhibits P13569 GCl in apical and basolateral membranes independent of phosphorylation . The results also suggest that > 10(-5) M bumetanide can not be used to specifically block the Na+-K+-2Cl- cotransporter . Prostaglandin-endoperoxide synthase genes P23219 and P35354 - novel modifiers of disease severity in cystic fibrosis patients . Cystic fibrosis ( CF ) is one of the most common autosomal recessive diseases among Caucasians caused by a mutation in the P13569 gene . However , the clinical outcome of CF pulmonary disease varies remarkably even in patients with the same P13569 genotype . This has led to a search for genetic modifiers located outside the P13569 gene . The aim of this study was to evaluate the effect of functional variants in prostaglandin-endoperoxide synthase genes ( P23219 and P35354 ) on the severity of lung disease in CF patients . To the best of our knowledge , it is the first time when analysis of P23219 and P35354 as potential CF modifiers is provided . The study included 94 CF patients homozygous for F508del mutation of P13569 . To compare their clinical condition , several parameters were recorded , e.g. a unique clinical score : disease severity status ( DSS ) . To analyse the effect of non- P13569 genetic polymorphisms on the clinical course of CF patients , the whole coding region of P23219 and selected P35354 polymorphisms were analysed . Statistical analysis of genotype-phenotype associations revealed a relationship between the heterozygosity status of identified polymorphisms and better lung function . These results mainly concern P35354 polymorphisms : -765G > C and 8473T > C . The P23219 and P35354 polymorphisms reducing P36551 protein levels had a positive effect on all analysed clinical parameters . This suggests an important role of these genes as protective modifiers of pulmonary disease in CF patients , due to inhibition of arachidonic acid conversion into prostaglandins , which probably reduces the inflammatory process . P05231 and P10145 increase the expression of glycosyltransferases and sulfotransferases involved in the biosynthesis of sialylated and/or sulfated Lewisx epitopes in the human bronchial mucosa . Bronchial mucins from patients suffering from CF ( cystic fibrosis ) exhibit glycosylation alterations , especially increased amounts of the sialyl-Lewis(x) ( NeuAcalpha2-3Galbeta1-4[Fucalpha1-3]GlcNAc-R ) and 6-sulfo-sialyl-Lewis(x) ( NeuAcalpha2-3Galbeta1-4[Fucalpha1-3][SO(3)H-6]GlcNAc-R ) terminal structures . These epitopes are preferential receptors for Pseudomonas aeruginosa , the bacteria responsible for the chronicity of airway infection and involved in the morbidity and early death of CF patients . However , these glycosylation changes can not be directly linked to defects in P13569 ( CF transmembrane conductance regulator ) gene expression since cells that secrete airway mucins express no or very low amounts of the protein . Several studies have shown that inflammation may affect glycosylation and sulfation of various glycoproteins , including mucins . In the present study , we show that incubation of macroscopically healthy fragments of human bronchial mucosa with P05231 ( interleukin-6 ) or P10145 results in a significant increase in the expression of alpha1,3/4-fucosyltransferases [ Q495W5 ( fucosyltransferase 11 gene ) and P21217 ] , alpha2-6- and alpha2,3-sialyltransferases [ Q9Y274 ( alpha2,3-sialyltransferase 6 gene ) and Q96JF0 ( alpha2,6-sialyltransferase 2 gene ) ] and GlcNAc-6-O-sulfotransferases [ Q8NCG5 ( carbohydrate sulfotransferase 4 gene ) and Q9GZX3 ] mRNA . In parallel , the amounts of sialyl-Lewis(x) and 6-sulfo-sialyl-Lewis(x) epitopes at the periphery of high-molecular-mass proteins , including Q99102 , were also increased . In conclusion , our results indicate that P05231 and -8 may contribute to the increased levels of sialyl-Lewis(x) and 6-sulfo-sialyl-Lewis(x) epitopes on human airway mucins from patients with CF . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . P13569 modulates neurosecretory function in pulmonary neuroendocrine cell-related tumor cell line models . The pulmonary neuroendocrine cell ( PNEC ) system consists of solitary cells and distinctive cell clusters termed neuroepithelial bodies ( P20929 ) localized in the airway epithelium . PNEC/ P20929 express a variety of bioactive substances , including amine ( serotonin , 5HT ) and neuropeptides . We have previously shown that P20929 cells are O(2) sensors expressing nicotinamide adenine diphosphate oxidase complex and O(2) sensitive K(+) channel . Recently , we demonstrated expression of functional cystic fibrosis transmembrane conductance regulator ( P13569 ) and Cl(-) conductances in P20929 cells of rabbit neonatal lung . Because PNEC/ P20929 are sparsely distributed and difficult to study in native lung , we investigated small-cell lung carcinoma ( SCLC ) and carcinoid tumor cell lines ( tumor counterparts of normal PNEC/ P20929 ) as models for PNEC/ P20929 . SCLC ( H146 , H345 ) and carcinoid ( H727 ) cell lines express neuroendocrine cell markers , including chromogranin A , neural cell adhesion molecule ( N- P62158 ) , 5HT , and tryptophan hydroxylase . We report that H146 , H345 , and H727 express P13569 messenger RNA ( reverse transcription polymerase chain reaction ) and protein ( immunoblotting ) and possess functional P13569 Cl(-) conductance , demonstrated by an iodide efflux assay inhibitable by transfection with antisense P13569 . Using an immunoassay to quantitate 5HT secretion , we also show that downregulation of P13569 abolishes hypoxia-induced 5HT release , and reduces secretory response to high potassium . Our findings suggest that P13569 may modulate neurosecretory activity of PNEC/ P20929 possessing O(2) sensor function . We propose that these tumor cell lines may be useful models for investigating the role of P13569 in PNEC/ P20929 functions in health and disease . 