It is a pilot study, conducting on 6 healthy women and aiming at defining new biomarkers related to wheat bran fermentation. For this, a breakfast, containing 5 biscuits enriched in 13-C wheat bran (which grown under 13-CO2 enriched atmosphere) is given to the subjects, then a following will be realized on 24h with breath, urine, fecal and blood samples. Analysis will be performed on these samples in order to determine potential biomarkers of fibers consumption to evaluate the metabolic effects of this consumption and to refine the relevance of nutritional recommendation regarding fibers for healthy subjects.
Study Type
INTERVENTIONAL
Allocation
NA
Purpose
BASIC_SCIENCE
Masking
NONE
Enrollment
6
The intervention will be done on 1 day, time during which, each subject will stay at the CRNH. A breakfast containing the fibers will be given to them, then during 24h a following will be done to get fecal, urinary, plasmatic and expiratory samples. Standardized lunch, dinner and collation will be served during the day.
Centre de Recherche en Nutrition Humaine Rhône-Alpes
Pierre-Bénite, France
Measurement of the 13-C isotopic enrichment kinetics
Measurement of the 13-C isotopic enrichment kinetics of plasmatic SCFAs after ingestion of 5 biscuits enriched with the 13C-labeled fiber, on 24h.The concentration and the 13C isotopic enrichment of SCFAs in the plasma will be determined according method and tools described by Ferchaud-Roucher and al, 2006 and Ahmed and el, 2016 at the Analyse center of Mass Spectrometry in Lyon
Time frame: 24 hours
Measurement of the 13-C isotopic enrichment kinetics in CO2 and CH4 from exhaled gases during 24h after fibers ingestion.
Gases will be taken thanks to the EasySamplerTM Breath Test Kit. The measurements will be done by gas chromatography thanks to a gas analyzer.
Time frame: 24 hours
Measurement of H2 kinetics in the exhaled breath during 24h.
H2 will be taken thanks to the EasySamplerTM Breath Test Kit. The measurements will be done by gas chromatography thanks to a gas analyzer.
Time frame: 24 hours
Measurement of the 13-C isotopic enrichment kinetics of SCFAs in urines after ingestion of 5 biscuits enriched with the 13C-labeled fiber, on 24h.
The concentration and the 13C isotopic enrichment of SCFAs in the urine will be determined according method and tools described by Ferchaud-Roucher and al, 2006 and Ahmed and el, 2016 at the Analyse center of Mass Spectrometry in Lyon
Time frame: 24 hours
Measurement of the 13-C isotopic enrichment kinetics of SCFAs in feces after ingestion of 5 biscuits enriched with the 13C-labeled fiber, on 48h or 72h.
The concentration and the 13C isotopic enrichment of SCFAs in the urine will be determined according method and tools described by Ferchaud-Roucher and al, 2006 and Ahmed and el, 2016 at the Analyse center of Mass Spectrometry in Lyon
Time frame: Hour 72
The appearance kinetic of 13C glucose in the plasma
Plasma samples will be prepared and analysed by coupling gas chromatography-combustion-isotopic mass spectrometry according to the method used at the CRNH Rhône-Alpes (Sauvinet et al., 2009)
Time frame: 24 hours
The platelets aggregation as cardimetabolic risk marker in the plasma
An immuno-enzymatic dosage of platelet thromboxan B2, after the chylomicrons (CM) incubation with platelets, will be done. The measurement of platelet aggregation by aggregometry will be done after the CM and collagen stimulated platelet incubation.
Time frame: Day -30
Sphingolipid molecular species concentrations in plasma and TGRL
Isolation of plasma and triglyceride-rich lipoproteins (TGRL) fractions by ultracentrifugation and measurement by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS)
Time frame: Day 0
Fatty acids in lipid classes (triglycerides, phospholipids, cholesteryl esters) of plasma and TGRL
Determination of molar percentages of fatty acids in lipid classes (triglycerides, phospholipids, cholesteryl esters) of plasma and TGRL, following analysis by gas chromatography.
Time frame: Day 0
apolipoprotein B48 concentrations in plasma and TGRL
apolipoprotein B48 concentrations in plasma and TGRLby ELISA technique
Time frame: Day 0
TGRL particle sizes
TGRL particle sizes by laser granulometry
Time frame: Day 0
Intensity of gastro-intestinal symptoms thanks to an analogue visual scale (AVS)
This AVS allows the measurement of 8 gastro-intestinal symptoms intensity, often described after fibers consumption on a 100 mm scale.
Time frame: 24 hours
Stool consistency
Stool consistency by Bristol Stool Chart (type1-7)
Time frame: one week (at each stool)
Stool frequency
By questionnaire
Time frame: one week
Gastro intestinal symptoms
By questionnaires and visual analogue scale (VAS) score (on a 90mm horizontal line; from no symptom (minimal) to serious symptom (maximum).
Time frame: one week
measurement of plasmatic PUFA concentration
Plasma samples will be prepared and analysed by gas-liquid chromatography-
Time frame: 24 hours
Metagnomic of gut microbiota
Stool samples will be prepared and analysed by 16SrDBA ilimina sequencing
Time frame: one week (at each stool)
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