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International audience; 3-Carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF) is a metabolite of furan fatty acid and a marker of fish oil intake. CMPF is described as a protein-bound uremic toxin and interacts with free oxygen radicals, which can induce cell damages. However, the clinical consequences of CMPF accumulation in haemodialysis patients remain poorly documented. The aims of this study are to investigate potential association between CMPF levels and (i) biochemical and nutritional parameters; (ii) cardiovascular events and (iii) mortality. Two hundred and fifty-two patients undergoing maintenance haemodialysis were included. Routine clinical biochemistry tests and assay for CMPF by HPLC technique were performed at the inclusion. Body composition parameters were measured using a bioimpedance spectroscopy method. The enrolled patients were prospectively monitored for cardiovascular events and mortality. CMPF level was positively correlated with nutritional parameters and lean mass and is significantly higher in patients without protein-energy wasting. However, the multivariate linear regression analysis indicated that CMPF level was not independently associated with albumin, prealbumin, creatinemia and body mass index. Elevated serum CMPF was not associated with mortality and cardiovascular morbidity. Our results indicate that CMPF is not a relevant uremic toxin in haemodialysis and in contrast could be a marker of healthy diet and omega 3 intakes.

International audience; RATIONALE: The first step in the diagnosis of oligosaccharidoses is to evidence abnormal oligosaccharides excreted in urine, usually performed by the poorly sensitive but efficient thin layer chromatography (TLC) method. Developing a tandem mass spectrometry (MS/MS) technique could be of great interest to replace TLC. METHODS: Abnormal underivatized oligosaccharides have been recently studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, allowing the unambiguous identification of oligosaccharidoses. Based on this previous work, we developed an advantageous and efficient liquid chromatography (LC)/MS/MS method using a more common triple quadrupole tandem mass spectrometer for oligosaccharides analysis. RESULTS: Oligosaccharidoses (n = 97) and control (n = 240) urine samples were analysed. A specific pattern was obtained for each oligosaccharidosis using this method. In urine, it allows not only the identification of all the oligosaccharidoses previously identified by TLC (fucosidosis, alphamannosidosis, aspartylglucosaminuria, GM1 gangliosidosis, sialidosis, galactosialidosis and Schindler disease), but also extends the field of diagnosis to mucolipidosis type II, Sandhoff disease, and beta-mannosidosis. The same technique was applied to 16 amniotic fluid supernatants from oligosaccharidosis-affected foetuses (n = 16) compared with 37 unaffected. All the affected foetuses could be clearly identified: sialidosis (n = 3), galactosialidosis (n = 4), aspartylglucosaminuria (n = 1), mucolipidosis type II (n = 4) or GM1 gangliosidosis (n = 4). This technique can be applied to early prenatal diagnosis as well as to the oligosaccharidosis screening in the case of non-immune hydrops fetalis. CONCLUSIONS: The method is quick and easy to run, with an LC analysis time of 13 min per sample. The quantitative validation could not be obtained in the absence of a specific standard and of a labelled internal standard for each compound. Even if this LC/MS/MS method is only qualitative, it is very specific and much more sensitive than TLC. It allows the urinary screening of oligosaccharidoses, even mild or late-onset forms, and the screening of antenatal forms in amniotic fluid. Copyright (C) 2017 John Wiley & Sons, Ltd.

