RESUMO
Allergic sensitization is commonly assessed in patients by performing the skin prick test (SPT) or determining specific immunoglobulin (IgE) levels in blood samples with the ImmunoCAP™ assay, which measures each allergen and sample separately. This paper explores the possibility to investigate respiratory allergies with a high throughput method, the Meso Scale Discovery (MSD) multiplex immunoassay, measuring IgE levels in low volumes of blood. The MSD multiplex immunoassay, developed and optimized with standards and allergens from Radim Diagnostics, was validated against the SPT and the ImmunoCAP assay. For 18 adults (15 respiratory allergy patients and three controls), blood collection and the SPT were performed within the same hour. Pearson correlations and Bland-Altman analysis showed high comparability of the MSD multiplex immunoassay with the SPT and the ImmunoCAP assay, except for house dust mite. The sensitivity of the MSD multiplexed assay was ≥78% for most allergens compared to the SPT and ImmunoCAP assay. Additionally, the specificity of the MSD multiplex immunoassay was ≥ 87% - the majority showing 100% specificity. Only the rye allergen had a low specificity when compared to the SPT, probably due to cross-reactivity. The reproducibility of the MSD multiplex immunoassay, assessed as intra- and interassay reproducibility and biological variability between different sampling moments, showed significantly high correlations (r = 0·943-1) for all tested subjects (apart from subject 13; r = 0·65-0·99). The MSD multiplex immunoassay is a reliable method to detect specific IgE levels against respiratory allergens in a multiplexed and high-throughput manner, using blood samples as small as from a finger prick.
Assuntos
Hipersensibilidade/diagnóstico , Imunoensaio/métodos , Doenças Respiratórias/diagnóstico , Adulto , Poluentes Atmosféricos/imunologia , Alérgenos/imunologia , Feminino , Ensaios de Triagem em Larga Escala , Humanos , Imunoglobulina E/metabolismo , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Adulto JovemRESUMO
Most nutritional studies on the development of children focus on mother-infant interactions. Maternal nutrition is critically involved in the growth and development of the fetus, but what about the father? The aim is to investigate the effects of paternal methyl-group donor intake (methionine, folate, betaine, choline) on paternal and offspring global DNA (hydroxy)methylation, offspring IGF2 DMR DNA methylation, and birth weight. Questionnaires, 7-day estimated dietary records, whole blood samples, and anthropometric measurements from 74 fathers were obtained. A total of 51 cord blood samples were collected and birth weight was obtained. DNA methylation status was measured using liquid chromatography-tandem mass spectrometry (global DNA (hydroxy)methylation) and pyrosequencing (IGF2 DMR methylation). Paternal betaine intake was positively associated with paternal global DNA hydroxymethylation (0.028% per 100 mg betaine increase, 95% CI: 0.003, 0.053, P=0.03) and cord blood global DNA methylation (0.679% per 100 mg betaine increase, 95% CI: 0.057, 1.302, P=0.03). Paternal methionine intake was positively associated with CpG1 (0.336% per 100 mg methionine increase, 95% CI: 0.103, 0.569, P=0.006), and mean CpG (0.201% per 100 mg methionine increase, 95% CI: 0.001, 0.402, P=0.049) methylation of the IGF2 DMR in cord blood. Further, a negative association between birth weight/birth weight-for-gestational age z-score and paternal betaine/methionine intake was found. In addition, a positive association between choline and birth weight/birth weight-for-gestational age z-score was also observed. Our data indicate a potential impact of paternal methyl-group donor intake on paternal global DNA hydroxymethylation, offspring global and IGF2 DMR DNA methylation, and prenatal growth.
Assuntos
Betaína/administração & dosagem , Peso ao Nascer/fisiologia , Colina/administração & dosagem , Metilação de DNA/fisiologia , Ácido Fólico/administração & dosagem , Metionina/administração & dosagem , Adulto , Bélgica/epidemiologia , Betaína/sangue , Colina/sangue , Feminino , Sangue Fetal/metabolismo , Ácido Fólico/sangue , Humanos , Masculino , Metionina/sangue , Pessoa de Meia-Idade , Obesidade/sangue , Obesidade/epidemiologiaRESUMO
Over the past 250 years, human life expectancy has increased dramatically and continues to do so in most countries worldwide. Genetic factors account for about one third of variation in life expectancy so that most inter-individual variation in lifespan is explained by stochastic and environmental factors. The ageing process is plastic and is driven by the accumulation of molecular damage causing the changes in cell and tissue function which characterise the ageing phenotype. Early life exposures mark the developing embryo, foetus and child with potentially profound implications for the individual's ageing trajectory. Maternal factors including age, smoking, socioeconomic status, infections, nutritional status and season of birth influence offspring life expectancy and the development of age-related diseases. Although the mechanistic processes responsible are poorly understood, many of these factors appear to affect foetal growth directly or via effects on placental development. Those born relatively small i.e. which did not achieve their genetic potential in utero, appear to be at greatest disadvantage especially if they become overweight or obese in childhood. Early life events and exposures which enhance ageing are likely to contribute to molecular damage and/or reduce the repair of such damage. Such molecular damage may produce immediate defects in cellular or tissue function that are retained into later life. In addition, there is growing evidence that early life exposures produce aberrant patterns of epigenetic marks that are sustained across the life-course and result in down-regulation of cell defence mechanisms.