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 . Microelectrode measurements of the effects of basolateral adenosine in polarized human intestinal epithelial cells in culture . Activation of the basolateral receptor for adenosine in HT-29cl.19A cells , by 100 microM adenosine , increased the equivalent short-circuit current ( DeltaIsc= 24+/-2 microA/cm2 ) , depolarized the intracellular potential ( DeltaVa= 26+/-2 mV ) and decreased the fractional apical membrane resistance ( DeltafRa=-0.48 ) . The changes in all parameters reached their peak values simultaneously . This suggests that the primary action of the adenosine-activated pathway is on only one membrane . DB00887 inhibited the transepithelial response and repolarized the cell potential . After preincubation with 100 microM forskolin , application of 300 microM adenosine caused a significant further change in Va , Isc , the transepithelial potential ( Vt ) and fRa . Together with the results from ion-replacement studies , the observations indicate that adenosine activates channels other than the cystic fibrosis transmembrane conductance regulator ( P13569 ) . The rank order of potencies of adenosine and adenosine analogues implies that the receptor is of the A2 subtype . Preincubation with 4-bromophenacyl bromide ( 4-BPB ) inhibited the effect of an adenosine analogue by 50 % , indicating that activation of phospholipase A2 may be involved in the adenosine-induced response . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . P13569 mediated chloride secretion in the avian renal proximal tubule . In primary cell cultures of the avian ( Gallus gallus ) renal proximal tubule parathyroid hormone and DB02527 activation generate a Cl(-)-dependent short circuit current ( I(SC) ) response , consistent with net transepithelial Cl(-) secretion . In this study we investigated the expression and physiological function of the Na-K-2Cl ( NKCC ) transporter and P13569 chloride channel , both associated with Cl(-) secretion in a variety of tissues , in these proximal tubule cells . Using both RT-PCR and immunoblotting approaches , we showed that NKCC and P13569 are expressed , both in proximal tubule primary cultures and in a proximal tubule fraction of non-cultured ( native tissue ) fragments . We also used electrophysiological methods to assess the functional contribution of NKCC and P13569 to forskolin-activated I(SC) responses in filter grown cultured monolayers . DB00887 ( 10 μM ) , a specific blocker of NKCC , inhibited forskolin activated I(SC) by about 40 % , suggesting that basolateral uptake of Cl(-) is partially mediated by NKCC transport . In monolayers permeabilized on the basolateral side with nystatin , forskolin activated an apical Cl(-) conductance , manifested as bidirectional diffusion currents in the presence of oppositely directed Cl(-) gradients . Under these conditions the apical conductance appeared to show some bias towards apical-to-basolateral Cl(-) current . Two selective P13569 blockers , P13569 Inhibitor 172 and GlyH-101 ( both at 20 μM ) inhibited the forskolin activated diffusion currents by 38-68 % , with GlyH-101 having a greater effect . These data support the conclusion that avian renal proximal tubules utilize an apical P13569 Cl(-) channel to mediate DB02527 -activated Cl(-) secretion . Interplay between inhibitory ferric and stimulatory curcumin regulates phosphorylation-dependent human cystic fibrosis transmembrane conductance regulator and ΔF508 activity . Curcumin potentiates cystic fibrosis transmembrane conductance regulator ( P13569 ) activation in an DB00171 -independent but phosphorylation-dependent manner . The underlying molecular mechanisms are unclear . Here , P29320 -293T cells cultured in an Fe(3+)-containing medium were transiently transfected with P13569 constructs , and the role of the inhibitory Fe(3+) bridge between intracellular loop 3 and the regulatory domain of P13569 in this pathway was investigated . The results showed that ethylenediaminetetraacetic acid ( DB00974 ) stimulated phosphorylation-dependent P13569 activation and the stimulation was suppressed by the deletion of the regulatory domain or the insertion of a C832A mutation that removes the Fe(3+)-binding interface . Furthermore , curcumin potentiation of P13569 was significantly weakened not only by Fe(3+)-insensitive mutations at the interface between the regulatory domain and intracellular loop 3 but also by N-ethylmaleimide or DB00974 pretreatment that removes Fe(3+) . More importantly , potentiation of P13569 was completely suppressed by