International audience; Folates, also known as B9 vitamins, serve as donors and acceptors in one-carbon (C1) transfer reactions. The latter are involved in synthesis of many important biomolecules, such as amino acids, nucleic acids and vitamin B5. Folates also play a central role in the methyl cycle that provides one-carbon groups for methylation reactions. The important functions fulfilled by folates make them essential in all living organisms. Plants, being able to synthesize folates de novo, serve as an excellent dietary source of folates for animals that lack the respective biosynthetic pathway. Unfortunately, the most important staple crops such as rice, potato and maize are rather poor sources of folates. Insufficient folate consumption is known to cause severe developmental disorders in humans. Two approaches are employed to fight folate deficiency: pharmacological supplementation in the form of folate pills and biofortification of staple crops. As the former approach is considered rather costly for the major part of the world population, biofortification of staple crops is viewed as a decent alternative in the struggle against folate deficiency. Therefore, strategies, challenges and recent progress of folate enhancement in plants will be addressed in this review. Apart from the ever-growing need for the enhancement of nutritional quality of crops, the world population faces climate change catastrophes or environmental stresses, such as elevated temperatures, drought, salinity that severely affect growth and productivity of crops. Due to immense diversity of their biochemical functions, folates take part in virtually every aspect of plant physiology. Any disturbance to the plant folate metabolism leads to severe growth inhibition and, as a consequence, to a lower productivity. Whereas today's knowledge of folate biochemistry can be considered very profound, evidence on the physiological roles of folates in plants only starts to emerge. In the current review we will discuss the implication of folates in various aspects of plant physiology and development.

International audience; A sensitive electrochemical molecularly imprinted sensor was developed for the detection of glyphosate (Gly), by electropolymerisation of p-aminothiophenol-functionalised gold nanoparticles in the presence of Gly as template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognise and bind Gly through hydrogen bonds between Gly molecules and aniline moieties. The performance of the developed sensor for the detection of Gly was investigated by linear sweep voltammetry using a hexacyanoferrate/hexacyanoferrite solution as redox probe, the electron transfer rate increasing when concentration of Gly increases, due to a p-doping effect. The molecularly imprinted sensor exhibits a broad linear range, between 1 pg/L and 1 mu g/L and a quantification limit of 0.8 pg/L. The selectivity of the proposed sensor was investigated towards the binding of Gly metabolite, aminomethylphosphonic acid, revealing excellent selectivity towards Gly. The developed sensor was successfully applied to detect Gly in tap water samples.

Hyperhomocysteinemia induces vascular endothelial dysfunction, an early hallmark of atherogenesis. While higher levels of circulating asymmetric dimethylarginine (ADMA) and symmetric dimethyl arginine (SDMA), endogenous inhibitors of nitric oxide synthesis, have been associated with increased cardiovascular risk, the role that ADMA and SDMA play in the initiation of hyperhomocysteinemia-induced endothelial dysfunction remains still controversial. In the present study, we studied the changes of circulating ADMA and SDMA in a rat model of acutely hyperhomocysteinemia-induced endothelial dysfunction. In healthy rats, endothelium-related vascular reactivity (measured as acetylcholine-induced transient decrease in mean arterial blood pressure), plasma ADMA and SDMA, total plasma homocysteine (tHcy), cysteine and glutathione were measured before and 2, 4 and 6 h after methionine loading or vehicle. mRNA expression of hepatic dimethylarginine dimethylaminohydrolase-1 (DDAH1), a key protein responsible for ADMA metabolism, was measured 6 h after the methionine loading or the vehicle. Expectedly, methionine load induced a sustained increase in tHcy (up to 54.9 +/- A 1.9 A mu M) and a 30 % decrease in vascular reactivity compared to the baseline values. Plasma ADMA and SDMA decreased transiently after the methionine load. Hepatic mRNA expression of DDAH1, cathepsin D, and ubiquitin were significantly lower 6 h after the methionine load than after the vehicle. The absence of an elevation of circulating ADMA and SDMA in this model suggests that endothelial dysfunction induced by acute hyperhomocysteinemia cannot be explained by an up-regulation of protein arginine methyltransferases or a down-regulation of DDAH1. In experimental endothelial dysfunction induced by acute hyperhomocysteinemia, down-regulation of the proteasome is likely to dampen the release of ADMA and SDMA in the circulation.

