Marie-cécile Alexandre-gouabau - Academia.edu (original) (raw)

Papers by Marie-cécile Alexandre-gouabau

Research paper thumbnail of Chylomicron β-carotene and retinyl palmitate responses are dramatically diminished when men ingest β-carotene with medium-chain rather than long-chain triglycerides

Journal of Nutrition

The effect of the ingestion of beta-carotene with medium-chain triglycerides (MCT) or long-chain ... more The effect of the ingestion of beta-carotene with medium-chain triglycerides (MCT) or long-chain triglycerides (LCT) on the bioavailability and the provitamin A activity of beta-carotene was investigated in humans. Sixteen healthy young men ingested, on two different days, a test meal containing 120 mg beta-carotene incorporated into 40 g LCT (LCT meal) or 40 g MCT (MCT meal). This meal was followed 6 h later by a beta-carotene-free meal containing 40 g LCT. Chylomicron beta-carotene, retinyl palmitate and triglycerides were measured every hour for 12.5 h after the first meal. No significant increase in chylomicron triglycerides was detected for the 6 h after the MCT meal intake, whereas a significant increase in chylomicron triglycerides was observed after the LCT meal intake. The chylomicron beta-carotene and retinyl palmitate responses to the MCT meal (0-6 h area under the curves, AUC) were significantly (P < 0.05) lower [AUC = 68.1 +/- 26.8 and 43. 4 +/- 10.4 nmol/(L.h), for ...

Research paper thumbnail of Processing of vitamin A and E in the human gastrointestinal tract

AJP Gastrointestinal and Liver Physiology

We aimed to provide basic data on the processing of vitamin A and E in the human gastrointestinal... more We aimed to provide basic data on the processing of vitamin A and E in the human gastrointestinal tract and to assess whether the size of emulsion fat globules affects the bioavailability of these vitamins. Eight healthy men received intragastrically two lipid formulas differing in their fat-globule median diameter (0.7 vs. 10. 1 microm. Formulas provided 28 mg vitamin A as retinyl palmitate and 440 mg vitamin E as all-rac alpha-tocopherol. Vitamins were measured in gastric and duodenal aspirates, as well as in chylomicrons, during the postprandial period. The gastric emptying rate of lipids and vitamin A and E was similar. The free retinol/total vitamin A ratio was not significantly modified in the stomach, whereas it was dramatically increased in the duodenum. The proportion of ingested lipid and vitamins was very similar in the duodenal content. The chylomicron response of lipids and vitamins was not significantly different between the two emulsions. Our main conclusions are as f...

Research paper thumbnail of The bioavailability of α- and β-carotene is affected by gut microflora in the rat

The British journal of nutrition

The present study examined whether the intestinal microflora could affect the bioavailability and... more The present study examined whether the intestinal microflora could affect the bioavailability and vitamin A activity of dietary α- and β-carotene in the rat. In the first set of experiments, we used conventional, germ-free (axenic), and human-flora-associated (heteroxenic) rats. In a second series, conventional rats were treated with either an antibiotic mixture or a potent inhibitor of gastric secretion (Omeprazole). All animals were first depleted of vitamin A over 4 weeks and then were fed on a sterilized diet supplemented with 14 mg β-carotene and 3 mg α-carotene/kg for 2 weeks. In both experiments, a reduction in the intestinal microflora resulted in an increased storage of β-carotene, α-carotene and vitamin A in the liver. Neither the nature of the metabolism of the intestinal microflora (aerobic or anaerobic) nor treatment with omeprazole, to modify intestinal pH, induced a significant effect on the measured variables. When incubated with 15 μmol β-carotene/l for 72 h, neithe...

Research paper thumbnail of The bioavailability of alpha- and beta-carotene is affected by gut microflora in the rat

British Journal Of Nutrition

The present study examined whether the intestinal microflora could affect the bioavailability and... more The present study examined whether the intestinal microflora could affect the bioavailability and vitamin A activity of dietary alpha- and beta-carotene in the rat. In the first set of experiments, we used conventional, germ-free (axenic), and human-flora-associated (heteroxenic) rats. In a second series, conventional rats were treated with either an antibiotic mixture or a potent inhibitor of gastric secretion (Omeprazole). All animals were first depleted of vitamin A over 4 weeks and then were fed on a sterilized diet supplemented with 14 mg beta-carotene and 3 mg alpha-carotene/kg for 2 weeks. In both experiments, a reduction in the intestinal microflora resulted in an increased storage of beta-carotene, alpha-carotene and vitamin A in the liver. Neither the nature of the metabolism of the intestinal microflora (aerobic or anaerobic) nor treatment with omeprazole, to modify intestinal pH, induced a significant effect on the measured variables. When incubated with 15 mumol beta-ca...

Research paper thumbnail of Rat intestinal β-carotene dioxygenase activity is located primarily in the cytosol of mature jejunal enterocytes

Journal of Nutrition

The purposes of this study were to determine the location of beta-carotene dioxygenase (EC 1.13.1... more The purposes of this study were to determine the location of beta-carotene dioxygenase (EC 1.13.11.21) activity within the rat gastrointestinal tract, within the villus and within enterocytes, and to identify the metabolites produced in each intestinal fraction. In Wistar female rats, maximal activity was detected in the cytosol (74-93% of the total cellular activity) of mature functional enterocytes harvested from the jejunum (67% of the intestinal activity). The specific activity, expressed in pmol of retinoids/(h x mg protein) rose from 49 +/- 3 in the stem cells to 199 +/- 12 in the mature functional cells (P < 0.05). Thus the intestinal beta-carotene cleavage activity might be regulated during the enterocyte maturation process. By using HPLC with diode array and radioactive detectors, retinal, and in the presence of NAD+, retinoic acid, were identified as the only metabolites produced. No beta-12'-, 10'-, and 8'-apo-carotenals were detected, even when various enz...

