Systems Metabolomics for Prediction of Metabolic Syndrome.

The evolution of human health is a continuum of transitions, involving multifaceted processes at multiple levels, and there is an urgent need for integrative biomarkers that can characterize and predict progression toward disease development. The objective of this work was to perform a systems metabolomics approach to predict metabolic syndrome (MetS) development. A case-control design was used within the French occupational GAZEL cohort (n = 112 males: discovery study; n = 94: replication/validation study). Our integrative strategy was to combine untargeted metabolomics with clinical, sociodemographic, and food habit parameters to describe early phenotypes and build multidimensional predictive models. Different models were built from the discriminant variables, and prediction performances were optimized either when reducing the number of metabolites used or when keeping the associated signature. We illustrated that a selected reduced metabolic profile was able to reveal subtle phenotypic differences 5 years before MetS occurrence. Moreover, resulting metabolomic markers, when combined with clinical characteristics, allowed improving the disease development prediction. The validation study showed that this predictive performance was specific to the MetS component. This work also demonstrates the interest of such an approach to discover subphenotypes that will need further characterization to be able to shift to molecular reclassification and targeting of MetS.

Feature Selection Methods for Early Predictive Biomarker Discovery Using Untargeted Metabolomic Data.

Untargeted metabolomics is a powerful phenotyping tool for better understanding biological mechanisms involved in human pathology development and identifying early predictive biomarkers. This approach, based on multiple analytical platforms, such as mass spectrometry (MS), chemometrics and bioinformatics, generates massive and complex data that need appropriate analyses to extract the biologically meaningful information. Despite various tools available, it is still a challenge to handle such large and noisy datasets with limited number of individuals without risking overfitting. Moreover, when the objective is focused on the identification of early predictive markers of clinical outcome, few years before occurrence, it becomes essential to use the appropriate algorithms and workflow to be able to discover subtle effects among this large amount of data. In this context, this work consists in studying a workflow describing the general feature selection process, using knowledge discovery and data mining methodologies to propose advanced solutions for predictive biomarker discovery. The strategy was focused on evaluating a combination of numeric-symbolic approaches for feature selection with the objective of obtaining the best combination of metabolites producing an effective and accurate predictive model. Relying first on numerical approaches, and especially on machine learning methods (SVM-RFE, RF, RF-RFE) and on univariate statistical analyses (ANOVA), a comparative study was performed on an original metabolomic dataset and reduced subsets. As resampling method, LOOCV was applied to minimize the risk of overfitting. The best k-features obtained with different scores of importance from the combination of these different approaches were compared and allowed determining the variable stabilities using Formal Concept Analysis. The results revealed the interest of RF-Gini combined with ANOVA for feature selection as these two complementary methods allowed selecting the 48 best candidates for prediction. Using linear logistic regression on this reduced dataset enabled us to obtain the best performances in terms of prediction accuracy and number of false positive with a model including 5 top variables. Therefore, these results highlighted the interest of feature selection methods and the importance of working on reduced datasets for the identification of predictive biomarkers issued from untargeted metabolomics data.

Higher calcium urinary loss induced by a calcium sulphate-rich mineral water intake than by milk in young women.

It is well known that the intestinal availability of Ca from Ca-rich mineral waters is equivalent to that of milk Ca. However, the effect of associated anions on Ca urinary loss needs to be addressed. The aim of the current study was to compare, under ordinary conditions of consumption, milk and a SO4-rich mineral water as the Ca provider in a large number of subjects consuming the same quantity of Ca from the two sources in a crossover study lasting for an extended period. Thirty-seven healthy women completed a 12-week protocol, divided into four periods of 3 weeks (W). In the first (W1-3) and third (W6-9) periods, dietary Ca intake was restricted to 600 mg/d. In the second (W4-6) and final (W10-12) periods, either 400 ml/d medium-fat milk or 1 litre of a Ca- and SO4-rich mineral water, each providing about 480 mg Ca/d, was added to the diet in a random manner. Dietary evaluation, blood and urinary measures were performed during the last week (W6 and W12) of each Ca supplementation period. The urinary excretion of Ca was higher (0.5 mmol/d more) with water than with milk (P<0.001). An examination of all the dietary factors known to influence calciuria suggested that the acidogenic action of SO4 was responsible for this increased calciuria. Thus, despite an equal Ca intake and assuming an unchanged intestinal absorption, these results suggest that Ca balance is better with milk consumption than with CaSO4-rich water.