International audience; The end-use value of durum wheat (Triticum turgidum L. subsp. durum (Desf.) Husn.) is mainly governed by its grain protein concentration and composition. Adjustment of variables to compensate for inter-annual and location variations in semolina quality leads to high cost for processors in the wheat industry. A better understanding of the mechanisms governing environmental variations of grain protein composition is thus required. Here, a field experiment was setup in a Mediterranean environment with the aim to analyze the effect of sowing date and nitrogen (N) fertilization on the dynamics of grain dry mass, water, protein composition and glutenin polymer size distribution for durum wheat cv. Creso. The results indicated that (1) grain dry mass accumulation was related to grain water concentration and stopped at 44.9% independently of the growing conditions and N supply; (2) during the grain filling period as well as at ripeness maturity, the quantity of the different protein fractions scaled with the quantity of N per grain; (3) SDS-extractable glutenin polymers were produced continuously until the same grain water concentration as the dry mass deposition was reached; (4) SDS-unextractable polymeric proteins were found as early as 7 days after anthesis and their rate of accumulation increased sharply when grain dry mass was 60% of its final value and proceeded at a constant rate until ripeness maturity, thus suggesting that the insolubilization of glutenin polymers is not directly related to the rapid loss of water after physiological maturity, but rather to the continuous dehydration of the grain. (C) 2014 Elsevier B.V. All rights reserved.

International audience; Carbohydrates have a wide variety of structures whose complexity and heterogeneity challenge the field of analytical chemistry. Tandem mass spectrometry, with its remarkable sensitivity and high information content, provides key advantages to addressing the structural elucidation of polysaccharides. Yet, classical fragmentation by collision-activated dissociation (CAD) in many cases fails to reach a comprehensive structural determination, especially when isomers have to be differentiated. In this work, for the first time, vacuum ultra-violet (VUV) synchrotron radiation is used as the activation process in tandem mass spectrometry of large oligosaccharides. Compared to low energy CAD (LE-CAD), photon activated dissociation brought more straightforward and valuable structural information. The outstanding feature was that complete series of informative ions were produced, with only minor neutral losses. Moreover, systematic fragmentation rules could be drawn thus facilitating the definitive assignments of fragment identities. As a result, most of the structures present in a complex mixture of oligogalacturonans could be comprehensively resolved, including many isomers differing in the position of methyl groups along the galacturonic acid backbone.

BACKGROUND Polyphenols have a favourable antioxidant potential on human health, suggesting that their high content in apple is responsible for the beneficial effects of apple consumption. They are also linked to the quality of apple juices and ciders since they are predominantly responsible for astringency, bitterness and colour. Major phenolic compounds were quantified by liquid chromatography in fruits and juices from a cider apple progeny harvested for 3 years. The total content of procyanidins and their average degree of polymerisation (DPn) were also determined in fruits by phloroglucinolysis. Variability and extraction yield of these compounds were determined. RESULTS The variability observed in the progeny was representative of the variability observed in many cider apple varieties. Hydroxycinnamic acids were the most extractable group, with an average extraction yield of 67%, whereas flavonols and anthocyanins were the least. CONCLUSION This study is the first to introduce variability and extraction yields of the main phenolic compounds in both fruits and juices of a cider apple progeny. This dataset will be used for an upcoming QTL mapping study, an original approach that has never been undertaken for cider apple. (c) 2013 Society of Chemical Industry

Because of their high toxicity, international regulatory institutions recommend monitoring specific polycyclic aromatic hydrocarbons (PAHs) in environmental and food samples. A fast, selective and sensitive method is therefore required for their quantitation in such complex samples. This article deals with the optimization, based on an experimental design strategy, of a cyclodextrin (CD) modified capillary zone electrophoresis separation method for the simultaneous separation of 19 PAHs listed as priority pollutants. First, using a central composite design, the normalized peak-start and peak-end times were modelled as functions of the factors that most affect PAH electrophoretic behavior: the concentrations of the anionic sulfobutylether-beta-CD and neutral methyl-beta-CD, and the percentage of MeOH in the background electrolyte. Then, to circumvent computational difficulties resulting from the changes in migration order likely to occur while varying experimental conditions, an original approach based on the systematic evaluation of the time intervals between all the possible pairs of peaks was used. Finally, a desirability analysis based on the smallest time interval between two consecutive peaks and on the overall analysis time, allowed us to achieve, for the first time in CE, full resolution of all 19 PAHs in less than 18 min. Using this optimized capillary electrophoresis method, a vegetable oil was successfully analyzed, proving its suitability for real complex sample analysis. (C) 2014 Elsevier B.V. All rights reserved.

