Wheat ati cm3, cm16 and 0.28 allergens produced in pichia pastoris display a different eliciting potential in food allergy to wheat

International audienceAlthough wheat is a staple food for most of the human population, some of its components trigger adverse reactions. Among wheat components, the alpha-amylase/trypsin inhibitors (ATI) are important triggers of several allergies and activators of innate immunity. ATI are a group of exogenous protease inhibitors and include several polypeptides. The three ATI polypeptides named CM3, CM16 and 0.28 are considered major allergens, and might also play a role in other common wheat-related pathologies, such as Non Celiac Wheat Sensitivity and even Celiac Disease. On this basis, we pointed to obtain high amounts of them in purity and to evaluate their allergenicity potential. We thus isolated the mRNA corresponding to the three ATI genes CM3, CM16 and 0.28 from 28 days post-anthesis wheat kernels and the corresponding cDNAs were used for heterologous expression in Pichia pastoris. The three purified proteins were tested in degranulation assay against human sera of patients with food allergy to wheat. A large range of degranulation values was observed for each protein according to the sera tested. All of the three purified proteins CM3, CM16 and 0.28 were active as allergens because they were able to induce basophils degranulation on wheat allergic patients' sera, with the highest values of beta-hexosaminidase release observed for CM3 protein

Assessing walking posture with geometric morphometrics: Effects of rearing environment in pigs

International audienceRearing social animals like pigs in isolation from conspecifics can have consequences on behaviour and physiology. The aim of this experiment was to determine whether rearing conditions affect body postures. We adapted a method for quantitative evaluation of postures based on geometric morphometrics, developed in horses, for pigs and applied it in different conditions. Forty eight 75-day old females were reared either alone in 2.25 m2 pens (IH, N = 24 animals and 4 groups) or in groups of four in 4.64 m2 pens (GH, N = 24) for two weeks. They were habituated to human handling (stroking, speaking) and marking on their backs every day, and tested individually once a day for 10 min in a corridor outside the home pen during the two subsequent weeks. We observed their behaviour and posture during the first exposure to the test (novelty), and the fourth and fifth (after habituation). On the sixth and seventh tests, a familiar stockperson was present in the corridor (human presence). Before each test, the animals were marked with seven landmarks along their length, corresponding to anatomical points and easily located. An experimenter took pictures of the animals walking along the corridor, and these pictures were transferred to tps software for analysis. GH animals were more often active in the rearing pen than IH (median (IQ) 15% of observations [12-20%] versus 2% [0-13%]; P < 0.05). All animals except one IH initiated contact with the handler during the last sessions of handling (Fisher's exact test, ns). Principal Component Analyses revealed significant effects of rearing and testing conditions on pigs’ behaviour and posture. Novelty led to fewer vocalisations and more exploration for IH than GH animals (P < 0.05), but there were no differences between treatments after habituation to the testing situation. The backs of IH animals were more rounded than those of GH (P < 0.05; dimension 1 of PCA), independently of the test condition. Human presence had no effect on posture. In conclusion, the method based on geometric morphometrics that we developed to study pig posture detected variations in walking posture in pigs associated with rearing conditions. Postures might reflect affective states in pigs, as shown in other species, but further studies are needed to verify thi

Évaluation de l'allergénicité des aliments : Application au diagnostic de 'allergie alimentaire

