interacts_with DB01240?
Cytochromes P450 are differently expressed in normal and varicose human saphenous veins : linkage with varicosis . The expression of cytochrome P450 ( CYP ) enzymes and cyclo-oxygenases ( P36551 ) was investigated in human saphenous veins by reverse transcription-polymerase chain reaction analysis . Non-varicose veins were obtained from patients undergoing aortocoronary bypass grafting , whereas varicose veins were obtained from patients undergoing stripping removal of varicose saphenous veins . In non-varicose veins , Q16678 , CYP2C , P05181 and Q02928 were detected , whereas P51589 , P20815 , P23219 and P35354 were detected almost exclusively in varicose veins . P78329 was not detectable . Except for Q02928 , the levels of individual CYP mRNA were higher in varicose veins than in control veins . Smooth muscle cell volume , determined by a colour image-analysis system , was increased approximately 1.5-fold in varicose veins . Because CYPs and COXs produce various vasoactive compounds , increased expression of these enzymes could be involved in the impairment of vascular tone and may contribute to varicose pathology . Then , CYP or P36551 modulators may be potentially active in the treatment of chronic venous insufficiency . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . Cyclooxygenase inhibition abrogates aeroallergen-induced immune tolerance by suppressing prostaglandin I2 receptor signaling . BACKGROUND : The prevalence of allergic diseases has doubled in developed countries in the past several decades . Cyclooxygenase ( P36551 ) -inhibiting drugs augmented allergic diseases in mice by increasing allergic sensitization and memory immune responses . However , whether P36551 inhibition can promote allergic airway diseases by inhibiting immune tolerance is not known . OBJECTIVE : To determine the role of the P36551 pathway and prostaglandin I2 ( DB01240 ) signaling through the P43119 ( IP ) in aeroallergen-induced immune tolerance . METHODS : Wild-type ( WT ) BALB/c mice and IP knockout mice were aerosolized with ovalbumin ( OVA ) to induce immune tolerance prior to immune sensitization with an intraperitoneal injection of OVA/alum . The P36551 inhibitor indomethacin or vehicle was administered in drinking water to inhibit enzyme activity during the sensitization phase . Two weeks after sensitization , the mice were challenged with OVA aerosols . Mouse bronchoalveolar lavage fluid was harvested for cell counts and TH2 cytokine measurements . RESULTS : WT mice treated with indomethacin had greater numbers of total cells , eosinophils , and lymphocytes , and increased P05113 and P35225 protein expression in BAL fluid compared to vehicle-treated mice . Similarly , IP knockout mice had augmented inflammation and TH2 cytokine responses compared to WT mice . In contrast , the DB01240 analog cicaprost attenuated the anti-tolerance effect of P36551 inhibition . CONCLUSION : P36551 inhibition abrogated immune tolerance by suppressing DB01240 IP signaling , suggesting that DB01240 signaling promotes immune tolerance and that clinical use of P36551 -inhibiting drugs may increase the risk of developing allergic diseases . Cellular and molecular biology of prostacyclin synthase . Q16647 ( PGIS ) cDNA comprises 1500 nucleotides coding for a 500 amino acid protein . It is a heme protein with spectral characteristics of cytochrome p450 ( CYP ) . It does not possess the typical CYP mono-oxygenase activity but catalyzes the rearrangement of prostaglandin H2 to form DB01240 . Analysis of its structure-function by molecular modeling and site-directed mutagenesis reveals a long substrate channel lined by hydrophobic residues . DB00151 -441 has been identified as the proximal axial ligand of heme . DB00135 -430 is nitrated by peroxynitrite which results in reduced PGIS catalytic activity , suggesting that DB00135 -430 is located close to the heme pocket . PGIS is constitutively expressed and may be upregulated by cytokines , reproductive hormones , and growth factors . It is physically colocalized with cyclooxygenases and phospholipases , and functionally coupled with these enzymes . PGIS coupling with P35354 has been shown to play an important role in vascular protection , embryo development and implantation , and cancer growth . Epoprostenol ( prostacyclin , DB01240 ) binding and activation of adenylate cyclase in platelets of diabetic and control subjects . 1 The binding of epoprostenol ( prostacyclin , DB01240 ) to isolated fractured human platelets has been studied using tritiated DB01240 . 2 High and low affinity binding sites for DB01240 have been identified ( Kd values = 16 and 382 nM ) . 3 Analysis of the prostacyclin-dependent activation of adenylate cyclase suggests that enzyme activation is mediated by the high affinity binding site . 4 Platelet P43119 binding and adenylate cyclase activation by DB01240 are unchanged in diabetic and normal human platelets . 5 This work suggests that hyperaggregability of diabetic platelets is not due to any alteration of platelet prostacyclin receptor numbers or their activation . Beneficial vasoactive endothelial effects of fluvastatin : focus on prostacyclin and nitric oxide . Statins are believed to exert beneficial effects against cardiovascular disease beyond correction of dyslipidemia . There are however still very sparse data on how individual statins interact with the production of vasoactive eicosanoids and nitric oxide ( NO ) in human vascular endothelial cells . Here we have determined how fluvastatin affects the mRNA expression of genes associated with vascular reactivity as well as the formation of two major vasodilators , prostacyclin ( DB01240 ) and NO , in human endothelial cells . Also , the influence of fluvastatin on arterial resistance was assessed in isolated small arteries . We show that the promoter activity of prostacyclin synthase ( Q16647 ) , the mRNA expression of Q16647 and endothelial nitric oxide synthase ( P29474 ) , and the production of DB01240 and NO are significantly induced by fluvastatin . Also , strong rapid dilatation ex vivo was observed , with the equal contribution of DB01240 and NO . Our findings in cell culture experiments and in isolated human arteries indicate that fluvastatin-evoked endothelium-derived vasodilator production may confer protection of the endothelial cells via both acute and long-term effects of fluvastatin treatment . If these effects take place in vivo , we suggest a protective pleiotropic role of fluvastatin on the cardiovascular system , particularly at the level of the vascular endothelium , to ameliorate the process of atherogenesis and in the acute manner to reduce vascular tone . Creating a genotype-based dosing algorithm for acenocoumarol steady dose . DB01418 is a commonly prescribed anticoagulant drug for the prophylaxis and treatment of venous and arterial thromboembolic disorders in several countries . In counterpart of warfarin , there is scarce information about pharmacogenetic algorithms for steady acenocoumarol dose estimation . The aim of this study was to develop an algorithm of prediction for acenocoumarol.The algorithm was created using the data from 973 retrospectively selected anticoagulated patients and was validated in a second independent cohort adding up to 2,683 patients . The best regression model to predict stable dosage in the Primary Cohort included clinical factors ( age and body mass index , BSA ) and genetic variants ( Q9BQB6 , P11712 * and P78329 polymorphisms ) and explained up to 50 % of stable dose . In the validation study the clinical algorithm yielded an adjusted R²=0.15 ( estimation´s standard error=4.5 ) and the genetic approach improved the dose forecast up to 30 % ( estimation´s standard error=4.6 ) . Again , the best model combined clinical and genetic factors ( R² = 0.48 ; estimation´s standard error=4 ) which provided the best results of doses estimates within 20 % of the real dose in patients taking lower ( ≤ 7 mg/week ) or higher ( ≥ 25 mg/week ) acenocoumarol doses . In conclusion , we developed a prediction algorithm using clinical data and three polymorphisms in Q9BQB6 , P11712 * and P78329 genes that provided a steady acenocoumarol dose for about 50 % of patients in the Validation Cohort . Such algorithm was especially useful to patients who need higher or lower acenocoumarol doses , those patients with higher time required until their stabilisation and are more prone to suffer a treatment derived complication . Two types of prostacyclin receptor coupling to stimulation of adenylate cyclase and phosphatidylinositol hydrolysis in a cultured mast cell line , BNu-2cl3 cells . DB01240 ( DB01240 ) -mediated signal transduction was examined in interleukin 3 ( P08700 ) -dependent BNu-2cl3 mast cells . DB01088 , a stable DB01240 analogue , induced the accumulation of intracellular DB02527 and IP3 , and an increase in the intracellular Ca2+ concentration . Pretreatment of the cells with a protein kinase C activator , 12-O-tetradecanoyl phorbol 13-acetate , suppressed the iloprost-induced IP3 accumulation and Ca2+ mobilization , but inversely potentiated the DB02527 accumulation , suggesting that neither of these signal transduction pathways of iloprosts is the result of a secondary effect of activation of the other . Removal of P08700 from the culture medium reduced the iloprost-induced IP3 accumulation and Ca2+ mobilization , while it had no effect on the iloprost-induced DB02527 accumulation at all . These results taken together suggest that BNu-2cl3 cells express two types of P43119 ; one couples to stimulation of adenylate cyclase , its expression being independent of P08700 , while the other couples to phosphatidylinositol hydrolysis , its expression being dependent on P08700 . 