International audience; In this work, we developed a surface functionalization way of silica monoliths with a rapid, simple, versatile, and localizable photografting step. The elaboration of a photoreactive layer at the surface of monoliths was first optimized. The functionalization with [γ-(methacryloyloxy)propyl]trimethoxysilane at 80°C in a hydro-organic solution containing triethylamine as catalyst allows reachng the highest density of methacrylate photoactive moieties on silica surfaces. These methacrylate reactive surfaces were subsequently photografted within few minutes with acrylate monomers bearing alkyl chains (C12 and C18). The photografting efficiency was determined by monitoring the retentive properties of monoliths in the RP mode. The retention factors are of the same order of magnitude as highly retentive columns obtained by modification of silica surface with long-alkyl chain silanes or by thermal polymerization of long-alkyl chain monomers. It was also verified that such grafting neither impaired the efficiency of the monolithic stationary phase (Hmin = 6-8 μm in nano-LC) nor its permeability (about 6 × 10(-14) m(2)). Further, it was also demonstrated that photografting is localizable in nonmasked defined areas. Results obtained in anion-exchange chromatography after photopolymerization of [2-(methacryloyloxy)ethyl]trimethylammonium chloride are presented as well to demonstrate the versatility of the developed approach.

International audience; A design of experiement approach is described for the optimization of the microscopic morphology of macro-mesoporous titania monoliths that were elaborated for the chromatographic enrichment of phosphorylated compounds. The monolithic titania gels were formed via an alkoxy-derived sol-gel route in association with a phase separation mechanism. The synthesis was performed at mild temperatures of gelation using starting mixtures of titanium n-propoxide, hydrochloric acid, N-methylformamide, water, and poly (ethylene oxide). The gelation temperature and the chemical compositions of N-methylformamide, water, and poly (ethylene oxide) were chosen as the most relevant experimental factors of the sol-gel process. Using the sizes of the skeletons and macropores as morphological descriptors of the dried porous monoliths, the statistical analyses simultaneously revealed the effects and interactions between the different factors. Crack-free TiO2 monolithic rods of 8 to 10 cm long with well-defined co-continuous macropores and micro-structured skeletons were obtained after selection of the sol-gel parameters and optimization of the drying and heat-treatment steps of the gels. The bimodal texture of the rods exhibited macropores of 1.5 mu m and mesopores centered at 5.2 nm with a total surface area of 140 m(2) g(-1). The ability of the macro-mesoporous titania rods to selectively bind phosphorylated compounds was demonstrated for O-phosphoamino acids (P-Ser, P-Thr, P-Tyr).

International audience; Phenolic compounds are a large class of plant secondary metabolites, showing a diversity of structures, from rather simple structures, e.g. phenolic acids, through polyphenols such as flavonoids, that comprise several groups, to polymeric compounds based on these different classes. Phenolic compounds are important for the quality of plant based foods: they are responsible for the colour of red fruits, juices and wines and substrates for enzymatic browning, and are also involved in flavour properties. In particular, astringency is ascribed to precipitation of salivary proteins by polyphenols, a mechanism possibly involved in defence against their anti-nutritional effects. Finally, phenolic compounds are considered to contribute to the health benefits associated to dietary consumption of fruits and vegetables. During food processing and storage, plant phenolics are converted to a variety of derived compounds. While methods to analyse lower molecular weight phenolic compounds are well developed, analysis of polymeric compounds remains a challenge. Indeed, strong interactions of polymeric phenolics with plant cell wall material limit their extraction. Besides, their polydispersity results in poor resolution and detection, especially of derived structures such as oxidation products. However, recent advances of the analytical techniques have allowed some progress in their structural characterisation. This review summarizes the current knowledge on methods to analyse polyphenols. It presents their reactions in foods and beverages and the resulting structures, and highlights some aspects related to their impact on colour, flavour and health properties, with examples taken mostly from wine research.

