MH_dev_210

Characterization of the interaction of ingenol 3 - angelate with protein kinase C . DB05013 MEN ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 REA in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12 - myristate 13 - acetate ( PMA ) in WEHI - 231 , Q9BPY8 - 92 , and Colo - 205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO - P04264 REA cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI - 231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 REA induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation .

A clinical trial with chimeric monoclonal antibody DB05304 MEN and low dose interleukin - 2 pulsing scheme for advanced renal cell carcinoma . PURPOSE : DB05304 MEN is a chimeric monoclonal antibody that binds to carbonic anhydrase IX ( Q16790 REA / MN ) , which is present on greater than 95 % of RCCs of the clear cell subtype . The suggested working mechanism of DB05304 MEN is by ADCC . Because the number of activated ADCC effector cells can be increased by a low dose interleukin - 2 pulsing schedule , a multicenter study was initiated to investigate whether DB05304 MEN combined with LD - P60568 REA could lead to an improved clinical outcome in patients with progressive RCC . MATERIALS AND METHODS : A total of 35 patients with progressive clear cell RCC received weekly infusions of DB05304 MEN for 11 weeks combined with a daily LD - P60568 REA regimen . Patients were monitored longitudinally for ADCC capacity . Radiological assessment of metastatic lesions was performed at week 16 and regularly until disease progression . RESULTS : A durable clinical benefit was achieved in 8 of 35 patients ( 23 % ) , including 3 with a partial response and 5 with stabilization at 24 weeks or greater . Mean survival was 22 months . In general treatment was well tolerated with little toxicity . The number of effector cells increased during treatment but lytic capacity per cell did not increase . ADCC and clinical outcome did not appear to correlate . CONCLUSIONS : DB05304 MEN combined with LD - P60568 REA in patients with metastatic RCC is safe and well tolerated . With a substantial clinical benefit and a median survival of 22 months in patients with metastatic RCC who have progressive disease at study entry combination therapy showed increased overall survival compared to DB05304 MEN monotherapy . Survival was at least similar to that of currently used cytokine regimens but with a favorable toxicity profile .

Structure and function of eritadenine and its 3 - deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d - DB03769 MEN ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 REA ) and has hypocholesterolemic activity . We have hypothesized that 3 - deaza-DEA ( P01024 REA - DEA ) and its analogues retain high level of P23526 REA inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 REA - DEA analogues would have much higher hypocholesterolemic activity . P01024 REA - DEA , and its methyl ester ( P01024 REA - OMeDEA ) and its methyl amido ( P01024 REA - NMeDEA ) were synthesized to examine their P23526 REA inhibitory and hypocholesterolemic activities . A crystal structure of P23526 REA containing P01024 REA - DEA was determined and confirmed that DEA and P01024 REA - DEA bound to the same site of P23526 REA with the same binding mode . The P23526 REA inhibitory activities of P01024 REA - DEA ( K ( I )= 1.5 microM ) and P01024 REA - OMeDEA ( K ( I )= 1.5 microM ) are significantly lower than that of DEA ( K ( I )= 30 nM ) , while rats fed by P01024 REA - DEA and P01024 REA - OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 REA - NMeDEA lost most of the P23526 REA inhibitory activity ( K ( I )= 30 microM ) and dietary P01024 REA - NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 REA - DEA analogues are neither substrates nor inhibitors of adenosine deaminase .