Effects of two conjugated linoleic Acid isomers on body fat mass in overweight humans.

OBJECTIVE: To examine the effects of two different conjugated linoleic acid (CLA) isomers at two different intakes on body composition in overweight humans. RESEARCH METHODS AND PROCEDURES: Eighty-one middle-aged, overweight, healthy men and women participated in this bicentric, placebo-controlled, double-blind, randomized study. For 6 weeks (run-in period), all subjects consumed daily a drinkable dairy product containing 3 g of high oleic acid sunflower oil. Volunteers were then randomized over five groups receiving daily either 3 g of high oleic acid sunflower oil, 1.5 g of cis-9,trans-11 (c9t11) CLA, 3 g of c9t11 CLA, 1.5 g of trans-10,cis-12 (t10c12) CLA, or 3 g of t10c12 CLA administrated as triacylglycerol in a drinkable dairy product for 18 weeks. Percentage body fat mass and fat and lean body mass were assessed at the end of the run-in and experimental periods by DXA. Dietary intake was also recorded. RESULTS: Body fat mass changes averaged 0.1 +/- 0.9 kg (mean +/- SD) in the placebo group and -0.3 +/- 1.4, -0.8 +/- 2.1, 0.0 +/- 2.3, and -0.9 +/- 1.7 kg in the 1.5-g c9t11, 3-g c9t11, 1.5-g t10c12, and 3-g t10c12 groups, respectively. Changes among the groups were not significantly different (p = 0.444). Also, lean body mass and dietary intake were not significantly different among the treatments. DISCUSSION: A daily consumption of a drinkable dairy product containing up to 3 g of CLA isomers for 18 weeks had no statistically significant effect on body composition in overweight, middle-aged men and women.

Daily protein intakes and eating patterns in young and elderly French.

An adequate level of protein intake is required to limit the gradual body protein loss observed during ageing. Different factors (cohort age, sex, life conditions) may modify protein intake and distribution. However the precise amounts, as well as their daily distribution which affects protein utilisation and N retention, are unknown in both young and elderly individuals. The hypothesis was tested that protein intake and its distribution over daily meals could be different between the young and elderly. The consumption of six different protein-rich food groups by 292 healthy individuals aged 20-30 and 65-75 years was determined throughout each day for 1 week. The data of the total protein intake and protein intakes at each meal were analysed by ANOVA for each sex separately, using age group as the independent factor. The average protein intake of men was lower in the older age group whilst the opposite trend was seen in women. The distribution of protein intake was different between the two age groups: 56.5 % of the daily protein was eaten at lunch by the elderly but only 47 % (P<0.0001) by the younger subjects. In the elderly subjects, those eating larger amounts consumed a greater proportion of protein-rich foods at dinner than those eating small amounts (30.4 v. 26.2 %, P=0.05). A high level of protein intake was related to a higher meat-product consumption in both the elderly and young individuals. In conclusion, the pattern of protein intake differs significantly between age groups and sexes.

Effect of short-chain fructooligosaccharides on intestinal calcium absorption and calcium status in postmenopausal women: a stable-isotope study.