International audience; Rye bran and aleurone, wheat bran and aleurone, and oat bran and cell wall concentrate were compared in their in vitro gut fermentation patterns of individual phenolic acids and short-chain fatty acids, preceded by enzymatic in vitro digestion mimicking small intestinal events. The formation of phenolic metabolites was the most pronounced from the wheat aleurone fraction. Phenylpropionic acids, presumably derived from ferulic acid (FA), were the major phenyl metabolites formed from all bran preparations. The processed rye, wheat, and oat bran fractions contained more water-extractable dietary fiber (DF) and had smaller particle sizes and were thus more easily fermentable than the corresponding brans. Rye aleurone and bran had the highest fermentation rate and extent probably due to high fructan and water-extractable arabinoxylan content. Oat samples also had a high content of water-extractable DF, beta-glucan, but their fermentation rate was lower. Enzymatic digestion prior to in vitro colon fermentation changed the structure of oat cell walls as visualized by microscopy and increased the particle size, which is suggested to have retarded the fermentability of oat samples. Wheat bran was the most slowly fermentable among the studied samples, presumably due to the high proportion of water-unextractable DF. The in vitro digestion reduced the fructan content of wheat samples, thus also decreasing their fermentability. Among the studied short-chain fatty acids, acetate dominated the profiles. The highest and lowest production of propionate was from the oat and wheat samples, respectively. Interestingly, wheat aleurone generated similar amounts of butyrate as the rye fractions even without rapid gas production.

International audience; In recent years, metal oxides such as titania have been commercially available as chromatographic beds that can potentially be used to achieve novel separations of polar compounds. For example beta blockers, which are more often encountered in environmental sciences, have a wide range of polarity, and their basic character leads to difficult sample treatment and separation on conventional silica-based sorbents. The contribution of titania to the selective analysis of nine beta blockers was evaluated in terms of retention mechanisms observed in hydrophilic interaction LC using acetonitrile/water mobile phases with various additives. The mobile phase additives enabled to control the beta blocker charge as well as the titania surface charge. Depending on their respective ionic state, various retention mechanisms were identified at low water contents (

International audience; Presently, few biomarker-based approaches are available for the evaluation of environmental exposure to persistent organic pollutants in dairy ruminants. In this study, goats (Capra hircus) were orally administered a mixture of pyrene, phenanthrene, and benzo[a]pyrene daily over a 40-d period (1 or 50 mg/d). Milk and urine 1-hydroxypyrene levels, ethoxyresorufin-O-deethylase (EROD) activity in peripheral blood lymphocytes (PBL) as well as urinary levels of 2- and 3-hydroxyphenanthrene were determined at 10-d intervals. 1-Hydroxypyrene excretion in milk and urine significantly increased and then achieved a plateau at 10 d. Transfer rates of 1-hydroxypyrene were calculated to be approximately 0.5 and 25% in milk and urine, respectively. Concentrations in milk and urine were proportional to the ingested doses. These results demonstrate that 1-hydroxypyrene in milk or urine may be used as a biomarker for evaluating the exposure of dairy ruminants to polycyclic aromatic hydrocarbons (PAHs) over an extended exposure period. Constitutive EROD activity in lymphocytes was 0.5 +/- 0.3 pmol resorufin/min/mg protein, and was significantly induced over the entire exposure time, before stabilizing after 40d at 6.30 +/- 1.3 and 18.89 +/- 1.12 pmol resorufin/min/mg protein for 1 mg/d and 50 mg/d doses, respectively. Induction kinetics were calculated using a logistic-like model and approximate dose-response curves were designed. We therefore propose EROD activity in PBL as a relevant, convenient, and noninvasive biomarker of subchronic exposure of dairy ruminants to CYP450 inducing PAH. Environ. Toxicol. Chem. 2011;30:1346-1353. (C) 2011 SETAC

