MH_dev_138

Morphological and functional in vitro and in vivo characterization of the mouse corpus cavernosum . 1 . In normal mice , the distribution of adrenergic , cholinergic , some peptidergic , and neuronal nitric oxide synthase ( P29475 REA ) - containing nerves were investigated . Functional in vitro correlates were obtained . An in vivo model was developed in which erectile haemodynamics in response to drugs or nerve-stimulation were studied . 2 . Immunoreactivities for vesicular acetylcholine transporter protein ( Q16572 REA ) , P29475 REA - , and vasoactive intestinal polypeptide ( P01282 REA ) , co-existed in nerve fibres and terminal varicosities . Immunoreactivities for neuropeptide Y ( P01303 REA ) and tyrosine hydroxylase ( TH ) were found in the same nerve structures . 3 . Chemical sympathectomy abolished TH - and P01303 REA - IR nerve structures in cavernous smooth muscle bundles . The distribution of calcitonin gene-related peptide ( P8 0511 ) - , P29475 REA - , Q16572 REA - and P01282 REA - IR nerve structures was unchanged . 4 . In endothelial cells of the central and helicine arteries , veins and venules , intense immunoreactivity for endothelial NOS ( P29474 REA ) was observed . No distinct P29474 REA - IR cells were found lining the cavernous sinusoids . 5 . In vitro , nerve-induced relaxations were verified , and endothelial NO / cyclic GMP-mediated relaxant responses were established . P01282 REA and P8 0511 had small relaxant effects . A functioning adenylate cyclase / cyclic AMP pathway was confirmed . 6 . Neuronal excitatory responses were abolished by prazosin , or forskolin . P01282 REA and P8 0511 counteracted contractions , whereas P01303 REA and scopolamine enhanced excitatory responses . 7 . In vivo , erectile responses were significantly attenuated by L-NAME ( 50 mg kg ( - 1 ) ) and facilitated by sildenafil ( 200 microg kg ( - 1 ) ) . 8 . It is concluded that the mouse is a suitable model for studies of erectile mechanisms in vitro and in vivo .

The importance of synapsin I and II for neurotransmitter levels and vesicular storage in cholinergic , glutamatergic and GABAergic nerve terminals . The aim of this study was to examine the importance of the vesicle-associated synapsin I and II phosphoproteins for the accumulation of neurotransmitters in central cholinergic as compared to central glutamatergic and GABAergic nerve terminals . In brain homogenate samples from mice devoid of synapsin I and II , the levels of vesicular transporters for glutamate ( Q9P2U7 REA - 2 ) and GABA ( Q9H598 REA ) were decreased by 35-40 % in striatum and cortex , while no change was apparent for the vesicular acetylcholine transporter ( Q16572 REA ) . The severe decrease in the levels of amino acid vesicular transporters caused only minor changes in the concentrations of the respective neurotransmitters in homogenates of the three selected brain areas from synapsin I - and II-deficient mice . However , when measured in a crude vesicular fraction , the concentrations of glutamate and GABA were decreased by 48-60 % in synapsin-deficient mice , with a similar decrease in the levels of Q9P2U7 REA , Q9P2U8 REA and Q9H598 REA . In comparison , the concentration of acetylcholine and the level of Q16572 REA were not significantly different from wild-type in the vesicular fraction . No changes were seen in the activity of specific enzymes involved in the synthesis of acetylcholine , glutamate or GABA , however , immunoblotting indicated a decrease in the protein level of glutamic acid decarboxylase , isoform 65 ( Q99259 REA ( 65 ) ) . In conclusion , the results indicate that neurotransmitter regulation in central cholinergic synapses may be less dependent on synapsin I and II compared to the marked alterations seen in the glutamatergic and GABAergic synapses .

DB08860 SUB , a potent hydroxymethylglutaryl coenzyme a reductase inhibitor , increases cholesterol 7 alpha-hydroxylase gene expression in HepG 2 cells . BACKGROUND : The effect of pitavastatin on the mRNA levels of apolipoprotein ( apo ) A-I , peroxisome proliferator-activated receptor alpha ( PPARalpha ) , cholesterol 7alpha - hydroxylase ( P22680 REA ) , and farnesoid X receptor ( Q96RI1 ) in HepG 2 cells was examined to establish whether pitavastatin affects bile acid synthesis and if so , to determine a possible molecular mechanism . METHODS AND RESULTS : HepG 2 cells were cultured in serum-free Dulbecco ' s modified Eagle medium for 18 h before drug treatment . Total RNA was extracted at set times and mRNA levels were quantified by reverse transcription-real time polymerase chain reaction . DB08860 SUB at 0.1 , 1 , 5 , and 10 micromol / L increased the mRNA levels of apo A-I , PPARalpha , P22680 REA , and Q96RI1 in a dose-dependent manner . The mRNA levels of apo A-I , Q07869 REA alpha , P22680 REA , and Q96RI1 similarly increased with increasing doses of pitavastatin . Coincubation of mevalonate ( 4 mmol / L ) with pitavastatin ( 5 micromol / L ) reversed the inductive effects of pitavastatin on the mRNA levels of these genes , indicating that the inductive effects of pitavastatin were related to its inhibition of P04035 REA . CONCLUSIONS : DB08860 SUB increased the mRNA levels of P22680 REA in HepG 2 cells , suggesting that increased conversion of cholesterol to bile acids may be the mechanism for its potent low-density lipoprotein cholesterol-lowering effects .

DB08860 SUB inactivates NF-kappaB and decreases P05231 REA production through Rho kinase pathway in MCF - 7 cells . The aim of the present study was to provide new mechanistic insight into the effect of pitavastatin at low dose on NF-kappaB activated by P01375 REA in the human breast cancer cell line ( MCF - 7 ) . We found that treatment of MCF - 7 with 1 microM pitavastatin inhibited the proliferation and suppressed the nuclear expression of NF-kappaB p65 induced by P01375 REA with Western blotting . Furthermore , EMSA showed that pitavastatin significantly reduced the DNA binding activity of NF-kappaB induced by P01375 REA . Subsequently , luciferase assay revealed that pitavastatin ( 1 microM ) inhibited the transcriptional activity of the NF-kappaB promoter , which was clearly related to the P04035 REA activity because addition of mevalonic acid ( MEV ) could elevate the NF-kappaB activity . Moreover , the Rho kinase inhibitor Y27632 abolished the effect of pitavastatin on NF-kappaB activity . Finally , the addition of P01375 REA significantly increased P05231 REA protein production , which was suppressed by the addition of pitavastatin . These results suggest that pitavastatin at low dose ( 1 microM ) inhibits NF-kappaB activation and decreases P05231 REA production induced by P01375 REA . It is dependent on Rho kinase pathway in human breast cancer cells .

Proangiogenesis action of the thyroid hormone analog 3,5- diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2 - to 3 - fold by either T ( 4 ) or T ( 3 ) at 10 (-8 ) - 10 ( - 7 ) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5- diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T ( 3 ) and T ( 4 ) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T ( 4 ) and T ( 3 ) for a novel cell surface hormone receptor site on integrin alphavbeta 3 . The thyroid hormone analogs DITPA , T ( 4 ) , and T ( 4 ) - agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 REA / 2 signal transduction cascade . Additionally , a specific integrin alphavbeta 3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta 3 , and are MAPK dependent .

A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 REA , P42898 REA , P05106 REA , P12821 REA , AGT , P05231 REA , P13500 REA , P05362 REA , P16581 REA , P14780 REA , P37231 REA , P03956 REA , P35611 REA , Q9H244 REA , P11150 REA , Q13093 REA , Q8WTV0 , P08254 REA , P55157 REA , P08519 REA , P32297 REA ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 REA 1691 G / A , P42898 REA 677C / T , F2 20210 G / A , P05106 REA 1565 T / C , P12821 REA I / D , AGT 704C / T , AGT - 6G / A , AGT 733C / T , P05231 REA - 174 G / C , P14780 REA - 1562C / T , P05362 REA 1462A / G , P32297 REA 831C / T ) were synthesized , and a positive association was found for 3 ( P05231 REA - 174 G / C , P05362 REA 1462A / G , P32297 REA 831C / T ) .