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 . Identification of the prostacyclin receptor by use of [15-3H1]19-(3-azidophenyl)-20-norisocarbacyclin , an irreversible specific photoaffinity probe . The prostaglandin I ( DB01240 ) receptor of mouse mastocytoma P-815 cells was characterized by photo-affinity labeling with the stable DB01240 analogue [ 15-3H1 ] -19-(3-azidophenyl)-20-norisocarbacyclin ( [ 3H ] APNIC ) used as a potential photoaffinity probe for the receptor . [3H]APNIC bound to the mastocytoma membrane with high affinity and in a saturable manner . Scatchard plot analysis indicated a single binding site with a Kd of 4.7 nM and a Bmax of 0.58 pmol/mg protein . The binding of [3H]APNIC was dose dependently inhibited by APNIC and iloprost , another stable DB01240 agonist , and to a much lesser extent by DB00917 . The binding of the radioligand showed sensitivity to the guanine nucleotide guanosine 5'-O-(3-thio-triphosphate) ( GTP gamma S ) . Photolysis of [3H]APNIC-prelabeled membranes resulted in incorporation of radiolabel into a protein of approximately 43 kDa . Photolabeling was inhibited by DB01240 agonists and other prostaglandins with specificity for the P43119 and was modulated by GTP gamma S . A protein of approximately 45 kDa was also labeled by [3H]APNIC in the membrane of porcine platelets , membranes that are known to be abundant in DB01240 receptors . These results demonstrate that [3H]APNIC specifically labels a protein that may represent the P43119 and that this radioprobe will be a useful reagent for further characterization and purification of the P43119 . P01375 inhibits flow and insulin signaling leading to NO production in aortic endothelial cells . Endothelial cells release nitric oxide ( NO ) acutely in response to increased " flow " or fluid shear stress ( FSS ) , and the increase in NO production is correlated with enhanced phosphorylation and activation of endothelial nitric oxide synthase ( P29474 ) . Both vascular endothelial growth factor and FSS activate endothelial protein kinase B ( P31749 ) by way of incompletely understood pathway(s) , and , in turn , P31749 phosphorylates P29474 at DB00133 -1179 , causing its activation . In this study , we found that either FSS or insulin stimulated insulin receptor substrate-1 ( P35568 ) tyrosine and serine phosphorylation and increased P35568 -associated phosphatidylinositol 3-kinase activity , phosphorylation of P31749 DB00133 -473 , phosphorylation of P29474 DB00133 -1179 , and NO production . Brief pretreatment of bovine aortic endothelial cells with tumor necrosis factor-alpha ( P01375 ) inhibited the above described FSS- or insulin-stimulated protein phosphorylation events and almost totally inhibited FSS- or insulin-stimulated NO production . These data indicate that FSS and insulin regulate P29474 phosphorylation and NO production by overlapping mechanisms . This study suggests one potential mechanism for the development of endothelial dysfunction in disease states with alterations in insulin regulation and increased P01375 levels . Inhibition of myoblast migration by prostacyclin is associated with enhanced cell fusion . Satellite cells are stem cells that are critical for the formation and growth of skeletal muscle during myogenesis . To differentiate and fuse , proliferating satellite cells or myoblasts must migrate and establish stable cell-cell contacts . However , the factors that regulate myoblast migration and fusion are not understood completely . We have identified DB01240 as a novel regulator of myogenesis in vitro . DB01240 is a member of the family of prostaglandins ( PG ) , autocrine/paracrine signaling molecules synthesized via the cyclooxygenase-1 and -2 pathways . Primary mouse muscle cells both secrete DB01240 and express the P43119 , IP , at various stages of myogenesis . Using genetic and pharmacological approaches , we show that DB01240 is a negative regulator of myoblast migration that also enhances cell fusion . Thus , DB01240 may act as a " brake " on migrating cells to facilitate cell-cell contact and fusion . Together , our results highlight the importance of the balance between positive and negative regulators in cell migration and myogenesis . This work may have implications for migration of other populations of adult stem cells and/or cells that undergo fusion . Diverse prostaglandin receptors activate distinct signal transduction pathways in rat myometrium . Attempts were made to identify prostaglandin ( PG ) receptors in rat myometrium , according to the differential rank order of potencies displayed by the natural PGs and their analogues , both at the level of second messenger generation and contraction . In estrogen-treated rat myometrium , PGs [ iloprost = DB01240 greater than DB00917 much greater than 16,16-dimethyl ( DM ) - DB00917 ; sulprostone = misoprostol = 0 ] induced adenosine 3',5'-cyclic monophosphate generation , indicating the contribution of a P43119 . The generation of inositol phosphates was stimulated by PGs ( PGF2 alpha greater than PGD2 much greater than DB00917 = DM- DB00917 much greater than iloprost greater than sulprostone = misoprostol = 0 ) , reflecting a PGF2 alpha-receptor-mediated process , which was insensitive to pertussis toxin ( PTX ) . Contractions caused by PGF2 alpha were closely correlated to PGF2 alpha-receptor activation associated with the phospholipase C pathway . By contrast , contractions evoked by DB00917 , equally mimicked by sulprostone and misoprostol , were abolished by PTX and were independent of phospholipase C activation . In the pregnant myometrium ( day 21 ) , the latter PGE-receptor-mediated mechanism also contributed to contractions caused by DB00917 ( less than microM concn ) . Phospholipase C activation was coupled not only to PGF2 alpha but also to PGE receptors and could be correlated with contractions induced by PGF2 alpha and DB00917 greater than microM concn ) . All PGs tested were coupled to inhibitory G protein-mediated adenylate cyclase inhibition , displaying an equipotency that did not allow characterization of the inhibitory PG receptors . [ Detection of a high-affinity prostaglandin I2 binding site in the human thyroid ] . This study is concerned with the identification and the pharmacological properties of P43119 binding sites on human thyroid membrane fractions . Scatchard analysis is not linear , revealing a high- and a low-affinity receptor binding site . ( 3 H ) DB01088 binding experiments were performed under various clinical conditions : in thyroid cancer the low-affinity binding sites disappear totally and the specific high-affinity binding sites are diminished according to the grade of differentiation of the cancer . An alteration in Bmax and Kd is also observed in cold nodules , in Hashimoto 's and Riedl 's thyroiditis and in hyperthyroidism , whereas hot nodules exhibit an increase in both the receptor subpopulations . The data provide evidence for specific DB01240 binding sites and support the suggestion of a direct regulatory key-role of DB01240 in thyroid intermediary metabolism . Roles of thromboxane A(2) and prostacyclin in the development of atherosclerosis in apoE-deficient mice . Production of thromboxane ( TX ) A2 and PG I2/prostacyclin ( DB01240 ) is increased in patients with atherosclerosis . However , their roles in atherogenesis have not been critically defined . To examine this issue , we cross-bred atherosclerosis-prone apoE-deficient mice with mice deficient in either the TXA receptor ( TP ) or the P43119 ( IP ) . Although they showed levels of serum cholesterol and triglyceride similar to those of apoE-deficient mice , apoE-/-TP-/- mice exhibited a significant delay in atherogenesis , and apoE-/-IP-/- mice exhibited a significant acceleration in atherogenesis compared with mice deficient in apoE alone . The plaques in apoE-/-IP-/- mice showed partial endothelial disruption and exhibited enhanced expression of P05362 and decreased expression of platelet endothelial cell adhesion molecule 1 ( P16284 ) in the overlying endothelial cells compared with those of apoE-/-TP-/- mice . Platelet activation with thrombin ex vivo revealed higher and lower sensitivity for surface P16109 expression in platelets of apoE-/-IP-/- and apoE-/-TP-/- mice , respectively , than in those of apoE-/- mice . Intravital microscopy of the common carotid artery revealed a significantly greater number of leukocytes rolling on the vessel walls in apoE-/-IP-/- mice than in either apoE-/-TP-/- or apoE-/- mice . We conclude that TXA2 promotes and DB01240 prevents the initiation and progression of atherogenesis through control of platelet activation and leukocyte-endothelial cell interaction . 