Research paper thumbnail of Mobilization of rétinol from cultured rat liver stellate cells does not require protéin secretion

Research paper thumbnail of Proteomics in the field of food and feed sciences

Proteomics has emerged recently as a powerful analytical method to investigate proteins. Its scop... more Proteomics has emerged recently as a powerful analytical method to investigate proteins. Its scope of use has considerably enlarged in the past years including in the fi eld of feed and food sciences. Through a series of studies conducted at I.N.R.A., we illustrate the interest of proteomic approaches to improve plant breeding, technological processing or nutrition studies. The objectives are to reach high-quality and healthy foods while ensuring a sustainable and competitive agriculture and a preserved environment.

Research paper thumbnail of Calibration Adjustment of the Mid-infrared Analyzer for an Accurate Determination of the Macronutrient Composition of Human Milk

Journal of Human Lactation, 2015

Human milk composition analysis seems essential to adapt human milk fortification for preterm neo... more Human milk composition analysis seems essential to adapt human milk fortification for preterm neonates. The Miris human milk analyzer (HMA), based on mid-infrared methodology, is convenient for a unique determination of macronutrients. However, HMA measurements are not totally comparable with reference methods (RMs). The primary aim of this study was to compare HMA results with results from biochemical RMs for a large range of protein, fat, and carbohydrate contents and to establish a calibration adjustment. Human milk was fractionated in protein, fat, and skim milk by covering large ranges of protein (0-3 g/100 mL), fat (0-8 g/100 mL), and carbohydrate (5-8 g/100 mL). For each macronutrient, a calibration curve was plotted by linear regression using measurements obtained using HMA and RMs. For fat, 53 measurements were performed, and the linear regression equation was HMA = 0.79RM + 0.28 (R(2) = 0.92). For true protein (29 measurements), the linear regression equation was HMA = 0.9RM + 0.23 (R(2) = 0.98). For carbohydrate (15 measurements), the linear regression equation was HMA = 0.59RM + 1.86 (R(2) = 0.95). A homogenization step with a disruptor coupled to a sonication step was necessary to obtain better accuracy of the measurements. Good repeatability (coefficient of variation &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 7%) and reproducibility (coefficient of variation &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 17%) were obtained after calibration adjustment. New calibration curves were developed for the Miris HMA, allowing accurate measurements in large ranges of macronutrient content. This is necessary for reliable use of this device in individualizing nutrition for preterm newborns.

Research paper thumbnail of Inflammation interferes with the assessment of vitamin A status in magnesium deficiency

Magnesium research : official organ of the International Society for the Development of Research on Magnesium, 2008

Hyporetinemia is observed in several pathological conditions including a primary deficiency of vi... more Hyporetinemia is observed in several pathological conditions including a primary deficiency of vitamin A and has also been reported to accompany inflammatory diseases. Experimental magnesium (Mg) deficiency in rodents is accompanied by an inflammatory syndrome. The present study was designed to determine whether the acute phase response in Mg-deficient rats can modify vitamin A status. Clinical symptoms of acute phase response were observed in Mg-deficient rats and were accompanied by a reduction in plasma retinol and of plasma retinol binding protein (RBP). Mg deficiency in rats resulted in hyporetinemia without a significant decrease in liver retinol reserves. Consequently, the data strongly suggest that the decrease in plasma retinol concentration, resulting from the level of its binding protein, is related to the inflammatory effect of Mg deficiency. These results point to the possible interference of Mg deficiency on the use of plasma retinol as an indicator of vitamin A status.

Research paper thumbnail of Use of UPLC-ESI-MS/MS to quantitate free amino acid concentrations in micro-samples of mammalian milk

SpringerPlus, 2013

Although free amino acids (FAA) account for a small fraction of total nitrogen in mammalian milk,... more Although free amino acids (FAA) account for a small fraction of total nitrogen in mammalian milk, they are more abundant in human milk than in most formulas, and may serve as a readily available source of amino acids for protein synthesis, as well as fulfill specific physiologic roles. We used reversed phase Ultra Performance Liquid Chromatography (UPLC) coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS) technique for FAA profiling in milks from three species (human, rat and cow) with a simple and rapid sample preparation. The derivatization procedure chosen, combined with UPLC-ESI-MS/MS allowed the quantitation of 21 FAA using labeled amino acids (Internal Standards) over a 10 min run time in micro-samples of mammalian milk (50 μL). The low limit of quantitation was 0.05 pmol/μL for most FAA with good repeatability and reproducibility (mean CV of 5.1%). Higher levels of total FAA were found in human (3032 μM) and rat milk (3460 μM) than in bovine milk (240 μM), with wide differences in the abundances of specific FAA between species. This robust analytical method could be applied to monitor FAA profile in human breast milk, and open the way to individualized adjustment of FAA content for the nutritional management of infants.

Research paper thumbnail of Effect of pre- and postnatal growth and post-weaning activity on glucose metabolism in the offspring

The Journal of endocrinology, 2015

Maternal caloric restriction during late gestation reduces birth weight, but whether long-term ad... more Maternal caloric restriction during late gestation reduces birth weight, but whether long-term adverse metabolic outcomes of intra-uterine growth retardation (IUGR) are dependent on either accelerated postnatal growth or exposure to an obesogenic environment after weaning is not established. We induced IUGR in twin-pregnant sheep using a 40% maternal caloric restriction commencing from 110 days of gestation until term (∼147 days), compared with mothers fed to 100% of requirements. Offspring were reared either as singletons to accelerate postnatal growth or as twins to achieve standard growth. To promote an adverse phenotype in young adulthood, after weaning, offspring were reared under a low-activity obesogenic environment with the exception of a subgroup of IUGR offspring, reared as twins, maintained in a standard activity environment. We assessed glucose tolerance together with leptin and cortisol responses to feeding in young adulthood when the hypothalamus was sampled for assess...