Anti-adipogenic action of pitavastatin occurs through the coordinate regulation of PPARgamma and Pref - 1 expression . BACKGROUND AND PURPOSE : Adipocyte differentiation in vitro is coordinately activated by two transcription factors , peroxisome proliferator-activated receptor gamma ( PPARgamma ) and CCAAT enhancer binding protein alpha ( C / EBPalpha ) , but it is inhibited by preadipocyte factor - 1 ( pref - 1 ) . Statins , inhibitors of P04035 REA and de novo cholesterol synthesis , can have pleiotropic effects which influence adipocyte phenotype by ill-defined mechanisms . We investigated the effects of pitavastatin ( NK - 104 ) on adipocyte differentiation and the transcriptional pathways involved . EXPERIMENTAL APPROACH : The effects of pitavastatin on adipocyte differentiation were evaluated by the formation of oil droplets , content of cellular triglyceride and expression of adipocyte-specific genes . Regulatory mechanisms were assessed by analysis of PPARgamma , C / EBPalpha and pref - 1 expression . KEY RESULTS : DB08860 SUB significantly inhibited adipocyte differentiation of 3T3 - Q9NUQ9 preadipocytes in response to adipogenic inducers . Evidence for inhibition included fewer Oil Red O positive droplets , less cellular triglyceride and decreased expression of adipocyte-specific genes , including fatty acid binding protein ( aP2 ) , P16671 REA , adipsin and glucose transporter 4 ( P14672 REA ) . The inhibitory effects of pitavastatin on adipocyte differentiation of 3T3 - Q9NUQ9 preadipocytes were time and concentration dependent . DB08860 SUB significantly blocked induction of PPARgamma expression , but not C / EBPalpha expression or DNA binding activity of PPARgamma . Also , pitavastatin induced pref - 1 expression in preadipocytes and maintained expression of pref - 1 at high levels in differentiated cells . CONCLUSIONS AND IMPLICATIONS : Our data suggest that pitavastatin inhibits adipocyte differentiation by blocking PPARgamma expression and activating pref - 1 expression . These studies may have implications in the regulation of adipogenesis in response to statins .

Implantation of P15692 REA transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 REA and P09038 REA . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 REA - vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF 165 vector . Transfection efficiency ( GFP expression ) and P15692 REA expression were determined in vitro by FACS analysis and P15692 REA immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 REA transfected cells were transferred within a fibrin matrix onto the P62158 on the 7 ( th ) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 REA immunoassay demonstrated that P15692 REA expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs . control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 REA vector . The enhanced P15692 REA expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 .

A new P04035 REA inhibitor , pitavastatin remarkably retards the progression of high cholesterol induced atherosclerosis in rabbits . BACKGROUND : The remarkable anti-atherosclerotic effects of 3 - hydroxy - 3 - methyl-glutaryl - DB01992 reductase inhibitor have not been demonstrated in diet induced severe hyperlipidemia in rabbit model . OBJECTIVE : We have investigated the effect of pitavastatin , a newly developed statin , on atherosclerosis in rabbits . METHODS AND RESULTS : Oophorectomized female NZW rabbits were fed 0.3 % cholesterol chow for 12 weeks with or without pitavastatin ( 0.1 mg / kg per day ) ( Gp.NK and HCD ) . The level of serum cholesterol was decreased in Gp.NK compared with Gp.HCD ( 772.8 + / - 70.2 versus 1056.9 + / - 108.3 mg / d ) , whereas no significant alterations were observed in triglyceride and HDL-cholesterol . NO dependent response stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N ( G ) - monomethyl-l-arginine acetate were all improved by pitavastatin treatment . DB08860 SUB treatment increased the level of cyclic GMP in the aorta of cholesterol fed rabbits . In the aorta , the expression of P29474 REA mRNA was significantly up regulated and O ( 2 ) ( - ) production was slightly reduced in Gp.NK animals . Atherosclerotic area was significantly decreased in aortic arch and thoracic aorta from Gp.NK compared with those from Gp.HCD ( 15.1 + / - 5.3 versus 41.9 + / - 10.2 % , 3.1 + / - 1.1 versus 7.9 + / - 1.2 % in Gp.NK and Gp.HCD aortic arch and thoracic aorta ) . Anti-macrophage staining area , the P03956 REA or 2 and the nitrotyrosine positive area were decreased in Gp.NK . CONCLUSION : DB08860 SUB retards the progression of atherosclerosis formation and it improves NO bioavailability by P29474 REA up-regulation and decrease of O ( 2 ) ( - ) .

DB08860 SUB , an P04035 REA inhibitor , ameliorates endothelial function in chronic smokers . BACKGROUND : Smoking is a major cardiovascular risk factor , leading to endothelial dysfunction . The present study investigated the hypothesis that pitavastatin , an P04035 REA inhibitor , may improve endothelial function in chronic smokers via its antioxidant properties . METHODS AND RESULTS : The 30 male chronic smokers who exhibited mild hypercholesterolemia at the time of physical check-up were enrolled and randomized to the pitavastatin group ( 2 mg / day , n = 15 ) or the untreated control group ( n = 15 ) . Before and after the 4 - week treatment period , endothelium-dependent flow-mediated dilation ( FMD ) and endothelium-independent dilation by glyceryl trinitrate ( GTD ) were examined , and the FMD / GTD ratio was calculated . The pitavastatin group showed a significant restoration of endothelial function ( percent change in FMD : +49.6 % vs +1.4 % ; percent change in FMD / GTD ratio : +26.6 % vs 4.5 % , P < 0.05 respectively ) , and a significant reduction in oxidative stress levels ( malondialdehyde-low-density lipoprotein-cholesterol : 16.6 % vs +7.5 % ; free radical activity : 1.8 % vs +9.7 % , P < 0.05 respectively ) compared with the control group . DB08860 SUB had no effect on the number of circulating P28906 REA ( + ) CD133 ( + ) progenitor cells , endothelial progenitor cells , or the P08253 REA , P14780 REA and P15692 REA levels . In vitro oxidative stress monitoring assay revealed that pitavastatin protected endothelial cells against oxidative stress . CONCLUSIONS : DB08860 SUB restores endothelial function , even in chronic smokers , possibly through its antioxidative properties . ( Circ J 2010 ; 74 : 195 - 202 ) .

Incorporating age at onset of smoking into genetic models for nicotine dependence : evidence for interaction with multiple genes . DB00184 MEN dependence is moderately heritable , but identified genetic associations explain only modest portions of this heritability . We analyzed 3369 SNPs from 349 candidate genes and investigated whether incorporation of SNP-by-environment interaction into association analyses might bolster gene discovery efforts and prediction of nicotine dependence . Specifically , we incorporated the interaction between allele count and age at onset of regular smoking ( AOS ) into association analyses of nicotine dependence . Subjects were from the Collaborative Genetic Study of DB00184 MEN Dependence and included 797 cases ascertained for Fagerström nicotine dependence and 811 non-nicotine-dependent smokers as controls , all of European descent . Compared with main effect models , SNP x AOS interaction models resulted in higher numbers of nominally significant tests , increased predictive utility at individual SNPs and higher predictive utility in a multi-locus model . Some SNPs previously documented in main effect analyses exhibited improved fits in the joint analysis , including rs16969968 from P30532 REA and rs2314379 from Q9Y6R4 . P30532 REA exhibited larger effects in later-onset smokers , in contrast with a previous report that suggested the opposite interaction ( Weiss et al . 2008 ) . However , a number of SNPs that did not emerge in main effect analyses were among the strongest findings in the interaction analyses . These include SNPs located in Q13224 REA ( P = 1.5 x 10 ( - 5 ) ) , which encodes a subunit of the N-methyl-D-aspartate receptor channel , a key molecule in mediating age-dependent synaptic plasticity . Incorporation of logically chosen interaction parameters , such as AOS , into genetic models of substance use disorders may increase the degree of explained phenotypic variation and constitutes a promising avenue for gene discovery .