BACKGROUND: An enhancing effect of short-chain fructooligosaccharides (scFOSs) on intestinal calcium absorption has been shown in animals and in some short-term human studies. However, the long-term effect of scFOSs on calcium absorption in humans is still unknown. OBJECTIVE: We investigated the long-term effect of a moderate daily dose (10 g) of scFOSs on intestinal calcium absorption in postmenopausal women. DESIGN: In a randomized, double-blind crossover protocol, 12 healthy, postmenopausal women received scFOSs or placebo for 5 wk. The treatments were separated by a 3-wk washout period. Subjects orally received (44)Ca (stable isotope) and a fecal marker. Feces were collected after the isotope intake for 5-7 d to measure unabsorbed isotope. Calcium-status indexes, calciotropic hormones, and bone turnover were also assessed. RESULTS: Mean (+/-SD) intestinal calcium absorption with scFOS treatment was not significantly different from that with placebo treatment (35.63 +/- 9.40% and 36.55 +/- 8.48%, respectively). However, a tendency for calcium absorption to be higher with scFOS treatment than with placebo treatment was observed in women who had been going through menopause for >6 y. CONCLUSIONS: scFOSs do not modify intestinal calcium absorption in postmenopausal women who do not receive hormonal replacement therapy. The results from a subgroup of women who had been going through menopause for >6 y (n = 6) suggest that scFOSs may influence calcium absorption in the late postmenopausal phase. The small number of subjects and the related P value warrant verification and further investigation with women in late menopause only.

Two polyol, low digestible carbohydrates improve the apparent absorption of magnesium but not of calcium in healthy young men.

The effects of nondigestible oligosaccharides including polyols on intestinal mineral absorption have been studied extensively in animal experiments, but their impact on mineral absorption in humans remains to be established. We investigated the effects of feeding two fermentable, low digestible carbohydrates, on the apparent absorption and balance of calcium (Ca) and magnesium (Mg) in humans. Nine healthy young men were given a control diet with dextrose or polyols, low digestible, fermentable carbohydrates (LHBC, HPFL) for 32-d periods according to a 3 x 3 Latin-square design. During the 18-d period of adaptation, the products were administered gradually in liquid form, up to a maximum of 100 g/d, which was then consumed for 14 d. Ca and Mg levels were measured in diets and in fecal and urine collections to assess apparent mineral absorption and balance. The relative apparent absorptions of Ca and Mg from the control diet were (means +/- SEM) 33.3 +/- 4.6 and 39.8 +/- 2.7%, respectively. Ingestion of both low digestible carbohydrates significantly increased the relative apparent absorption of Mg by about 25%. LHBC, but not HPFL, ingestion increased urinary Mg excretion. Apparent absorption, urinary excretion and balance of Ca were not altered by the ingestion of either low digestible carbohydrate. Ingestion of the low digestible, fermentable carbohydrates, with balanced diets, improved apparent Mg absorption without significant effects on apparent absorption or retention of Ca in healthy young men. Further human studies are therefore still needed to confirm the effects of these products in other populations.

Alterations of sensory perceptions in healthy elderly subjects during fasting and refeeding. A pilot study.

BACKGROUND: Sensory perception losses may contribute to age-related malnutrition by affecting food selection and consumption. OBJECTIVE: To determine the effects of a 36-hour fast followed by a 6-hour refeeding period on sensory perceptions in 7 healthy elderly subjects (65-80 years of age) and 6 healthy young subjects (18-35 years of age). METHODS: Self-perceived hunger and olfactory ratings were recorded on visual analogue scales in response to three different classes of odorant stimuli (salt, sweet and sour). Odorant stimuli were administered three times during the study, twice during the fasting period (12 and 24 h fasted) and once at the end of the re-nutrition period. RESULTS: A significant difference was found between the two groups for the self-perceived hunger ratings in response to the sour stimuli. A significant difference was observed between the two groups for olfactory ratings as regards the salt and sour odorant stimuli. Among the metabolic changes associated with fasting and refeeding, blood glucose was significantly related (r(2) = 0.97, p = 0.001) to the perception of hunger in the control group subjects, but no such relationship was found for the elderly subjects (r(2) = 0.16, p = NS). CONCLUSION: (1) Self-perceived hunger and olfactory ratings are specifically affected in healthy elderly. (2) Nutritional status can modulate sensory perceptions in elderly and young during the transition from fasting to refeeding.