MH_dev_245

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

Identification of a lung adenocarcinoma cell line with Q16204 - P07949 REA fusion gene and the effect of P07949 REA inhibitors in vitro and in vivo . Rearrangements of the proto-oncogene P07949 REA are newly identified potential driver mutations in lung adenocarcinoma ( LAD ) . However , the absence of cell lines harboring P07949 REA fusion genes has hampered the investigation of the biological relevance of P07949 REA and the development of P07949 REA - targeted therapy . Thus , we aimed to identify a P07949 REA fusion positive LAD cell line . Eleven LAD cell lines were screened for P07949 REA fusion transcripts by reverse transcription-polymerase chain reaction . The biological relevance of the Q16204 - P07949 REA gene products was assessed by cell growth , survival and phosphorylation of P27361 REA / 2 and AKT with or without the suppression of P07949 REA expression using RNA interference . The efficacy of P07949 REA inhibitors was evaluated in vitro using a culture system and in an in vivo xenograft model . Expression of the Q16204 - P07949 REA fusion gene in LC - 2 / ad cells was demonstrated by the mRNA and protein levels , and the genomic break-point was confirmed by genomic DNA sequencing . Mutations in P01116 REA and P00533 REA were not observed in the LC - 2 / ad cells . Q16204 - P07949 REA was constitutively active , and the introduction of a siRNA targeting the P07949 REA 3 ' region decreased cell proliferation by downregulating P07949 REA and P27361 REA / 2 phosphorylation . Moreover , treatment with P07949 REA - inhibitors , including vandetanib , reduced cell viability , which was accompanied by the downregulation of the AKT and P27361 REA / 2 signaling pathways . DB05294 MEN exhibited anti-tumor effects in the xenograft model . Endogenously expressing Q16204 - P07949 REA contributed to cell growth . The inhibition of kinase activity could be an effective treatment strategy for LAD . LC - 2 / ad is a useful model for developing fusion P07949 REA - targeted therapy .

Metabolism of risperidone to 9 - hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9 - hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 REA , P05177 REA , P10632 REA , P11712 REA - arg 144 , P11712 REA - cys 144 , P33261 REA , P10635 REA , P08684 REA and P20815 REA supplemented with an NADPH-generating system . DB01267 SUB was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9 - hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol ( - 1 ) CYP min ( - 1 ) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9 - hydroxyrisperidone is highly correlated with P10635 REA and 3A activities . Thus , both P10635 REA and 3A4 are involved in the 9 - hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 REA ) and ketoconazole ( inhibitor of P08684 REA ) can inhibit the formation of 9 - hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9 - hydroxyrisperidone in rat . The formation of 9 - hydroxyrisperidone is highly correlated with testosterone 6beta - hydroxylase activities , suggesting that inducible CYP 3A contributes significantly to the metabolism of risperidone in rat .

Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 REA - induced survival . This inhibition can not be overcome by increasing concentrations of P05113 REA and is not due to the blocking of Na + channels by lidocaine . Here we report that one class of K + channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 REA . In contrast , increasing concentrations of P08700 REA or granulocyte-macrophage P04141 REA overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 - sensitive K + channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [ 3H ] glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 REA ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases .

Role of xanthine oxidoreductase and NAD ( P ) H oxidase in endothelial superoxide production in response to oscillatory shear stress . Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis . Because oxidative stress contributes to atherosclerosis , we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon . Bovine aortic endothelial cells were exposed to static , laminar ( 15 dyn / cm2 ) , and oscillatory shear stress ( + / - 15 dyn / cm2 ) . Oscillatory shear increased superoxide ( O2 . - ) production by more than threefold over static and laminar conditions as detected using electron spin resonance ( P03372 REA ) . This increase in O2 * - was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources . DB05262 MEN also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence . DB02134 - dependent O2 * - production was increased in homogenates of endothelial cells exposed to oscillatory shear stress . This was associated with decreased xanthine dehydrogenase ( P47989 REA ) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase ( XO ) to P47989 REA . We also studied endothelial cells lacking the p47phox subunit of the NAD ( P ) H oxidase . These cells exhibited dramatically depressed O2 * - production and had minimal XO protein and activity . Transfection of these cells with p47phox restored XO protein levels . Finally , in bovine aortic endothelial cells , prolonged inhibition of the NAD ( P ) H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2 * - production in response to oscillatory shear stress . These data suggest that the NAD ( P ) H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress .

Association of P35462 REA and Q13224 REA with impulse control and related behaviors in Parkinson ' s disease . We aimed to assess whether allelic variants of dopamine receptor , glutamate receptor , and serotonin transporter genes are associated with the appearance of impulse control and related behaviors ( ICRB ) in Parkinson ' s disease ( PD ) with dopamine replacement therapy ( P29323 REA ) . We surveyed ICRB in consecutive Korean patients with PD who were treated with stable P29323 REA using modified Minnesota Impulsive Disorders Interview over a period of 4 months . In the 404 patients who completed the interview and the 559 Korean healthy normal controls , genotyping was performed for variants of the P35462 REA p . S9G , P14416 REA Taq 1A , Q13224 REA c . 366C > G , c . 2664C > T and c . - 200T > G , and the promoter region of the serotonin transporter gene ( 5 - HTTLPR ) . Behavioral abnormalities suggestive of ICRB including compulsive buying , gambling , sexual behavior and eating , and punding , were present in 14.4 % of the patients . Variants of P14416 REA and 5 - HTTLPR were not associated with the risk of developing ICRB . However , the AA genotype of P35462 REA p . S9G and the CC genotype of Q13224 REA c . 366C > G were more frequent in patients with ICRB than in nonaffected patients ( odds ratio [ OR ] = 2.21 , P = 0.0094 ; and 2.14 , P = 0.0087 , after adjusting for age and sex ) . After controlling for clinical variables in the multivariate analysis , carriage of either AA genotype of P35462 REA or CC genotype of Q13224 REA was identified as an independent risk factor for ICRB ( adjusted OR : 2.57 , P = 0.0087 ) . Variants of P35462 REA p . S9G and Q13224 REA c . 366C > G may be associated with the appearance of ICRB in PD .

Genetic polymorphisms in the oxidative stress pathway and susceptibility to non-Hodgkin lymphoma . Oxidative damage caused by reactive oxygen species ( ROS ) and other free radicals is involved in a number of pathological conditions including cancer . In a population-based case-control study of non-Hodgkin lymphoma ( Q9NZ71 ) ( n = 518 cases , 597 controls ) among women in Connecticut , we analyzed one or more single nucleotide polymorphisms ( SNPs ) in ten candidate genes ( P14550 REA , Q04828 REA , P42330 REA , P13498 REA , P07203 REA , P05164 REA , P35228 REA , NOS 3 , O15527 REA , and P04179 REA ) that mediate oxidative stress directly or indirectly in the NADPH oxidase-dependent respiratory burst . Odds ratios ( OR ) and 95 % confidence intervals ( CI ) were adjusted for age and race . Polymorphisms in P14550 REA and P13498 REA were significantly associated with increased risk of Q9NZ71 . There was a 1.7- fold ( 95 % CI = 1.2- 2.4 , P = 0.0047 ) increased risk of Q9NZ71 for individuals who were variant homozygous for the P14550 REA ( IVS 5 + 282T > C ) SNP . The effect was most pronounced for risk of diffuse large B-cell lymphoma , but risk estimates were non-significantly elevated for other common B-cell histologies and T-cell lymphomas as well . In addition , individuals variant homozygous for the P13498 REA ( Ex4 + 11C > T ) SNP had a 1.6- fold ( 95 % CI = 1.1- 2.4 , P = 0.019 ) increased risk of Q9NZ71 that was particularly pronounced for T-cell lymphoma ( OR = 3.5 , 95 % CI = 1.3- 9.6 , P = 0.013 ) , but was also associated with non-significant increased risks for each of the common B-cell histologies . These results suggest that SNPs in genes related to the oxidative stress pathway may be associated with increased risk of Q9NZ71 .