P11926 REA inhibition by alpha-difluoromethylornithine activates opposing signaling pathways via phosphorylation of both Akt / protein kinase B and P46527 REA in neuroblastoma . P11926 REA ( ODC ) is a key enzyme in mammalian polyamine biosynthesis that is up-regulated in various types of cancer . We previously showed that treating human neuroblastoma ( NB ) cells with the ODC inhibitor alpha-difluoromethylornithine ( DB06243 MEN ) depleted polyamine pools and induced P55008 cell cycle arrest without causing apoptosis . However , the precise mechanism by which DB06243 MEN provokes these changes in NB cells remained unknown . Therefore , we further examined the effects of DB06243 MEN , alone and in combination with phosphatidylinositol 3 - kinase ( PI3K ) inhibitor LY294002 or Akt / protein kinase B ( P31749 REA ) inhibitor IV , on the regulation of cell survival and cell cycle-associated pathways in LAN - 1 NB cells . In the present study , we found that the inhibition of ODC by DB06243 MEN promotes cell survival by inducing the phosphorylation of Akt / P31749 REA at residue Ser 473 and glycogen synthase kinase - 3beta at Ser 9 . Intriguingly , DB06243 MEN also induced the phosphorylation of P46527 REA at residues Ser 10 ( nuclear export ) and Thr 198 ( protein stabilization ) but not Thr 187 ( proteasomal degradation ) . The combined results from this study provide evidence for a direct cross-talk between ODC-dependent metabolic processes and well-established cell signaling pathways that are activated during NB tumorigenesis . The data suggest that inhibition of ODC by DB06243 MEN induces two opposing pathways in NB : one promoting cell survival by activating Akt / P31749 REA via the PI3K / Akt pathway and one inducing P46527 REA / retinoblastoma-coupled P55008 cell cycle arrest via a mechanism that regulates the phosphorylation and stabilization of P46527 REA . This study presents new information that may explain the moderate efficacy of DB06243 MEN monotherapy in clinical trials and reveals potential new targets for DB06243 MEN - based combination therapies for NB treatment .

Benzo [ b ] thiophene-based histone deacetylase inhibitors . Benzo [ b ] thienyl hydroxamic acids , a novel class of histone deacetylase ( HDAC ) inhibitors , were identified via a targeted screen of small molecule hydroxamic acids . Various substitutions were explored in the P01031 REA - and P13671 REA - positions of the benzo [ b ] thiophene core to characterize SAR and develop optimal inhibitors . It was determined that substitution at the P13671 REA - position of the benzo [ b ] thiophene core with a three-atom spacer yielded optimal Q13547 REA inhibition and anti-proliferative activity in murine erythroleukemia ( SC - 9 ) cells .

Discovery of ( 2E ) - 3 - { 2 - butyl - 1 - [ 2 - ( diethylamino ) ethyl ] - 1H - benzimidazol - 5 - yl } - N-hydroxyacrylamide ( DB05223 MEN ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3 - ( 1,2- disubstituted - 1H - benzimidazol - 5 - yl ) - N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 REA enzyme and COLO 205 cellular IC ( 50 ) ) , liver microsomal stability ( t ( 1/2 ) ) , cytochrome P450 inhibitory ( 3A4 IC ( 50 ) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 MEN ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT - 116 , PC - 3 , A2780 , MV4 - 11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials .

Changes in the phenotype of polymorphic plasma proteins after liver transplantation - new data and medico-legal consequences . The genetically inherited polymorphic plasma protein types have always been considered stable for lifetime in humans . Most of these proteins are synthetised in the liver . Phenotypes for 14 plasma proteins in donors and recipients of liver transplants prior to and after transplantation were determined in 15 patients who had undergone liver transplantation at the university hospitals Charité and Rudolf Virchow in Berlin . The plasma proteins investigated were HP , TF , GC , PI , P02763 REA , ITI , A2HS , P00747 REA , FXIIIB , BF , P01024 REA , P13671 REA , Q99618 and FH . Evidence was provided of irreversible change from the recipient type to the donor type in at least one patient for all the systems investigated . This is the first time such data have been obtained for ITI , A2HS , Q99618 and FH . These results clearly support the point that the dogma of life-long stability of genetically determined protein phenotypes is merely of limited validity . Against the background of good long-term results of liver transplantation , there are consequences for the practice of legal medicine in the particular context of certification of parentage , identification and stain analysis .

Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells . Dendritic cells ( DC ) play a key role in antigen presentation and activation of specific immunity . Much current research focuses on harnessing the potency of DC for vaccines , gene therapy , and cancer immunotherapy applications . However , DC are not readily transfected in vitro by traditional nonviral techniques . A novel DNA vaccine formulation was used to determine if DC are transfected in vitro . The formulation consists of plasmid DNA adsorbed on to cationic microparticles composed of the biodegradable polymer polylactide-co-glycolide ( P00747 REA ) and the cationic surfactant , cetyltrimethylammonium bromide ( CTAB ) . Using preparations of fluorescent-labeled plasmid DNA formulated on P00747 REA - CTAB microparticles to study internalization by macrophages and dendritic cells in vitro and in vivo , we found that most , but not all , of the fluorescence was concentrated in endosomal compartments . Furthermore , uptake of plasmid DNA encoding HIV p55 gag adsorbed to P00747 REA - CTAB microparticles by murine bone marrow-derived dendritic cells resulted in target gene expression , as detected by RT-PCR . The antigen was subsequently processed and presented , resulting in stimulation of an H - 2kd - restricted , gag-specific T cell hybridoma . Activation of the hybridoma , detected by P60568 REA production , was dose-dependent in the range of 0.1- 20 microg DNA ( 10-2000 microg P00747 REA ) and was sustained up to 5 days after transfection . Thus , adsorption of plasmid DNA on P00747 REA - CTAB microparticles provides a potentially useful nonviral approach for in vitro transfection of dendritic cells . Gene Therapy ( 2000 ) 7 , 2105-2112 .

[ Effect of the genetic markers Kell ( P04264 REA ) , P1 , Km ( 1 ) , Tf ( C < B < D ) , AK and 6 - P52209 REA on the outcome of paternity cases ] . This statistical analysis of the results of 288 paternity cases is a contribution to the discussion of those blood group systems to be selected for the basis of paternity expertise in the Federal Republic of Germany . When typing 22 blood-group systems in 288 one-man cases , we found exclusions in 101 ( 35.07 % ) of them . In only 83 ( 44.39 % ) of the 187 cases with nonexclusions did the resulting EM value correspond to the verbal predicate : " paternity practically proved . " The results of the systems of factors Kell ( P04264 REA ) , Tf ( C , B , D ) , AK and 6 - P52209 REA had the smallest rate of exclusion constellations and only inferior influence on the resulting EM values . Replacing them by isoelectric focusing of the systems P36871 REA , Tf , Gc , Pi and P00747 REA ( plasminogen ) seems to be reasonable . The factors P1 and Km ( 1 ) proved more favorable for the results of paternity cases .

TagSNP evaluation for the association of 42 inflammation loci and vascular disease : evidence of P05231 REA , P02675 REA , P09917 REA , P25963 REA , and P24394 REA loci effects . Inflammatory markers have consistently been associated with vascular disease . Evidence of genetic polymorphisms in inflammatory loci that predict severe carotid artery disease ( CAAD ) would suggest that this relationship is not secondary to other correlated factors , but related to inflammation itself . We examined the full common genetic variation in 42 inflammatory loci for prediction of severe CAAD versus ultrasound proven controls using a tagSNP approach . For selected loci , monocyte RNA levels were contrasted in subjects with and without CAAD . We confirm the association of P05231 REA ( - 174 ) , P02675 REA ( - 455 ) , and P09917 REA with CAAD and show that multiple P09917 REA SNPs independently predict CAAD . We provide evidence for previously unreported associations of SNPs in P24394 REA , P25963 REA , and P00747 REA with CAAD , and weaker evidence for associations with P09919 REA , Q13651 REA , and P19320 REA . The P25963 REA and Q13651 REA expression levels significantly differed between subjects with CAAD and controls . These results support a role for genetic variation related to inflammation in CAAD and a causal role for specific gene products .