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 . [ Thrombocyte prostacyclin receptors in gestational hypertension and pre-eclampsia ] . OBJECTIVE : To determine prostacyclin ( DB01240 ) receptor characteristics in pregnancies complicated by hypertension and to assess any relation to the clinical outcome . METHODS : Radioligand binding studies with [ 3H ] - DB01088 were performed to measure receptor capacity ( Bmax ) and affinity ( Kd-1 ) using platelet membranes from patients with preeclampsia , gestational hypertension or normal pregnancy . RESULTS : P43119 capacity did not differ between the patient groups . In contrast , P43119 affinity was diminished in gestational hypertension and considerably reduced in preeclampsia compared to normal pregnancy . A similar pattern was found in fetal growth ( normal pregnancy > gestational hypertension > preeclampsia ) . Furthermore , the rate of low Apgar scores and acidosis was increased in preeclampsia . CONCLUSIONS : In preeclampsia reduced platelet P43119 affinity was found as well as poor pregnancy outcome in comparison with normal pregnancy , whereas these differences were less pronounced in gestational hypertension . This suggests a role of DB01240 and its receptor in gestational hypertension and in particular in preeclampsia . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Long-term amphetamine treatment exacerbates inflammatory lung reaction while decreases airway hyper-responsiveness after allergic stimulus in rats . Asthma is an allergic lung disease can be modulated by drugs that modify the activity of central nervous system ( CNS ) such as amphetamine ( P49418 ) . P49418 is a highly abused drug that exerts potent effects on behavior and immunity . In this study we investigated the mechanism involved in the effects of long-term P49418 treatment on the increased magnitude of allergic lung response . We evaluated mast cells degranulation , cytokines release , airways responsiveness and , expression of adhesion molecules . Male Wistar rats were treated with P49418 or vehicle ( PBS ) for 21 days and sensitized with ovalbumin ( OVA ) one week after the first injection of vehicle or P49418 . Fourteen days after the sensitization , the rats were challenged with an OVA aerosol , and 24h later their parameters were analyzed . In allergic rats , the treatment with P49418 exacerbated the lung cell recruitment due increased expression of P05362 , P16284 and Mac-1 in granulocytes and macrophages recovered from bronchoalveolar lavage . Elevated levels of P05112 , but decreased levels of P22301 were also found in samples of lung explants after P49418 treatment . Conversely , the ex-vivo tracheal hyper-responsiveness to methacholine ( DB06709 ) was reduced by P49418 treatment , whereas the force contraction of tracheal segments due to in vitro antigen challenge remained unaltered . Our findings suggest that lung inflammation and airway hyper-responsiveness due to OVA challenge are under the distinct control of P49418 during long-term treatment . Our data strongly indicate that P49418 positively modulates allergic lung inflammation via the increase of P05362 , P16284 , Mac-1 and P05112 . P49418 also abrogates the release of the anti-inflammatory cytokine P22301 . DB01088 has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 + T cells was measured by ELISA . The expression of the transcription factor FoxP3 after co-culture of DCs with P01730 + CD45RA+ T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 , P05231 , P10145 and IL-12p70 in mDCs , while it enhanced P22301 production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA+ T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP3 expression and P22301 production . Additionally , iloprost inhibited the MIP-3beta-induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans . GM- P04141 priming drives bone marrow-derived macrophages to a pro-inflammatory pattern and downmodulates DB00917 in response to O60603 ligands . In response to pathogen recognition by Toll-like receptors ( TLRs ) on their cell surface , macrophages release lipid mediators and cytokines that are widely distributed throughout the body and play essential roles in host responses . Granulocyte macrophage colony-stimulating factor ( GM- P04141 ) is important for the immune response during infections to improve the clearance of microorganisms . In this study , we examined the release of mediators in response to O60603 ligands by bone marrow-derived macrophages ( BMDMs ) primed with GM- P04141 . We demonstrated that when stimulated with O60603 ligands , non-primed BMDMs preferentially produced PGE(2) in greater amounts than Q06643 (4) . However , GM- P04141 priming shifted the release of lipid mediators by BMDMs , resulting in a significant decrease of PGE(2) production in response to the same stimuli . The decrease of PGE(2) production from primed BMDMs was accompanied by a decrease in PGE-synthase mRNA expression and an increase in P01375 -α and nitric oxide ( NO ) production . Moreover , some GM- P04141 effects were potentiated by the addition of IFN-γ . Using a variety of O60603 ligands , we established that PGE(2) release by GM- P04141 -primed BMDMs was dependent on O60603 co-receptors ( Q15399 , Q9Y2C9 ) , P08571 , MyD88 and the nuclear translocation of NFκB but was not dependent on peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) activation . Indeed , GM- P04141 priming enhanced O60603 , O00206 and MyD88 mRNA expression and phospho-IκBα formation . These findings demonstrate that GM- P04141 drives BMDMs to present a profile relevant to the host during infections . Induction of prostacyclin receptor expression in human erythroleukemia cells . We have identified both high-affinity ( KD = 36 +/- 3 nM ) and low-affinity ( KD = 2.1 +/- 0.8 microM ) prostacyclin ( DB01240 ) -receptor sites on human erythroleukemia ( HEL ) cells using the radiolabelled prostacyclin analogue . [3H]iloprost . The addition of the phorbol ester , TPA , to the culture medium caused a 5-10-fold increase in the number of both the low- and the high-affinity sites , without any change in their affinity constants . DB01088 stimulated HEL cell membrane adenylate cyclase activity 5-fold . This stimulation was potentiated in the presence of GTP , indicating a conventional P43119 -G2-adenylate cyclase system . HEL cells represent a source of prostacyclin receptor mRNA which may be of value in expression cloning of this receptor . On the multiplicity of platelet prostaglandin receptors . I . Evaluation of competitive antagonism by aggregometry . Methods for the evaluation of competitive interactions at receptors associated with platelet activation and inhibition using aggregometry of human PRP have been developed . The evidence supports the suggestion that PGE1 and DB01240 share a common receptor for inhibition of platelet reactivity , but only a portion ( if any ) of the aggregation stimulation associated with DB00917 is the result of DB00917 binding ( without efficacy ) to this receptor . DB00917 ( at .3-20 microM ) is an effective antagonist of PGE1 , DB01240 , and PGD2 producing a shift of about one order of magnitude in the IC50-values obtained from complete aggregation inhibition dose response curves . The antagonism of PGD2 inhibition is particularly notable , 80 nM DB00917 levels are detectable . This and other actions of DB00917 indicate another platelet receptor for DB00917 . PGE1 acts at both the DB00917 and P43119 . Other substances showing DB01240 -like actions only at high doses ( 1-30 microM ) , display additive responses with DB01240 indicative of decreased affinity for the I2/E1 receptor and the absence of DB00917 -like aggregation stimulation activity . DB01240 methyl ester has intrinsic inhibitory action not associated with in situ ester hydrolysis . The methyl ester is dissaggregatory showing particular specificity for inhibition of release and second wave aggregation . Human prostacyclin receptor . DB01240 , a member of the eicosanoid family of lipid mediators , is the major product of arachidonic acid metabolism formed in the marcovascular endothelium . It is a potent vasodilator , antithrombotic , and antiplatelet agent that mediates it effects through a membrane-associated receptor termed the IP . Cloning of the cDNA for IP , from human and other species , indicated its membership of the G protein-coupled receptor superfamily and has allowed detailed examination of the signaling and regulatory pathways utilized by this receptor . This article examines the current state of knowledge of the IP , its signaling and regulation , and its biological role in vivo and examines the possible existence of multiple P43119 sites . Q01094 :DP-1 induces p53 and overrides survival factors to trigger apoptosis . The E2F DNA binding activity consists of a heterodimer between E2F and DP family proteins , and these interactions are required for association of E2F proteins with P06400 and the P06400 -related proteins P28749 and Q08999 , which modulate E2F transcriptional activities . Q01094 expression is sufficient to release fibroblasts from G0 and induce entry into S phase , yet it also initiates apoptosis . To investigate the mechanisms of E2F-induced apoptosis , we utilized interleukin-3 ( P08700 ) -dependent 32D.3 myeloid cells , a model of hematopoietic progenitor programmed cell death . In the absence of P08700 , Q01094 alone was sufficient to induce apoptosis , and p53 levels were diminished . DP-1 alone was not sufficient to induce cell cycle progression or alter rates of death following P08700 withdrawal . However , overexpression of both Q01094 and DP-1 led to the rapid death