Engineering human T cells for resistance to methotrexate and mycophenolate mofetil as an in vivo cell selection strategy . Gene transfer and drug selection systems that enforce ongoing transgene expression in vitro and in vivo which are compatible with human pharmaceutical drugs are currently underdeveloped . Here , we report on the utility of incorporating human enzyme muteins that confer resistance to the lymphotoxic / immunosuppressive drugs methotrexate ( MTX ) and mycophenolate mofetil ( DB00688 MEN ) in a multicistronic lentiviral vector for in vivo T lymphocyte selection . We found that co-expression of human dihydrofolate reductase ( P00374 REA ( FS ) ; L22F , F31S ) and inosine monophosphate dehydrogenase II ( P12268 REA ( IY ) ; T333I , S351Y ) conferred T cell resistance to the cytocidal and anti-proliferative effects of these drugs at concentrations that can be achieved clinically ( up to 0.1 µM MTX and 1.0 µM DB00603 ) . Furthermore , using a immunodeficient mouse model that supports the engraftment of central memory derived human T cells , in vivo selection studies demonstrate that huEGFRt ( + ) P00374 REA ( FS + ) P12268 REA ( IY + ) T cells could be enriched following adoptive transfer either by systemic administration of MTX alone ( 4.4 - fold ) , DB00688 MEN alone ( 2.9- fold ) , or combined MTX and DB00688 MEN ( 4.9- fold ) . These findings demonstrate the utility of both P00374 REA ( FS ) / MTX and P12268 REA ( IY ) / DB00688 MEN for in vivo selection of lentivirally transduced human T cells . Vectors incorporating these muteins in combination with other therapeutic transgenes may facilitate the selective engraftment of therapeutically active cells in recipients .

Oxidative stress in susceptibility to breast cancer : study in Spanish population . BACKGROUND : Alterations in the redox balance are involved in the origin , promotion and progression of cancer . Inter-individual differences in the oxidative stress regulation can explain a part of the variability in cancer susceptibility.The aim of this study was to evaluate if polymorphisms in genes codifying for the different systems involved in oxidative stress levels can have a role in susceptibility to breast cancer . METHODS : We have analyzed 76 single base polymorphisms located in 27 genes involved in oxidative stress regulation by SNPlex technology . First , we have tested all the selected SNPs in 493 breast cancer patients and 683 controls and we have replicated the significant results in a second independent set of samples ( 430 patients and 803 controls ) . Gene-gene interactions were performed by the multifactor dimensionality reduction approach . RESULTS : Six polymorphisms rs1052133 ( O15527 REA ) , rs406113 and rs974334 ( GPX 6 ) , rs2284659 ( P08294 REA ) , rs4135225 ( P10599 REA ) and rs207454 ( P47989 REA ) were significant in the global analysis . The gene-gene interactions demonstrated a significant four-variant interaction among rs406113 ( GPX 6 ) , rs974334 ( GPX 6 ) , rs105213 ( O15527 REA ) and rs2284659 ( P08294 REA ) ( p-value = 0.0008 ) with high-risk genotype combination showing increased risk for breast cancer ( OR = 1.75 [ 95 % CI ; 1.26- 2.44 ] ) . CONCLUSIONS : The results of this study indicate that different genotypes in genes of the oxidant / antioxidant pathway could affect the susceptibility to breast cancer . Furthermore , our study highlighted the importance of the analysis of the epistatic interactions to define with more accuracy the influence of genetic variants in susceptibility to breast cancer .

