Time window-dependent effect of perinatal maternal protein restriction on insulin sensitivity and energy substrate oxidation in adult male offspring

Epidemiological and experimental evidence suggests that a suboptimal environment during perinatal life programs offspring susceptibility to the development of metabolic syndrome and Type 2 diabetes. We hypothesized that the lasting impact of perinatal protein deprivation on mitochondrial fuel oxidation and insulin sensitivity would depend on the time window of exposure. To improve our understanding of underlying mechanisms, an integrative approach was used, combining the assessment of insulin sensitivity and untargeted mass spectrometry-based metabolomics in the offspring. A hyperinsulinemic-euglycemic clamp was performed in adult male rats born from dams fed a low-protein diet during gestation and/or lactation, and subsequently exposed to a Western diet (WD) for 10 wk. Metabolomics was combined with targeted acylcarnitine profiling and analysis of liver gene expression to identify markers of adaptation to WD that influence the phenotype outcome evaluated by body composition analysis. At adulthood, offspring of protein-restricted dams had impaired insulin secretion when fed a standard diet. Moreover, rats who demonstrated catch-up growth at weaning displayed higher gluconeogenesis and branched-chain amino acid catabolism, and lower fatty acid β-oxidation compared with control rats. Postweaning exposure of intrauterine growth restriction-born rats to a WD exacerbated incomplete fatty acid β-oxidation and excess fat deposition. Control offspring nursed by protein-restricted mothers showed peculiar low-fat accretion through adulthood and preserved insulin sensitivity even after WD-exposure. Altogether, our findings suggest a testable hypothesis about how maternal diet might influence metabolic outcomes (insulin sensitivity) in the next generation such as mitochondrial overload and/or substrate oxidation inflexibility dependent on the time window of perinatal dietary manipulation

Detection of hazardous food contaminants by transcriptomics fingerprinting

Unprecedented biotechnological advances in the past decade have delivered powerful transcriptomics methods that provide new opportunities for a risk-based and, hence, more effective control of food quality and safety. The fundamental hypothesis underlying the application of a transcriptomics or other “-omics” strategies is that toxicant-specific changes of gene expression will reveal the presence of chemical hazards with possibly adverse health effects. Indeed, living cells display wide adaptive plasticity by responding with characteristic expression signatures to various stimuli (e.g., proliferation signals, genotoxic or cytotoxic insults, oxidative bursts and other types of chemical stress). Flexible transcriptomics platforms that can be scaled to suit different applications in food quality and safety involve oligonucleotide microarrays (also known as “DNA chips”), focused polymerase chain reaction (PCR) arrays and, more recently, high-throughput “next generation” sequencing technologies. In this review, we illustrate the potential impact of transcriptomics for the detection of hazardous food constituents or contaminants, including dioxins, xenoestrogens, mycotoxins and Maillard reaction products. We also discuss the predictive power of “-omics” fingerprinting with respect to the identification of emerging contaminants and the significance of these methods for risk assessment at all stages of the food-production chain

Multi-functional sample preparation procedure for measuring phytoestrogens in milk, cereals, and baby-food by liquid-chromatography tandem mass spectrometry with subsequent determination of their estrogenic activity using transcriptomic assay

A method dedicated to the determination of a multiple range of phytoestrogens as endocrine disruptor compounds in infant food products was developed, with as double objective the specific measurement of 13 parameters and the evaluation of the estrogenic potency associated to this quantitative profile. A combined enzymatic and acidic chemical hydrolysis followed by a double purification on two successive C(18) and SiOH Solid Phase Extraction cartridges permitted to efficiently purify milk, cereals and baby-food samples while eliminating naturally occurring estrogen hormones. A specific liquid chromatography-tandem mass spectrometric measurement authorised unambiguous identification and quantification of the target compounds. The proposed methodology was fully validated and applied to a set of around 30 real samples, demonstrating the presence of phytoestrogens at levels globally ranging from several microgkg(-1) (ppb) to several tens mgkg(-1) (ppm). The prepared sample extracts were proven to be suitable and compatible with the evaluation of their induced biological transcriptional activity on MCF-7 cell lines. Because permitting to cope with difficult issues such as low-dose and mixture effects, this proposed methodology may appear of particular interest for further exposure assessment studies and hazard characterisation investigations related to this class of endocrine disruptor compounds

Ibuprofen disrupts steroidogenesis in the human fetal testis.

International audienc

Assessment of two complementary liquid chromatography coupled to high resolution mass spectrometry metabolomics strategies for the screening of anabolic steroid treatment in calves

Anabolic steroids are banned in food producing livestock in Europe. Efficient methods based on mass spectrometry detection have been developed to ensure the control of such veterinary drug residues. Nevertheless, the use of “cocktails” composed of mixtures of low amounts of several substances as well as the synthesis of new compounds of unknown structure prevent efficient prevention. New analytical tools able to detect such abuse are today mandatory. In this context, metabolomics may represent new emerging strategies for investigating the global physiological effects associated to a family of substances and therefore, to suspect the administration of steroids. The purpose of the present study was to set up, assess and compare two complementary mass spectrometry-based metabolomic strategies as new tools to screen for steroid abuse in cattle and demonstrate the feasibility of such approaches. The protocols were developed in two European laboratories in charge of residues analysis in the field of food safety. Apart from sample preparation, the global process was different in both laboratories from LC-HRMS fingerprinting to multivariate data analysis through data processing and involved both LC-Orbitrap-XCMS and UPLC-ToF-MS-MetAlign strategies. The reproducibility of both sample preparation and MS measurements were assessed in order to guarantee that any differences in the acquired fingerprints were not caused by analytical variability but reflect metabolome modifications upon steroids administration. The protocols were then applied to urine samples collected on a large group of animals consisting of 12 control calves and 12 calves administrated with a mixture of 17ß-estradiol 3-benzoate and 17ß-nandrolone laureate esters according to a protocol reflecting likely illegal practices. The modifications in urine profiles as indicators of steroid administration have been evaluated in this context and proved the suitability of the approach for discriminating anabolic treated animals from control ones. Such an approach may therefore open a new way for the screening of anabolic steroid administration through targeted monitoring of relevant biomarkers highlighted as a result of the metabolomics study