Identification of differentially methylated CpG Sites in fibroblasts from Keloid Scars

As a part of an abnormal healing process of dermal injuries and irritation, keloid scars arise on the skin as benign fibroproliferative tumors. Although the etiology of keloid scarring remains unsettled, considerable recent evidence suggested that keloidogenesis may be driven by epigenetic changes, particularly, DNA methylation. Therefore, genome-wide scanning of methylated cytosine-phosphoguanine (CpG) sites in extracted DNA from 12 keloid scar fibroblasts (KF) and 12 control skin fibroblasts (CF) (six normal skin fibroblasts and six normotrophic fibroblasts) was conducted using the Illumina Human Methylation 450K BeadChip in two replicates for each sample. Comparing KF and CF used a Linear Models for Microarray Data (Limma) model revealed 100,000 differentially methylated (DM) CpG sites, 20,695 of which were found to be hypomethylated and 79,305 were hypermethylated. The top DM CpG sites were associated with TNKS2, FAM45B, LOC723972, GAS7, RHBDD2 and CAMKK1. Subsequently, the most functionally enriched genes with the top 100 DM CpG sites were significantly (p ≤ 0.05) associated with SH2 domain binding, regulation of transcription, DNA-templated, nucleus, positive regulation of protein targeting to mitochondrion, nucleoplasm, Swr1 complex, histone exchange, and cellular response to organic substance. In addition, NLK, CAMKK1, LPAR2, CASP1, and NHS showed to be the most common regulators in the signaling network analysis. Taken together, these findings shed light on the methylation status of keloids that could be implicated in the underlying mechanism of keloid scars formation and remission

Methylome-wide association study of central adiposity implicates genes involved in immune and endocrine systems

Aim: We conducted a methylome-wide association study to examine associations between DNA methylation in whole blood and central adiposity and body fat distribution, measured as waist circumference, waist-to-hip ratio and waist-to-height ratio adjusted for body mass index, in 2684 African-American adults in the Atherosclerosis Risk in Communities study. Materials & methods: We validated significantly associated cytosine-phosphate-guanine methylation sites (CpGs) among adults using the Women's Health Initiative and Framingham Heart Study participants (combined n = 5743) and generalized associations in adolescents from The Raine Study (n = 820). Results & conclusion: We identified 11 CpGs that were robustly associated with one or more central adiposity trait in adults and two in adolescents, including CpG site associations near TXNIP, ADCY7, SREBF1 and RAP1GAP2 that had not previously been associated with obesity-related traits

Effects of 59Fe, 65Zn and of three soil types on dry matter yield, chemical composition and nitrogen fixation in Phaseolus vulgaris L. cv. carioca

