interacts_with DB06209?
Structure of the human Q9H244 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 -like receptors and Q9H244 -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 ) 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 , AZD1283 . The structure reveals a distinct straight conformation of helix V , which sets P2Y12R apart from all other known class A GPCR structures . With AZD1283 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 AZD1283-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 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 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 and soluble P29965 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 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 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 does not increase gastric muscle contractility in newborn rats . Feeding intolerance resulting from delayed gastric emptying is common in premature neonates . DB01233 ( 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 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 , 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 ) 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 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 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 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 genotyping test for predicting clopidogrel on-treatment platelet reactivity . DB00758 is a prodrug which is converted into active metabolite by cytochrome P450 isoenzyme , P33261 . Numerous polymorphisms of P33261 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 genotyping assay is more likely to bring personalized antiplatelet therapy into real practice . We assessed the feasibility of the Verigene 2C19/ P35520 Nucleic Acid Test , a fully automated microarray-based assay , compared to bidirectional sequencing , and performed VerifyNow Q9H244 assay to evaluate the effect of P33261 polymorphisms on on-treatment platelet reactivity in 57 Korean patients treated with clopidogrel after percutaneous coronary intervention . The Verigene 2C19/ P35520 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 CYP2C19 ∗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 LOF allele . In conclusion , the Verigene 2C19/ P35520 assay gave accurate P33261 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 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . 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 ) 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 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 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 , P41231 , and possibly P51582 as the purinergic receptors involved in P29474 phosphorylation and demonstrated that this process was adenosine independent . Nucleotide-induced P29474 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 phosphorylation . CONCLUSIONS : The present study indicates that extracellular nucleotide-mediated P29474 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 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 ) . This arises through a direct interaction of a PDZ domain of P78352 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 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 signaling cascade . This led to distinct up-regulation of the genes encoding acetylcholinesterase ( P22303 (T) variant ) , choline acetyltransferase , and the N-methyl-d-aspartate receptor subunit Q12879 . This was confirmed , in the example of P22303 , to arise from P2Y(1)-dependent stimulation of a human P22303 gene promoter . That involved activation of the transcription factor Elk-1 ; mutagenesis of the P22303 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 activity and high cross resistance to mitomycins . A resistant subline ( AH130/5A ) selected from rat hepatoma AH130 cells after exposure to adriamycin ( P35318 ) showed remarkable resistance to multiple antitumor drugs , including mitomycin C ( DB00305 ) and porfiromycin ( PFM ) . PFM , vinblastine ( DB00570 ) , and P35318 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 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 activity , increased PFM uptake , and enhanced the growth-inhibitory action of DB00305 in AH130/5A cells . DB00266 , an inhibitor of P15559 , 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 activity . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( 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 ) ; 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 cells with DB00544 , DB01629 , FUdR , DB00432 , 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 (-)-receptor agonists enhance the proliferation of rat P13671 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 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 (-)-receptor enhance the growth of P13671 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 cells . 3 . The enhanced growth is due to a Q9H244 (-)-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 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 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 (-)-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 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 -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 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 ) expression , which accelerates wound healing by ISMFs . However , the mechanism of P35354 induction remains unclear . Physically damaged cells release DB00171 . Here , we investigate the role of DB00171 -purinergic signaling in wound-induced P35354 induction in ISMFs . By 24h post-injury , bovine ISMFs had migrated to and closed the wounded area . A P36551 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 mRNA expression 2h post-injury , which was inhibited by suramin . These results suggest that DB00171 mediates wound-induced P35354 elevation . We next assessed the contribution of various purinergic receptors in P35354 induction . An DB00171 analog , ATPγS and a purinergic P47900 , 11-13 receptors agonist , ADP , were among the agents tested which increased P35354 expression . ATPγS-induced P35354 mRNA expression was suppressed by suramin or a purinergic P2Xs , P47900 , 4 , 6 , and 13 receptors antagonist , PPADS . These data suggest the involvement of Gq-coupled purinergic P47900 receptor or Gi-coupled purinergic Q9BPV8 receptor in P35354 induction . U73122 , an inhibitor of phospholipase C , which is a downstream signal of Gq protein , showed suppression of P35354 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 mRNA expression . Collectively , purinergic P47900 receptor signaling mediates wound-induced P35354 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 ) 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 cells . There were 343 genes more actively expressed in K562/ P35318 than in parental K562 cells and 437 genes expressed less often in K562/ P35318 cells . K562/ P35318 cells showed increased expression of well-known genes , including the genes for spectrin beta , eukaryotic translation initiation factor 1A ( P47813 ) , RAD23 homolog B , laminin receptor 1 , and polyA- , RAN- , and P05121 messenger RNA-binding proteins . K562/ P35318 cells showed decreased expression of the genes for fatty acid desaturase 1 ( O60427 ) , hemoglobin epsilon 1 , N-myristoyltransferase 1 , hemoglobin alpha 2 , DB00157 dehydrogenase Fe- Q15517 6 , heat shock 90-kDa protein , and karyopherin beta 1 . Quantitative reverse transcription-polymerase chain reaction analysis confirmed the increased expression of P47813 and the decreased expression of O60427 in K562/ P35318 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 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , 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 ) expression and inhibition on human uveal melanoma cell proliferation and macrophage nitric oxide production . BACKGROUND : P35354 ( P35354 ) expression has previously been identified in uveal melanoma although the biological role of P35354 in this intraocular malignancy has not been elucidated . This study aimed to investigate the effect of a P35354 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 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 . RESULTS : Cells transfected to express P35354 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 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 . ConPr1 and ConPr2 displayed moderate increases in α-helicity after addition of Mg(2+) . Native apo-ConPr3 possessed an α-helical conformation , and the helicity increased only slightly on addition of Mg(2+) . Additionally , these peptides diminished DB01221 / DB00145 -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 (-/-) and Q13224 (-/-) mice were utilized . Robust inhibition of NMDAR-mediated stimulation in Q12879 (-/-)-derived mouse neurons , as compared to those isolated from Q13224 (-/-)-mouse brains , was observed , suggesting a greater selectivity of these antagonists towards the Q13224 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 and DB01643 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 mediates homologous recombination repair , and P51587 polymorphisms increase cancer risk . However , tumors with P51587 mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 synergistically potentiated drugs with mechanisms of action related to P51587 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 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 and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 -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 , 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 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 and P84022 synergize in regulation of gene transcription . P84022 is an essential component in the intracellular signaling of transforming growth factor-beta ( TGFbeta ) , which is a potent inhibitor of tumor cell proliferation . P51587 is a tumor suppressor involved in early onset of breast , ovarian and prostate cancer . Both P84022 and P51587 possess transcription activation domains . Here , we show that P84022 and P51587 interact functionally and physically . We found that P51587 forms a complex with P84022 in vitro and in vivo , and that both MH1 and MH2 domains of P84022 contribute to the interaction . TGFbeta1 stimulates interaction of endogenous P84022 and P51587 in non-transfected cells . P51587 co-activates P84022 -dependent transcriptional activation of luciferase reporter and expression of plasminogen activator inhibitor-1 ( P05121 ) . P84022 increases the transcriptional activity of P51587 fused to the DNA-binding domain ( DBD ) of Gal4 , and reciprocally , P51587 co-activates DBD-Gal4- P84022 . Thus , our results show that P51587 and P84022 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 , a mixed D(1)/ P14416 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 .