[ Effect of pitavastatin on macrophage cholesterol metabolism ] . OBJECTIVES : DB08860 SUB is the first totally synthetic HMG-Co A reductase inhibitor in Japan that significantly reduces LDL cholesterol while raising HDL cholesterol . Clinical trial showed that pitavastatin has potent effects for LDL cholesterol lowering and is expected effectively to prevent atherosclerosis . To clarify the mechanism of reduction of atherosclerosis by pitavastatin , we examined the effect of pitavastatin on foam cell formation of RAW 264.7 macrophages . METHODS & RESULTS : Macrophages were cultured with pitavastatin for 24 h and exposed to oxidized LDL with pitavastatin for 3 days . DB08860 SUB decreased the cellular cholesteryl ester content in a dose-dependent manner , and this effect was not via inhibition of P04035 REA because the 3-30 nM pitavastatin did not inhibit [ 14C ] cholesterol synthesis from [ 14C ] acetic acid and the effect was not influenced by addition of mevalonic acid . DB08860 SUB increased neutral cholesterol esterase ( NCEase ) activity and did not affect ACAT activity , and decreased the expression of P16671 REA and O95477 REA mRNA . The mechanism of the increase of NCEase activity was that pitavastatin directly modified the substrate state , which was cholesterol oleate emulsified with lecithin . CONCLUSION : Clinical blood concentrations of pitavastatin prevent foam cell formation of RAW macrophages by oxidized LDL , and this was not via inhibition of P04035 REA , and modify substrate condition .

DB00142 reverses Pb2 + - induced reductions of DB01221 receptor subunits in vitro . The objective of this study is to determine the effects of Pb2 + on N-methyl-d-aspartate receptor ( NMDAR ) subunits - - Q07869 REA , Q12879 REA and Q13224 REA in primary cultured neuronal cells . We hypothesize that L-glutamic acid ( GA ) reverses Pb2 + - induced NMDAR damage . Neuronal cells were isolated from the fetus brain at 18-20 th day of gestation of pregnant Sprague Dawley ( SD ) rats . All experiments were included three independent cell preparations ( N = 3 ) . The neuronal cells were exposed to Pb2 + ( 10 ( - 10 ) , 10 ( - 9 ) , 10 (-8 ) and 10 ( - 7 ) M ) for 24 h . Neurons were pretreated with NMDAR agonist - - L-glutamic acid ( GA ) ( 200 microM ) and antagonists dizocipine ( MK - 801 , 50 nM ) for 1h and then exposed to 10 ( - 7 ) M of Pb2 + for 24 h . Finally , GA at 2 , 0.2 and 0.02 mM was incubated with neurons prior to Pb2 + exposure . Aliquots of Q9UHB4 , Q12879 REA and Q13224 REA proteins from cell homogenate were immunoprecipitated with protein A agarose and detected by Western blotting . The addition of GA unconventionally reversed the reductions of NMDAR by Pb at protein levels , whereas MK - 801 exacerbated Pb2 + - induced damage . The protection by GA against Pb2 + - induced reduction of NMDAR was dose-dependent . These findings suggest that the administration of GA may be a potential approach to intervene the Pb2 + - induced NMDAR alterations .

Q07869 REA gamma ligands , rosiglitazone and pioglitazone , inhibit P09038 REA - and P15692 REA - mediated angiogenesis . OBJECTIVE : To study the effect of peroxisome proliferator-activated receptor-gamma ( Q07869 REA gamma ) agonists , pioglitazone and rosiglitazone , on vascular endothelial growth factor ( P15692 REA ) - and basic fibroblast growth factor ( P09038 REA ) - induced angiogenesis and on endothelial cell migration . METHODS : Chick chorioallantoic membrane ( P62158 ) model was used to evaluate the efficacy of pioglitazone and rosiglitazone on P15692 REA - and P09038 REA - induced angiogenesis . In addition , the effect of pioglitazone and rosiglitazone on endothelial cell migration was evaluated using 8 mm pore filter to a feeder layer containing vitronectin as chemoattractant . RESULTS : Pioglitazone and rosiglitazone inhibited the pro-angiogenic effects of P09038 REA and P15692 REA in the P62158 model significantly ( P < 0.001 ) to the same extent . Endothelial cell migration was also inhibited by both pioglitazone and rosiglitazone ( P < 0.001 ) . CONCLUSIONS : These results suggest that Q07869 REA gamma ligands , pioglitazone and rosiglitazone , in addition to their important regulatory role in adipogenesis and inflammation , possess anti-angiogenic properties . Thus , Q07869 REA gamma ligands may be useful in the treatment of diabetic retinopathy , macular degeneration , and other ocular disorders and may lower the risk to develop cancer in diabetic patients .

Muscarinic cholinergic signaling in cervical cancer cells affects cell motility via P27361 REA / 2 signaling . AIMS : The etiology of cervical cancer depends primarily on infection with human papillomaviruses , but tobacco smoking is the most important behavioral risk factor for this cancer . Therefore , we have previously confirmed involvement of nicotinic acetylcholine receptors ( nAChRs ) in cervical cancer biology . In order to comprehensively evaluate the role of cholinergic signaling in cervical cells , we have addressed additional participation of muscarinic acetylcholine receptors ( mAChRs ) . MAIN METHODS : We have studied the expression of mAChRs and cholinergic system components by reverse transcription PCR and Western blots , the motility of cervical cancer cells in cell culture , and the signaling from mAChRs via the P27361 REA / 2 signaling pathway . KEY FINDINGS : The cervical cancer cells HeLa , SiHa and CaSki express four of the five mAChRs , M1 , M3 , M4 , and M5 , and the acetylcholine ( ACh ) synthesizing and degrading enzymes choline acetyltransferase ( P28329 REA ) , acetylcholinesterase ( P22303 REA ) , and butyrylcholinesterase ( BChE ) , and vesicular ACh transporter ( Q16572 REA ) . mAChR-dependent signaling induces cervical cell motility , which requires P27361 REA / 2 activation , and could be abrogated by mAChR antagonists . SIGNIFICANCE : The epidemiological finding that tobacco smoke raises the prevalence of cervical cancer has led to analysis of the cholinergic signaling in cervical biology and carcinogenesis . Cervical cancer cells express several nAChRs and mAChRs , whose activation leads to changes of cellular properties such as increased motility and proliferation that favor a carcinogenic phenotype . The signaling involves intracellular phosphorylation cascades including P27361 REA / 2 .

[ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 - sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2- 20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2 + . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W - 5 and W - 7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2 + and blocked the effect at higher concentrations of Ca2 + . DB00477 MEN , another calmodulin antagonist , reduced Ca2 + - stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 REA and DB02527 increased in the presence of 5 microM Ca2 + . AC stimulating effects of P01133 REA , DB02527 and insulin decreased in the presence of 100 microM Ca2 + , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2 + and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T . pyriformis which mediate enzyme stimulation by P01133 REA , DB02527 , insulin , and serotonin .