of cells even in the presence of survival factors . In the presence of P08700 , levels of endogenous wild-type p53 increased in response to Q01094 , and coexpression of DP-1 further augmented p53 levels . These results provide evidence that E2F is a functional link between the tumor suppressors p53 and P06400 . However , induction of p53 alone was not sufficient to trigger apoptosis , suggesting that the ability of E2F to override survival factors involves additional effectors . DB05229 sodium , a stable prostacyclin analogue , elicits dilation of isolated porcine retinal arterioles : roles of P29474 and potassium channels . PURPOSE : DB01240 ( DB01240 ) is usually described as an endoEDRFsthelium-derived relaxing factor , but the vasoreactivity to DB01240 in the retinal arterioles and the underlying mechanisms are not fully understood . We examined the effects of DB01240 on the retinal microcirculation using beraprost sodium ( BPS ) , a stable DB01240 analogue , and the signaling mechanisms involved in this vasomotor activity . METHODS : Porcine retinal arterioles were isolated , cannulated , and pressurized without flow in vitro . Video microscopic techniques recorded the diametric responses to BPS . RESULTS : DB05229 sodium elicited dose-dependent ( 0.1 pM-0.1 μM ) vasodilation of the retinal arterioles that was abolished by the P43119 ( IP ) antagonist CAY10441 . DB05229 sodium-induced vasodilation decreased by 50 % after the endothelium was removed and was inhibited by the nitric oxide ( NO ) synthase inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) comparable with denudation . Inhibition of soluble guanylyl cyclase by 1H-1,2,4-oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) and blockage of protein kinase A ( PKA ) by Rp-8-Br-cAMPS were comparable to L-NAME . DB05229 sodium-induced vasodilation was also inhibited by the nonselective potassium channel inhibitor , tetraethylammonium , and the adenosine triphosphate-sensitive potassium ( KATP ) channel blocker , glibenclamide . Residual vasodilation in the presence of glibenclamide decreased further with subsequent application of ODQ . CONCLUSIONS : DB05229 sodium , a stable DB01240 analogue , causes vasodilation of the retinal arterioles mediated via the IP receptor . The current findings suggest that BPS elicits endothelium-dependent and -independent dilation of the retinal arterioles mediated by NO induced by activation of PKA in the endothelium and the KATP channel activation in the vascular smooth muscle , respectively . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . P35354 -derived prostacyclin confers atheroprotection on female mice . Female gender affords relative protection from cardiovascular disease until the menopause . We report that estrogen acts on estrogen receptor subtype alpha to up-regulate the production of atheroprotective prostacyclin , DB01240 , by activation of cyclooxygenase 2 ( P35354 ) . This mechanism restrained both oxidant stress and platelet activation that contribute to atherogenesis in female mice . Deletion of the P43119 removed the atheroprotective effect of estrogen in ovariectomized female mice . This suggests that chronic treatment of patients with selective inhibitors of P35354 could undermine protection from cardiovascular disease in premenopausal females . DB01240 primes pregnant human myometrium for an enhanced contractile response in parturition . An incomplete understanding of the molecular events that regulate the myometrial transition from the quiescent pregnant state to the active contractile state during labor has hindered the development of improved therapies for preterm labor . During myometrial activation , proteins that prime the smooth muscle for contraction are upregulated , allowing maximal responsiveness to contractile agonists and thereby producing strong phasic contractions . Upregulation of one such protein , P35354 , generates PGs that induce contractions . Intriguingly , the predominant myometrial PG produced just prior to labor is prostacyclin ( DB01240 ) , a smooth muscle relaxant . However , here we have shown that activation of P43119 ( IP ) upregulated the expression of several contractile proteins and the gap junction protein connexin 43 through DB02527 /PKA signaling in human myometrial tissue in organ and cell culture . Functionally , these IP-dependent changes in gene expression promoted an enhanced contractile response to oxytocin in pregnant human myometrial tissue strips , which was inhibited by the IP antagonist RO3244794 . Furthermore , contractile protein induction was dependent on the concentration and time of exposure to the DB01240 analog iloprost and was blocked by both RO3244794 and PKA knockdown . We therefore propose that DB01240 -mediated upregulation of contractile proteins and connexin 43 is a critical step in myometrial activation , allowing for a maximal contractile response . Our observations have important implications regarding activation of the myometrium prior to the onset of labor . Role of prostacyclin in the cardiovascular response to thromboxane A2 . Thromboxane ( Tx ) A2 is a vasoconstrictor and platelet agonist . DB00945 affords cardioprotection through inhibition of TxA2 formation by platelet cyclooxygenase ( P23219 ) . DB01240 ( DB01240 ) is a vasodilator that inhibits platelet function . Here we show that injury-induced vascular proliferation and platelet activation are enhanced in mice that are genetically deficient in the P43119 ( IP ) but are depressed in mice genetically deficient in the TxA2 receptor ( TP ) or treated with a TP antagonist . The augmented response to vascular injury was abolished in mice deficient in both receptors . Thus , DB01240 modulates platelet-vascular interactions in vivo and specifically limits the response to TxA2 . This interplay may help explain the adverse cardiovascular effects associated with selective P35354 inhibitors , which , unlike aspirin and nonsteroidal anti-inflammatory drugs ( NSAIDs ) , inhibit DB01240 but not TxA2 . Strategies for the analysis of oncogene overexpression . Studies of the neu oncogene in breast carcinoma . The development of a consistent strategy for the analysis of oncogene expression at the cellular level is essential for understanding the roles of these genes in the development and progression of human neoplasia . Detection of the neu oncogene products in breast carcinoma was selected as a model for analysis of oncogene expression . Fifty-two primary human breast carcinomas were evaluated by quantitation of neu DNA amplification and mRNA expression and by localization of neu mRNA and protein ( p 185 ) at the cellular level by in situ hybridization ( ISH ) and immunohistochemistry ( IHC ) . The specificity and sensitivity of the molecular and immunologic probes for neu were established with the use of genetically engineered cell lines that overexpressed either neu or epidermal growth factor receptor ( P00533 ) . Twenty-nine percent of breast carcinomas demonstrated neu DNA amplification and mRNA overexpression , and there was close correlation between the level of neu mRNA expression and detection of neu gene products by ISH and IHC . Thirty-two percent of carcinomas demonstrated neu mRNA overexpression by ISH . The immunohistochemical method using Q96RJ0 monoclonal antibody for p185 was exquisitely sensitive in acetone-fixed frozen sections and provided an excellent approach for judging overexpression as confirmed by the various molecular analyses . All areas of nonmalignant breast epithelium stained weakly , and a wide range of staining intensity was observed in malignant breast epithelium , with 31 % of carcinomas judged to be p185 overexpressors . Heterogeneous expression of p185 was seen in some carcinomas . This study provides a strategic approach for the evaluation of oncogene expression in human tumors . Chromosomal localization of the human prostanoid receptor gene family . Prostaglandins ( PGD2 , DB00917 , PGF2 alpha , and DB01240 ) and thromboxane A2 ( TXA2 ) are biologically active molecules derived from the metabolism of arachidonic acid by cyclooxygenases . They produce a wide variety of physiological and pathophysiological effects mediated through specific G protein-coupled cell surface receptors . In this study , we have mapped the chromosomal positions of the human genes that encode the DB00917 receptor subtypes ( P34995 , PTGER2 , and P43115 ) , the PGF2 alpha receptor ( P43088 ) , the P43119 ( P43119 ) , and the TXA2 receptor ( P21731 ) using in situ hybridization . The P34995 , P21731 , and P43119 genes mapped to chromosome 19 at positions 19p13.1 , 19p13.3 , and 19q13.3 , respectively . The P43088 and P43115 genes mapped to chromosome 1 at positions 1p31.1 and 1p31.2 , respectively , and PTGER2 gene mapped to chromosome band 5p13.1 . Altered expression of inflammation-related genes in human carotid atherosclerotic plaques . OBJECTIVE : Inflammation is a pivotal process in atherosclerosis development and progression , but the underlying molecular mechanisms remain largely obscure . We have conducted an extensive expression study of atherosclerotic plaques to identify the inflammatory pathways involved in atherosclerosis . METHODS : We studied 11 human carotid plaques , their respective adjacent regions and 7 control arteries from different subjects . Expression of 92 genes was studied by TaqMan low-density array human inflammation panel . Human aortic endothelial and smooth muscle cells were used for in vitro experiments . RESULTS : The mRNA levels of 44/92 genes ( 48 % ) differed significantly between the tissues examined ( 13 up-regulated and 31 down-regulated ) . Dysregulated genes encode molecules belonging to different functional classes although most of them encode enzymes involved in the eicosanoid synthesis pathway . The expression of Q16647 and P43119 genes was decreased in human aortic endothelial and smooth muscle cells stimulated with oxLDL and P01375 -α . CONCLUSIONS : This study not only reveals several dysregulated genes in human lesions but also focuses the role played by the genes involved in the eicosanoid synthesis pathway during atherosclerotic development . The decrease of Q16647 and P43119 expression after oxLDL treatment mirrors the decreased mRNA levels in atherosclerotic lesions versus control arteries , which suggests that oxidation is important for Q16647 and P43119 regulation in human vessel cells during atherosclerosis development . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . [ Polarized light as an epigenetic factor in inhibition of inflammation ; a genome-wide expression analysis in recurrent respiratory diseases of children ] . Whole-body polarized light therapy has been primarily investigated in various clinical observations and in a few in vitro model systems . AIMS : In the present study , clinical and molecular effects of whole-body polarized light treatment on children suffering from recurrent respiratory infection were studied . METHODS : Incidence and duration of respiratory symptoms as well as the length of appropriate antibiotic therapy have been measured . Simultaneously , genome-wide gene expression pattern was examined by whole genome cDNA microarray in peripheral lymphocytes of children . RESULTS : Twenty of twenty five children showed a marked clinical improvement , while in five of twenty five had poor or no changes . Gene expression pattern of the peripheral lymphocytes of the patients was compared in favorable and poor responders . Lymphocytes of the children with a documented improved clinical response to polarized light therapy showed a decrease in the expression of chemokine genes , such as P09341 , P19875 , P10145 and in that of the tumor necrosis alpha ( TNFα ) gene . On the contrary , a rapid elevation was found in the expression of gene encoding for P78329 , a leukotriene-B(4)-metabolizing enzyme . In children with poor clinical response to polarized light therapy , no similar changes were detected in the gene expression pattern of the lymphocytes . CONCLUSIONS : Improved clinical symptoms and modified gene expression profile of lymphocytes reveals anti-inflammatory effect upon whole body polarized light irradiation . Decreased susceptibility to renovascular hypertension in mice lacking the prostaglandin I2 receptor IP . Persistent reduction of renal perfusion pressure induces renovascular hypertension by activating the renin-angiotensin-aldosterone system ; however , the sensing mechanism remains elusive . Here we investigated the role of DB01240 in renovascular hypertension in vivo , employing mice lacking the P43119 ( IP-/- mice ) . In WT mice with a two-kidney , one-clip model of renovascular hypertension , the BP was significantly elevated . The increase in BP in IP-/- mice , however , was significantly lower than that in WT mice . Similarly , the increases in plasma renin activity , renal renin mRNA , and plasma aldosterone in response to renal artery stenosis were all significantly lower in IP-/- mice than in WT mice . All these parameters were measured in mice lacking the four DB00917 receptor subtypes individually , and we found that these mice had similar responses to WT mice . DB01240 is produced by P35354 and a selective inhibitor of this enzyme , SC-58125 , also significantly reduced the increases in plasma renin activity and renin mRNA expression in WT mice with renal artery stenosis , but these effects were absent in IP-/- mice . When the renin-angiotensin-aldosterone system was activated by salt depletion , SC-58125 blunted the response in WT mice but not in IP-/- mice . These results indicate that DB01240 derived from P35354 plays a critical role in regulating the release of renin and consequently renovascular hypertension in vivo . Dopamine modulating drugs influence striatal (+)-[11C]DTBZ binding in rats : Q05940 binding is sensitive to changes in vesicular dopamine concentration . Binding of (+)-[11C]DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of (+)-[11C]DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , +14 % ) or DB01576 ( d- P49418 , 20 mg/kg , +12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , -16 % ) or levodopa ( -20 % ) . We suggest that in vivo (+)-[11C]DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed . Anti-inflammatory activity of Taraxacum officinale . Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases . The dried plant was extracted with 70 % ethanol to generate its ethanol extract ( TEE ) . For some experiments , ethyl acetate ( EA ) , n-butanol ( BuOH ) and aqueous ( Aq ) fractions were prepared in succession from TEE . TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl ( DPPH ) assay , a diminishing effect on intracellular reactive oxygen species ( ROS ) level , and an anti-angiogenic activity in the chicken chorioallantoic ( P62158 ) assay . In the carrageenan-induced air pouch model , TEE inhibited production of exudate , and significantly diminished nitric oxide ( NO ) and leukocyte levels in the exudate . It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice . Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated macrophages were also assessed . Among the fractions , the n-butanol fraction ( BuOH ) was identified to be most effective in the P62158 assay . Collectively , Taraxacum officinale contains anti-angiogenic , anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and P35354 expression and/or its antioxidative activity . Pecam-1 is a modulator of stat family member phosphorylation and localization : lessons from a transgenic mouse . P16284 ( CD31 ) is a member of the immunoglobin ( Ig ) superfamily of cell adhesion molecules whose expression is restricted to hematopoietic and vascular cells . P16284 can recruit adapter and signaling molecules via its immunoreceptor tyrosine activation motif ( ITAM ) , suggesting that P16284 plays a role in signal transduction pathways . To study the involvement of P16284 in signaling cascades in vivo , we used the major histocompatibility ( MHC ) I gene promoter to target ectopic P16284 expression in transgenic mice . We noted an attenuation of mammary gland development at early stages of virgin ductal branching morphogenesis . STAT5a , a modulator of milk protein gene expression during lactation , was localized to the nuclei of ductal epithelial cells of 6-week-old virgin P16284 transgenics , but not in control mice . This correlated with decreases in ductal epithelial cell proliferation and induction of P38936 , an inhibitor of cell cycle progression . Using in vitro model systems we demonstrated P16284 /STAT5a association and found that residue Y701 in P16284 's cytoplasmic tail is important for P16284 / P42229 association and that P16284 modulates increases in STAT5a tyrosine phosphorylation levels . We suggest that by serving as a scaffolding , P16284 can bring substrates ( STAT5a ) and enzymes ( a kinase ) into close proximity , thereby modulating phosphorylation levels of selected proteins , as previously noted for beta-catenin . Interactions between prostaglandin E2 and inhibitors of platelet aggregation which act through cyclic AMP . Prostaglandin ( PG ) E2 potentiates platelet aggregation at low concentrations ( 10(-8)-10(-6) M ) . It also inhibits aggregation at a higher concentration ( 10(-5) M ) , probably by acting through cyclic adenosine 3',5'-monophosphate ( cyclic AMP ) . The mechanism of this biphasic effect of DB00917 and its implications for thrombosis are not clearly understood . Using a sensitive cyclic AMP assay , in conjunction with platelet aggregation studies , we have examined the interactions between DB00917 and other inhibitors of platelet aggregation which act through cyclic AMP . Low concentrations of DB00917 reversed the inhibition of platelet aggregation and increase in cyclic AMP levels induced by DB01240 , PGD2 and adenosine ( which stimulate adenylate cyclase ( AC ) through separate and specific platelet receptors ) . In contrast , low concentrations of DB00917 added to the inhibition of platelet aggregation and increase in cyclic AMP levels induced by forskolin ( which stimulates AC directly ) and AH-P 719 and DN-9693 ( which inhibit cyclic AMP phosphodiesterase ( PDE ] . These results suggest that the biphasic effect of DB00917 may be mediated by interaction with two separate platelet receptors . Low concentrations appear to potentiate aggregation by acting at a receptor which is directly coupled to an inhibitory guanine nucleotide-binding protein ( Gi ) , possibly the putative PG endoperoxide receptor . High concentrations of DB00917 appear to inhibit aggregation by acting at an additional receptor , probably the P43119 . The ease with which DB00917 reverses the effects of DB01240 , PGD2 and adenosine , but adds