452 research outputs found
More Than 200 Genes Required for Methane Formation from H2 and CO2 and Energy Conservation Are Present in Methanothermobacter marburgensis and Methanothermobacter thermautotrophicus
The hydrogenotrophic methanogens Methanothermobacter marburgensis and Methanothermobacter thermautotrophicus can easily be mass cultured. They have therefore been used almost exclusively to study the biochemistry of methanogenesis from H2 and CO2, and the genomes of these two model organisms have been sequenced. The close relationship of the two organisms is reflected in their genomic architecture and coding potential. Within the 1,607 protein coding sequences (CDS) in common, we identified approximately 200 CDS required for the synthesis of the enzymes, coenzymes, and prosthetic groups involved in CO2 reduction to methane and in coupling this process with the phosphorylation of ADP. Approximately 20 additional genes, such as those for the biosynthesis of F430 and methanofuran and for the posttranslational modifications of the two methyl-coenzyme M reductases, remain to be identified
Viktimisierungsstudie Sachsen (VisSa) - Studie zur Betroffenheit von Frauen durch sexualisierte Gewalt, hÀusliche/partnerschaftliche Gewalt und Stalking: Abschlussbericht
Um belastbare Zahlen zur Situation von gewaltbetroffenen Frauen in Sachsen zu erheben, hat das SĂ€chsische Staatsministerium der Justiz und fĂŒr Demokratie, Europa und Gleichstellung (SMJusDEG) die »Dunkelfeldstudie zur Viktimisierung von Frauen durch hĂ€usliche Gewalt, Stalking und sexualisierte Gewalt« (VisSa-Studie) in Auftrag gegeben.
Redaktionsschluss: 01.01.202
Optimization of biotinyl-tyramide-based in situ hybridization for sensitive background-free applications on formalin-fixed, paraffin-embedded tissue specimens
BACKGROUND: Over the past five years in situ hybridization techniques employing tyramide amplification reagents have been developed and promise the potential detection of low/single-copy nucleic acid sequences. However the increased sensitivity that tyramide amplification brings about may also lead to problems of background staining that confound data interpretation. METHODS: In this study those factors enabling background-free biotinyl-tyramide based in situ hybridization assay of formalin-fixed paraffin-embedded tissues have been examined. SiHa, HeLa and CaSki cell lines known to contain HPV integrated into the cell genome, and archival cervical pre-invasive lesions and carcinomas have been successfully assessed using biotinylated HPV and centromeric probes. RESULTS: The single most important factor both for sensitivity and clean background was a tissue unmasking regimen that included treatment with 10 mM sodium citrate pH 6.0 at 95°C followed by digestion with pepsin/0.2 M HCl. Concentrations both of probe and primary streptavidin-peroxidase conjugate and pH of hybridization mix and stringency washes were also critical for sensitivity. Certain probes were more associated with background staining than others. This problem was not related to probe purity or size. In these instances composition of hybridization mix solution was especially critical to avoid background. 3-amino-9-ethylcarbazole was preferred over 3,3'-diaminobenzidene as a chromogen because background was cleaner and the 1â2 copies of HPV16 integrated in SiHa cells were readily demonstrable. HPV detection on metaphase spreads prepared from SiHa cells was only successful when a fluorescent detection method was combined with tyramide reagent. 'Punctate' and 'diffuse' signal patterns were identified amongst tissues consistent with the former representing integration and 'diffuse' representing episomal HPV. Only punctate signals were detected amongst the cell lines and were common amongst high-grade pre-invasive lesions and carcinomas. However it remains to be determined why single/low-copy episomal HPV in basal/parabasal cells of low-grade lesions is not also detectable using tyramide-based techniques and whether every punctate signal represents integration. CONCLUSIONS: A tyramide-based in situ hybridization methodology has been established that enables sensitive, background-free assay of clinical specimens. As punctate signals characterize HPV in high-grade cervical lesions the method may have potential for clinical applications
The Ty1 integrase protein can exploit the classical nuclear protein import machinery for entry into the nucleus
Like its retroviral relatives, the long terminal repeat retrotransposon Ty1 in the yeast Saccharomyces cerevisiae must traverse a permanently intact nuclear membrane for successful transposition and replication. For retrotransposition to occur, at least a subset of Ty1 proteins, including the Ty1 integrase, must enter the nucleus. Nuclear localization of integrase is dependent upon a C-terminal nuclear targeting sequence. However, the nuclear import machinery that recognizes this nuclear targeting signal has not been defined. We investigated the mechanism by which Ty1 integrase gains access to nuclear DNA as a model for how other retroelements, including retroviruses like HIV, may utilize cellular nuclear transport machinery to import their essential nuclear proteins. We show that Ty1 retrotransposition is significantly impaired in yeast mutants that alter the classical nuclear protein import pathway, including the Ran-GTPase, and the dimeric import receptor, importin-α/ÎČ. Although Ty1 proteins are made and processed in these mutant cells, our studies reveal that an integrase reporter is not properly targeted to the nucleus in cells carrying mutations in the classical nuclear import machinery. Furthermore, we demonstrate that integrase coimmunoprecipitates with the importin-α transport receptor and directly binds to importin-α. Taken together, these data suggest Ty1 integrase can employ the classical nuclear protein transport machinery to enter the nucleus
Gljivice kao sastavni dio bioaerosola u nastambama za muzne krave i nesilice konzumnih jaja
The air of animal dwellings can contain great amounts of bioaerosol composed of dust, bacteria, fungi, and endotoxins. The composition may depend on animal species, building construction, animal accommodation, and microclimate parameters, to name just a few factors. Pathogens contained may be a serious threat to
animal and human health.