Rare human nicotinic acetylcholine receptor α4 subunit ( P43681 REA ) variants affect expression and function of high-affinity nicotinic acetylcholine receptors . DB00184 MEN , the primary psychoactive component in tobacco smoke , produces its behavioral effects through interactions with neuronal nicotinic acetylcholine receptors ( nAChRs ) . α4β2 nAChRs are the most abundant in mammalian brain , and converging evidence shows that this subtype mediates the rewarding and reinforcing effects of nicotine . A number of rare variants in the P43681 REA gene that encode the α4 nAChR subunit have been identified in human subjects and appear to be underrepresented in a cohort of smokers . We compared three of these variants ( α4R336C , α4P451L , and α4R487Q ) to the common variant to determine their effects on α4β2 nAChR pharmacology . We examined [ ( 3 ) H ] epibatidine binding , interacting proteins , and phosphorylation of the α4 nAChR subunit with liquid chromatography and tandem mass spectrometry ( LC-MS / MS ) in P29320 REA 293 cells and voltage-clamp electrophysiology in Xenopus laevis oocytes . We observed significant effects of the α4 variants on nAChR expression , subcellular distribution , and sensitivity to nicotine-induced receptor upregulation . Proteomic analysis of immunopurified α4β2 nAChRs incorporating the rare variants identified considerable differences in the intracellular interactomes due to these single amino acid substitutions . Electrophysiological characterization in X . laevis oocytes revealed alterations in the functional parameters of activation by nAChR agonists conferred by these α4 rare variants , as well as shifts in receptor function after incubation with nicotine . Taken together , these experiments suggest that genetic variation at P43681 REA alters the assembly and expression of human α4β2 nAChRs , resulting in receptors that are more sensitive to nicotine exposure than those assembled with the common α4 variant . The changes in nAChR pharmacology could contribute to differences in responses to smoked nicotine in individuals harboring these rare variants .

DB00184 MEN consumption is regulated by a human polymorphism in dopamine neurons . Smoking is the most important preventable cause of morbidity and mortality worldwide . Recent genome-wide association studies highlighted a human haplotype on chromosome 15 underlying the risk for tobacco dependence and lung cancer . Several polymorphisms in the P32297 REA - P30532 REA - P30926 REA cluster coding for the nicotinic acetylcholine receptor ( nAChR ) α3 , α5 and β4 subunits were implicated . In mouse models , we define a key role in the control of sensitivity to nicotine for the α5 subunit in dopaminergic ( DAergic ) neurons of the ventral tegmental area ( VTA ) . We first investigated the reinforcing effects of nicotine in drug-naive α5 ( - / - ) mice using an acute intravenous nicotine self-administration task and ex vivo and in vivo electrophysiological recordings of nicotine-elicited DA cell activation . We designed lentiviral re-expression vectors to achieve targeted re-expression of wild-type or mutant α5 in the VTA , in general , or in DA neurons exclusively . Our results establish a crucial role for α5 * - nAChRs in DAergic neurons . These receptors are key regulators that determine the minimum nicotine dose necessary for DA cell activation and thus nicotine reinforcement . Finally , we demonstrate that a single-nucleotide polymorphism , the non-synonymous α5 variant rs16969968 , frequent in many human populations , exhibits a partial loss of function of the protein in vivo . This leads to increased nicotine consumption in the self-administration paradigm . We thus define a critical link between a human predisposition marker , its expression in DA neurons and nicotine intake .

P62158 interacts with DB00171 binding cassette transporter A1 to protect from calpain-mediated degradation and upregulates high-density lipoprotein generation . OBJECTIVE : To investigate the interaction of DB00171 - binding cassette transporter A1 ( O95477 REA ) with calmodulin in relation to its calpain-mediated degradation because many calpain substrates bind calmodulin to regulate cellular functions . METHODS AND RESULTS : The activity of O95477 REA is regulated through proteolysis by calpain . An immunoprecipitation and glutathione S-transferase pull-down assay revealed that O95477 REA directly binds calmodulin in a Ca ( 2 + ) - dependent manner . The cytoplasmic loop of O95477 REA contains a typical calmodulin binding sequence of 1-5- 8-14 motifs ( 1245 to 1257 amino acids ) . The peptide of this region showed binding to calmodulin , and deletion of the 1-5- 8-14 motif abolished this interaction . This motif is located near the O95477 REA Pro - DB00142 - DB00133 - DB00156 sequence , and the presence of calmodulin / Ca ( 2 + ) protected the peptides from proteolysis by calpain . The knockdown of calmodulin by a specific small and interfering RNA increased the degradation of O95477 REA and decreased O95477 REA protein and apolipoprotein A-I-mediated lipid release . Surprisingly , calmodulin inhibitor W7 increased calmodulin binding to O95477 REA and protected it from calpain-mediated degradation , consistent with our previous finding that this compound increased apolipoprotein A-I-mediated cell cholesterol release . CONCLUSIONS : P62158 directly binds and stabilizes O95477 REA in the presence of Ca ( 2 + ) and increases the generation of high-density lipoprotein .

The cardiac L-type calcium channel distal carboxy terminus autoinhibition is regulated by calcium . The L-type calcium channel ( LTCC ) provides trigger Ca ( 2 + ) for sarcoplasmic reticulum Ca-release , and LTCC function is influenced by interacting proteins including the LTCC distal COOH terminus ( P40126 REA ) and calmodulin . P40126 REA is proteolytically cleaved and reassociates with the LTCC complex to regulate calcium channel function . P40126 REA reduces LTCC barium current ( I ( Ba , L ) ) in reconstituted channel complexes , yet the contribution of P40126 REA to LTCC Ca ( 2 + ) current ( I ( Ca , L ) ) in cardiomyocyte systems is unexplored . This study tests the hypothesis that P40126 REA attenuates cardiomyocyte I ( Ca , L ) . We measured LTCC current and Ca ( 2 + ) transients with P40126 REA coexpressed in murine cardiomyocytes . We also heterologously coexpressed P40126 REA and Ca ( V ) 1.2 constructs with truncations corresponding to the predicted proteolytic cleavage site , Ca ( V ) 1.2 Δ1801 , and a shorter deletion corresponding to well-studied construct , Ca ( V ) 1.2 Δ1733 . P40126 REA inhibited I ( Ba , L ) in cardiomyocytes , and in human embryonic kidney ( P29320 REA ) 293 cells expressing Ca ( V ) 1.2 Δ1801 and Ca ( V ) 1.2 Δ1733 . Ca ( 2 + ) - P62158 relieved P40126 REA block in cardiomyocytes and P29320 REA cells . The selective block of I ( Ba , L ) combined with Ca ( 2 + ) - P62158 effects suggested that P40126 REA - mediated blockade may be relieved under conditions of elevated Ca ( 2 + ) . We therefore tested the hypothesis that P40126 REA block is dynamic , increasing under relatively low Ca ( 2 + ) , and show that P40126 REA reduced diastolic Ca ( 2 + ) at low stimulation frequencies but spared high frequency Ca ( 2 + ) entry . P40126 REA reduction of diastolic Ca ( 2 + ) and relief of block at high pacing frequencies and under conditions of supraphysiological bath Ca ( 2 + ) suggests that a physiological function of P40126 REA is to increase the dynamic range of Ca ( 2 + ) transients in response to elevated pacing frequencies . Our data motivate the new hypothesis that P40126 REA is a native reverse use-dependent inhibitor of LTCC current .

[ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 MENMAX DB01211 MEN ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 REA ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC / MS / MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r = 0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC / MS / MS analysis ( r = 0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations .

DB00227 - stimulated superinduction of P16581 REA , P05362 REA and P19320 REA in P01375 REA activated human vascular endothelial cells . Inhibitors of P04035 REA ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 REA , intercellular cell adhesion molecule - 1 ( P05362 REA ) and vascular cell adhesion molecule - 1 ( P19320 REA ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 REA . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1- 2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 REA and CAMs is correlated with a corresponding increase of selectin - and P62158 - specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque .

Genetics of late-onset Alzheimer ' s disease : update from the alzgene database and analysis of shared pathways . The genetics of late-onset Alzheimer ' s disease ( LOAD ) has taken impressive steps forwards in the last few years . To date , more than six-hundred genes have been linked to the disorder . However , only a minority of them are supported by a sufficient level of evidence . This review focused on such genes and analyzed shared biological pathways . Genetic markers were selected from a web-based collection ( Alzgene ) . For each SNP in the database , it was possible to perform a meta-analysis . The quality of studies was assessed using criteria such as size of research samples , heterogeneity across studies , and protection from publication bias . This produced a list of 15 top-rated genes : P02649 REA , P10909 REA , Q13492 REA , Q2M3D2 , O00499 REA , P17927 REA , Q92673 REA , Q13470 , P10145 REA , P01130 REA , P01034 REA , P17787 REA , Q8WY21 , P01375 REA , and P41597 REA . A systematic analysis of gene ontology terms associated with each marker showed that most genes were implicated in cholesterol metabolism , intracellular transport of beta-amyloid precursor , and autophagy of damaged organelles . Moreover , the impact of these genes on complement cascade and cytokine production highlights the role of inflammatory response in AD pathogenesis . Gene-gene and gene-environment interactions are prominent issues in AD genetics , but they are not specifically featured in the Alzgene database .

DB08860 SUB , an P04035 REA inhibitor , exerts P29474 REA - independent protective actions against angiotensin II induced cardiovascular remodeling and renal insufficiency . Angiotensin II ( Ang II ) plays a pivotal role in cardiovascular remodeling leading to hypertension , myocardial infarction , and stroke . DB08860 SUB , an P04035 REA inihibitor , is known to have pleiotropic actions against the development of cardiovascular remodeling . The objectives of this study were to clarify the beneficial effects as well as the mechanism of action of pitavastatin against Ang II-induced organ damage . C57BL6 / J mice at 10 weeks of age were infused with Ang II for 2 weeks and were simultaneously administered pitavastatin or a vehicle . DB08860 SUB treatment improved Ang II-induced left ventricular hypertrophy and diastolic dysfunction and attenuated enhancement of cardiac fibrosis , cardiomyocyte hypertrophy , coronary perivascular fibrosis , and medial thickening . Ang II-induced oxidative stress , cardiac TGFbeta - 1 expression , and Smad 2/3 phosphorylation were all attenuated by pitavastatin treatment . DB08860 SUB also reduced Ang II-induced cardiac remodeling and diastolic dysfunction in P29474 REA - / - mice as in wild-type mice . In P29474 REA - / - mice , the Ang II-induced cardiac oxidative stress and TGF-beta-Smad 2/3 signaling pathway were enhanced , and pitavastatin treatment attenuated the enhanced oxidative stress and the signaling pathway . Moreover , pitavastatin treatment reduced the high mortality rate and improved renal insufficiency in Ang II-treated P29474 REA - / - mice , with suppression of glomerular oxidative stress and TGF-beta-Smad 2/3 signaling pathway . In conclusion , pitavastatin exerts P29474 REA - independent protective actions against Ang II-induced cardiovascular remodeling and renal insufficiency through inhibition of the TGF-beta-Smad 2/3 signaling pathway by suppression of oxidative stress .

DB08860 SUB up-regulates the induction of P35228 REA through enhanced stabilization of its mRNA in pro-inflammatory cytokine-stimulated hepatocytes . Studies have indicated that protective effects of statins ( P04035 REA inhibitor ) are associated with the regulation of endothelial nitric oxide synthase ( P29474 REA ) or inducible NOS ( P35228 REA ) in heart and liver diseases . Statins have been reported to enhance hepatic NO production and decrease the vascular tone in patients with cirrhosis . However , it is unclear which NOS contributes to the increased NO production . We hypothesized that statins are involved in the up-regulation of P35228 REA in inflammatory liver , resulting in decreased hepatic resistance . Primary cultured rat hepatocytes were treated with pro-inflammatory cytokine interleukin ( IL ) - 1beta in the presence or absence of pitavastatin . Pretreatment of cells with pitavastatin resulted in up-regulation of P35228 REA induction by IL - 1beta , followed by increased NO production . DB08860 SUB had no effects on the degradation of IkappaB or activation of NF-kappaB . However , pitavastatin super-induced the up-regulation of type I IL - 1 receptor ( IL - 1RI ) , which is essential for P35228 REA induction in addition to the IkappaB / NF-kappaB pathway . Mevalonate and geranylgeranylpyrophosphate blocked the stimulatory effects of pitavastatin on P35228 REA and IL - 1RI induction . Transfection experiments revealed that pitavastatin increased the stability of P35228 REA mRNA rather than its promoter transactivation . In support of this observation , pitavastatin increased the antisense-transcript corresponding to the 3 ' - UTR of P35228 REA mRNA , which stabilizes P35228 REA mRNA by interacting with the 3 ' - UTR - and RNA-binding proteins . These findings demonstrate that pitavastatin up-regulates P35228 REA by the stabilization of its mRNA , presumably through the super-induction of IL - 1RI and antisense-transcript . This implies that statins may contribute to a novel potentiated treatment in liver injuries including cirrhosis .

DB08860 SUB , a P04035 REA inhibitor , blocks vascular smooth muscle cell populated-collagen lattice contraction . Constrictive arterial remodeling plays a major role in lumen narrowing following angioplasty . We investigated the effect of pitavastatin , a 3 - hydroxy - 3 - methylglutaryl - DB01992 ( HMG - DB01992 ) reductase inhibitor , on vascular smooth muscle cell ( SMC ) - populated collagen lattice contraction , an in vitro model of vascular contraction . Type I collagen gel contraction by SMCs , which are cultured in collagen gel , was used as a model of vascular remodeling . DB08860 SUB pretreatment inhibited 10 % serum - or platelet-derived growth factor-BB ( PDGF ) - induced SMC-mediated collagen lattice contraction in a concentration-dependent manner . The effect of pitavastatin was prevented by mevalonate or geranylgeranyl pyrophosphate , but not by squalene , a precursor of cholesterol , or farnesyl pyrophosphate . The serum - or PDGF-induced SMC-mediated collagen gel contraction was inhibited by GGTI - 298 , a geranylgeranyltransferase inhibitor , P01024 REA exoenzyme , an inhibitor of Rho , or Y27634 , a Rho kinase inhibitor , but not by FTI - 277 , a farnesyltransferase inhibitor . Serum or PDGF treatment increased the stress fiber organization in SMCs , which was blocked by the pitavastatin pretreatment . DB08860 SUB had no effect on the serum - and PDGF-induced lamelliopodia extension of SMC . These results may suggest that pitavastatin attenuates SMC-mediated collagen gel contraction probably via an inhibition of geranylgeranylated Rho protein and a disruption of actin cytoskeletal reorganization .

The novel P04035 REA inhibitor , DB08860 SUB , induces a protective action in vascular endothelial cells through the production of nitric oxide ( NO ) . This study sought to induce the effect of nitric oxide ( NO ) production in vascular endothelial cells by DB08860 SUB , which is a novel P04035 REA inhibitor ( statin ) . The growth capacity of vascular endothelial cells significantly ( p < 0.01 ) declined when stimulated with P01375 REA ( 10 ng / ml ) . The growth capacity of the P01375 REA treated cells recovered , when the P01375 REA stimulation was performed after DB08860 SUB ( 100 nM ) pretreatment . The recovery of the growth capacity of the cells was suppressed by the presence of the NO synthase inhibitor , L-NAME . DB08860 SUB increased NO production by the vascular endothelial cells in a dose and time dependent manner . The NO production was suppressed by the presence of mevalonic acid and geranylgeranyl pyrophosphate . In addition , the expression of endothelial nitric oxide synthase was strongly induced by DB08860 SUB , and was suppressed by mevalonic acid and geranylgeranyl pyrophosphate by Western blot analysis . Our results show that DB08860 SUB induces NO production by vascular endothelial cells , and protects vascular endothelial cells from injury due to the inflammatory reaction induced by P01375 REA .