Inactivation of caspase - 1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin - 1 ( IL - 1 ) - beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL - 1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL - 1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE / caspase - 1 ) or through caspase - 1 gene deletion . METHODS : P29466 REA was selectively blocked by using pralnacasan or DB05507 MEN . IL - 1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL - 1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 REA inhibition reduced the release of IL - 1beta in organotypic slices exposed to LPS + DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 MEN ( intraperitoneal , 25-200 mg / kg ) to rats blocked seizure-induced production of IL - 1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase - 1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase - 1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL - 1beta .

Analyses of cross species polymerase chain reaction products to infer the ancestral state of human polymorphisms . In numerous population genetic and disease association studies decisions about the ancestry of polymorphic alleles are often made based on the relative frequency of the alleles in the extant populations with the most frequent allele being deemed as ancestral . However , the frequency of an allele in a population is generally not a perfect indicator of its ancestral status . A more accurate method to assess ancestral / derived status of polymorphic alleles involves identification of shared alleles between species . We used this strategy to examine genomic regions homologous to several human polymorphisms in four species of non-human primates . Cross species polymerase chain reaction ( CS-PCR ) , with primers designed from human sequence , was used to investigate regions of interest . Nineteen polymorphisms at six loci ( P14416 REA , HOXB @ , PAH , D4S10 , P10745 REA , and P07949 REA ) were examined either by restriction fragment length analysis of PCR products ( PCR-RFLP ) or by direct sequencing . At seventeen of the eighteen PCR-RFLPs , non-human primates were monomorphic and identical to each other for either lack of restriction enzyme site or presence of the site . Thus , at these seventeen polymorphic sites the shared alleles are most likely to be the ancestral ones in humans . In several cases we have used sequence data to further demonstrate that the nucleotide at the site of the polymorphism is conserved between species confirming the hypothesis of a single ancestral allele . However , not all human alleles can be simply resolved into ancestral and derived ; sequence data from one PCR-RFLP ( in an intron of the PAH locus ) and a single strand conformational polymorphism ( SSCP ) in the 3 ' untranslated region ( UTR ) of the P14416 REA gene illustrate this point .

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

Genetic markers associated with abstinence length in alcohol-dependent subjects treated with acamprosate . DB00659 MEN supports abstinence in some alcohol-dependent subjects , yet predictors of response are unknown . To identify response biomarkers , we investigated associations of abstinence length with polymorphisms in candidate genes in glycine and glutamate neurotransmission pathways and genes previously implicated in acamprosate response . Association analyses were conducted in the discovery sample of 225 alcohol-dependent subjects treated with acamprosate for 3 months in community-based treatment programs in the United States . Data from 110 alcohol-dependent males treated with acamprosate in the study PREDICT were used for replication of the top association findings . Statistical models were adjusted for relevant covariates , including recruitment site and baseline clinical variables associated with response . In the discovery sample , shorter abstinence was associated with increased intensity of alcohol craving and lower number of days between the last drink and initiation of acamprosate treatment . After adjustment for covariates , length of abstinence was associated with the Q13224 REA rs2058878 ( P= 4.6 × 10 ( - 5 ) ) . In the replication sample , shorter abstinence was associated with increased craving , increased depressive mood score and higher alcohol consumption . Association of abstinence length with Q13224 REA rs2058878 was marginally significant ( P= 0.0675 ) ; as in the discovery sample , the minor A allele was associated with longer abstinence . Furthermore , rs2300272 , which is in strong linkage disequilibrium with rs2058878 , was also associated with abstinence length ( P= 0.049 ) . This is the first report of a replicated association of genetic markers with the length of abstinence in acamprosate-treated alcoholics . Investigation of the underlying mechanisms of this association and its usefulness for individualized treatment selection should follow .