International audience; Cation-exchange chromatography was applied for the separation of short synthetic peptide standards, with various charges and hydrophobicities. The methodology to develop simple, reproducible and accurate models, based on physicochemical peptide properties, was described. A multilinear regression method was used for the calculation of the models, and descriptors were chosen according to the observed phenomena. The predictive and interpretative ability of the chromatographic models was evaluated considering cross-validated data (root mean-squared error of calibration, root mean-squared error, root mean-squared error of cross-validation and the Fisher ratio). Hydrophobic coefficients for amino acids were calculated with or without consideration of peptide sequences. A simple model, with only two parameters (charge and hydrophobic coefficient) was built. It enabled an accurate prediction of short peptide elution (up to nine residues). As the model was intended for further characterization of complex mixtures of unknown peptides, some mixtures were analyzed to investigate possible interactions between molecules. Peptides eluted in exactly the same pattern as when injected alone, supporting the use of these models for complex mixtures of small peptides.

The present work is aimed at gaining conclusive mechanistic insights into the radiation-induced formation of the 5'R and 5'S diastereomers of both adenine and guanine 5',8-cyclo-2'-deoxyribonucleosides, with emphasis on the delineation of the inhibitory effect of O(2) in isolated and cellular DNA. The levels of purine 5',8-cyclo-2'-deoxyribonucleosides as assessed by HPLC-MS/MS were found to decrease steadily with the increase of O(2) concentration, the 5',8-cyclo-2'-deoxyguanosine being produced more efficiently than the 5',8-cyclo-2'-deoxyadenosine for low O(2) concentrations. A high stereoselectivity was observed in the intramolecular addition of the C5' radical to the C8 of the purine leading, after the creation of the C5'-C8 bond and a subsequent oxidation step, to the predominant formation of the 5'R diastereomer for both purine 5',8-cyclonucleosides. The reduced formation yield of the 4 tandem lesions in the presence of O(2) explains, at least partly, the low efficiency of radiation-induced yields of the purine 5',8-cyclo-2'-deoxyribonucleosides in cellular DNA, which are about two orders of magnitude lower than the previously reported data obtained from HPLC-MS analysis.

International audience; The effects of Trametes hirsuta laccase and Pentopan Mono BG xylanase and their combination on oat, wheat, and mixed oat-wheat doughs and the corresponding breads were investigated. Laccase treatment decreased the content of water-extractable arabinoxylan (WEAX) in oat dough due to oxidative cross-linking of feruloylated arabinoxylans. Laccase treatment also increased the proportion of water-soluble polysaccharides (WSNSP) apparently due to the beta-glucanase side activity present in the laccase preparation. As a result of the laccase treatment, the firmness of fresh oat bread was increased. Xylanase treatment doubled the content of WEAX in oat dough and slightly increased the amount of WSNSP. Increased stiffness of the dough and firmness of the fresh bread were detected, probably because of the increased WEAX content, which decreased the amount of water available for P-glucan. The combination of laccase and xylanase produced slight hydrolysis of P-glucan by the P-glucanase side activity of laccase and enhanced the availability of AX for xylanase with concomitant reduction of the amount and molar mass of WSNSP. Subsequently, the volume of oat bread was increased. Laccase treatment tightened wheat dough, probably due to cross-linking of WEAX to higher molecular weight. In oat-wheat dough, laccase slightly increased the proportion of WSNSP between medium to low molecular weight and increased the specific volume of the bread. Xylanase increased the contents of WEAX and WSNSP between medium to low molecular weight in oat-wheat dough, which increased the softness of the dough, as well as the specific volume and softness of the bread. The results thus indicate that a combination of laccase and xylanase was beneficial for the textures of both oat and oat-wheat breads.