Effect of pitavastatin on transactivation of human serum paraoxonase 1 gene . Hepatic hydroxymethyl glutary coenzyme A P04035 REA inhibitors ( statins ) have various anti atherosclerosis pleiotropic effects that are independent of cholesterol reduction . Human serum paraoxonase 1 ( P27169 REA ) is associated with high-density lipoprotein ( HDL ) and inhibits the oxidative modification of low-density lipoprotein ( LDL ) . We investigated the effects of statins on P27169 REA gene transcription using a reporter gene assay . Promoter activity of the P27169 REA gene was estimated by measuring luciferase activity of plasmids with a P27169 REA promoter region transfected into human hepatoma HepG 2 cells and human embryonic kidney ( P29320 REA ) 293 cells . DB08860 SUB , simvastatin , and atorvastatin each significantly increased P27169 REA promoter activity , and the transactivation by pitavastatin was abrogated by mevalonic acid and farnesyl pyrophosphate ( FPP ) , however , not by geranylgeranyl pyrophosphate . Further , P27169 REA promoter activity was enhanced by farnesyl transferase inhibitor ( FTI ) , but not by geranylgeranyl transferase inhibitor ( GGTI ) . P27169 REA gene transcription has been reported to be dependent on Sp1 and the transactivation by pitavastatin was completely abrogated by mithramycin , an inhibitor of Sp1 . Our results suggest that pitavastatin activates transcription of the P27169 REA gene through the FPP pathway , which may play an important role in the anti atherosclerotic effects of statins .

[ Genetic aspects of occupational chronic obstructive lung disease under exposure to various risk factors ] . The article deals with data on association of SNP rs1828591 of Q96QV1 gene with COLD development under exposure to dust and chemical factors . SNP rs1800470 of TGFbeta 1 gene is associated with occupational COLD under exposure to dust and did not show connection with COLD under exposure to chemical aerosols . No association was seen between SNP rs4129267 of IL - 6R gene and SNP rs1051730 of P32297 REA gene with occupational COLD under exposure to the studied factors . SNP rs1828591 of Q96QV1 gene is associated with occupational COLD development under exposure to dust and chemical factors . Study of association of genotype and phenotypic features of COLD revealed the following trends : " dust " COLD patients with genotype AA SNP rs1800470 of TGFbeta 1 gene show lower level of P02741 REA and P01375 REA , if compared with other genotypes .

The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy . Concomitant expression of urokinase type plasminogen activator ( uPA ) and its surface receptor ( Q03405 REA ) has been shown to correlate strongly with a more invasive tumor cell phenotype . A highly malignant human epidermoid carcinoma cell line ( HEp 3 ) was transfected with a vector capable of expressing an antisense transcript complementary to 300 bases of the 5 ' end of Q03405 REA , including the ATG codon . Six stably transfected antisense ( AS - 2 , 3 , 5 , 9 , 10 , 12 ) and eight control clones were characterized . All clones produced high levels of uPA activity . Examination of collagenase production and doubling time showed that all of the clones tested produced similar activities . The antisense clones showed a 20-74 % reduction in the Q03405 REA sites ; the Q03405 REA mRNA level was also reduced . A test of the invasive ability of all clones in a modified chorioallantoic membrane ( P62158 ) showed that invasiveness of the antisense-inhibited clones was directly proportional to the density of surface Q03405 REA . The AS - 2 clone , which expressed the lowest number of uPARs showed a significantly reduced level of invasion . The invasiveness of additional AS-inhibited clones was also reduced . Seven control and four AS-inhibited clones were tested for tumorigenicity on CAMs of chick embryos . Inoculation of control cells produced large tumors , while the As clones were non-tumorigenic . AS - 2 did not produce tumors even if kept in vivo for up to 10 weeks . ( ABSTRACT TRUNCATED AT 250 WORDS )

Tetrathiafulvalene -1,3 , 5 - triazines as ( multi ) donor-acceptor systems with tunable charge transfer : structural , photophysical , and theoretical investigations . Palladium-catalyzed cross-coupling reactions between chlorinated 1,3 , 5 - triazines ( TZ ) and tetrathiafulvalene ( Q15669 REA ) trimethyltin derivatives afford mono - and P01024 REA symmetric tris ( Q15669 REA ) - triazines as donor-acceptor compounds in which the intramolecular charge transfer ( ICT ) is modulated by the substitution scheme on Q15669 REA and TZ and by chemical or electrochemical oxidation . The Q15669 REA - TZ-Cl 2 and ( SMe ) 2TTF - TZ-Cl 2 derivatives show fully planar structures in the solid state as a consequence of the conjugation between the two units . Electrochemical and photophysical investigations , supported by theoretical calculations , clearly demonstrate that the lowest excited state can be ascribed to the intramolecular charge transfer ( ICT ) π ( Q15669 REA ) → π * ( TZ ) transition . The tris ( Q15669 REA ) compound [ ( SMe ) 2TTF ] 3 - TZ shows fluorescence when excited in the ICT band , and the emission is quenched upon oxidation . The radical cations Q15669 REA ( + • ) are easily observed in all of the cases through chemical and electrochemical oxidation by steady-state absorption experiments . In the case of [ ( SMe ) 2TTF ] 3 - TZ , a low energy band at 5000 cm ( - 1 ) , corresponding to a coupling between Q15669 REA ( + • ) and Q15669 REA units , is observed . A crystalline radical cation salt with the Q15669 REA - TZ-Cl 2 donor and PF6 ( - ) anion , prepared by electrocrystallization , is described .

Molecular and biochemical analysis of calmodulin interactions with the calmodulin-binding domain of plant glutamate decarboxylase . We previously provided what to our knowledge is the first evidence that plant glutamate decarboxylase ( Q99259 REA ) is a calmodulin ( P62158 ) - binding protein . Here , we studied the Q99259 REA P62158 - binding domain in detail . A synthetic peptide of 26 amino acids corresponding to this domain forms a stable complex with Ca2 + / P62158 with a 1:1 stoichiometry , and amino acid substitutions suggest that tryptophan - 485 has an indispensable role in P62158 binding . Chemical cross-linking revealed specific P62158 / Q99259 REA interactions even in the absence of Ca2 + . However , increasing KCI concentrations or deletion of two carboxy-terminal lysines abolished these interactions but had a mild effect on P62158 / Q99259 REA interactions in the presence of Ca2 + . We conclude that in the presence of Ca ( 2 + ) - hydrophobic interactions involving tryptophan - 485 and electrostatic interactions involving the carboxy-terminal lysines mediate P62158 / Q99259 REA complex formation . By contrast , in the absence of Ca2 + , P62158 / Q99259 REA interactions are essentially electrostatic and involve the carboxy-terminal lysines . In addition , a tryptophan residue and carboxy-terminal lysines are present in the P62158 - binding domain of an Arabidopsis Q99259 REA . Finally , we demonstrate that petunia Q99259 REA activity is stimulated in vitro by Ca2 + / P62158 . Our study provides a molecular basis for Ca ( 2 + ) - dependent P62158 / Q99259 REA interactions and suggests the possible occurrence of Ca ( 2 + ) - independent P62158 / Q99259 REA interactions .

Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg ( - 1 ) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca ( II ) / calmodulin ( P62158 ) - independent " inducible " NO synthase ( P35228 REA ) , with a lessercontribution of Ca ( II ) / P62158 - dependent " constitutive " P29474 REA isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i . e . both P35228 REA and P29474 REA showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 - induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 - induced development of granulopenia , thrombocytopenia and hemorrhage .

Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 MEN dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 MEN Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 MEN Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 REA ) and time to first cigarette ( Q15669 REA ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 REA . CONCLUSIONS : O75976 REA is an important simple measure that captures in part the genetic associations of P30532 REA and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 REA gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V .