Research paper thumbnail of Human choriocarcinoma cell line JEG-3 produces and secretes active retinoids from retinol

Molecular Human Reproduction, 2002

Vitamin A (retinol) and its active derivatives (the retinoids) are essential for growth and devel... more Vitamin A (retinol) and its active derivatives (the retinoids) are essential for growth and development of the mammalian fetus. Maternally-derived retinol has to pass through the placenta to reach the developing fetus. Despite its apparent importance, little is known about placental metabolism of retinol, and particularly placental production and/or secretion of active retinoids. It has been previously considered that retinoids are recruited from the uterine environment to influence placental development and function during gestation. We have studied retinoid metabolism in the human choriocarcinoma cell line JEG-3 and demonstrate, for the first time, that active retinoids are produced endogenously by the JEG-3 cell line from retinol. These retinoids induce gene expression from a retinoic acidresponsive enhancer element reporter plasmid and modulate placental transglutaminase activity. Furthermore, retinoids are secreted from JEG-3, as shown by the activation of retinoic acid-responsive β lacZ reporter cells grown in conditioned media. These results suggest that there could be an active role for trophoblast-derived retinoids during human development.

Research paper thumbnail of Simple method for clinical determination of 13 carotenoids in human plasma using an isocratic high-performance liquid chromatographic method

Journal of Chromatography B: Biomedical Sciences and Applications, 2001

Research paper thumbnail of Vitamin A contained in the lipid droplets of rat liver stellate cells is substrate for acid retinyl ester hydrolase

Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1995

V~ronique Az~/is-Braesco a,*, Isabelle Dodeman b l Serge Delpal b 9 ~

Research paper thumbnail of Oxidative stress status and antioxidant status are apparently not related to carotenoid status in healthy subjects

Journal of Laboratory and Clinical Medicine, 1998

Several lines of evidence suggest that carotenoids may have a beneficial effect on health as a re... more Several lines of evidence suggest that carotenoids may have a beneficial effect on health as a result of their antioxidant properties. In addition to beta-carotene, five other carotenoids are recovered in noticeable amounts from human plasma and tissues. Although the effect of beta-carotene on in vivo lipid peroxidation has been documented, few data are available on the effects of the other carotenoids. We evaluated the ability of the main human carotenoids to reduce lipid peroxidation by determining the correlations between plasma carotenoid concentration and plasma antioxidant capacity (in 79 healthy volunteers) and between carotenoid status and breath pentane excretion (in a subgroup of 24 subjects). Carotenoid intake was assessed by means of a 3-day food recall. Carotenoid status was evaluated by measurement of beta-carotene, lycopene, lutein/zeaxanthin, and alpha-carotene in plasma and buccal mucosal cells. Antioxidant status was evaluated by measurement of the total antioxidant capacity of the plasma. Oxidative stress status was evaluated by breath pentane measurements. Food recall data and the carotenoid concentrations in plasma and buccal mucosal cells showed that the subjects had normal carotenoid intake and normal carotenoid status. The total antioxidant capacity of the plasma was not related to the concentration of any specific carotenoid. The level of expired air pentane was not related to the carotenoid status of the subjects. These results show that normal concentrations of carotenoids in plasma and tissues are not correlated with these clinical markers of antioxidant and oxidative stress status.

Research paper thumbnail of PAV-1, a new rat hepatic stellate cell line converts retinol into retinoic acid, a process altered by ethanol

The International Journal of Biochemistry & Cell Biology, 2002

During liver fibrogenesis or long term culture, hepatic stellate cells (HSCs) evolved from &a... more During liver fibrogenesis or long term culture, hepatic stellate cells (HSCs) evolved from &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;quiescent&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; to activated phenotype called &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;myofibroblast-like&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;, a transition prevented by retinoic acid (RA). Little is known about RA generation by HSCs. Our study aimed to check the ability of these cells to produce RA from retinol (Rol) and the alterations of this metabolic step by ethanol. To study this metabolic pathway, primary cultures of HSCs represent the most physiological model but technically suffer several drawbacks. To circumvent these problems, an immortalized rat HSC line (named PAV-1) has been established. We validated PAV-1 cell line as a convenient model to study retinoids metabolism by HSCs. Then, we showed that PAV-1 cells express Rol-binding proteins (RBPs), enzymes and nuclear receptors involved in RA signaling pathway. We also demonstrated in situ generation of functional all-trans-RA (ATRA), using transient transfections with a RA-sensitive reporter gene, in situ modulation of tissue transglutaminase (tTG) activity and HPLC experiments. This production was Rol dose-dependent; 4-methylpyrazole, citral, and ethanol-inhibited which argues in favor of an enzymatic process.In conclusion, we first demonstrate in situ RA generation from Rol in a newly immortalized rat HSC line, named PAV-1. Inhibition of RA production by ethanol in PAV-1 and recent data, suggesting fundamental role of RA to prevent fibrosis development in the liver, allow us to hypothesize that Rol metabolism could be a primary target for ethanol during development of hepatic fibrosis.