to the effects of AH-P 719 and DN-9693 , suggests that PDE inhibitors might offer greater potential than these AC stimulators as an anti-thrombotic strategy. ( ABSTRACT TRUNCATED AT 250 WORDS ) Q16647 gene transfer inhibits neointimal formation by suppressing Q07869 delta expression . OBJECTIVE : DB01240 ( P06744 (2) ) is a potent ligand of peroxisome proliferator-activated receptor delta ( Q07869 delta ) that regulates cell growth and differentiation . The aim of this study was to elucidate how endogenous P06744 (2) overexpression affects the expressions of Q07869 delta and mitogen-activated protein kinases ( MAPKs ) in the development of neointimal formation in experimental angioplasty with adenovirus-mediated P06744 (2) synthase ( Ad-PGIS ) gene transfer . METHODS AND RESULTS : In human aortic smooth muscle cells , protein blotting analysis showed that P06744 (2) overproduction decreased the levels of phosphorylated p38 MAPK ( P-p38 MAPK ) ( 2.0-fold versus 0.83-fold relative to control ) . Immunohistochemical analysis in balloon-injured arteries revealed diffuse expression of Q07869 delta in the neointima of control vessels , with no expression in uninjured vessels . The level of Q07869 delta expression was lower in Ad-PGIS-treated arteries than in control vessels , with the Q07869 delta localized in the neointima adjacent to endothelium . Staining of P-p38 MAPK showed a similar pattern to Q07869 delta among the three groups . Morphometric analysis at day 14 revealed that Ad-PGIS reduced the intima-to-media ratio by up to 59 % . CONCLUSIONS : Ad-PGIS gene transfer reduced Q07869 delta expression and inhibited neointimal formation after balloon injury in accordance with the reduction in the phosphorylation of p38 MAPK . Exploration of the antiplatelet activity profile of betulinic acid on human platelets . Betulinic acid , a natural pentacyclic triterpene acid , presents a diverse mode of biological actions including antiretroviral , antibacterial , antimalarial , and anti-inflammatory activities . The potency of betulinic acid as an inhibitor of human platelet activation was evaluated , and its antiplatelet profile against in vitro platelet aggregation , induced by several platelet agonists ( adenosine diphosphate , thrombin receptor activator peptide-14 , and arachidonic acid ) , was explored . Flow cytometric analysis was performed to examine the effect of betulinic acid on P16109 membrane expression and O95456 binding to activated platelets . Betulinic acid potently inhibits platelet aggregation and also reduced O95456 binding and the membrane expression of P16109 . Principal component analysis was used to screen , on the chemical property space , for potential common pharmacophores of betulinic acid with approved antithrombotic drugs . A common pharmacophore was defined between the NMR-derived structure of betulinic acid and prostacyclin agonists ( DB01240 ) , and the importance of its carboxylate group in its antiplatelet activity was determined . The present results indicate that betulinic acid has potential use as an antithrombotic compound and suggest that the mechanism underlying the antiplatelet effects of betulinic acid is similar to that of the P43119 agonists , a hypothesis that deserves further investigation . Protein kinase Cdelta and calmodulin regulate epidermal growth factor receptor recycling from early endosomes through Arp2/3 complex and cortactin . The intracellular trafficking of the epidermal growth factor receptor ( P00533 ) is regulated by a cross-talk between calmodulin ( P62158 ) and protein kinase Cdelta ( PKCdelta ) . On inhibition of P62158 , PKCdelta promotes the formation of enlarged early endosomes and blocks P00533 recycling and degradation . Here , we show that PKCdelta impairs P00533 trafficking due to the formation of an F-actin coat surrounding early endosomes . The PKCdelta-induced polymerization of actin is orchestrated by the Arp2/3 complex and requires the interaction of cortactin with PKCdelta . Accordingly , inhibition of actin polymerization by using cytochalasin D or by overexpression of active cofilin , restored the normal morphology of the organelle and the recycling of P00533 . Similar results were obtained after down-regulation of cortactin and the sequestration of the Arp2/3 complex . Furthermore we demonstrate an interaction of cortactin with P62158 and PKCdelta , the latter being dependent on P62158 inhibition . In summary , this study provides the first evidence that P62158 and PKCdelta organize actin dynamics in the early endosomal compartment , thereby regulating the intracellular trafficking of P00533 . DB01240 is a specific effector of adipose cell differentiation . Its dual role as a DB02527 - and Ca(2+)-elevating agent . The mitogenic-adipogenic activity of carbaprostacyclin ( cPGI2 ) , a stable analogue of prostacyclin ( DB01240 ) , has been proposed to be related to its ability to elicit DB02527 production and to activate the protein kinase A cascade ( Négrel , R. , Gaillard , D. , and Ailhaud , G. ( 1989 ) Biochem. J. 257 , 399-405 ) . In the present study , cPGI2 has been compared with other activators of the DB02527 pathway , namely isoproterenol , forskolin and 8-bromo- DB02527 , with respect to adipose cell differentiation . Carbaprostacyclin behaved as a much more potent and efficient effector of mouse Ob1771 preadipocyte differentiation than the latter agents . Moreover , cPGI2 also exerted a specific amplifying mitogenic-adipogenic role , as compared with isoproterenol in rat as well as human adipose precursor cells in primary culture , suggesting that the prostanoid was able to generate an additional second messenger . The fact that ionomycin was able to potentiate and amplify the differentiation induced by 8-bromo- DB02527 led us to give evidence , using preadipocytes preloaded with the fluorescent calcium chelator Indo-1 , that cPGI2 , besides its ability to activate adenyl cyclase , was also able to induce a transient increase in intracellular free calcium . This phenomenon was independent of DB02527 production or inositol phospholipid breakdown and appeared to be mediated after binding to a single class of P43119 . The potential to generate simultaneously two synergistic intracellular signals allows us to ascribe to DB01240 a key and specific role in the differentiation of adipose precursor cells in vitro that would likely lead in vivo to the recruitment of " dormant " preadipocytes to become adipocytes . Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma : a case report . Uterine carcinosarcoma ( malignant mixed Mullerian tumor ) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells . We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage ( IB due to FIGO ) uterine carcinosarcoma of the homologous type with superficial ( 3mm ) myo-invasion . The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies . Positive immunostaining for steroid receptors ( estrogen-alpha and progesterone receptors ) , cytokeratin , and P00533 was detected only in the carcinomatous area , whereas beta-catenin , BCL-2 , P35354 , p16(INK4a) , P60484 , Q8IUH3 , and vimentin were immunoreactive in both components . P10275 , CD10 , desmin , HER-2/neu , and P04637 were found to be negative either in the carcinomatous or in the sarcomatous area . Tumor proliferative activity was higher in the carcinomatous ( 25 % ) than in the sarcomatous ( 2 % ) component . Based on these findings , immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy . Developmental regulation of prostacyclin synthase and prostacyclin receptors in the ovine uterus and conceptus during the peri-implantation period . This study documents the expression of prostacyclin ( DB01240 ) synthase ( Q16647 ) and DB01240 receptors in the trophoblast and uterus of the ewe at the time of maternal recognition of pregnancy ( i.e. days 7 , 9 , 12 , 14 and 17 ) . The membrane receptor for DB01240 ( P43119 ) and the nuclear receptors , i.e. peroxisome proliferator-activated receptors ( Q07869 ) and their heterodimer partners the retinoid X receptors ( RXR ) , were analysed . In the endometrium , Q16647 transcript and protein were expressed at day 9 of pregnancy and levels declined from days 12 to 17 . Immunohistochemistry and in situ hybridization indicated that Q16647 was mainly located in the luminal epithelium of the endometrium . Endometrial P43119 , Q07869 , P37231 and P48443 expression was regulated during the peri-implantation period whereas Q03181 , P19793 and P28702 were consistently expressed . In the trophoblast , Q16647 transcript levels rose as development progressed and peaked at day 17 . P43119 and Q07869 transcripts peaked before day 12 and then declined and became nearly undetectable by day 17 , whereas Q03181 and P37231 transcript levels rose steadily from days 12 to 17 . Because the PPARs and the RXRs display different expression profiles , we suggest that different heterodimers may form and support distinct functions as development proceeds . Our results also underline the importance of Q16647 and Q03181 in the trophoblast and P43119 in the uterus , suggesting that DB01240 is of both uterine and trophoblastic origin and is involved in a complex signalling pathway at around the time of implantation in the ewe . Structure-activity relationships associated with 3,4,5-triphenyl-1H-pyrazole-1-nonanoic acid , a nonprostanoid prostacyclin mimetic . A series of phenylated pyrazoloalkanoic acid derivatives were synthesized and evaluated as inhibitors of ADP-induced human platelet aggregation . 3,4,5-Triphenyl-1H-pyrazole-1-nonanoic acid ( 8d ) , with an IC50 of 0.4 microM , was the most potent inhibitor identified in this study . Biochemical studies determined that 8d increased intraplatelet DB02527 accumulation and stimulated platelet membrane-bound adenylate cyclase in a concentration-dependent fashion . Displacement of [3H]iloprost by 8d from platelet membranes indicated that the platelet prostacyclin ( DB01240 ) receptor is the locus of biological action . Structure-activity studies demonstrated that the minimum structural requirements for binding to the platelet P43119 and inhibition of ADP-induced platelet aggregation within this series are a vicinally diphenylated pyrazole substituted with an omega-alkanoic acid side chain eight or nine atoms long . Potency depended upon both side-chain length and its topological relationship with the two phenyl rings . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . Effects of inhaled prostacyclin analogue on chronic hypoxic pulmonary hypertension . Inhaled DB01240 has been reported to elicit pulmonary vasodilation , but whether it is also effective in treating chronic hypoxic pulmonary hypertension is still uncertain . We designed this study to address the in vivo effectiveness of inhaled DB05229 , a stable DB01240 analogue , on pulmonary vascular tone during hypoxic exposure in normoxic ( N ) and chronically hypoxic ( CH ) rats . Pulmonary vasodilation was observed by low-dose inhaled DB05229 in N rats , but not in CH rats . It was not until higher doses of DB05229 were given that pulmonary vasodilation was obtained in CH rats . When the agent was continuously administered by an intravascular route at the inhaled dose , it elicited no vasodilation in N rats . On the contrary , it elicited profound vasodilation in CH rats , although a concomitant systemic hypotension was observed . The P43119 mRNA expression was unchanged in the lungs of CH rats compared with that of N rats . We conclude that low doses of aerosolized DB05229 may reduce pulmonary vascular tone in rats without preexisting lung diseases . In contrast , when hypoxic pulmonary hypertension is present , the threshold of DB05229 inhalation was elevated to provoke pulmonary vasodilation . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . DB01240 ( P06744 ) receptor binding and cyclic AMP synthesis activities of PGI1 analogues , SM-10906 and its methyl ester , SM-10902 , in mastocytoma P-815 cells . The prostacyclin I1 ( PGI1 ) analogue , SM-10906 and its methyl ester , SM-10902 , have been compared with the DB01240 analogue , iloprost , with respect to binding to the P43119 , stimulation of adenylate cyclase activity and inhibition of thrombin-induced Ca2+ mobilization in mastocytoma P-815 cells . SM-10906 displaced [3H]iloprost binding to the membrane fraction , the IC50 value being 100 nM , but showed very low affinity for the prostaglandin E ( PGE ) receptor . SM-10906 dose-dependently stimulated GTP-dependent adenylate cyclase activity in the membrane fraction , the EC50 value being 35 nM . Furthermore , SM-10906 prevented a thrombin-induced increase in the intracellular Ca2+ concentration , the IC50 value being 300 nM . These IC50 and EC50 values are much lower than those of SM-10902 . These results demonstrate that SM-10906 , a stable PGI1 derivative , is an agonist for the [3H]iloprost-binding ( DB01240 ) receptor , and that it prevents thrombin-induced Ca2+ mobilization through stimulation of the adenylate cyclase system in mastocytoma cells . On the other hand , a methyl ester derivative of PGI1 , SM-10902 , was inactive in the binding assay , but it seems to be a partial agonist for adenylate cyclase activity [ corrected ] . 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 . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . P43119 up-regulates the expression of angiogenic genes in human endometrium via cross talk with epidermal growth factor Receptor and the extracellular signaling receptor kinase 1/2 pathway . DB01240 ( P06744 ) is a member of the prostanoid family of lipid mediators that mediates its effects through a seven-transmembrane G protein-coupled receptor ( IP receptor ) . Recent studies have ascertained a role for prostanoid-receptor signaling in angiogenesis . In this study we examined the temporal-spatial expression of the IP receptor within normal human endometrium and additionally explored the signaling pathways mediating the role of IP receptor in activation of target angiogenic genes . Quantitative RT-PCR analysis demonstrated the highest endometrial expression of the IP receptor during the menstrual phase compared with all other stages of the menstrual cycle . Immunohistochemical analysis localized the site of IP receptor expression to the glandular epithelial compartment with stromal and perivascular cell immunoreactivity . Expression of the immunoreactive IP receptor protein was greatest during the proliferative and early secretory phases of the menstrual cycle . To explore the role of the IP receptor in glandular epithelial cells , we used the Ishikawa endometrial epithelial cell line . Stimulation of Ishikawa cells and human endometrial biopsy explants with 100 nm iloprost ( a P06744 analog ) rapidly activated P27361 /2 signaling and induced the expression of proangiogenic genes , basic fibroblast growth factor , angiopoietin-1 , and angiopoietin-2 , in an epidermal growth factor receptor ( P00533 ) -dependent manner . Furthermore , P00533 colocalized with IP receptor in the glandular epithelial compartment . These data suggest that P06744 -IP interaction within glandular epithelial cells can promote the expression of proangiogenic genes in human endometrium via cross talk with the P00533 . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . DB01240 -deficient mice develop ischemic renal disorders , including nephrosclerosis and renal infarction . BACKGROUND : DB01240 ( DB01240 ) is a short-lived endogenous inhibitor of platelet aggregation and a potent vasodilator and regulator of the growth of vascular smooth muscle cells . To study the role of DB01240 in the vascular system in vivo , DB01240 -deficient ( PGID ) mice were established by genetic disruption of the DB01240 synthase gene . METHODS AND RESULTS : DB01240 synthase-null mice were generated by replacing the exons of DB01240 synthase gene that encodes for the catalytic site of the enzyme with a neomycin resistance gene . In these mice , DB01240 levels in the plasma , kidneys , and lungs were reduced , whereas thromboxane and prostaglandin E2 levels became elevated . Blood pressure and the amounts of urea nitrogen and creatinine in plasma of the PGID mice were significantly higher than those of wild-type mice ( P < 0.05 ) . They developed progressive morphological abnormalities in the kidneys , accompanied by atrophy , surface irregularity , fibrosis , cyst , arterial sclerosis , and hypertrophy of vessel walls . Thickening of the thoracic aortic media and adventitia were observed in aged PGID mice . Importantly , these phenotypes have not been reported in P43119 -deficient mice . CONCLUSIONS : DB01240 deficiency resulted in the development of vascular disorders with the thickening of vascular walls and interstitial fibrosis , especially in mouse kidneys . The findings demonstrated in vivo that DB01240 is important in the homeostasis of blood vessels . Our established PGID mice are useful for studies on the initiation and development of vascular diseases , such as ischemic renal disorders with arterial sclerosis and infarction , and also for studies on the novel signaling pathway of DB01240 . CNS-specific prostacyclin ligands as neuronal survival-promoting factors in the brain . DB01240 ( DB01240 ) is a critical regulator of the cardiovascular system , via dilatation of vascular smooth muscle and inhibition of platelet aggregation ( Moncada , S. 1982 , Br. J. Pharmacol. , 76 , 3 ) . Our previous studies demonstrated that a novel subtype of P43119 , which is clearly distinct from a peripheral subtype in terms of ligand specificity , is expressed in the rostral region of the brain , e.g. cerebral cortex , hippocampus , thalamus and striatum , and that ( 15R ) -16-m-17,18,19,20-tetranorisocarbacyclin ( 15R- Q8NDX1 ) and 15-deoxy-16-m-17,18,19,20-tetranorisocarbacyclin ( 15-deoxy- Q8NDX1 ) specifically bind to the central nervous system ( CNS ) -specific P43119 . Here , we report that these CNS-specific P43119 ligands , including DB01240 itself , prevented the neuronal death . They prevented apoptotic cell death of hippocampal neurons induced by high ( 50 % ) oxygen atmosphere , xanthine + xanthine oxidase , and serum deprivation . IC50s for neuronal death were approximately 30 and 300 nM for 15-deoxy- Q8NDX1 and 15R- Q8NDX1 , respectively , which well correlated with the binding potency for the CNS-specific P43119 . 