The aim of our study was to analyse the fungi aerosol content in a stable housing dairy cows and in a coop for laying hens over the three autumn months of 2007. The air was sampled on Petri dishes with Sabouraud glucose agar. After laboratory treatment, we identifi ed the most common fungi. Their count in the stable ranged from 3.98x103 CFU m-3 to 5.11x104 CFU m-3 and in the coop from 6.89 x104 CFU m-3 to 1.13x105 CFU m-3. The difference between the two animal dwellings was statistically different at the level of p<0.05. In both dwellings, the most common were the fungi Aspergillus sp., Penicillium sp., and yeasts, followed by Cladosporium sp., Fusarium sp., Mucor sp., Scopulariopsis sp., Alternaria sp., and Rhizopus sp.
Our results are entirely in line with values reported in literature and are at the lower end of the range. They call for further investigation that would eventually lead to setting air quality standards for animal dwellings and to developing reliable monitoring systems in order to ensure safe food and safe environment.U zraku nastambi za drĆŸanje ĆŸivotinja stvaraju se znatne koliÄine bioaerosola. Njega Äine praĆĄina, bakterije, gljivice, endotoksini i plinovi. Brojnost im ovisi o graÄevinsko-tehniÄkim znaÄajkama nastambi, naseljenosti ĆŸivotinjama, naÄinu drĆŸanja, temperaturno-vlaĆŸnim odnosima u staji i aktivnostima oko hranjenja, muĆŸnje,
skupljanja jaja i drugih poslova. Ove Äestice, ako su patogene, mogu biti ozbiljna prijetnja za zdravlje ljudi.
Mjerenja su obavljana u staji muznih krava te u objektu za nesilice, 2007. godine, tijekom tri jesenja mjeseca.
Zrak je uzorkovan na Petrijeve zdjelice sa Sabouraudovim glukoznim agarom, ureÄajem MAS 100. Nakon obrade u laboratoriju, prema osnovnim i mikromorfoloĆĄkim osobinama poraslih kolonija identifi cirani su najÄeĆĄÄe zastupljeni rodovi gljivica.
Srednja vrijednost broja gljivica u zraku staje za muzne krave kretala se od 3,98x103 CFU m-3 do 5,11x104 CFU m-3. Broj gljivica u zraku objekta za nesilice kretao se od 6,89x104 CFU m-3 do
1,13x105 CFU m-3. Ove vrijednosti statistiÄki su se znaÄajno razlikovale na razini p<0,05.
U obje pretraĆŸivane nastambe najÄeĆĄÄe su bili zastupljeni rodovi Aspergillus, Penicillium i kvasnice. U manjem postotku utvrÄene su gljivice iz rodova Cladosporium sp., Fusarium sp., Mucor sp., Scopulariopsis sp., Alternaria sp. i Rhizopus sp.
Rezultati ovih istraĆŸivanja o kvantitativnom i kvalitativnom sastavu gljivica u zraku pretraĆŸenih nastambi potpuno su u skladu s vrijednostima zabiljeĆŸenim u literaturi te se nalaze na donjim granicama opisanih raspona. UtvrÄeni broj i rodovi gljivica ukazuju na nuĆŸnost daljnjih istraĆŸivanja te potrebu postavljanja standardnih vrijednosti glede kvalitete zraka u nastambama za ĆŸivotinje, kao i razvoj vjerodostojnog sustava praÄenja navedenih Äimbenika, s ciljem stvaranja sigurne hrane i sigurnog okoliĆĄa
Common Variants at 10 Genomic Loci Influence Hemoglobin A(1C) Levels via Glycemic and Nonglycemic Pathways
OBJECTIVE Glycated hemoglobin (HbA1c), used to monitor and diagnose diabetes, is influenced by average glycemia over a 2- to 3-month period. Genetic factors affecting expression, turnover, and abnormal glycation of hemoglobin could also be associated with increased levels of HbA1c. We aimed to identify such genetic factors and investigate the extent to which they influence diabetes classification based on HbA1c levels.