Research paper thumbnail of Retinol mobilization from cultured rat hepatic stellate cells does not require retinol binding protein synthesis and secretion

The International Journal of Biochemistry & Cell Biology, 2001

Retinol mobilization from retinyl esters stores of hepatic stellate cells (HSCs) is a key step in... more Retinol mobilization from retinyl esters stores of hepatic stellate cells (HSCs) is a key step in the regulation of mammalian retinol homeostasis, but the precise mechanisms of such a mobilization are still poorly understood. Using primary cultures of HSCs, we first demonstrated that HSCs expressed immunoreactivity against retinol-binding-protein (RBP) when cultured in a medium containing RBP but were unable to synthesize RBP transcripts and proteins. Using pulse and chase-type experiments, we demonstrated that radioactive retinol was released in culture medium without binding proteins. Inhibition of protein secretion by brefeldin A did not modify quantitatively retinol release. This data ruled out, for the first time, the direct involvement of RBP in retinol mobilization from HSCs. Moreover, HSCs co-cultured with primary isolated hepatocytes displayed an increase of retinol transfer from HSCs to hepatocytes when they established direct physical contacts, as compared with co-cultures without contact. Based on this latter data, a mechanism of retinol mobilization from HSCs via the hepatocytes using retinol transfer through cellular membranes is proposed.

Research paper thumbnail of Esterification of Vitamin A by the Human Placenta Involves Villous Mesenchymal Fibroblasts

Pediatric Research, 2000

Vitamin A (retinol) and its active derivatives (retinoic acids) are essential for growth and deve... more Vitamin A (retinol) and its active derivatives (retinoic acids) are essential for growth and development of the mammalian fetus. Maternally derived retinol must pass the placenta to reach the developing fetus. Despite its apparent importance, little is known concerning placental transfer and metabolism of retinol, and particularly of placental production and storage of retinyl esters. To elucidate this metabolic pathway, we incubated, in the presence of retinol, 1) human full-term placental explants and 2) primary cultures of major cells types contributing to placental function: trophoblasts and villous mesenchymal fibroblasts. We used HPLC to determine the types and concentrations of retinyl esters produced by these explants and cells. About 14% of total cellular retinol in placental explants was esterified. The most abundant esters were myristate and palmitate. Primary cell cul-tures showed that fibroblasts efficiently produced retinyl esters, but trophoblasts did not. In both types of experiments, no retinyl esters were detected in the culture medium, suggesting that retinyl esters were produced for storage purpose. These results suggest that villous mesenchymal fibroblasts are primary sites of retinol esterification and storage in the placenta. (Pediatr Res 48: 565-572, 2000) Abbreviations RA, retinoic acid RBP, retinol binding protein HBSS, Hanks' balanced salt solution DMEM, Dulbecco's modified Eagle medium ABSTRACT 565 mesenchymal fibroblasts) were tested to determine their individual capacity for esterification.

Research paper thumbnail of Recovery of Exfoliated Cells From the Gastrointestinal Tract of Premature Infants: A New Tool to Perform “Noninvasive Biopsies?”

Pediatric Research, 2007

To gain insight into specific gene expression in the gastrointestinal (GI) tract of preterm infan... more To gain insight into specific gene expression in the gastrointestinal (GI) tract of preterm infants, we adapted a method to isolate exfoliated epithelial cells. Gastric residual fluid aspirates (n = 89) or stool samples (n = 10) were collected from 96 neonates (gestational age, 24-36 wk). Cells were characterized by microscopic observation, cytokeratin-18 immunodetection, and expression of transcripts. The human origin of cellular DNA was confirmed by amplification of specific X and Y chromosome sequences. Isolation yielded 100-500 cells per sample for gastric aspirates (n = 8) and 10-20 cells for fecal samples (n = 5). Epithelial origin was confirmed by immunodetection of cytokeratin 18. Analyses of reverse transcribed products, using two independent methods, from 15 gastric fluid and two stool samples showed that 18S-rRNA and transcripts of beta-actin, glyceraldehyde-3-phosphate dehydrogenase (gapdh), and period1 were in quantities corresponding to at least 10 cells. On 59 aspirates, we found beta-actin transcripts (all but one), cytokeratin 18 (eight positive of eight samples), SLC26-A7-1 (13 positive of 19 samples), period2 (17 positive of 17 samples), and clock (25 positive of 26 samples). Exfoliated cells can be recovered from gastric aspirates and fecal samples and serve as a tool to investigate the impact of therapeutic and nutritional regimens on the maturation of GI functions.

Research paper thumbnail of Age-associated B vitamin deficiency as a determinant of chronic diseases

Nutrition Research Reviews, 2004

The number of elderly individuals is growing rapidly worldwide and degenerative diseases constitu... more The number of elderly individuals is growing rapidly worldwide and degenerative diseases constitute an increasing problem in terms of both public health and cost. Nutrition plays a role in the ageing process and there has been intensive research during the last decade on B vitaminrelated risk factors in vascular and neurological diseases and cancers. Data from epidemiological studies indicate that subclinical deficiency in most water-soluble B vitamins may occur gradually during ageing, possibly due to environmental, metabolic, genetic, nutritional and pathological determinants, as well as to lifestyle, gender and drug consumption. Older adults have distinct absorption, cell transport and metabolism characteristics that may alter B vitamin bioavailability. Case-control and longitudinal studies have shown that, concurrent with an insufficient status of certain B vitamins, hyperhomocysteinaemia and impaired methylation reactions may be some of the mechanisms involved before a degenerative pathology becomes evident. The question that arises is whether B vitamin inadequacies contribute to the development of degenerative diseases or result from ageing and disease. The present paper aims to give an overview of these issues at the epidemiological, clinical and molecular levels and to discuss possible strategies to prevent B vitamin deficiency during ageing.