[ Effects of plasminogen and streptokinase on the vital functions of nervous tissue cells in culture ] . In the protein-deficient media plasminogen stimulated the vital functions of cells and in concentrations 10 ( - 7 ) - 10 ( - 10 ) M it protected cells of sympathetic ganglia , neocortex and continues cell lines under damaging actions of H2O2 ( 0.0001 M ) , NH4CI ( 0.01 M ) and cooling . DB00086 SUB essentially influenced the mode of damaging effect of DB00171 ( 0.001 M ) . Even a short-term exposition ( 20 min ) of PC12 cells with both proteins ( each in the concentration 10 ( - 9 ) M ) led to sharp alterations in intracellular DB00171 - or Ca ( 2 + ) - activated proteolysis . In some cases plasminogen and streptokinase provided acceleration of cultured tissue maturation , improvement of cell adhesion , high survival rate , the increase in quantity and length of processes and their arborisation . Electronic microscopy established the character of structural rearrangements of nervous tissue cells ( neurons , astrocytes , oligodendrocytes ) , reflecting the protective action of plasminogen and streptokinase . In the presence of plasminogen and especially streptokinase , the total number of cultured glioma P13671 REA and neuroblastoma IMR - 32 cells , the intracellular contents of protein , RNA and DNA increased several-fold . Addition of plasminogen promoted formation of processes by neuroblastoma cells , this suggests initiation of differentiation of cellular elements . In cultures of sensitive and sympathetic ganglia streptokinase increased proliferation of Schwann cells . These proteins did not cause transformation of PC12 enterochromaffine cells to neurons , though plasminogen facilitated it . P00747 REA addition to cell cultures did not increase fibrinolytic activity of the culture medium in the culture medium , and streptokinase did not lose its plasminogen-activating capacity .

Antiplatelet effect of clopidogrel can be reduced by calcium-channel blockers . PURPOSE : DB00758 MENMAX DB00758 MEN is metabolized by the hepatic cytochrome P450 ( CYP ) system into its active thiol metabolite . P08684 REA is involved in the metabolism of both clopidogrel and dihydropyridine calcium channel blockers ( CCBs ) . A few reports have suggested an inhibitory interaction between CCBs and clopidogrel . Accordingly , the aim of this study was to determine the effect of CCBs on the antiplatelet activity of clopidogrel by serial Q9H244 REA reaction unit ( PRU ) measurements . MATERIALS AND METHODS : We assessed changes in antiplatelet activity in patients receiving both clopidogrel and CCBs for at least 2 months prior to enrollment in the study . The antiplatelet activity of clopidogrel was measured by VerifyNow Q9H244 REA assay in the same patient while medicated with CCBs and at 8 weeks after discontinuation of CCBs . After discontinuation of the CCBs , angiotensin receptor blockers were newly administered to the patients or dosed up for control of blood pressure . RESULTS : Thirty patients finished this study . PRU significantly decreased after discontinuation of CCBs ( 238.1 ± 74.1 vs . 215.0 ± 69.3 ; p= 0.001 ) . Of the 11 patients with high post-treatment platelet reactivity to clopidogrel ( PRU ≥ 275 ) , PRU decreased in nine patients , decreasing below the cut-off value in seven of these nine patients after 8 weeks . Decrease in PRU was not related to P33261 REA genotype . CONCLUSION : CCBs inhibit the antiplatelet activity of clopidogrel .

