Practical and Robust NMR-Based Metabolic Phenotyping of Gut Health in Early Life.

While substantial efforts have been made to optimize and standardize fecal metabolomics for studies in adults, the development of a standard protocol to analyze infant feces is, however, still lagging behind. Here, we present the development of a hands-on and robust protocol for proton 1H NMR spectroscopy of infant feces. The influence of extraction solvent, dilution ratio, homogenization method, filtration, and duration of centrifugation on the biochemical composition of infant feces was carefully evaluated using visual inspection of 1H NMR spectra in combination with multivariate statistical modeling. The optimal metabolomics protocol was subsequently applied on feces from seven infants collected at 8 weeks, 4, and 9 months of age. Interindividual variation was exceeding the variation induced by different fecal sample preparation methods, except for filtration. We recommend extracting fecal samples using water with a dilution ratio of 1:5 feces-to-water to homogenize using bead beating and to remove particulates using centrifugation. Samples collected from infants aged 8 weeks and 4 months showed elevated concentrations of milk oligosaccharide derivatives and lactic acid, whereas short-chain fatty acids (SCFAs) and branched-chain amino acids (BCAAs) were higher in the 9 month samples. The established protocol enables hands-on and robust analyses of the infant gut metabolome. The wide-ranging application of this protocol will facilitate interlaboratory comparison of infants' metabolic profiles and finally aid in a better understanding of infant gut health.

Reduction of smoking in Dutch adolescents over the past decade and its health gains: a repeated cross-sectional study.

BACKGROUND: Smoking is the main preventable lifestyle-related risk factor threatening human health. In this study, time trends in smoking behaviour between 1996 and 2005 among adolescents enrolled in secondary school were assessed. METHODS: In 1996, 2001 and 2005, a survey was conducted in the south-eastern region of the Netherlands. All students in second and fourth year of secondary education (1996: n = 20 000; 2001: n = 27 500; 2005: n = 24 000) were asked to complete a questionnaire about their smoking behaviour. A simulation model was used to estimate lifetime health gains related to the observed trends. RESULTS: In 1996, 2001 and 2005, the number of questionnaires analysed were 13 554 (68%), 20 767 (76%) and 17 896 (75%), respectively. The results show a decrease in 'ever smoking' as well as 'current smoking' between 1996 and 2005. Among second year high school students, current smoking prevalence decreased from 22.2% in 1996 to 8.0% in 2005 (P(trend) < 0.001). Among fourth year students, current smoking declined from 37.5% in 1996 to 22.0% in 2005 (P(trend) < 0.001). Time trends were not influenced by gender or educational level. Model projections show that if these students not take up smoking later in life, 11 500 new cases of COPD, 3400 new cases of lung cancer and 1800 new cases of myocardial infarction could be prevented for the Dutch 13-year-olds. CONCLUSION: This study found that, in the past decade, smoking prevalence among adolescents has declined by almost 50%, potentially resulting in a considerable reduction in new cases of COPD or lung cancer.