Combined adenine phosphoribosyltransferase and P34059 REA deficiency . We describe a Czech patient with combined adenine phosphoribosyltransferase ( P07741 REA ) deficiency ( 2,8- dihydroxyadenine urolithiasis ) and P34059 REA ( P34059 REA ) deficiency ( mucopolysaccharidosis Type IVA , Morquio disease A ) . DB00173 MEN and its extremely insoluble derivative , 2,8- dihydroxyadenine , were identified in the urine , and P07741 REA deficiency was confirmed in erythrocytes . There was excessive excretion of keratan sulfate in the urine , and P34059 REA deficiency was confirmed in leukocytes . P34059 REA and P07741 REA are both located on chromosome 16q24 . 3 , suggesting that the patient had a deletion involving both genes . PCR amplification of genomic DNA indicated that a novel junction was created by the fusion of sequences distal to P34059 REA exon 2 and proximal to P07741 REA exon 3 , and that the size of the deleted region was approximately 100 kb . The deletion breakpoints were localized within P34059 REA intron 2 and P07741 REA intron 2 . Several other genes , including the alpha subunit of cytochrome B ( P13498 REA ) , which is deleted or mutated in the autosomal form of chronic granulomatous disease , are located in the 16q24 . 3 region , but PCR amplification showed that this gene was present in the proband . A patient with hemizygosity for P34059 REA deficiency and P07741 REA deficiency has been reported from Japan recently . These findings indicate that : ( i ) P07741 REA is located telomeric to P34059 REA ; ( ii ) P34059 REA and P07741 REA are transcribed in the same orientation ( centromeric to telomeric ) ; and ( iii ) combined P07741 REA / P34059 REA deficiency may be more common than hitherto realized .

Functional polymorphisms of folate-metabolizing enzymes in relation to homocysteine concentrations in systemic lupus erythematosus . OBJECTIVE : To determine if functional polymorphisms of folate / homocysteine pathway enzymes are associated with homocysteine concentrations and / or coronary artery calcification ( CAC ) scores in patients with systemic lupus erythematosus ( SLE ) and controls . METHODS : We investigated 163 SLE patients and 160 controls . Functional polymorphisms in 6 genes in the folate / homocysteine pathway were genotyped : 5,10- methylenetetrahydrofolate reductase ( P42898 REA ) 677C > T , P42898 REA 1298A > C , cystathionine ss-synthase ( P35520 REA ) 844ins68 , methionine synthase ( Q99707 REA ) 2756A > G , methionine synthase reductase ( Q9UBK8 ) 66A > G , thymidylate synthase ( P04818 REA ) 1494del6 , and dihydrofolate reductase ( P00374 REA ) c . 86 + 60_78 . RESULTS : Homocysteine levels were higher in African American SLE patients than Caucasian patients and African American controls . Genotype distributions were significantly different in African American and Caucasian controls for 6 of the 7 polymorphisms . Genotype distributions for each polymorphism did not differ significantly between SLE patients and controls even after stratification by race . Glomerular filtration rate was strongly negatively correlated to homocysteine levels , and was therefore adjusted for as a covariate in the models of the effects of the polymorphisms on homocysteine levels . In SLE patients none of the 7 polymorphisms was associated with homocysteine concentrations . In Caucasian controls only P42898 REA 677C > T and 1298A > C showed effects on homocysteine similar to what would be expected from the literature . There were no genotypic associations with median CAC scores in SLE patients or controls with and without stratification by race . CONCLUSION : Polymorphisms in folate / homocysteine metabolizing enzymes do not predict higher homocysteine levels or CAC scores in patients with SLE .

Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 REA ) subunits of K ( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K ( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K ( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K ( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 REA varies between tissues : Q09428 REA in beta-cells , SUR 2A in cardiac muscle , and SUR 2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 REA . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 MENMAX DB00222 MEN , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K ( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K ( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K ( DB00171 ) channel subunits . While K ( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR 2 - type K ( DB00171 ) channels ( e . g . , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K ( DB00171 ) channels are to be avoided .