Research paper thumbnail of Chylomicron β-carotene and retinyl palmitate responses are dramatically diminished when men ingest β-carotene with medium-chain rather than long-chain triglycerides

Journal of Nutrition

The effect of the ingestion of beta-carotene with medium-chain triglycerides (MCT) or long-chain ... more The effect of the ingestion of beta-carotene with medium-chain triglycerides (MCT) or long-chain triglycerides (LCT) on the bioavailability and the provitamin A activity of beta-carotene was investigated in humans. Sixteen healthy young men ingested, on two different days, a test meal containing 120 mg beta-carotene incorporated into 40 g LCT (LCT meal) or 40 g MCT (MCT meal). This meal was followed 6 h later by a beta-carotene-free meal containing 40 g LCT. Chylomicron beta-carotene, retinyl palmitate and triglycerides were measured every hour for 12.5 h after the first meal. No significant increase in chylomicron triglycerides was detected for the 6 h after the MCT meal intake, whereas a significant increase in chylomicron triglycerides was observed after the LCT meal intake. The chylomicron beta-carotene and retinyl palmitate responses to the MCT meal (0-6 h area under the curves, AUC) were significantly (P < 0.05) lower [AUC = 68.1 +/- 26.8 and 43. 4 +/- 10.4 nmol/(L.h), for ...

Research paper thumbnail of Processing of vitamin A and E in the human gastrointestinal tract

AJP Gastrointestinal and Liver Physiology

We aimed to provide basic data on the processing of vitamin A and E in the human gastrointestinal... more We aimed to provide basic data on the processing of vitamin A and E in the human gastrointestinal tract and to assess whether the size of emulsion fat globules affects the bioavailability of these vitamins. Eight healthy men received intragastrically two lipid formulas differing in their fat-globule median diameter (0.7 vs. 10. 1 microm. Formulas provided 28 mg vitamin A as retinyl palmitate and 440 mg vitamin E as all-rac alpha-tocopherol. Vitamins were measured in gastric and duodenal aspirates, as well as in chylomicrons, during the postprandial period. The gastric emptying rate of lipids and vitamin A and E was similar. The free retinol/total vitamin A ratio was not significantly modified in the stomach, whereas it was dramatically increased in the duodenum. The proportion of ingested lipid and vitamins was very similar in the duodenal content. The chylomicron response of lipids and vitamins was not significantly different between the two emulsions. Our main conclusions are as f...

Research paper thumbnail of The bioavailability of α- and β-carotene is affected by gut microflora in the rat

The British journal of nutrition

The present study examined whether the intestinal microflora could affect the bioavailability and... more The present study examined whether the intestinal microflora could affect the bioavailability and vitamin A activity of dietary α- and β-carotene in the rat. In the first set of experiments, we used conventional, germ-free (axenic), and human-flora-associated (heteroxenic) rats. In a second series, conventional rats were treated with either an antibiotic mixture or a potent inhibitor of gastric secretion (Omeprazole). All animals were first depleted of vitamin A over 4 weeks and then were fed on a sterilized diet supplemented with 14 mg β-carotene and 3 mg α-carotene/kg for 2 weeks. In both experiments, a reduction in the intestinal microflora resulted in an increased storage of β-carotene, α-carotene and vitamin A in the liver. Neither the nature of the metabolism of the intestinal microflora (aerobic or anaerobic) nor treatment with omeprazole, to modify intestinal pH, induced a significant effect on the measured variables. When incubated with 15 μmol β-carotene/l for 72 h, neithe...

Research paper thumbnail of The bioavailability of alpha- and beta-carotene is affected by gut microflora in the rat

British Journal Of Nutrition

The present study examined whether the intestinal microflora could affect the bioavailability and... more The present study examined whether the intestinal microflora could affect the bioavailability and vitamin A activity of dietary alpha- and beta-carotene in the rat. In the first set of experiments, we used conventional, germ-free (axenic), and human-flora-associated (heteroxenic) rats. In a second series, conventional rats were treated with either an antibiotic mixture or a potent inhibitor of gastric secretion (Omeprazole). All animals were first depleted of vitamin A over 4 weeks and then were fed on a sterilized diet supplemented with 14 mg beta-carotene and 3 mg alpha-carotene/kg for 2 weeks. In both experiments, a reduction in the intestinal microflora resulted in an increased storage of beta-carotene, alpha-carotene and vitamin A in the liver. Neither the nature of the metabolism of the intestinal microflora (aerobic or anaerobic) nor treatment with omeprazole, to modify intestinal pH, induced a significant effect on the measured variables. When incubated with 15 mumol beta-ca...

Research paper thumbnail of Rat intestinal β-carotene dioxygenase activity is located primarily in the cytosol of mature jejunal enterocytes

Journal of Nutrition

The purposes of this study were to determine the location of beta-carotene dioxygenase (EC 1.13.1... more The purposes of this study were to determine the location of beta-carotene dioxygenase (EC 1.13.11.21) activity within the rat gastrointestinal tract, within the villus and within enterocytes, and to identify the metabolites produced in each intestinal fraction. In Wistar female rats, maximal activity was detected in the cytosol (74-93% of the total cellular activity) of mature functional enterocytes harvested from the jejunum (67% of the intestinal activity). The specific activity, expressed in pmol of retinoids/(h x mg protein) rose from 49 +/- 3 in the stem cells to 199 +/- 12 in the mature functional cells (P < 0.05). Thus the intestinal beta-carotene cleavage activity might be regulated during the enterocyte maturation process. By using HPLC with diode array and radioactive detectors, retinal, and in the presence of NAD+, retinoic acid, were identified as the only metabolites produced. No beta-12'-, 10'-, and 8'-apo-carotenals were detected, even when various enz...