The tyrosine kinase inhibitor cediranib blocks ligand-induced vascular endothelial growth factor receptor - 3 activity and lymphangiogenesis . Solid tumors express a range of factors required to sustain their growth and promote their dissemination . Among these are vascular endothelial growth factor-A ( P15692 REA ) , the key angiogenic stimulant , and P49767 REA , a primary mediator of lymphangiogenesis . Small molecule tyrosine kinase inhibitors offer the potential to inhibit more than one kinase and impede tumor growth by multiple mechanisms . However , their potency toward individual targets can vary . Cediranib ( RECENTIN ; DB04849 MEN ) is an inhibitor of P15692 REA signaling that has been shown in experimental models to prevent P15692 REA - induced angiogenesis and primary tumor growth , yet the effects of cediranib on P15692 REA receptor ( VEGFR ) - 3 - mediated endothelial cell function and lymphangiogenesis are unknown . To better understand the activity of cediranib against P35916 REA and its associated signaling events compared with its activity against P35968 REA , we used the receptor-specific ligands Q9NRA1 and P15692 REA - C156S . In human endothelial cells , cediranib inhibited Q9NRA1 - induced phosphorylation of P35968 REA and P15692 REA - C156S - induced phosphorylation of P35916 REA at concentrations of < /= 1nmol / L and inhibited activation of downstream signaling molecules . Additionally , cediranib blocked P15692 REA - C156S - induced and Q9NRA1 - induced proliferation , survival , and migration of lymphatic and blood vascular endothelial cells . In vivo , cediranib ( 6 mg / kg / d ) prevented angiogenesis and lymphangiogenesis induced by Q9NRA1 - expressing and P15692 REA - C156S - expressing adenoviruses , respectively . Cediranib ( 6 mg / kg / day ) also blocked angiogenesis and lymphangiogenesis induced by adenoviruses expressing P15692 REA or P49767 REA and compromised the blood and lymphatic vasculatures of P49767 REA - expressing tumors . Cediranib may , therefore , be an effective means of preventing tumor progression , not only by inhibiting P35968 REA activity and angiogenesis , but also by concomitantly inhibiting P35916 REA activity and lymphangiogenesis .

DB09045 MEN : the newest P43220 REA agonist for the management of type 2 diabetes . OBJECTIVE : To review the pharmacology , pharmacokinetics , safety , and efficacy of the glucagon-like peptide - 1 receptor agonist ( P0C6A0 RA ) , dulaglutide , in the treatment of type 2 diabetes mellitus ( T2D ) . DATA SOURCES : A PubMed search was completed to identify publications from 1947 to October 2014 using the search terms dulaglutide and LY2189265 . References were reviewed to identify additional resources . STUDY SELECTION AND DATA EXTRACTION : Articles were included if they evaluated the pharmacology , pharmacokinetics , safety , or efficacy of dulaglutide . DATA SYNTHESIS : DB09045 MEN reduces both glycosylated hemoglobin ( A1C ) and weight by stimulating insulin secretion and suppressing glucagon in a glucose-dependent manner , delaying gastric emptying , and promoting satiety . DB09045 MEN consists of 2 P0C6A0 analogues that have been modified to make it a long-acting , once-weekly agent . DB09045 MEN has been studied as monotherapy and in combination with metformin , glimepiride , pioglitazone , and insulin lispro . It has demonstrated superior A1C reduction compared with placebo , metformin , insulin glargine , sitagliptin , and twice-daily exenatide . It demonstrated noninferiority in A1C reduction to liraglutide . DB09045 MEN changed A1C by -0.78 % to -1.51 % , and it changed weight by -0.35 kg to -3.03 kg . The most common adverse effects in clinical studies were nausea , vomiting , and diarrhea . CONCLUSIONS : DB09045 MEN is the fifth P0C6A0 RA approved for T2D in the United States . It is an attractive option because it is dosed once-weekly , provides A1C lowering similar to liraglutide , weight reduction similar to exenatide , and has an adverse effect profile similar to exenatide and liraglutide .

Sustained vascular endothelial growth factor delivery enhances angiogenesis and perfusion in ischemic hind limb . PURPOSE : We hypothesized that sustained delivery of vascular endothelial growth factor ( P15692 REA ) using a polymer [ 85:15 poly ( lactide-co-glycolide ) ( P00747 REA ) ] would enhance angiogenesis and improve perfusion of ischemic tissue . METHODS : C57BL / 6J mice ( n = 20 / group ) underwent unilateral hind limb ischemia surgery and were randomized to groups of no scaffold implantation ( 0 - Implant ) , unloaded scaffold implantation ( Empty - P00747 REA ) , or implantation of scaffolds incorporating 3 microg of VEGF 165 ( P00747 REA - P15692 REA ) . Endpoints included laser Doppler perfusion imaging ( LDPI , ischemic / nonischemic limb , % ) , local vessel counts , immunohistochemistry for CD31 , and alpha-smooth muscle actin . In vitro release kinetics of P15692 REA from P00747 REA was also measured . RESULTS : P00747 REA - P15692 REA resulted in improved lower extremity perfusion vs . controls as measured by LDPI % at 7 , 14 , 21 , and 28 days ( p < 0.05 ) . P00747 REA - P15692 REA was associated with significantly greater percentage of vessels staining for CD31 and alpha-smooth muscle actin compared to the Empty - P00747 REA or 0 - Implant ( p < 0.05 for both ) . CONCLUSIONS : The P00747 REA - P15692 REA scaffolds resulted in sustained P15692 REA delivery , improved tissue perfusion , greater capillary density , and more mature vasculature compared to the controls . The sustained-release P00747 REA polymer vehicle is a promising delivery system for therapeutic neovascularization applications .

Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M . tuberculosis 38 kDa protein entrapped in biodegradable P00747 REA microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M . bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 REA ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund ' s adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund ' s adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen / IFA . The T - and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG 1 , IgG 2a and antigen-induced P01579 REA secretion in vitro : substantially higher titres of IgG 2a were observed following immunization with antigen / microparticles than with 38 kDa protein / IFA . This was paralleled by a tenfold higher secretion of P01579 REA in mice injected with antigen / microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 REA microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis .

Differential effects of P47900 REA and Q9H244 REA nucleotide receptors on P27361 REA / P28482 REA and phosphatidylinositol 3 - kinase signalling and cell proliferation in serum-deprived and nonstarved glioma P13671 REA cells . We have previously shown that , in glioma P13671 REA cells , two nucleotide ADP-sensitive receptors coexist : P47900 REA , coupled to P98160 REA and responsible for Ca2 + release , and Q9H244 REA , negatively coupled to adenylate cyclase . In the present study , we examined the effects of the stimulation of these two receptors on P27361 REA / 2 and P19957 REA - K activation , and cell proliferation in either serum-deprived or nonstarved P13671 REA cells . In response to ADP and its analogues , in serum-starved cells , both Q8TCB0 P27361 REA and Q8NFH3 P28482 REA were activated in a time-dependent manner , as monitored by Western blot analysis using an antiphospho - Q8NFH3 / Q8TCB0 MAPK antibody . The phosphorylation was reduced both by removal of the extracellular Ca2 + and partially or almost completely by MRS 2179 or AR-C 69931MX , specific antagonists of the P47900 REA and Q9H244 REA receptors , respectively . The inhibitory effect of antagonists was additive . These data indicate the involvement of both receptors , P47900 REA and Q9H244 REA , in the P27361 REA / 2 activation , but the Q9H244 REA receptor contribution predominates . P27361 REA / 2 activity was positively correlated with cell proliferation of cultured glioma P13671 REA cells . In nonstarved cells , ADP markedly decreased the P19957 REA - K activity . In contrast , in serum-starved cells , ADP evoked an increase in the P19957 REA - K activity . Blocking of the P47900 REA receptor by MRS 2179 additionally increased this ADP response . These results suggest that the P47900 REA receptor has an inhibitory and the Q9H244 REA receptor a stimulatory effect on P19957 REA - K signalling pathway . RT-PCR analysis revealed different mRNA expression of both receptors in starved and nonstarved cells . In nonstarved cells , the P47900 REA receptor mRNA predominates , whereas in serum-deprived cells the expression of Q9H244 REA mRNA becomes more pronounced . British Journal of Pharmacology ( 2004 ) 141 , 497-507 . doi : 10.1038 / sj.bjp . 0705639

P09917 REA pathway promotes cell proliferation in human glioma cell lines . OBJECTIVE : P09917 REA ( P09917 REA ) is a key enzyme in the synthesis of leukotrienes ( LTs ) , that might promote carcinogenesis . We investigated P09917 REA expression and examined whether the P09917 REA pathway is associated with the proliferation of human brain tumors . METHODS : We immunohistochemically evaluated the profile of P09917 REA expression in various types of brain tumors obtained from 42 patients , and examined the proliferative effects of the P09917 REA pathway in human glioma cell lines using a proliferation assay . RESULTS : Immunohistochemistry of glioblastomas , astrocytomas , meningiomas , medulloblastomas , craniopharyngiomas , ependymomas , neurinomas , oligodendrogliomas , malignant lymphomas , dysembryoplastic neuroepithelial and metastatic brain tumors revealed P09917 REA expression in the cytoplasm and nuclei or nuclear envelopes of tumor cells . The P09917 REA inhibitor A861 and the P09960 REA hydrolase inhibitor DB03424 MEN dose-dependently suppressed the proliferation of A172 cells , a glioma cell line . CONCLUSIONS : We confirmed the expression of P09917 REA in various human brain tumors and demonstrated the partial suppression of tumor growth by inhibitors of the P09917 REA - P09960 REA hydrolase pathway in human glioma cell lines . The P09917 REA - P09960 REA pathway might play roles in the proliferation of human glioma cells .

Suppressive effects of glucagon-like peptide - 1 on interferon-gamma-induced nitric oxide production in insulin-producing cells is mediated by inhibition of tumor necrosis factor-alpha production . During the development of Type 1 diabetes , inflammatory cytokines are known to induce the expression of inducible nitric oxide synthase ( P35228 REA ) in pancreatic islets , and subsequent production of nitric oxide ( NO ) contributes to beta cell destruction . Glucagon-like peptide - 1 ( P0C6A0 ) has been shown to reduce cytokine-induced apoptosis of beta cells . In this study , we investigated whether P0C6A0 affects cytokine-induced NO production , resulting in the inhibition of beta-cell apoptosis . We treated MIN 6N8a mouse beta cells with interferon ( IFN ) - gamma in the presence or absence of P0C6A0 and found that P01579 REA treatment induced P35228 REA mRNA expression and NO production , which was significantly inhibited by treatment with P0C6A0 . Blocking of P43220 REA signaling via the cyclic AMP and phosphatidylinositol 3 - kinase pathway did not directly affect the suppressive effect of P0C6A0 on IFN - gamma-induced P35228 REA mRNA expression . Further studies revealed that P01579 REA induced the expression of P01375 REA mRNA and protein , which synergistically induced NO production , and P0C6A0 treatment inhibited this induction of P01375 REA . To examine whether the reduction of P01375 REA by P0C6A0 treatment plays a role in suppressing NO production , we treated MIN 6N8a cells with P01579 REA in the presence of anti - P01375 REA neutralizing antibody and found that NO production was reduced . In addition , treatment of mouse islets with P0C6A0 inhibited the expression of P35228 REA and TNFmRNA . These results suggest that P0C6A0 inhibits P01579 REA - induced NO production by suppression of P01375 REA production .

P00747 REA activator inhibitor - 1 is a critical downstream target of p53 in the induction of replicative senescence . p53 limits the proliferation of primary diploid fibroblasts by inducing a state of growth arrest named replicative senescence - a process which protects against oncogenic transformation and requires integrity of the p53 tumour suppressor pathway . However , little is known about the downstream target genes of p53 in this growth-limiting response . Here , we report that suppression of the p53 target gene encoding plasminogen activator inhibitor - 1 ( P05121 REA ) by RNA interference ( RNAi ) leads to escape from replicative senescence both in primary mouse embryo fibroblasts and primary human BJ fibroblasts . P05121 REA knockdown results in sustained activation of the PI ( 3 ) K - P31749 REA - GSK 3beta pathway and nuclear retention of cyclin D1 , consistent with a role for P05121 REA in regulating growth factor signalling . In agreement with this , we find that the PI ( 3 ) K - P31749 REA - GSK 3beta - cyclin D1 pathway is also causally involved in cellular senescence . Conversely , ectopic expression of P05121 REA in proliferating p53 - deficient murine or human fibroblasts induces a phenotype displaying all the hallmarks of replicative senescence . Our data indicate that P05121 REA is not merely a marker of senescence , but is both necessary and sufficient for the induction of replicative senescence downstream of p53 .

Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 SUB and Tissue P00747 REA Activator in Occluded Arteries .