DB00134 - dependence phenotype in the de novo pathway in P38398 REA and P51587 REA mutation carriers with and without breast cancer . DB00134 - dependence phenotype ( P16444 REA ) refers to the reduced ability of cells to proliferate when methionine is restricted and / or replaced by its immediate precursor homocysteine . P16444 REA is a characteristic of human tumors in vivo , human tumor cell lines , and normal somatic tissue in some individuals . It was hypothesized that P16444 REA is a risk factor for developing breast cancer in BRCA ( P38398 REA and P51587 REA ) germline mutation carriers . To test the hypothesis , human peripheral blood lymphocytes of BRCA carriers with and without breast cancer and healthy non-carrier relatives ( controls ) were cultured for 9 days in medium containing either 0.1 mmol / L L-methionine or 0.2 mmol / L D , L-homocysteine , with the ratio of viable cell growth in both types of medium after 9 days used to calculate the methionine-dependence index ( MDI ) , a measure of P16444 REA . We also tested whether P16444 REA was associated with common polymorphisms in methionine metabolism . Viable cell growth , MDI , and polymorphism frequency in Q9UBK8 ( A66G and C524T ) and P42898 REA ( A1298C and A1793G ) did not differ among the study groups ; however , MDI tended to be higher in BRCA carriers with breast cancer than those without and was significantly increased in P42898 REA 677T allele carriers relative to wild-type carriers ( P= 0.017 ) . The presence of Q99707 REA A2756G mutant allele and P42898 REA C677T mutant allele in carriers was associated with increased breast cancer risk [ odds ration , 3.2 ( P= 0.16 ; 95 % confidence interval , 0.76- 13.9 ) and 3.9 ( P= 0.09 ; 95 % confidence interval , 0.93- 16.3 ) , respectively ] . The results of this study support the hypothesis that defects in methionine metabolism may be associated with breast cancer risk in BRCA carriers .

Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis / metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 REA ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 REA ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 REA ) at 3 and 12 months . P10632 REA * 4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 REA levels ( P= 0.003 and P= 0.007 ) and were 2.2- and 2.5- fold more likely to develop P42126 REA and CND ( P= 0.039 and P= 0.041 ) , respectively . P34913 REA 55Arg , P51589 REA * 7 , P10632 REA * 1B , and P10632 REA g . 36785A allele carriers had lower EET and DHET P04141 REA levels . P10632 REA g . 25369T and P10632 REA g . 36755A allele carriers had higher EET levels . Patients with P10632 REA * 2C and P34913 REA 404del variants had worse long-term outcomes while those with P34913 REA 287Gln , P51589 REA * 7 , and P11712 REA g . 816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3 - month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis / metabolic pathway and the pathophysiology of aSAH .

Genetic variants in apoptosis and immunoregulation-related genes are associated with risk of chronic lymphocytic leukemia . To identify low-penetrance susceptibility alleles for chronic lymphocytic leukemia ( CLL ) , we performed a case-control study genotyping 768 single-nucleotide polymorphisms ( SNP ) in 692 cases of CLL and 738 controls . We investigated nonsynonymous SNPs , SNPs with potential functional effect , and tag SNPs in regulatory gene regions in a total of 172 genes involved in cancer biology . After adjustment for multiple testing , we found a strong association between CLL risk and six genetic variants : P51946 REA ( rs2266690 , V270A ) , O14727 REA ( rs17028658 , 3 ' region ) , Q14005 REA ( rs4505265 , first intron ) , Q14790 REA ( rs1045485 , D302H ) , P35228 REA ( rs2779251 , promoter ) , and P32248 REA ( rs3136687 , intron 1 ) . We found association with CLL susceptibility and 22 haplotypes in O14727 REA , P05231 REA , O14836 REA , Q14005 REA , P42574 REA , P32248 REA , P01374 REA / P01375 REA , Q07812 REA , P10415 REA , P48061 REA , CASP 10 / Q14790 REA , P29466 REA , P13500 REA , Q16611 REA , and P01583 REA candidate genes . Finally , we evaluated using public data sets the potential functional effect on gene expression levels of the CLL associated genetic variants detected in regulatory regions . Minor alleles for O14727 REA and Q14005 REA were associated with lower mRNA levels ; no expression differences were observed for P32248 REA , whereas P35228 REA could not be assessed . This study suggests that common genetic variation in apoptosis - and immunoregulation-related genes is associated with the CLL risk .