The aim of this work was to study in greenhouse conditions the effects of two levels of iron and zinc on yield and chemical composition of common bean (Phaseolus vulgaris L.) and on atmospheric nitrogen fixation, in three soils, classified as Terra Roxa Estruturada (TRE), Latossol Vermelho Escuro (LVE), and Podzolico Vermelho Amarelo (PVA). The coefficient of utilization of these micronutrients by this crop and its distribution in above-ground parts and roots were also assessed. The rates for iron were 1.5 and 3.0 ppm, and for zinc, 2.5 and 5.0 ppm. It was applied 7.5 µCi of 59Fe/kg of soil with the lower rate of the stable iron, and 5.0 and 10.0 µCi of Zn/kg of soil in the pots corresponding to the lower and higher rate of the stable zinc, respectively. The plants were harveste at the age of 60 days and nitrogen, phosphorus, potassium, iron and zinc contents were determined. Immediately after harvest, symbiotic nitrogen fixation was assessed, using the acetylene reduction method. The detection of 59Fe and 65zn radioactivity were carried out on nitric percloric extract, by gamma ray spectrometry. The behavior of common bean presented high variation among the three soils, for all the variables. There was no influence of treatments of iron and zinc on dry matter of above ground part and root and also on the weight and number of nodules. The rate of 3.0 ppm of iron decreased the capacity of nodules to fix atmospheric nitrogen in relation to rate of 1.5 ppm, while the rate of 5.0 ppm of zinc increased this capacity, in relation to the rate of 2.5 ppm. There was significative effect of treatments on nitrogen, potassium and zinc contents in above ground part and on nitrogen and zinc contents in the root. The absorption of zinc from the fertilizer and the percentagem of zinc in the plant derived from fertilizer were diretly influenced by rate of zinc The higher coefficient of utilization of zinc from the fertilizer was 4.0%.No presente trabalho, conduzido em casa de vegetação, procuramos estudar os efeitos dos micronutrientes ferro e zinco na produção de materia seca, composição química do feijoeiro (Phaseolus vulgaris L.) e na fixação do nitrogênio atmosférico, em três solos, classificados como Terra Roxa Estruturada (TRE), Latossol Vermelho Escuro (LVE) e Podzólico Vermelho Amarelo (PVA). Procuramos também determinar os índices de aproveitamento destes micronutrientes pelo feijoeiro e sua distribuição na parte aérea e na raiz. O delineamento experimental foi um fatorial 3x7, sendo três solos e sete tratamentos por solo, com três repetições. Nos tratamentos, foram utilizados duas doses de ferro e duas doses de zinco em separado ou combinando as doses menores e maiores destes micronutrientes (Fe1Zn1, Fe2Zn2). As doses de ferro foram 1,5 e 3,0 ppm e as de zinco foram 2,5 e 5,0 ppm. Foram aplicados 7,5 µCi de 59Fe/kg de solo nos vasos correspondentes à dose menor de ferro e 5,0 e 10,0 µCi de 65Zn/kg de solo nos vasos correspondentes respectivamente à dose menor e maior de zinco. Todos os tratamentos receberam uma adubação básica. O comportamento do feijoeiro apresentou grande variação entre os três tipos de solos, para todas as variáveis. Não houve influência dos tratamentos de ferro e zinco na produção de parte aérea e raiz e nem no peso e numero dos nodulos. A dose de 3,0 ppm de ferro diminuiu a capacidade dos nódulos de fixarem nitrogênio atmosférico em relação à dose de 1,5 ppm enquanto que a dose de 5,0 ppm de zinco aumentou esta capacidade, em relação à dose de 2,5 ppm. Houve um efeito significativo dos tratamentos na concentração de nitrogênio, potássio, ferro e zinco na parte aérea e na concentração de nitrogênio, e zinco na raiz. A absorção de zinco dos fertilizantes e a percentagem do zinco na planta proveniente do adubo foram influenciadas diretamente pelas doses de zinco. O maior coeficiente de aproveitamento do zinco do adubo foi de 4,0%

Genomic and phenotypic insights from an atlas of genetic effects on DNA methylation

DNA methylation quantitative trait locus (mQTL) analyses on 32,851 participants identify genetic variants associated with DNA methylation at 420,509 sites in blood, resulting in a database of >270,000 independent mQTLs.Characterizing genetic influences on DNA methylation (DNAm) provides an opportunity to understand mechanisms underpinning gene regulation and disease. In the present study, we describe results of DNAm quantitative trait locus (mQTL) analyses on 32,851 participants, identifying genetic variants associated with DNAm at 420,509 DNAm sites in blood. We present a database of >270,000 independent mQTLs, of which 8.5% comprise long-range (trans) associations. Identified mQTL associations explain 15-17% of the additive genetic variance of DNAm. We show that the genetic architecture of DNAm levels is highly polygenic. Using shared genetic control between distal DNAm sites, we constructed networks, identifying 405 discrete genomic communities enriched for genomic annotations and complex traits. Shared genetic variants are associated with both DNAm levels and complex diseases, but only in a minority of cases do these associations reflect causal relationships from DNAm to trait or vice versa, indicating a more complex genotype-phenotype map than previously anticipated.Molecular Epidemiolog

Identification of novel sarcoma risk genes using a two-stage genome wide DNA sequencing strategy in cancer cluster families and population case and control cohorts