Research paper thumbnail of Mobilization of rétinol from cultured rat liver stellate cells does not require protéin secretion

Research paper thumbnail of Proteomics in the field of food and feed sciences

Proteomics has emerged recently as a powerful analytical method to investigate proteins. Its scop... more Proteomics has emerged recently as a powerful analytical method to investigate proteins. Its scope of use has considerably enlarged in the past years including in the fi eld of feed and food sciences. Through a series of studies conducted at I.N.R.A., we illustrate the interest of proteomic approaches to improve plant breeding, technological processing or nutrition studies. The objectives are to reach high-quality and healthy foods while ensuring a sustainable and competitive agriculture and a preserved environment.

Research paper thumbnail of Calibration Adjustment of the Mid-infrared Analyzer for an Accurate Determination of the Macronutrient Composition of Human Milk

Journal of Human Lactation, 2015

Human milk composition analysis seems essential to adapt human milk fortification for preterm neo... more Human milk composition analysis seems essential to adapt human milk fortification for preterm neonates. The Miris human milk analyzer (HMA), based on mid-infrared methodology, is convenient for a unique determination of macronutrients. However, HMA measurements are not totally comparable with reference methods (RMs). The primary aim of this study was to compare HMA results with results from biochemical RMs for a large range of protein, fat, and carbohydrate contents and to establish a calibration adjustment. Human milk was fractionated in protein, fat, and skim milk by covering large ranges of protein (0-3 g/100 mL), fat (0-8 g/100 mL), and carbohydrate (5-8 g/100 mL). For each macronutrient, a calibration curve was plotted by linear regression using measurements obtained using HMA and RMs. For fat, 53 measurements were performed, and the linear regression equation was HMA = 0.79RM + 0.28 (R(2) = 0.92). For true protein (29 measurements), the linear regression equation was HMA = 0.9RM + 0.23 (R(2) = 0.98). For carbohydrate (15 measurements), the linear regression equation was HMA = 0.59RM + 1.86 (R(2) = 0.95). A homogenization step with a disruptor coupled to a sonication step was necessary to obtain better accuracy of the measurements. Good repeatability (coefficient of variation &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 7%) and reproducibility (coefficient of variation &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 17%) were obtained after calibration adjustment. New calibration curves were developed for the Miris HMA, allowing accurate measurements in large ranges of macronutrient content. This is necessary for reliable use of this device in individualizing nutrition for preterm newborns.

Research paper thumbnail of Inflammation interferes with the assessment of vitamin A status in magnesium deficiency

Magnesium research : official organ of the International Society for the Development of Research on Magnesium, 2008

Hyporetinemia is observed in several pathological conditions including a primary deficiency of vi... more Hyporetinemia is observed in several pathological conditions including a primary deficiency of vitamin A and has also been reported to accompany inflammatory diseases. Experimental magnesium (Mg) deficiency in rodents is accompanied by an inflammatory syndrome. The present study was designed to determine whether the acute phase response in Mg-deficient rats can modify vitamin A status. Clinical symptoms of acute phase response were observed in Mg-deficient rats and were accompanied by a reduction in plasma retinol and of plasma retinol binding protein (RBP). Mg deficiency in rats resulted in hyporetinemia without a significant decrease in liver retinol reserves. Consequently, the data strongly suggest that the decrease in plasma retinol concentration, resulting from the level of its binding protein, is related to the inflammatory effect of Mg deficiency. These results point to the possible interference of Mg deficiency on the use of plasma retinol as an indicator of vitamin A status.

Research paper thumbnail of Use of UPLC-ESI-MS/MS to quantitate free amino acid concentrations in micro-samples of mammalian milk

SpringerPlus, 2013

Although free amino acids (FAA) account for a small fraction of total nitrogen in mammalian milk,... more Although free amino acids (FAA) account for a small fraction of total nitrogen in mammalian milk, they are more abundant in human milk than in most formulas, and may serve as a readily available source of amino acids for protein synthesis, as well as fulfill specific physiologic roles. We used reversed phase Ultra Performance Liquid Chromatography (UPLC) coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS) technique for FAA profiling in milks from three species (human, rat and cow) with a simple and rapid sample preparation. The derivatization procedure chosen, combined with UPLC-ESI-MS/MS allowed the quantitation of 21 FAA using labeled amino acids (Internal Standards) over a 10 min run time in micro-samples of mammalian milk (50 μL). The low limit of quantitation was 0.05 pmol/μL for most FAA with good repeatability and reproducibility (mean CV of 5.1%). Higher levels of total FAA were found in human (3032 μM) and rat milk (3460 μM) than in bovine milk (240 μM), with wide differences in the abundances of specific FAA between species. This robust analytical method could be applied to monitor FAA profile in human breast milk, and open the way to individualized adjustment of FAA content for the nutritional management of infants.

Research paper thumbnail of Effect of pre- and postnatal growth and post-weaning activity on glucose metabolism in the offspring

The Journal of endocrinology, 2015

Maternal caloric restriction during late gestation reduces birth weight, but whether long-term ad... more Maternal caloric restriction during late gestation reduces birth weight, but whether long-term adverse metabolic outcomes of intra-uterine growth retardation (IUGR) are dependent on either accelerated postnatal growth or exposure to an obesogenic environment after weaning is not established. We induced IUGR in twin-pregnant sheep using a 40% maternal caloric restriction commencing from 110 days of gestation until term (∼147 days), compared with mothers fed to 100% of requirements. Offspring were reared either as singletons to accelerate postnatal growth or as twins to achieve standard growth. To promote an adverse phenotype in young adulthood, after weaning, offspring were reared under a low-activity obesogenic environment with the exception of a subgroup of IUGR offspring, reared as twins, maintained in a standard activity environment. We assessed glucose tolerance together with leptin and cortisol responses to feeding in young adulthood when the hypothalamus was sampled for assess...