International audience; The hypertrophy and hyperplasia of infected roots into clubs are the intrinsic characteristics of clubroot, one of the economically most important diseases in Brassica crops worldwide. Polyamines, arginine (Arg)-derived metabolites, have long been recognized as cell proliferation and differentiation regulators in plants and consequently are suitable candidates for potential gall development factors. Furthermore, Arg catabolism, through arginase, which is strongly connected to polyamine metabolism, would play an important role in response to wound trauma and pathogen infection. In this study, we exploited the Arabidopsis (Arabidopsis thaliana)-Plasmodiophora brassicae pathosystem to investigate the involvement of polyamine metabolism and Arg catabolism in host responses to the pathogen infection and in partial clubroot resistance mechanisms. We demonstrated at the transcriptional, enzymatic, and metabolic levels that polyamine metabolism and Arg catabolism are induced during the later stages of disease in compatible Arabidopsis-P. brassicae interactions. However, susceptible and partially resistant plants showed strikingly different Arg metabolism signatures. Susceptible plants were characterized by a transient agmatine production, a massive induction of arginase, and a strong accumulation of proline. The potential functions of this marked activation of the arginase pathway in the P. brassicae pathogenicity strategy are discussed. Partially resistant plants showed a continuous agmatine production and a weaker arginase pathway activity than the susceptible genotype. Results suggest that the symptom severity was strongly associated to the differential regulation of root polyamine metabolism and Arg catabolism. Further work using arginase transgenic plants will provide insight into the physiological function of the arginase pathway in partial clubroot resistance.

International audience; In a previous paper we reported that beta-D-glucans isolated from Saccharomyces cerevisiae could adsorb zearalenone. reduce its bioavailability in the digestive tract, and protect animals against its adverse effects. We have now investigated, in vitro, the kinetics of the interaction between other mycotoxins and beta-D-glucans from several sources at three pH values found along the digestive tract (3.0, 6.0, and 8.0). Acid and neutral conditions gave the highest affinity rates for aflatoxins B1 > deoxynivalenol > ochratoxin A and involved both the (1 -> 3)-beta-D-glucans and the (1 -> 6)-beta-D-glucans. Alkaline conditions, owing to their destructuring action on glucans, were favorable only for the adsorption of patulin. Using molecular mechanics, we found that hydroxyl, ketone, and lactone groups are involved in the formation of both hydrogen bonds and van der Waals interactions between aflatoxins B 1, deoxynivalenol and patulin, and beta-D-glucans. Differences in the binding capacity of the mycotoxins are due to their specific physical and chemical characteristics.

International audience; Analysis of the interaction between the host immune system and the intracellular parasite Mycobacterium leprae has identified a 35 kDa protein as a dominant antigen, The native 35 kDa protein was purified from the membrane fraction of M. leprae and termed MMPI (major membrane protein I). As the purified protein was not amenable to N-terminal sequencing, partial proteolysis was used to establish the sequences of 21 peptides, A fragment of the 35 kDa proteinen-coding gene was amplified by the polymerase chain reaction from M. leprae chromosomal DNA with oligonucleotide primers derived from internal peptide sequences and the whole gene was subsequently isolated from a M. leprae cosmid library. The nucleotide sequence of the gene revealed an open reading frame of 307 amino acids containing most of the peptide sequences derived from the native 35 kDa protein, The calculated subunit mass was 33.7 kDa, but the native protein exists as a multimer of 950 kDa, Database searches revealed no identity between the 35 kDa antigen and known protein sequences. The gene was expressed in Mycobacterium smegmatis under the control of its own promoter or at a higher level using an 'up-regulated' promoter derived from Mycobacterium fortuitum. The gene product reacted with monoclonal antibodies raised to the native protein, Using the bacterial alkaline phosphatase reporter system, we observed that the 35 kDa protein was unable to be exported across the membrane of recombinant M. smegmatis. The 35 kDa protein-encoding gene is absent from members of the Mycobacterium tuberculosis complex, but homologous sequences were detected in Mycobacterium avium, Mycobacterium haemophilum and M. smegmatis, The avaibility of the recombinant 35 kDa protein will permit dissection of both antibody- and T-cell-mediated immune responses in leprosy patients.