[ The dynamics of cytokines and their related substances in skin wounds ] . Molecular-pathological study on cytokines and their related substances in skin wound healing was performed from the view points of forensic pathology ; 1 . The dynamics of inflammatory cytokines such as interleukin ( IL ) - 1 alpha , P01584 REA , P05231 REA and tumor necrosis factor-alpha ( P01375 REA alpha ) during murine wound healing was examined at the levels of protein and mRNA . As for the results , the production of those inflammatory cytokines could be considered as one of the local vital reactions after wounding , and it was clarified that those inflammatory cytokines were one of the biological markers for forensic wound age determination . Practically , our immunohistochemical and morphometrical study with human skin wounds with different wound ages suggested that the ratios of P01583 REA - positive cells , considerably exceeding 30 % , indicated a postinfliction interval of 1 day or less . 2 . In mouse skin incision , a significant increase in P22301 REA mRNA occurred between 30 and 180 min . The increased expression of P22301 REA mRNA could be considered a vital reaction in skin wounds . Furthermore , it was clarified that P22301 REA may have an important regulatory role in limiting the extent and duration of inflammatory response during skin wound healing . 3 . Fas and P48023 REA ( Fas L ) were immunohistochemically detectable on macrophages and fibroblasts in skin wound healing process . Morphometrically , the number of cell expressing Fas and / or Fas L reached a peak at 6 days after wounding . Therefore , it is considered that apoptosis through Fas and Fas L may play an important role for reducing the cellularity during proliferative phase of wound healing in mice .

Phytoestrogens induce apoptosis through a mitochondria / caspase pathway in human breast cancer cells . OBJECTIVE : To explore the effect and pathway of phytoestrogens on the growth of breast cancer cell line MCF - 7 . METHODS : MCF - 7 cells ( human estrogen receptor-positive and progesterone receptor-positive breast cancer cells ) were cultured in serum-free medium for 24 h and then treated with genistein , resveratrol , and quercetin ( 10 ( - 10 ) - 10 ( - 4 ) mol / l ) . After further incubation for 24 , 48 , 72 , and 92 h , the cells were harvested and extracted for 3 - ( 4,5- dimethylthiazol - 2 - yl ) -2,5- diphenyltetrazolium bromide ( MTT ) assay . According to the above results , the proteins involving proliferative and apoptotic pathways were evaluated by Western blot analysis . RESULTS : DB01645 MEN , resveratrol , and quercetin significantly inhibited cellular proliferation in a dose - and time-dependent manner . Significantly elevated caspase - 3 activity was noted after treatment with genistein ( 10 ( - 9 ) - 10 ( - 7 ) mol / l ) , as well as with resveratrol and quercetin ( 10 ( - 9 ) - 10 ( - 5 ) mol / l ) . Significant reduction of PI3K and AKT protein and significant increase of P48023 REA , Fas-associated protein with death domain , cytochrome C , truncated Bid , caspase - 9 , and caspase - 3 were all noted after genistein , resveratrol , and quercetin treatment . 17β - Estradiol induced completely opposite results . P03372 REA ( ER ) α expression was significantly increased with 17β - estradiol , whereas ERβ expression was significantly elevated in the cultures with genistein , resveratrol , and quercetin . CONCLUSIONS : These data demonstrate that genistein , resveratrol , and quercetin have antiproliferative effects on breast cancer cells . Their induction of apoptosis involves the activation of both the intrinsic and extrinsic apoptotic pathways , which may be related to the differential affinity to ERα and ERβ . Whether phytoestrogens have similar effects on normal breast cells remains to be examined .