Research paper thumbnail of Human choriocarcinoma cell line JEG-3 produces and secretes active retinoids from retinol

Molecular Human Reproduction, 2002

Vitamin A (retinol) and its active derivatives (the retinoids) are essential for growth and devel... more Vitamin A (retinol) and its active derivatives (the retinoids) are essential for growth and development of the mammalian fetus. Maternally-derived retinol has to pass through the placenta to reach the developing fetus. Despite its apparent importance, little is known about placental metabolism of retinol, and particularly placental production and/or secretion of active retinoids. It has been previously considered that retinoids are recruited from the uterine environment to influence placental development and function during gestation. We have studied retinoid metabolism in the human choriocarcinoma cell line JEG-3 and demonstrate, for the first time, that active retinoids are produced endogenously by the JEG-3 cell line from retinol. These retinoids induce gene expression from a retinoic acidresponsive enhancer element reporter plasmid and modulate placental transglutaminase activity. Furthermore, retinoids are secreted from JEG-3, as shown by the activation of retinoic acid-responsive β lacZ reporter cells grown in conditioned media. These results suggest that there could be an active role for trophoblast-derived retinoids during human development.

Research paper thumbnail of Simple method for clinical determination of 13 carotenoids in human plasma using an isocratic high-performance liquid chromatographic method

Journal of Chromatography B: Biomedical Sciences and Applications, 2001

Research paper thumbnail of Vitamin A contained in the lipid droplets of rat liver stellate cells is substrate for acid retinyl ester hydrolase

Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1995

V~ronique Az~/is-Braesco a,*, Isabelle Dodeman b l Serge Delpal b 9 ~

Research paper thumbnail of Oxidative stress status and antioxidant status are apparently not related to carotenoid status in healthy subjects

Journal of Laboratory and Clinical Medicine, 1998

Several lines of evidence suggest that carotenoids may have a beneficial effect on health as a re... more Several lines of evidence suggest that carotenoids may have a beneficial effect on health as a result of their antioxidant properties. In addition to beta-carotene, five other carotenoids are recovered in noticeable amounts from human plasma and tissues. Although the effect of beta-carotene on in vivo lipid peroxidation has been documented, few data are available on the effects of the other carotenoids. We evaluated the ability of the main human carotenoids to reduce lipid peroxidation by determining the correlations between plasma carotenoid concentration and plasma antioxidant capacity (in 79 healthy volunteers) and between carotenoid status and breath pentane excretion (in a subgroup of 24 subjects). Carotenoid intake was assessed by means of a 3-day food recall. Carotenoid status was evaluated by measurement of beta-carotene, lycopene, lutein/zeaxanthin, and alpha-carotene in plasma and buccal mucosal cells. Antioxidant status was evaluated by measurement of the total antioxidant capacity of the plasma. Oxidative stress status was evaluated by breath pentane measurements. Food recall data and the carotenoid concentrations in plasma and buccal mucosal cells showed that the subjects had normal carotenoid intake and normal carotenoid status. The total antioxidant capacity of the plasma was not related to the concentration of any specific carotenoid. The level of expired air pentane was not related to the carotenoid status of the subjects. These results show that normal concentrations of carotenoids in plasma and tissues are not correlated with these clinical markers of antioxidant and oxidative stress status.

Research paper thumbnail of PAV-1, a new rat hepatic stellate cell line converts retinol into retinoic acid, a process altered by ethanol

The International Journal of Biochemistry & Cell Biology, 2002

During liver fibrogenesis or long term culture, hepatic stellate cells (HSCs) evolved from &a... more During liver fibrogenesis or long term culture, hepatic stellate cells (HSCs) evolved from &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;quiescent&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; to activated phenotype called &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;myofibroblast-like&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;, a transition prevented by retinoic acid (RA). Little is known about RA generation by HSCs. Our study aimed to check the ability of these cells to produce RA from retinol (Rol) and the alterations of this metabolic step by ethanol. To study this metabolic pathway, primary cultures of HSCs represent the most physiological model but technically suffer several drawbacks. To circumvent these problems, an immortalized rat HSC line (named PAV-1) has been established. We validated PAV-1 cell line as a convenient model to study retinoids metabolism by HSCs. Then, we showed that PAV-1 cells express Rol-binding proteins (RBPs), enzymes and nuclear receptors involved in RA signaling pathway. We also demonstrated in situ generation of functional all-trans-RA (ATRA), using transient transfections with a RA-sensitive reporter gene, in situ modulation of tissue transglutaminase (tTG) activity and HPLC experiments. This production was Rol dose-dependent; 4-methylpyrazole, citral, and ethanol-inhibited which argues in favor of an enzymatic process.In conclusion, we first demonstrate in situ RA generation from Rol in a newly immortalized rat HSC line, named PAV-1. Inhibition of RA production by ethanol in PAV-1 and recent data, suggesting fundamental role of RA to prevent fibrosis development in the liver, allow us to hypothesize that Rol metabolism could be a primary target for ethanol during development of hepatic fibrosis.