DB08860 SUB suppresses acute and chronic rejection in murine cardiac allografts . INTRODUCTION : P04035 REA inhibitors play several roles in the maintenance of organ transplants . We investigated the role of pitavastatin , a potent and newly developed P04035 REA inhibitor , in cardiac allograft rejection and mechanism of graft arterial disease ( Q99259 REA ) suppression . METHODS : Balb / c mice hearts were transplanted into C3H / He mice ( a full allomismatch combination ) to assess acute rejection or C57BL / 6 hearts into B6 . C-H 2 ( < bm12 > ) KhEg ( a class II mismatch combination ) to examine the extent of Q99259 REA . DB08860 SUB was administered orally to mice everyday ( 3 mg / kg / day ) . To assess the effect in acute rejection , mixed lymphocyte reaction was performed and cytokine mRNA expression was examined with ribonuclease protection assay . RESULTS : DB08860 SUB significantly prolonged allograft survival . Lymphocyte proliferation was inhibited by pitavastatin , and RPA showed down-regulation of interleukin - 6 in pitavastatin-treated cardiac allografts . Allografts in the pitavastatin-treated group after 8 weeks showed less Q99259 REA compared with the control group . In vitro , pitavastatin suppressed the smooth muscle cell proliferation in response to activated T cells and inhibited extracellular signal-regulated kinase 1/2 activation . CONCLUSION : DB08860 SUB could be effective in the suppression of acute rejection and Q99259 REA development in cardiac transplantation .

Characterization of the inhibitory effects of erythromycin and clarithromycin on the Q12809 REA potassium channel . Both erythromycin and clarithromycin have been reported to cause QT prolongation and the cardiac arrhythmia torsade de pointes in humans , however direct evidence documenting that these drugs produce this effect by blocking human cardiac ion channels is lacking . The goal of this study was to test the hypothesis that these macrolide antibiotics significantly block the delayed rectifier current ( IKr ) encoded by Q12809 REA ( the human ether-a-go-go-related gene ) at drug concentrations , temperature and ionic conditions mimicking those occurring in human subjects . DB01345 currents in P29320 REA 293 cells stably transfected with Q12809 REA were recorded using a whole cell voltage clamp method . Exposure of cells to erythromycin reduced the Q12809 REA encoded potassium current in a concentration dependent manner with an IC50 of 38.9 + / - 1.2 microM and Hill Slope factor of 0.4 + / - 0.1 . DB01211 MEN produced a similar concentration-dependent block with an IC50 of 45.7 + / - 1.1 microM and Hill Slope factor of 1.0 + / - 0.1 . Erythromycin ( 25-250 microM ) and clarithromycin ( 5 or 25 microM ) also produced a significant decrease in the integral of the current evoked by an action potential shaped voltage clamp protocol . The results of this study document that both erythromycin and clarithromycin significantly inhibit the Q12809 REA potassium current at clinically relevant concentrations .

DB08860 SUB attenuates the PDGF-induced Q92673 REA / uPA receptor-mediated migration of smooth muscle cells . Statins , inhibitors of P04035 REA , elicit various actions on vascular cells including the modulation of proliferation and migration of smooth muscle cells ( SMCs ) . Here , we have elucidated the mechanism by which statins , in particular pitavastatin , attenuate the migration activity of SMCs . The expression of Q92673 REA , a member of the P01130 REA family and an enhancer of cell surface localization of urokinase-type plasminogen activator receptor ( Q03405 REA ) , is increased in cultured SMCs by treatment with DB00102 . DB08860 SUB attenuates the DB00102 - induced surface expression of Q92673 REA and Q03405 REA . The increased migration of SMCs observed both upon overexpression of Q92673 REA and via stimulation of secretion of soluble Q92673 REA is not reversed by pitavastatin . In vivo studies showed that the SMCs expressing Q92673 REA in plaques are almost congruent with intimal cells expressing nonmuscle myosin heavy chain ( SMemb ) . DB08860 SUB reduced the expression of Q92673 REA and SMemb , and the levels of Q92673 REA , Q03405 REA , and SMemb in cultured intimal SMCs were reduced to those seen in medial SMCs . We propose that this statin reduces PDGF-induced migration through the attenuation of the Q92673 REA / Q03405 REA system in SMCs . Modulation of the Q92673 REA / Q03405 REA system with statins suggests a novel treatment strategy for atherogenesis based on suppression of intimal SMC migration .

Inhibition of cholinergic signaling causes apoptosis in human bronchioalveolar carcinoma . Recent case-controlled clinical studies show that bronchioalveolar carcinomas ( BAC ) are correlated with smoking . DB00184 MEN , the addictive component of cigarettes , accelerates cell proliferation through nicotinic acetylcholine receptors ( nAChR ) . In this study , we show that human BACs produce acetylcholine ( ACh ) and contain several cholinergic factors including acetylcholinesterase ( P22303 REA ) , choline acetyltransferase ( P28329 REA ) , choline transporter 1 ( Q9GZV3 , Q9GZV3 ) , vesicular acetylcholine transporter ( Q16572 REA , Q16572 REA ) , and nACh receptors ( AChRs , CHRNAs ) . DB00184 MEN increased the production of ACh in human BACs , and ACh acts as a growth factor for these cells . DB00184 MEN - induced ACh production was mediated by α7 - , α3β2 - , and β3 - nAChRs , P28329 REA and Q16572 REA pathways . We observed that nicotine upregulated P28329 REA and Q16572 REA . Therefore , we conjectured that Q16572 REA antagonists , such as vesamicol , may suppress the growth of human BACs . Vesamicol induced potent apoptosis of human BACs in cell culture and nude mice models . Vesamicol did not have any effect on P01133 REA or insulin-like growth factor-II-induced growth of human BACs . siRNA-mediated attenuation of Q16572 REA reversed the apoptotic activity of vesamicol . We also observed that vesamicol inhibited Akt phosphorylation during cell death and that overexpression of constitutively active Akt reversed the apoptotic activity of vesamicol . Taken together , our results suggested that disruption of nicotine-induced cholinergic signaling by agents such as vesamicol may have applications in BAC therapy .

Red blood cell surface adhesion molecules : their possible roles in normal human physiology and disease . Human erythrocytes express a relatively large number of known adhesion receptors , despite the fact that red blood cells ( RBCs ) are generally considered to be nonadhesive for endothelial cell surfaces . Some of these adhesion receptors are expressed by many other tissues , while others have more limited tissue distribution . Some adhesion receptors , including P16671 REA and VLA - 4 , are only expressed by immature erythroid cells , while others are present on mature erythrocytes . The structure and function of these proteins is reviewed here . LW , P16671 REA , P19256 REA , and CD147 have been shown in other tissues to mediate cell-cell interaction . Other receptors , such as P16070 REA , VLA - 4 , and B - P62158 / LU , can mediate adhesion to components of extracellular matrix . In addition , their roles in normal erythropolesis , as well as in the pathophysiology of human disease , are summarized . The most convincing evidence for a pathophysiologic role for any of these receptors on erythrocytes comes from studies of cells from patients homozygous for hemoglobin S , as RBC adhesion is thought to contribute to vaso-occlusion . Thus , receptors such as B - P62158 / LU may become targets for future therapy aimed at preventing or ameliorating this thrombotic process .

Separation and purification of the tonoplast ATPase and pyrophosphatase from plants with constitutive and inducible Crassulacean acid metabolism . Tonoplast vesicles were isolated from Kalanchoe daigremontiana Hamet et Pierrer de la Bâthie and Mesembryanthemum crystallinum L . , exhibiting constitutive and inducible crassulacean acid metabolism ( P62158 ) , respectively . Membrane-bound proteins were detergent-solubilized with 2 % of Triton X - 100 . During P62158 induction in M . crystallinum , ATPase activity increases four-fold , whereas pyrophosphatase activity decreases somewhat . With all plants , ATPase and pyrophosphatase could be separated by size-exclusion chromatography ( SEC , Sephacryl S 400 ) , and the ATPase was further purified by diethylaminoethyl-ion-exchange chromatography . Sodium-dodecyl-sulfate electrophoresis of the SEC fractions from K . daigremontiana containing maximum ATPase activity separates several protein bands , indicating subunits of 72 , 56 , 48 , 42 , 28 , and 16 kDa . Purified ATPase from M . crystallinum in the P01024 REA and P62158 states shows a somewhat different protein pattern . With M . crystallinum , an increase in DB00171 - hydrolysis and changes in the subunit composition of the native enzyme indicate that the change from the P01024 REA to the P62158 state is accompanied by de-novo synthesis and by structural changes of the tonoplast ATPase .

DB08860 SUB inhibits vascular smooth muscle cell proliferation by inactivating extracellular signal-regulated kinases 1/2 . We recently reported that lysophosphatidylcholine ( lysoPC ) acts on vascular smooth muscle cells ( VSMCs ) to produce a mitogenic response through the activation of extracellular signal-regulated kinases 1/2 ( P27361 REA / 2 ) . In this study , we examined the role of P04035 REA inhibitors on lysoPC-induced VSMC proliferation . DB08860 SUB , a new P04035 REA inhibitor , suppressed lysoPC-induced DNA synthesis in primary cultured rat VSMCs . Since lysoPC-induced P27361 REA / 2 activation contributes to smooth muscle cell proliferation , we explored the effect of pitavastatin on P27361 REA / 2 activation . DB08860 SUB inhibited lysoPC-induced P27361 REA / 2 phosphorylation and P27361 REA / 2 activation . The other P04035 REA inhibitors , atrovastatin and fluvastatin , also inhibited lysoPC-induced P27361 REA / 2 phosphorylation . DB08860 SUB also inhibited lysoPC-induced c-fos mRNA expression . To gain insight into the mechanism of the inhibitory effect of pitavastatin on P27361 REA / 2 activation by lysoPC , we examined the role of the mevalonate pathways . Mevalonate and farnesylpyrophosphate reduced the inhibition of P27361 REA / 2 phosphorylation by pitavastatin . These studies demonstrate that pitavastatin may inhibit lysoPC-induced VSMC proliferation , at least in part , by inactivating P27361 REA / 2 , which is linked to mevalonate metabolism .

DB08860 SUB prevents DB01221 - induced retinal ganglion cell death by suppressing leukocyte recruitment . Excitotoxicity is a major cause of retinal ganglion cell ( RGC ) death during ischemic diseases such as vessel occlusion and diabetic retinopathy . However , the underlying mechanisms are not well understood . Statins , inhibitors of the P04035 REA , have neuroprotective effects in addition to their original role in lowering cholesterol . We hypothesize that pitavastatin , a recently introduced potent statin , is protective against N-methyl-d-aspartic acid ( DB01221 ) - induced RGC death . DB08860 SUB , administered by gavage , abolished DB01221 - induced loss of RGCs . To elucidate the mechanisms underlying the neuroprotective effect of pitavastatin , we investigated its impact on inflammation . DB01221 increased the expression of interleukin - 1beta and P01375 REA , and endothelial adhesion molecules , including P05362 REA , and induced leukocyte accumulation in the retinal vessels . DB08860 SUB significantly reduced DB01221 - induced leukocyte accumulation and up-regulation of endothelial adhesion molecules , whereas cytokine expression was unaffected . Systemic blockade of P05362 REA in wild-type mice or absence of P05107 REA in gene-deficient ( P05107 REA ( - / - ) ) mice significantly suppressed DB01221 - induced leukocyte accumulation and RGC death . These findings suggest a novel and causative role for inflammatory leukocyte recruitment in DB01221 - induced excitotoxicity . Furthermore , we show the novel neuroprotective effect of statins against excitotoxicity-induced RGC death . Statins or other anti-inflammatory agents may thus have therapeutic benefits in excitotoxicity-associated neuronal diseases through blockade of leukocyte recruitment .

The effect of prenatal nicotine on mRNA of central cholinergic markers and hematological parameters in rat fetuses . A number of studies have demonstrated the influence of nicotine on fetal development . This study determined the expression of choline acetyltransferase ( P28329 REA ) , vesicular acetylcholine transporter ( Q16572 REA ) , and high-affinity choline transporter ( Q9GZV3 ) in the forebrain and hindbrain following chronic prenatal nicotine exposure in the rat fetus ( maternal rats were subcutaneously injected with nicotine at different gestation periods ) . We also measured the effect of chronic nicotine exposure on fetal blood pO ( 2 ) , pCO ( 2 ) , pH , Na ( + ) and K ( + ) concentrations , as well as lactic acid levels . Maternal nicotine exposure during pregnancy was associated with a decrease in fetal pO ( 2 ) coupled with a significant increase in pCO ( 2 ) and lactic acid as well as restricted fetal growth . Additionally , maternal nicotine administration also reduced P28329 REA , Q16572 REA , and Q9GZV3 mRNA levels in the fetal brain . DB00184 MEN - induced fetal hypoxic responses and reduced cholinergic marker expression in the brain were more severe when nicotine was started in early gestation . Our results provide new information about the effects of repeated exposure to nicotine in utero on the expression of central P28329 REA , Q16572 REA , and Q9GZV3 in the rat fetus . These results indicate that repeated hypoxic episodes or / and a direct effect of nicotine on the central cholinergic system during pregnancy may contribute to brain developmental problems in fetal origin .

Immunohistochemistry in histoid leprosy . BACKGROUND : Histoid leprosy is a rare form of multibacillary leprosy as the result of secondary or even primary resistance to dapsone . The etiopathogenesis has not been clarified up to now . METHODS : An immunohistochemical study was carried out for the expression of various markers on epidermal and dermal cell populations using sections of frozen skin specimens from 5 patients with histoid leprosy as compared to specimens from 7 tuberculoid and 7 lepromatous patients . RESULTS : Dendritic epidermal cells , identified by monoclonal antibodies against CD1 , HLA-DR , P08575 REA , and P16671 REA , were found reduced in histoid leprosy as compared to both tuberculoid and lepromatous groups . A gradual reduction of keratinocytic HLA-DR expression from tuberculoid to lepromatous to histoid leprosy was observed . The pattern of P16671 REA , P01730 REA , and CD8 expression of lymphomonocytic cells in the dermis of histoid lesions was similar to that of tuberculoid leprosy , but without the formation of an organized granuloma . P08575 REA + cells as well as activated lymphocytic cells , expressed by the activation immunophenotype ( CD1 , HLA-DR , CD25 , CD71 , P01133 REA - R ) were found frequently in all groups . CONCLUSIONS : The in situ immunohistochemical findings support a modified hypersensitivity reaction of the cellular type that results in an inhibition of the lesional expansion , but not in the destruction of the bacilli within the histoid lesion .

A common haplotype of the nicotine acetylcholine receptor alpha 4 subunit gene is associated with vulnerability to nicotine addiction in men . DB00184 MEN is the major addictive substance in cigarettes , and genes involved in sensing nicotine are logical candidates for vulnerability to nicotine addiction . We studied six single-nucleotide polymorphisms ( SNPs ) in the P43681 REA gene and four SNPs in the P17787 REA gene with respect to nicotine dependence in a collection of 901 subjects ( 815 siblings and 86 parents ) from 222 nuclear families with multiple nicotine-addicted siblings . The subjects were assessed for addiction by both the Fagerstrom Test for DB00184 MEN Dependence ( FTND ) and the Revised Tolerance Questionnaire ( RTQ ) . Because only 5.8 % of female offspring were smokers , only male subjects were included in the final analyses ( 621 men from 206 families ) . Univariate ( single-marker ) family-based association tests ( FBATs ) demonstrated that variant alleles at two SNPs , rs1044396 and rs1044397 , in exon 5 of the P43681 REA gene were significantly associated with a protective effect against nicotine addiction as either a dichotomized trait or a quantitative phenotype ( i . e . , age-adjusted FTND and RTQ scores ) , which was consistent with the results of the global haplotype FBAT . Furthermore , the haplotype-specific FBAT showed a common ( 22.5 % ) P43681 REA haplotype , GCTATA , which was significantly associated with both a protective effect against nicotine addiction as a dichotomized trait ( Z = -3.04 , P < . 005 ) and significant decreases of age-adjusted FTND ( Z = -3.31 , P < . 005 ) or RTQ scores ( Z = -2.73 , P= . 006 ) . Our findings provide strong evidence suggesting a common P43681 REA haplotype might be protective against vulnerability to nicotine addiction in men .