RESEARCH DESIGN AND METHODS We studied associations with HbA1c in up to 46,368 nondiabetic adults of European descent from 23 genome-wide association studies (GWAS) and 8 cohorts with de novo genotyped single nucleotide polymorphisms (SNPs). We combined studies using inverse-variance meta-analysis and tested mediation by glycemia using conditional analyses. We estimated the global effect of HbA1c loci using a multilocus risk score, and used net reclassification to estimate genetic effects on diabetes screening.
RESULTS Ten loci reached genome-wide significant association with HbA1c, including six new loci near FN3K (lead SNP/P value, rs1046896/P = 1.6 Ă 10â26), HFE (rs1800562/P = 2.6 Ă 10â20), TMPRSS6 (rs855791/P = 2.7 Ă 10â14), ANK1 (rs4737009/P = 6.1 Ă 10â12), SPTA1 (rs2779116/P = 2.8 Ă 10â9) and ATP11A/TUBGCP3 (rs7998202/P = 5.2 Ă 10â9), and four known HbA1c loci: HK1 (rs16926246/P = 3.1 Ă 10â54), MTNR1B (rs1387153/P = 4.0 Ă 10â11), GCK (rs1799884/P = 1.5 Ă 10â20) and G6PC2/ABCB11 (rs552976/P = 8.2 Ă 10â18). We show that associations with HbA1c are partly a function of hyperglycemia associated with 3 of the 10 loci (GCK, G6PC2 and MTNR1B). The seven nonglycemic loci accounted for a 0.19 (% HbA1c) difference between the extreme 10% tails of the risk score, and would reclassify âŒ2% of a general white population screened for diabetes with HbA1c.
CONCLUSIONS GWAS identified 10 genetic loci reproducibly associated with HbA1c. Six are novel and seven map to loci where rarer variants cause hereditary anemias and iron storage disorders. Common variants at these loci likely influence HbA1c levels via erythrocyte biology, and confer a small but detectable reclassification of diabetes diagnosis by HbA1c
Genome-wide association study identifies a variant in HDAC9 associated with large vessel ischemic stroke
Genetic factors have been implicated in stroke risk but few replicated associations have been reported. We conducted a genome-wide association study (GWAS) in ischemic stroke and its subtypes in 3,548 cases and 5,972 controls, all of European ancestry. Replication of potential
signals was performed in 5,859 cases and 6,281 controls. We replicated reported associations between variants close to PITX2 and ZFHX3 with cardioembolic stroke, and a 9p21 locus with large vessel stroke. We identified a novel association for a SNP within the histone deacetylase 9(HDAC9) gene on chromosome 7p21.1 which was associated with large vessel stroke including additional replication in a further 735 cases and 28583 controls (rs11984041, combined P =
1.87Ă10â11, OR=1.42 (95% CI) 1.28-1.57). All four loci exhibit evidence for heterogeneity of effect across the stroke subtypes, with some, and possibly all, affecting risk for only one subtype. This suggests differing genetic architectures for different stroke subtypes
A comprehensive 1000 Genomes-based genome-wide association meta-analysis of coronary artery disease
Existing knowledge of genetic variants affecting risk of coronary artery disease (CAD) is largely based on genome-wide association studies (GWAS) analysis of common SNPs. Leveraging phased haplotypes from the 1000 Genomes Project, we report a GWAS meta-analysis of 185 thousand CAD cases and controls, interrogating 6.7 million common (MAF>0.05) as well as 2.7 million low frequency (0.005<MAF<0.05) variants. In addition to confirmation of most known CAD loci, we identified 10 novel loci, eight additive and two recessive, that contain candidate genes that newly implicate biological processes in vessel walls. We observed intra-locus allelic heterogeneity but little evidence of low frequency variants with larger effects and no evidence of synthetic association. Our analysis provides a comprehensive survey of the fine genetic architecture of CAD showing that genetic susceptibility to this common disease is largely determined by common SNPs of small effect siz
New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk.
Levels of circulating glucose are tightly regulated. To identify new loci influencing glycemic traits, we performed meta-analyses of 21 genome-wide association studies informative for fasting glucose, fasting insulin and indices of beta-cell function (HOMA-B) and insulin resistance (HOMA-IR) in up to 46,186 nondiabetic participants. Follow-up of 25 loci in up to 76,558 additional subjects identified 16 loci associated with fasting glucose and HOMA-B and two loci associated with fasting insulin and HOMA-IR. These include nine loci newly associated with fasting glucose (in or near ADCY5, MADD, ADRA2A, CRY2, FADS1, GLIS3, SLC2A2, PROX1 and C2CD4B) and one influencing fasting insulin and HOMA-IR (near IGF1). We also demonstrated association of ADCY5, PROX1, GCK, GCKR and DGKB-TMEM195 with type 2 diabetes. Within these loci, likely biological candidate genes influence signal transduction, cell proliferation, development, glucose-sensing and circadian regulation. Our results demonstrate that genetic studies of glycemic traits can identify type 2 diabetes risk loci, as well as loci containing gene variants that are associated with a modest elevation in glucose levels but are not associated with overt diabetes
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