Research paper thumbnail of Retinol mobilization from cultured rat hepatic stellate cells does not require retinol binding protein synthesis and secretion

The International Journal of Biochemistry & Cell Biology, 2001

Retinol mobilization from retinyl esters stores of hepatic stellate cells (HSCs) is a key step in... more Retinol mobilization from retinyl esters stores of hepatic stellate cells (HSCs) is a key step in the regulation of mammalian retinol homeostasis, but the precise mechanisms of such a mobilization are still poorly understood. Using primary cultures of HSCs, we first demonstrated that HSCs expressed immunoreactivity against retinol-binding-protein (RBP) when cultured in a medium containing RBP but were unable to synthesize RBP transcripts and proteins. Using pulse and chase-type experiments, we demonstrated that radioactive retinol was released in culture medium without binding proteins. Inhibition of protein secretion by brefeldin A did not modify quantitatively retinol release. This data ruled out, for the first time, the direct involvement of RBP in retinol mobilization from HSCs. Moreover, HSCs co-cultured with primary isolated hepatocytes displayed an increase of retinol transfer from HSCs to hepatocytes when they established direct physical contacts, as compared with co-cultures without contact. Based on this latter data, a mechanism of retinol mobilization from HSCs via the hepatocytes using retinol transfer through cellular membranes is proposed.

Research paper thumbnail of Esterification of Vitamin A by the Human Placenta Involves Villous Mesenchymal Fibroblasts

Pediatric Research, 2000

Vitamin A (retinol) and its active derivatives (retinoic acids) are essential for growth and deve... more Vitamin A (retinol) and its active derivatives (retinoic acids) are essential for growth and development of the mammalian fetus. Maternally derived retinol must pass the placenta to reach the developing fetus. Despite its apparent importance, little is known concerning placental transfer and metabolism of retinol, and particularly of placental production and storage of retinyl esters. To elucidate this metabolic pathway, we incubated, in the presence of retinol, 1) human full-term placental explants and 2) primary cultures of major cells types contributing to placental function: trophoblasts and villous mesenchymal fibroblasts. We used HPLC to determine the types and concentrations of retinyl esters produced by these explants and cells. About 14% of total cellular retinol in placental explants was esterified. The most abundant esters were myristate and palmitate. Primary cell cul-tures showed that fibroblasts efficiently produced retinyl esters, but trophoblasts did not. In both types of experiments, no retinyl esters were detected in the culture medium, suggesting that retinyl esters were produced for storage purpose. These results suggest that villous mesenchymal fibroblasts are primary sites of retinol esterification and storage in the placenta. (Pediatr Res 48: 565-572, 2000) Abbreviations RA, retinoic acid RBP, retinol binding protein HBSS, Hanks' balanced salt solution DMEM, Dulbecco's modified Eagle medium ABSTRACT 565 mesenchymal fibroblasts) were tested to determine their individual capacity for esterification.

Research paper thumbnail of Recovery of Exfoliated Cells From the Gastrointestinal Tract of Premature Infants: A New Tool to Perform “Noninvasive Biopsies?”

Pediatric Research, 2007

To gain insight into specific gene expression in the gastrointestinal (GI) tract of preterm infan... more To gain insight into specific gene expression in the gastrointestinal (GI) tract of preterm infants, we adapted a method to isolate exfoliated epithelial cells. Gastric residual fluid aspirates (n = 89) or stool samples (n = 10) were collected from 96 neonates (gestational age, 24-36 wk). Cells were characterized by microscopic observation, cytokeratin-18 immunodetection, and expression of transcripts. The human origin of cellular DNA was confirmed by amplification of specific X and Y chromosome sequences. Isolation yielded 100-500 cells per sample for gastric aspirates (n = 8) and 10-20 cells for fecal samples (n = 5). Epithelial origin was confirmed by immunodetection of cytokeratin 18. Analyses of reverse transcribed products, using two independent methods, from 15 gastric fluid and two stool samples showed that 18S-rRNA and transcripts of beta-actin, glyceraldehyde-3-phosphate dehydrogenase (gapdh), and period1 were in quantities corresponding to at least 10 cells. On 59 aspirates, we found beta-actin transcripts (all but one), cytokeratin 18 (eight positive of eight samples), SLC26-A7-1 (13 positive of 19 samples), period2 (17 positive of 17 samples), and clock (25 positive of 26 samples). Exfoliated cells can be recovered from gastric aspirates and fecal samples and serve as a tool to investigate the impact of therapeutic and nutritional regimens on the maturation of GI functions.

Research paper thumbnail of Age-associated B vitamin deficiency as a determinant of chronic diseases

Nutrition Research Reviews, 2004

The number of elderly individuals is growing rapidly worldwide and degenerative diseases constitu... more The number of elderly individuals is growing rapidly worldwide and degenerative diseases constitute an increasing problem in terms of both public health and cost. Nutrition plays a role in the ageing process and there has been intensive research during the last decade on B vitaminrelated risk factors in vascular and neurological diseases and cancers. Data from epidemiological studies indicate that subclinical deficiency in most water-soluble B vitamins may occur gradually during ageing, possibly due to environmental, metabolic, genetic, nutritional and pathological determinants, as well as to lifestyle, gender and drug consumption. Older adults have distinct absorption, cell transport and metabolism characteristics that may alter B vitamin bioavailability. Case-control and longitudinal studies have shown that, concurrent with an insufficient status of certain B vitamins, hyperhomocysteinaemia and impaired methylation reactions may be some of the mechanisms involved before a degenerative pathology becomes evident. The question that arises is whether B vitamin inadequacies contribute to the development of degenerative diseases or result from ageing and disease. The present paper aims to give an overview of these issues at the epidemiological, clinical and molecular levels and to discuss possible strategies to prevent B vitamin deficiency during ageing.