sufficient Fe(3+) . Finally , the insertion of Fe(3+)-insensitive H950R/S768R increased the curcumin-independent activity of ΔF508 but weakened its curcumin potentiation . Thus , Fe(3+) homeostasis in epithelia may play a critical role in regulating P13569 activity , and targeting Fe(3+)-chelating potentiators may direct new therapies for cystic fibrosis . Methoxsalen stimulates electrogenic Cl- secretion in the mouse jejunum . We used the short-circuit current ( I(sc) ) and patch-clamp techniques to investigate the effects of methoxsalen ( MTX ) on the electrogenic Cl- secretion of the mouse jejunum . MTX stimulated a sustained increase in Isc that was dose dependent . DB00887 inhibited MTX-stimulated Isc in a dose-dependent manner consistent with activation of Cl- secretion . MTX failed to stimulate I(sc) following maximal activation of the DB02527 pathway by forskolin , but did increase Isc after a submaximal dose of forskolin . DB01016 , a blocker of the cystic fibrosis transmembrane conductance regulator ( P13569 ) , reduced the MTX-stimulated increase of Isc by 59 +/- 6 % . The DB02527 -dependent K+ channel blocker 293B did not alter the MTX-activated I(sc) ; however , clotrimazole , an intermediate Ca2(+)-activated K+ channel ( IK(Ca) ) blocker , reduced the MTX-stimulated I(sc) . MTX did not alter Na(+)-glucose cotransport across the mouse jejunum . In cell-attached membrane patches , MTX increased the open probability of the basolateral IK(Ca) channel of isolated crypts . These data suggest that the P13569 and IK(Ca) channels participate in the MTX-activated , sustained Cl- secretory response of the mouse jejunum . Reduction of neuronal and inducible nitric oxide synthase gene expression in patients with cystic fibrosis . As a consequence of diminished nitric oxide synthase ( NOS ) protein concentration , the airway concentration of nitric oxide ( NO ) is reduced in patients with cystic fibrosis ( CF ) . This appears to lead to a reduced elimination of such microorganisms as Pseudomonas aeruginosa . The objective of this study was to analyze whether inducible ( P35228 ) , endothelial ( P29474 ) and neuronal ( P29475 ) NOS are reduced at mRNA level and if so whether this is caused directly by the defective CF transmembrane conductance regulator ( P13569 ) . Nasal polyps from three patients with CF and four otherwise healthy patients were obtained . The expression of the three NOS isoenzymes was quantified using real-time PCR . The P35228 expression was assessed in colon carcinoma cells ( CaCo ) transfected with a normal and a mutated ( DeltaF508 ) P13569 . In CF patients , P35228 mRNA expression was 10-to 20-fold and P29475 gene expression was one-fifth to one-tenth that in control patients ( P < 0.001 ) . In CaCo cells , P35228 gene expression under basal and endotoxin-stimulated conditions did not differ between cells transfected with a mutated P13569 and those transfected with an intact P13569 . This observation suggests that cystic fibrosis is associated with reduced P35228 and P29475 gene expression in nasopharyngeal tissue , possibly disturbing the barrier against infective agents already at the site of entrance . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Role of chloride transport proteins in the vasorelaxant action of nitroprusside in isolated rat aorta . Chloride ions play a key role in smooth muscle contraction , but little is known concerning their role in smooth muscle relaxation . Here we investigated the effect of chloride transport inhibitors on the vasorelaxant responses to nitroprusside in isolated and endothelium-denuded rat aorta , precontracted with phenylephrine 1 muM . Incubation of aortic rings in NO(3)(-) media strongly potentiated the vasorelaxant responses to nitroprusside . DB00887 , DIDS ( 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid ) and acetazolamide strongly potentiated the vasorelaxant responses to nitroprusside ( by 70-100 % ) . EC(50) were 2.3+/-0.5 microM for bumetanide , 26+/-15 microM for DIDS and 510+/-118 microM for acetazolamide ( n=6 for condition ) . Niflumic acid , a selective inhibitor of ClCa ( calcium-activated chloride channels ) , potentiated nitroprusside relaxation to a similar extent as chloride transport inhibitors , in a non-additive manner . Zinc and nickel ions , both modestly potentiated nitroprusside vasorelaxation ( by 20-30 % ) . Cobaltum had negligible effect on nitroprusside vasorelaxation . P15085 ( p-chlorophenoxy-acetic acid ) , an inhibitor of volume-sensitive chloride channels ( ClC ) , slightly potentiated nitroprusside vasorelaxation ( by 15 % ) , and the cystic fibrosis transmembrane conductance regulator ( P13569 ) chloride channel inhibitors P13569 (inh)172 ( 5-[(4-Carboxyphenyl)methylene]-2-thioxo-3- [ (3-trifluoromethyl)phenyl-4-thiazolidinone ) , DPC ( diphenylamine-2,2'-dicarboxylic acid ) and glibenclamide were without significant effect . In conclusion , inhibition of chloride transport proteins strongly potentiates the vasorelaxant responses to nitroprusside in isolated rat aorta . This effect seems mediated by chloride depletion and inhibition of a chloride channel activated by both , calcium and cyclic GMP ( cGMP ) . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate .