MH_dev_257

Structure of the human Q9H244 REA receptor in complex with an antithrombotic drug . P2Y receptors ( P2YRs ) , a family of purinergic G-protein-coupled receptors ( GPCRs ) , are activated by extracellular nucleotides . There are a total of eight distinct functional P2YRs expressed in human , which are subdivided into P47900 REA - like receptors and Q9H244 REA - like receptors . Their ligands are generally charged molecules with relatively low bioavailability and stability in vivo , which limits our understanding of this receptor family . P2Y12R regulates platelet activation and thrombus formation , and several antithrombotic drugs targeting P2Y12R - - including the prodrugs clopidogrel ( Plavix ) and prasugrel ( DB06209 SUB ) that are metabolized and bind covalently , and the nucleoside analogue ticagrelor ( DB08816 ) that acts directly on the receptor - - have been approved for the prevention of stroke and myocardial infarction . However , limitations of these drugs ( for example , a very long half-life of clopidogrel action and a characteristic adverse effect profile of ticagrelor ) suggest that there is an unfulfilled medical need for developing a new generation of P2Y12R inhibitors . Here we report the 2.6 Å resolution crystal structure of human P2Y12R in complex with a non-nucleotide reversible antagonist , AZD 1283 . The structure reveals a distinct straight conformation of helix V , which sets P2Y12R apart from all other known class A GPCR structures . With AZD 1283 bound , the highly conserved disulphide bridge in GPCRs between helix III and extracellular loop 2 is not observed and appears to be dynamic . Along with the details of the AZD 1283 - binding site , analysis of the extracellular interface reveals an adjacent ligand-binding region and suggests that both pockets could be required for dinucleotide binding . The structure provides essential insights for the development of improved P2Y12R ligands and allosteric modulators as drug candidates .

Glycoprotein IIb / IIIa and Q9H244 REA receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 REA release and procoagulant responses . Glycoprotein IIb / IIIa ( P08514 REA / IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 REA antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 REA antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 REA and soluble P29965 REA levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and / or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 REA / IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 REA / IIIa antagonists and inhibitors of both Q9H244 REA receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies .

DB01233 MEN does not increase gastric muscle contractility in newborn rats . Feeding intolerance resulting from delayed gastric emptying is common in premature neonates . DB01233 MEN ( MCP ) , the most frequently used prokinetic drug in neonates , enhances gastric muscle contractility through inhibition of dopamine receptors . Although its therapeutic benefit is established in adults , limited data are available to support its clinical use in infants . Hypothesizing that developmentally dependent differences are present , we comparatively evaluated the effect of MCP on fundus muscle contractility in newborn , juvenile , and adult rats . The muscle strips were either contracted with electrical field stimulation ( O43281 ) to induce cholinergic nerve-mediated acetylcholine release or carbachol , a cholinergic agonist acting directly on the muscarinic receptor . Although in adult rats MCP increased O43281 - induced contraction by 294 ± 122 % of control ( P < 0.01 ) , no significant effect was observed in newborn fundic muscle . MCP had no effect on the magnitude of the carbachol-induced and / or bethanechol-induced gastric muscle contraction at any age . In response to dopamine , an 80.7 ± 5.3 % relaxation of adult fundic muscle was observed , compared with only a 8.4 ± 8.7 % response in newborn tissue ( P < 0.01 ) . P14416 REA expression was scant in neonates and significantly increased in adult gastric tissue ( P < 0.01 ) . In conclusion , the lack of MCP effect on the newborn fundic muscle contraction potential relates to developmental differences in dopamine D2 receptor expression . To the extent that these novel data can be extrapolated to neonates , the therapeutic value of MCP as a prokinetic agent early in life requires further evaluation .

DB00360 MEN , a cofactor for NOS , improves endothelial dysfunction during chronic alcohol consumption . We sought to investigate mechanisms that may account for impaired nitric oxide synthase ( NOS ) - dependent dilatation of cerebral arterioles during alcohol consumption . Our goals were to examine 1 ) the effect of exogenous application of a cofactor for NOS , i . e . , tetrahydrobiopterin ( BH4 ) on the reactivity of pial arterioles during alcohol consumption ; and 2 ) endothelial NOS ( P29474 REA ) protein in nonalcohol-fed and alcohol-fed rats . Sprague-Dawley rats were fed liquid diets with or without alcohol for 2-3 mo . We measured in vivo diameter of pial arterioles in response to NOS-dependent agonists ( ACh and ADP ) and a NOS-independent agonist ( nitroglycerin ) before and during application of BH4 . Blood vessels were then harvested for Western blot analysis of P29474 REA protein . In nonalcohol-fed rats , ACh and ADP produced vasodilatation , which was impaired in alcohol-fed rats . Vasodilatation to nitroglycerin was similar in both groups of rats . Application of BH4 did not alter vasodilatation in nonalcohol-fed rats but improved impaired vasodilatation in alcohol-fed rats . Also , P29474 REA protein in cerebral cortex microvessels , the basilar artery , and aorta was not different between nonalcohol-fed and alcohol-fed rats . Thus impaired NOS-dependent vasodilatation during alcohol consumption does not appear to be related to an alteration in P29474 REA protein but may be related to a deficiency and / or alteration in the utilization of BH4 .

Feasibility of a microarray-based point-of-care P33261 REA genotyping test for predicting clopidogrel on-treatment platelet reactivity . DB00758 is a prodrug which is converted into active metabolite by cytochrome P450 isoenzyme , P33261 REA . Numerous polymorphisms of P33261 REA are reported , and a strong link exists between loss-of-function ( LOF ) or gain-of-function polymorphisms , clopidogrel metabolism , and clinical outcome . Hence , a fully automated point-of-care P33261 REA genotyping assay is more likely to bring personalized antiplatelet therapy into real practice . We assessed the feasibility of the Verigene 2C19 / P35520 REA Nucleic Acid Test , a fully automated microarray-based assay , compared to bidirectional sequencing , and performed VerifyNow Q9H244 REA assay to evaluate the effect of P33261 REA polymorphisms on on-treatment platelet reactivity in 57 Korean patients treated with clopidogrel after percutaneous coronary intervention . The Verigene 2C19 / P35520 REA assay identified ∗ 2 , ∗ 3 , and ∗ 17 polymorphisms with 100 % concordance to bidirectional sequencing in 180 minutes with little hands-on time . Patients were classified into 4 groups : extensive ( ∗ 1 / ∗ 1 ; n = 12 , 21.1 % ) , intermediate ( ∗ 1 / ∗ 2 , ∗ 1 / ∗ 3 ; n = 33 , 57.9 % ) , poor ( ∗ 2 / ∗ 2 , ∗ 2 / ∗ 3 , and ∗ 3 / ∗ 3 ; n = 11 , 19.3 % ) , and ultrarapid metabolizers ( ∗ 1 / ∗ 17 ; n = 1 , 1.8 % ) . The prevalence of the CYP 2C19 ∗ 2 , ∗ 3 , and ∗ 17 alleles was 36.0 % , 12.3 % , and 0.9 % . Platelet reactivity showed gene dose response according to the number of P33261 REA LOF allele . In conclusion , the Verigene 2C19 / P35520 REA assay gave accurate P33261 REA genotype results which were in well match with the differing on-treatment platelet reactivity .

Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 REA , Q16678 REA , P11712 REA , P33261 REA , P05181 REA , P05093 REA , P11511 REA , P35869 REA , P03372 REA , Q92731 REA , ERRRG , P06401 REA , P07099 REA , P34913 REA , P37059 REA , P37058 REA , P28161 REA , P21266 REA , GSTT 2 , P09211 REA , NAT 1 , NAT 2 , P21964 REA , P07327 REA , P00325 REA , P00326 REA , P05091 REA , P35228 REA , NOS 3 , P01583 REA , P01584 REA , O15527 REA , P36639 REA [ P36639 REA ] , P14416 REA , P35462 REA , P21917 REA , P31645 REA , P04150 REA [ GCCR ] , P42898 REA , and P15559 REA . In the present study , the Japanese allele frequencies were verified by using nationwide population samples .

Mechanism of purinergic activation of endothelial nitric oxide synthase in endothelial cells . BACKGROUND : Decreased endothelial nitric oxide ( NO ) synthase ( P29474 REA ) activity and NO production are critical contributors to the endothelial dysfunction and vascular complications observed in many diseases , including diabetes mellitus . Extracellular nucleotides activate P29474 REA and increase NO generation ; however , the mechanism of this observation is not fully clarified . METHODS AND RESULTS : To elucidate the signaling pathway ( s ) leading to nucleotide-mediated P29474 REA phosphorylation at DB00133 - 1177 , human umbilical vein endothelial cells were treated with several nucleotides , including DB00171 , UTP , and ADP , in the presence or absence of selective inhibitors . These experiments identified P47900 REA , P41231 REA , and possibly P51582 REA as the purinergic receptors involved in P29474 REA phosphorylation and demonstrated that this process was adenosine independent . Nucleotide-induced P29474 REA phosphorylation and activity were inhibited by BAPTA-AM ( an intracellular free calcium chelator ) , rottlerin ( a protein kinase Cdelta inhibitor ) , and protein kinase Cdelta siRNA . In contrast , blockade of AMP-activated protein kinase , calcium / calmodulin-dependent kinase II , calcium / calmodulin-dependent kinase kinase , serine / threonine protein kinase B , protein kinase A , extracellular signal-regulated kinase 1/2 , and p38 mitogen-activated protein kinase did not affect nucleotide-mediated P29474 REA phosphorylation . CONCLUSIONS : The present study indicates that extracellular nucleotide-mediated P29474 REA phosphorylation is calcium and protein kinase Cdelta dependent . This newly identified signaling pathway opens new therapeutic avenues for the treatment of endothelial dysfunction .

DB00171 induces synaptic gene expressions in cortical neurons : transduction and transcription control via P47900 REA receptors . Studies in vertebrate neuromuscular synapses have revealed previously that DB00171 , via P2Y receptors , plays a critical role in regulating postsynaptic gene expressions . An equivalent regulatory role of DB00171 and its P2Y receptors would not necessarily be expected for the very different situation of the brain synapses , but we provide evidence here for a brain version of that role . In cultured cortical neurons , the expression of P2Y ( 1 ) receptors increased sharply during neuronal differentiation . Those receptors were found mainly colocalized with the postsynaptic scaffold postsynaptic density protein 95 ( P78352 REA ) . This arises through a direct interaction of a PDZ domain of P78352 REA with the C-terminal PDZ-binding motif , D-T-S-L of the P2Y ( 1 ) receptor , confirmed by the full suppression of the colocalization upon mutation of two amino acids therein . This interaction is effective in recruiting P78352 REA to the membrane . Specific activation of P2Y ( 1 ) ( G-protein-coupled ) receptors induced the elevation of intracellular Ca ( 2 + ) and activation of a mitogen-activated protein kinase / P04049 REA signaling cascade . This led to distinct up-regulation of the genes encoding acetylcholinesterase ( P22303 REA ( T ) variant ) , choline acetyltransferase , and the N-methyl-d-aspartate receptor subunit Q12879 REA . This was confirmed , in the example of P22303 REA , to arise from P2Y ( 1 ) - dependent stimulation of a human P22303 REA gene promoter . That involved activation of the transcription factor Elk - 1 ; mutagenesis of the P22303 REA promoter revealed that Elk - 1 binding at its specific responsive elements in that promoter was induced by P2Y ( 1 ) receptor activation . The combined findings reveal that DB00171 , via its P2Y ( 1 ) receptor , can act trophically in brain neurons to regulate the gene expression of direct effectors of synaptic transmission .

Establishment by adriamycin exposure of multidrug-resistant rat ascites hepatoma AH130 cells showing low P15559 REA activity and high cross resistance to mitomycins . A resistant subline ( AH130 / 5A ) selected from rat hepatoma AH130 cells after exposure to adriamycin ( P35318 REA ) showed remarkable resistance to multiple antitumor drugs , including mitomycin C ( DB00305 ) and porfiromycin ( PFM ) . PFM , vinblastine ( DB00570 ) , and P35318 REA accumulated in AH130 / 5A far less than in the parent AH130 ( AH130 / P ) cells . AH130 / 5A cells showed overexpression of P-glycoprotein ( A6NDG6 ) , an increase in glutathione S-transferase activity , and a decrease in P15559 REA and glutathione peroxidase activity . The resistance to DB00305 and DB00570 of AH130 / 5A cells was partly reversed by H - 87 , an inhibitor of A6NDG6 . Buthionine sulfoximine , an inhibitor of glutathione synthase , did not affect the action of DB00305 . tert-Butylhydroquinone induced P15559 REA activity , increased PFM uptake , and enhanced the growth-inhibitory action of DB00305 in AH130 / 5A cells . DB00266 MENMAX DB00266 MEN , an inhibitor of P15559 REA , decreased PFM uptake and reduced the growth-inhibitory action of DB00305 in AH130 / P cells . These results indicated that the adriamycin treatment of hepatoma cells caused multifactorial multidrug resistance involving a decrease in P15559 REA activity .

Detection of thymidylate synthase modulators by a novel screening assay . P04818 REA ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5 - fluorouracil ( DB00544 ) , 5 - fluorouridine ( DB01629 ) , 5 - fluoro - 2 ' - deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 MEN ) ; to the nonpyrimidine TS-inhibitors AG - 331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5 - azacytidine , 8 - thioguanosine ) . Except for 5 - azacytidine and 8 - thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630 - P13671 REA cells with DB00544 , DB01629 , FUdR , DB00432 MEN , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly .

Q9H244 REA ( - ) - receptor agonists enhance the proliferation of rat P13671 REA glioma cells through activation of the Q8NFH3 / 44 mitogen-activated protein kinase . 1 . Extracellularly added P ( 1 ) , P ( 3 ) - di ( adenosine - 5 ' ) triphosphate ( Ap ( 3 ) A ) , P ( 1 ) , P ( 4 ) - di ( adenosine - 5 ' ) tetraphosphate ( Ap ( 4 ) A ) , DB00171 , ADP , AMP and adenosine are growth inhibitory for rat P13671 REA glioma cells . Analysis of nucleotide hydrolysis and the use of nucleotidase inhibitors demonstrated that the latter inhibition is due to hydrolysis of the nucleotides to adenosine . 2 . Agonists of the Q9H244 REA ( - ) - receptor enhance the growth of P13671 REA cells if their hydrolysis to adenosine is inhibited by pyridoxalphosphate - 6 - azophenyl - 2 ' , 4 ' - disulfonic acid ( PPADS ) . In these conditions , the potency to stimulate cell growth parallels the ranking of the receptor agonists , i . e . 2 - methylthioadenosine - 5 ' - diphosphate ( 2MeSADP ) > Ap ( 3 ) A > Ap ( 4 ) A . DB00171 and ADP are still hydrolysed in the presence of PPADS and have no proliferative effect on P13671 REA cells . 3 . The enhanced growth is due to a Q9H244 REA ( - ) - receptor-mediated activation of Q8NFH3 / 44 mitogen-activated protein kinase ( MAPK ) as shown by immunoblotting and protein kinase assays for active MAPK and the use of the MAPK / extracellular signal-regulated kinase kinase ( MEK ) inhibitor PD98059 . 4 . The UTP-induced enhancement of the growth of P13671 REA cells is due to activation of MAPK by a PPADS sensitive nucleotide receptor . 5 . In conclusion , the effect of nucleotides on the growth of P13671 REA cells is determined by ecto-nucleotidases and by activation of nucleotide receptors . Hydrolysis of nucleotides to adenosine induces growth inhibition while inhibition of the hydrolysis of agonists of the Q9H244 REA ( - ) - receptor enhances cell growth by activation of MAPK .

Membranous glomerulonephritis with the use of etanercept in ankylosing spondylitis . OBJECTIVE : To report a case of membranous glomerulonephritis with the use of etanercept in a patient with ankylosing spondylitis . CASE SUMMARY : A 60 - year-old female with severe ankylosing spondylitis and no history of renal disease was started on etanercept 50 mg subcutaneously once a week after standard treatment modalities failed . She had not been on any nephrotoxic drugs , including nonsteroidal antiinflammatory drugs , for over 1 year . After 2 months of etanercept therapy , the patient presented with anasarca , proteinuria , hypoalbuminemia , and normal serum creatinine . Renal biopsy showed features classic for membranous glomerulonephritis ( MGN ) . DB00005 MEN was discontinued , with resolution of anasarca and proteinuria over the following 3 months . DISCUSSION : Very few cases of MGN in patients with ankylosing spondylitis have been reported in the literature ; additionally , MGN following etanercept therapy has not been described in patients with ankylosing spondylitis so far . The exact pathogenesis of etanercept-induced MGN in ankylosing spondylitis is not known . P01375 REA - α antagonists have been associated with the formation of autoantibodies and induction of lupus-like ailments in patients with rheumatoid arthritis . It is possible that a similar dysregulation of the immune system might have accounted for the development of MGN in this patient . The Naranjo probability scale showed that this patient ' s MGN was probably associated with etanercept therapy . CONCLUSIONS : In addition to the well-described serious adverse drug events , including serious infections , the use of etanercept in ankylosing spondylitis can be associated with the development of MGN . Health care providers , especially rheumatologists , should be aware of this potentially serious adverse drug reaction .

Purinergic P47900 REA receptor signaling mediates wound stimuli-induced cyclooxygenase - 2 expression in intestinal subepithelial myofibroblasts . Intestinal subepithelial myofibroblasts ( ISMFs ) are crucial for barrier formation against inflammatory stimuli . Physical injury induces cyclooxygenase - 2 ( P35354 REA ) expression , which accelerates wound healing by ISMFs . However , the mechanism of P35354 REA induction remains unclear . Physically damaged cells release DB00171 . Here , we investigate the role of DB00171 - purinergic signaling in wound-induced P35354 REA induction in ISMFs . By 24h post-injury , bovine ISMFs had migrated to and closed the wounded area . A P36551 REA inhibitor , indomethacin or a purinergic P2 receptor antagonist , suramin , inhibited wound healing . However , additional treatment with indomethacin did not influence wound healing in suramin-treated ISMFs . RT-PCR showed an increase in P35354 REA mRNA expression 2h post-injury , which was inhibited by suramin . These results suggest that DB00171 mediates wound-induced P35354 REA elevation . We next assessed the contribution of various purinergic receptors in P35354 REA induction . An DB00171 analog , ATPγS and a purinergic P47900 REA , 11-13 receptors agonist , ADP , were among the agents tested which increased P35354 REA expression . ATPγS-induced P35354 REA mRNA expression was suppressed by suramin or a purinergic P2Xs , P47900 REA , 4 , 6 , and 13 receptors antagonist , PPADS . These data suggest the involvement of Gq-coupled purinergic P47900 REA receptor or Gi-coupled purinergic Q9BPV8 receptor in P35354 REA induction . U73122 , an inhibitor of phospholipase C , which is a downstream signal of Gq protein , showed suppression of P35354 REA mRNA expression . However , pertussis toxin , a Gi inhibitor , did not show suppression . We also revealed that inhibitors of p38 MAPK and PKC inhibited ATPγS-induced P35354 REA mRNA expression . Collectively , purinergic P47900 REA receptor signaling mediates wound-induced P35354 REA expression through p38 MAPK and PKC pathways in ISMFs .

Monitoring the expression profiles of doxorubicin-resistant K562 human leukemia cells by serial analysis of gene expression . We examined the expression profiles of doxorubicin-resistant K562 cells by serial analysis of gene expression ( Q9NXZ1 ) to identify novel and / or partially characterized genes that might be related to drug resistance in human leukemia . Q9NXZ1 complementary DNA ( cDNA ) libraries were constructed from K562 and doxorubicin-resistant K562 ( K562 / P35318 REA ) cells , and concatamer sequences were analyzed with Q9NXZ1 2000 software . We used 9792 tags in the identification of 1076 different transcripts , 296 of which were similarly expressed in K562 and K562 / P35318 REA cells . There were 343 genes more actively expressed in K562 / P35318 REA than in parental K562 cells and 437 genes expressed less often in K562 / P35318 REA cells . K562 / P35318 REA cells showed increased expression of well-known genes , including the genes for spectrin beta , eukaryotic translation initiation factor 1A ( P47813 REA ) , RAD 23 homolog B , laminin receptor 1 , and polyA - , RAN - , and P05121 REA messenger RNA-binding proteins . K562 / P35318 REA cells showed decreased expression of the genes for fatty acid desaturase 1 ( O60427 REA ) , hemoglobin epsilon 1 , N-myristoyltransferase 1 , hemoglobin alpha 2 , DB00157 dehydrogenase Fe - Q15517 REA 6 , heat shock 90 - kDa protein , and karyopherin beta 1 . Quantitative reverse transcription-polymerase chain reaction analysis confirmed the increased expression of P47813 REA and the decreased expression of O60427 REA in K562 / P35318 REA cells . Prior to this investigation , such differences in the expression of these genes in doxorubicin-resistant leukemia cells were unknown . Although we do not provide any evidence in the present report for the potential roles of these genes in drug resistance , Q9NXZ1 may provide a perspective into our understanding of drug resistance in human leukemia that is different from that provided by cDNA microarray analysis .

DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 REA , Q9H244 REA , and Q9BPV8 receptors ; the DB00171 / UTP-specific P41231 REA receptor ; and the DB00171 - selective Q96G91 REA receptor . ADP ( 0.05- 50 muM ) induced calcium flux that was completely blocked by a P47900 REA receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 REA and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 REA - and Q9H244 REA - selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 REA in response to the O60603 REA ligand , peptidoglycan , and blocked the production of P01375 REA , P10145 REA , and MIP - 1beta in response to leukotriene D ( 4 ) . These effects were mimicked by two DB00171 analogues , adenosine 5 ' - O - ( 3 - thiotriphosphate ) and 2 ' , 3 ' - O - ( 4 - benzoyl-benzoyl ) adenosine 5 ' - triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5 ' - O - ( 3 - thiotriphosphate ) , and 2 ' , 3 ' - O - ( 4 - benzoyl-benzoyl ) adenosine 5 ' - triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G ( s ) - coupled ADP / DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs .

The effects of a cyclooxygenase - 2 ( P35354 REA ) expression and inhibition on human uveal melanoma cell proliferation and macrophage nitric oxide production . BACKGROUND : P35354 REA ( P35354 REA ) expression has previously been identified in uveal melanoma although the biological role of P35354 REA in this intraocular malignancy has not been elucidated . This study aimed to investigate the effect of a P35354 REA inhibitor on the proliferation rate of human uveal melanoma cells , as well as its effect on the cytotoxic response of macrophages . METHODS : Human uveal melanoma cell lines were transfected to constitutively express P35354 REA and the proliferative rate of these cells using two different methods , with and without the addition of Amfenac , was measured . DB00435 production by macrophages was measured after exposure to melanoma-conditioned medium from both groups of cells as well as with and without Amfenac , the active metabolite of DB06802 MEN . RESULTS : Cells transfected to express P35354 REA had a higher proliferation rate than those that did not . The addition of Amfenac significantly decreased the proliferation rate of all cell lines . DB00435 production by macrophages was inhibited by the addition of melanoma conditioned medium , the addition of Amfenac partially overcame this inhibition . CONCLUSION : Amfenac affected both P35354 REA transfected and non-transfected uveal melanoma cells in terms of their proliferation rates as well as their suppressive effects on macrophage cytotoxic activity .

Antagonist properties of Conus parius peptides on N-methyl-D-aspartate receptors and their effects on CREB signaling . Three members of a family of small neurotoxic peptides from the venom of Conus parius , conantokins ( Con ) Pr1 , Pr2 , and Pr3 , function as antagonists of N-methyl-D-aspartate receptors ( NMDAR ) . We report structural characterizations of these synthetic peptides , and also demonstrate their antagonistic properties toward ion flow through NMDAR ion channels in primary neurons . ConPr 1 and ConPr 2 displayed moderate increases in α-helicity after addition of Mg ( 2 + ) . Native apo-ConPr 3 possessed an α-helical conformation , and the helicity increased only slightly on addition of Mg ( 2 + ) . Additionally , these peptides diminished DB01221 / DB00145 MEN - mediated currents and intracellular Ca ( 2 + ) ( iCa ( 2 + ) ) influx in mature rat primary hippocampal neurons . Electrophysiological data showed that these peptides displayed slower antagonistic properties toward the NMDAR than conantokins from other species of cone snails , e . g . , ConT and ConG . Furthermore , to demonstrate selectivity of the C . parius-derived conantokins towards specific NMDAR subunits , cortical neurons from Q12879 REA ( - / - ) and Q13224 REA ( - / - ) mice were utilized . Robust inhibition of NMDAR-mediated stimulation in Q12879 REA ( - / - ) - derived mouse neurons , as compared to those isolated from Q13224 REA ( - / - ) - mouse brains , was observed , suggesting a greater selectivity of these antagonists towards the Q13224 REA subunit . These C . parius conantokins mildly inhibited NMDAR-induced phosphorylation of CREB at DB00133 ( 133 ) , suggesting that the peptides modulated iCa ( 2 + ) entry and , thereby , activation of CREB , a transcription factor that is required for maintaining long-term synaptic activity . Our data mechanistically show that while these peptides effectively antagonize NMDAR-directed current and iCa ( 2 + ) influx , receptor-coupled CREB signaling is maintained . The consequence of sustained CREB signaling is improved neuronal plasticity and survival during neuropathologies .

Inhibition of P51587 REA and DB01643 MEN Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 REA mediates homologous recombination repair , and P51587 REA polymorphisms increase cancer risk . However , tumors with P51587 REA mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 REA ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 REA synergistically potentiated drugs with mechanisms of action related to P51587 REA function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality . " TS knockdown induced complementary lethality to TS-targeting drugs ( 5 - FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 REA and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 REA and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 REA - targeting antisense oligdeoxynucleotide ( ASO ) " BR - 1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi : 10.1038 / mtna . 2013.7 published online 12 March 2013 .

Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 SUB , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 REA receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk : benefit ratio will likely play a major role in directing the best place for therapy with this new agent .

P51587 REA and P8 4022 synergize in regulation of gene transcription . P8 4022 is an essential component in the intracellular signaling of transforming growth factor-beta ( TGFbeta ) , which is a potent inhibitor of tumor cell proliferation . P51587 REA is a tumor suppressor involved in early onset of breast , ovarian and prostate cancer . Both P8 4022 and P51587 REA possess transcription activation domains . Here , we show that P8 4022 and P51587 REA interact functionally and physically . We found that P51587 REA forms a complex with P8 4022 in vitro and in vivo , and that both MH1 and MH2 domains of P8 4022 contribute to the interaction . TGFbeta 1 stimulates interaction of endogenous P8 4022 and P51587 REA in non-transfected cells . P51587 REA co-activates P8 4022 - dependent transcriptional activation of luciferase reporter and expression of plasminogen activator inhibitor - 1 ( P05121 REA ) . P8 4022 increases the transcriptional activity of P51587 REA fused to the DNA-binding domain ( DBD ) of Gal 4 , and reciprocally , P51587 REA co-activates DBD-Gal 4 - P8 4022 . Thus , our results show that P51587 REA and P8 4022 form a complex and synergize in regulation of transcription .

Pharmacological Q9BWK5 mapping of age-associated changes in basal ganglia circuitry of awake rhesus monkeys . While the pathophysiological changes induced by the loss of dopamine innervation in the basal ganglia by Parkinson ' s disease ( PD ) are well studied , little is known about functional changes in the neural circuitry of this area during normal aging . Here we report the first survey of age-associated changes in the basal ganglia of behaviorally characterized , awake rhesus monkeys , using pharmacological Q9BWK5 to map responses to dopaminergic stimulation . DB00714 MEN , a mixed D ( 1 ) / P14416 REA agonist , evoked little change in the substantia nigra ( SN ) of aged animals while significantly reducing activation in young adult monkeys . Compared to young animals , both apomorphine and DB01576 ( which increases synaptic dopamine levels ) significantly increased activation of the aged rhesus globus pallidus externa ( GPe ) . In addition , the aged animals showed decreased activity in the putamen in response to DB01576 administration . Although the responses in the SN and putamen of the aged monkeys differed from those in animal models of PD , the apomorphine-evoked activation of their GPe corresponded with apomorphine-induced increases in neuronal activity seen in Parkinson ' s patients and animal models . Given the major role of the GPe in regulating motor behavior , the altered responses in the aged GPe may contribute significantly to the motor slowing and movement dysfunctions characterizing advanced age .