6-Keto-PGF1alpha ( a stable metabolite of DB01240 ) , peripheral nervous system-specific DB01240 ligands and other prostaglandins ( PGs ) than DB01240 did not show such neuroprotective effects . In vivo , 15R- Q8NDX1 protected P00915 pyramidal neurons against ischaemic damage in gerbils . These results indicate that CNS-specific DB01240 ligands have neuronal survival-promoting activity both in vitro and in vivo , and may represent a new type of therapeutic drug for neurodegeneration . SynGAP- O75970 -CaMKII synaptic complexes regulate p38 Q96HU1 kinase activity and DB01221 receptor-dependent synaptic AMPA receptor potentiation . The synapse contains densely localized and interacting proteins that enable it to adapt to changing inputs . We describe a Ca2+-sensitive protein complex involved in the regulation of AMPA receptor synaptic plasticity . The complex is comprised of MUPPI , a multi-PDZ domain-containing protein ; SynGAP , a synaptic P20936 ; and the Ca2+/calmodulin-dependent kinase CaMKII . In synapses of hippocampal neurons , SynGAP and CaMKII are brought together by direct physical interaction with the PDZ domains of O75970 , and in this complex , SynGAP is phosphorylated . Ca2+ P62158 binding to CaMKII dissociates it from the O75970 complex , and Ca2+ entering via the NMDAR drives the dephosphorylation of SynGAP . Specific peptide-induced SynGAP dissociation from the O75970 -CaMKII complex results in SynGAP dephosphorylation accompanied by O75791 MAPK inactivation , potentiation of synaptic AMPA responses , and an increase in the number of AMPAR-containing clusters in hippocampal neuron synapses . siRNA-mediated SynGAP knockdown confirmed these results . These data implicate SynGAP in NMDAR- and CaMKII-dependent regulation of AMPAR trafficking . Pharmacophore definition and three-dimensional quantitative structure-activity relationship study on structurally diverse prostacyclin receptor agonists . DB01240 is an endogenous mediator that shows potent platelet inhibitory activity and powerful relaxation of peripheral resistance vessels . P43119 agonists are valuable drugs in the treatment of various vascular diseases spanning primary pulmonary hypertension to Raynaud 's syndrome . Although agonists from various structural classes were synthesized , a common pharmacophore was never defined . Therefore , an attempt was made to integrate the different agonists into a single model . A dataset of structurally diverse prostacyclin receptor agonists was tested for its affinity to the human platelet prostacyclin receptor . The dataset included prostanoid and nonprostanoid ligands comprising iloprost , cicaprost , and BMY45778 . Extensive conformational analyses were performed for both classes of compounds because of the absence of rigid templates . The search and superimposition procedure yielded a pharmacophore that aligns the essential carboxylate group of the agonists as well as demonstrates that different functional groups in prostanoid and nonprostanoid agonists can be arranged in a uniform conformation . A three-dimensional quantitative structure-activity relationship study was performed using the programs O75791 and GOLPE . This analysis yielded a cross-validated correlation coefficient of 0.77 . With this model , it is possible to predict the affinity of untested compounds . DB01240 -IP signaling and prostaglandin E2-EP2/EP4 signaling both mediate joint inflammation in mouse collagen-induced arthritis . Prostaglandin (PG)I2 ( prostacyclin [ P06744 ] ) and DB00917 are abundantly present in the synovial fluid of rheumatoid arthritis ( RA ) patients . Although the role of DB00917 in RA has been well studied , how much DB01240 contributes to RA is little known . To examine this issue , we backcrossed mice lacking the P43119 ( IP ) to the DBA/1J strain and subjected them to collagen-induced arthritis ( CIA ) . IP-deficient ( IP-/- ) mice exhibited significant reduction in arthritic scores compared with wild-type ( WT ) mice , despite anti-collagen antibody production and complement activation similar to WT mice . IP-/- mice also showed significant reduction in contents of proinflammatory cytokines , such as interleukin ( IL ) -6 in arthritic paws . Consistently , the addition of an IP agonist to cultured synovial fibroblasts significantly enhanced P05231 production and induced expression of other arthritis-related genes . On the other hand , loss or inhibition of each PGE receptor subtype alone did not affect elicitation of inflammation in CIA . However , a partial but significant suppression of CIA was achieved by the combined inhibition of EP2 and EP4 . Our results show significant roles of both DB01240 -IP and DB00917 -EP2/EP4 signaling in the development of CIA , and suggest that inhibition of DB00917 synthesis alone may not be sufficient for suppression of RA symptoms . P08700 interleukin-5 , and granulocyte-macrophage colony-stimulating factor expression in nasal polyps . PURPOSE : Nasal polyps ( NP ) are grape-like clusters of chronically inflamed tissue . Little is known about the underlying cells and cytokines involved in nasal polyposis . For the present study , we hypothesize that elevated tissue levels of interleukin-3 ( P08700 ) , interleukin-5 ( P05113 ) , and granulocyte-macrophage colony-stimulation factor ( GM- P04141 ) contribute to eosinophil recruitment and activation in NP . MATERIALS AND METHODS : To begin to test this hypothesis , we evaluated P08700 , P05113 , and GM- P04141 levels and distributions in nasal polyp specimens obtained intraoperatively from 13 patients and two normal controls . For these studies , nasal polyp levels were determined by enzyme-linked immunosorbent assay ( ELISA ) , and P08700 , P05113 , and GM- P04141 distribution was determined by immunohistochemistry . RESULTS : Immunohistochemical staining of the NP indicated that in all 13 patient samples , P08700 , P05113 , and GM- P04141 were associated with infiltrating cells , primarily eosinophils , in the NP . Quantitation of P08700 , P05113 , and GM- P04141 in NP tissue homogenates indicated that P08700 , P05113 , and GM- P04141 levels were evaluated in the NP tissues when compared with control tissues . Additionally , elevation of individual cytokines correlated with previous polypectomy ( P08700 ) , steroid use ( P08700 , P05113 , and GM- P04141 ) , asthma ( P05113 ) , and age ( GM- P04141 ) . CONCLUSION : These data support our hypothesis that P08700 , P05113 , and GM- P04141 are likely to play a key role in eosinophil recruitment/activation and NP formation and support recently advanced theories that cytokines play a key role in the pathogenesis of this disease . Different expression of PGE synthase , P43088 , P01375 , Fas and oxytocin in the bovine corpus luteum of the estrous cycle and pregnancy . Functional differences between the corpus luteum ( CL ) of pregnancy and CL of the cycle in cows were examined . Messenger RNA and protein levels of prostaglandin ( PG ) E synthase ( O14684 ) , PGF2α receptor ( PGFR ) , tumor necrosis factor-α ( P01375 ) and Fas were found to be higher in the CL of pregnancy than in CL of the cycle . DB00107 ( OT ) mRNA and protein levels were lower in the CL of pregnancy . Messenger RNA levels of progesterone receptor ( PR ) , luteinizing hormone receptor ( LHR ) , DB00917 receptor ( PGER ) , P49763 synthase ( P42330 ) , P01375 receptor type I ( TNFRI ) and P01375 receptor type II ( TNFRII ) did not differ between the cycle and pregnancy . DB00917 and PGF2α production by cultured bovine endometrial tissues was decreased by a supernatant derived from the homogenized CL of pregnancy but not by that of the CL of the cycle , suggesting that specific substances in the CL of pregnancy affect endometrial PG production in cows . Collectively , O14684 , PGFR , P01375 , Fas or OT may contribute to differences between the CL of pregnancy and CL of the estrous cycle in cows . Effects of inhalational anesthetics on alpha2-adrenergic signaling in isolated platelets . ( 1 ) The hypothesis that inhalational anesthetics affect G-protein linked alpha2 adrenergic signaling pathway was investigated using human platelets as a model system . ( 2 ) Alpha2 receptor stimulation by UK-14304 , a potent and selective agonist , inhibits DB02527 production induced by prostaglandin I2 ( DB01240 ) . ( 3 ) Brief stimulation ( 30 s ) with DB01240 raised DB02527 levels in platelets by 25-fold ; UK-14304 suppressed the DB01240 stimulus by 80 % . ( 4 ) Halothane at fractional minimum alveolar concentration ( MAC ) through super physiological levels ( 16 MAC ) had no effect on basal or prostacyclin stimulated levels of DB02527 , nor did it have any effect on the inhibition of DB02527 production by UK-14304 . Moreover , isoflurane , enflurane and sevoflurane had no significant effect on DB02527 production at 1.5 or 8 MAC . The results suggest alpha2 and DB01240 signaling pathways are not sensitive to volatile anesthetics including the alpha2 or P43119 /G-protein complex , G-protein/adenylyl cyclase complex and adenylyl cyclase itself . ( 5 ) The possibility that halothane and related anesthetics act more distally in the pathway , on DB02527 -dependent protein kinase ( PKA ) , was investigated by measuring the phosphorylation pattern of endogenous platelet proteins by PKA . ( 6 ) An increase in the [32P]phosphate incorporation was observed in platelets exposed to either , low doses of DB01240 or isobutylmethylxanthine ( DB07954 ) . Halothane , isoflurane , enflurane or sevoflurane further increased the level of [ 32P ] -incorporation . The apparent increase in PKA activity suggests that at least in platelets , volatile anesthetics activate PKA-dependent pathways which should antagonize alpha2 adrenergic signaling .