Background Although familial clustering of cancers is relatively common, only a small proportion of familial cancer risk can be explained by known cancer predisposition genes. Methods In this study we employed a two-stage approach to identify candidate sarcoma risk genes. First, we conducted whole exome sequencing in three multigenerational cancer families ascertained through a sarcoma proband (n = 19) in order to prioritize candidate genes for validation in an independent case-control cohort of sarcoma patients using family-based association and segregation analysis. The second stage employed a burden analysis of rare variants within prioritized candidate genes identified from stage one in 560 sarcoma cases and 1144 healthy ageing controls, for which whole genome sequence was available. Results Variants from eight genes were identified in stage one. Following gene-based burden testing and after correction for multiple testing, two of these genes, ABCB5 and C16orf96, were determined to show statistically significant association with cancer. The ABCB5 gene was found to have a higher burden of putative regulatory variants (OR = 4.9, p-value = 0.007, q-value = 0.04) based on allele counts in sarcoma cases compared to controls. C16orf96, was found to have a significantly lower burden (OR = 0.58, p-value = 0.0004, q-value = 0.003) of regulatory variants in controls compared to sarcoma cases. Conclusions Based on these genetic association data we propose that ABCB5 and C16orf96 are novel candidate risk genes for sarcoma. Although neither of these two genes have been previously associated with sarcoma, ABCB5 has been shown to share clinical drug resistance associations with melanoma and leukaemia and C16orf96 shares regulatory elements with genes that are involved with TNF-alpha mediated apoptosis in a p53/TP53-dependent manner. Future genetic studies in other family and population cohorts will be required for further validation of these novel findings

Field-based detection and monitoring of uranium in contaminated groundwater using two immunosensors

Field-based monitoring of environmental contaminants has long been a need for environmental scientists. Described herein are two kinetic exclusion-based immunosensors, a field portable sensor (FPS) and an inline senor, that were deployed at the Integrated Field Research Challenge Site of the U.S. Department of Energy in Rifle, CO. Both sensors utilized a monoclonal antibody that binds to a U(VI)-dicarboxyphenanthroline complex (DCP) in a kinetic exclusion immunoassay format. These sensors were able to monitor changes of uranium in groundwater samples from {approx} 1 {micro}M to below the regulated drinking water limit of 126 nM (30 ppb). The FPS is a battery-operated sensor platform that can determine the uranium level in a single sample in 5-10 min, if the instrument has been previously calibrated with standards. The average minimum detection level (MDL) in this assay was 0.33 nM (79 ppt), and the MDL in the sample (based on a 1:200?1:400 dilution) was 66?132 nM (15.7?31.4 ppb). The inline sensor, while requiring a grounded power source, has the ability to autonomously analyze multiple samples in a single experiment. The average MDL in this assay was 0.12 nM (29 ppt), and the MDL in the samples (based on 1:200 or 1:400 dilutions) was 24?48 nM (5.7?11.4 ppb). Both sensor platforms showed an acceptable level of agreement (r{sup 2} = 0.94 and 0.76, for the inline and FPS, respectively) with conventional methods for uranium quantification

Epigenetic age acceleration in adolescence associates with BMI, inflammation and risk score for middle age cardiovascular disease

Background‘Accelerated ageing’, assessed by adult DNA methylation predicts cardiovascular disease (CVD). Adolescent accelerated aging might predict CVD earlier. We investigated whether epigenetic age acceleration (assessed age 17-years) associated with adiposity/CVD-risk measured (ages 17, 20, 22-years), and projected CVD by middle-age.MethodsDNA methylation measured in peripheral blood provided 2 estimates of epigenetic age acceleration; intrinsic (IEAA, (preserved across cell types) and extrinsic (EEAA, dependent on cell admixture and methylation levels within each cell type).Adiposity was assessed by anthropometry, ultrasound and DEXA (ages 17, 20, 22 years). CVD-risk factors (lipids, HOMA-IR, blood pressure, inflammatory markers) were assessed at age 17-years. CVD development by age 47 years was calculated by Framingham algorithms. Results are presented as regression coefficients/5-year epigenetic age acceleration (IEAA/EEAA) for adiposity, CVD-risk factors and CVD development.ResultsIn 995 participants (49.6% female, age 17.3+/-0.6 years), EEAA (/5-years) was associated with increased BMI of 2.4% (95%CI 1.2-3.6%) and 2.4% (0.8-3.9%) at 17 and 22 years, respectively. EEAA was associated with increases of 23% (3-33%) in hsCRP, 10% (4-17%) in interferon-gamma induced protein (IP-10) and 4% (2-6%) in tumour necrosis factor receptor 2 (sTNFR2), adjusted for BMI and HOMA-IR. EEAA(/5-years) results in a 4% increase in hard endpoints of CVD by 47 years old and a 3% increase, after adjustment for conventional risk factors.ConclusionsAccelerated epigenetic age in adolescence was associated with inflammation, BMI measured 5 years later, and probability of middle-age CVD. Irrespective whether this is cause or effect, assessing epigenetic age might refine disease prediction