The aim of this thesis consists in reviewing the advanced knowledge about the evaluation of food allergenicity and the different applications to the diagnosis of food allergy. Definition, characteristics and classification of food allergens precede the description of physical and chemical factors involved in the modification and cross-reactivity of food allergens. This review leads to the current tools used for the in vivo and in vitro diagnosis of food allergy as well as the recent procedures which have been developed for the detection of allergen traces in industrial food. A close collaboration between clinicians and biologists improves the diagnosis procedures. This cooperation is illustrated by various clinical situations in which new tools (i.e. specific IgE to recombinant allergens, food allergen traces identification in meals or drugs) have improved the diagnostic procedures and consequent therapeutic measures.Cette thèse actualise les connaissances de l'évaluation de l'allergénicité des aliments et son application au diagnostic de l'allergie alimentaire. Après la définition, les caractéristiques et la classification des allergènes alimentaires, les phénomènes physico-chimiques modifiant l'allergénicité des aliments ainsi que les réactivités croisées sont décrites. Ainsi sont introduits les outils cliniques et biologiques utiles au diagnostic de l'allergie alimentaire et à la détection des traces d'allergènes alimentaires. Une collaboration étroite entre cliniciens et chercheurs biologistes, permet d'optimiser la prise en charge diagnostique et thérapeutique de l'allergie alimentaire. Cette démarche se concrétise par la mise à disposition et l'utilisation de divers outils (développement d'allergènes recombinants, dosage de contaminants alimentaires dans des médicaments ou aliments, ...) et est illustrée par diverses mises en situation clinique réelles

Evaluation of Food allergenicity : Application to the diagnosis of food allergy

Cette thèse actualise les connaissances de l'évaluation de l'allergénicité des aliments et son application au diagnostic de l'allergie alimentaire. Après la définition, les caractéristiques et la classification des allergènes alimentaires, les phénomènes physico-chimiques modifiant l'allergénicité des aliments ainsi que les réactivités croisées sont décrites. Ainsi sont introduits les outils cliniques et biologiques utiles au diagnostic de l'allergie alimentaire et à la détection des traces d'allergènes alimentaires. Une collaboration étroite entre cliniciens et chercheurs biologistes, permet d’optimiser la prise en charge diagnostique et thérapeutique de l’allergie alimentaire. Cette démarche se concrétise par la mise à disposition et l'utilisation de divers outils (développement d’allergènes recombinants, dosage de contaminants alimentaires dans des médicaments ou aliments, ...) et est illustrée par diverses mises en situation clinique réelles.The aim of this thesis consists in reviewing the advanced knowledge about the evaluation of food allergenicity and the different applications to the diagnosis of food allergy. Definition, characteristics and classification of food allergens precede the description of physical and chemical factors involved in the modification and cross-reactivity of food allergens. This review leads to the current tools used for the in vivo and in vitro diagnosis of food allergy as well as the recent procedures which have been developed for the detection of allergen traces in industrial food. A close collaboration between clinicians and biologists improves the diagnosis procedures. This cooperation is illustrated by various clinical situations in which new tools (i.e. specific IgE to recombinant allergens, food allergen traces identification in meals or drugs) have improved the diagnostic procedures and consequent therapeutic measures

Testing optimal methods to compare horse postures using geometric morphometrics

International audienceThe study of animal behavior, especially regarding welfare, needs the development of tools to identify, quantify and compare animal postures with interobserver reliability. While most studies subjectively describe animal postures, or quantify only limited parts of the body, the usage of geometric morphometrics has allowed for the description of horses’ and pigs’ upper body outline and the comparison of postures from different populations thanks to robust statistical analysis. We have attempted here to optimize the geometric morphometrics (GM) method already used in horses by introducing the outline analysis with sliding semilandmarks (SSL), by eliminating the balance movement of the neck and by focusing only on parts of the upper line. For this purpose, photographs of 85 horses from 11 riding schools, known for differing in terms of housing and working conditions, were analyzed with previous and new GM methods and these results were compared with each other. Using SSL and eliminating the neck movement appeared to better discriminate the horse populations than the previous GM method. Study of parts of the dorsum proved efficient too. This new methodology should now be used to examine if posture could be an indicator of horse welfare state, and similar studies should be performed in other species in order to validate the same methodology

Could posture reflect welfare state? A study using geometric morphometrics in riding school horses

International audienceDespite the fact that animal posture is known to reflect emotional state, the presence of chronic postures associated with poor welfare has not been investigated with an objective tool for measuring, quantifying and comparing postures. The use of morphometric geomet-rics (GM) to describe horse posture (profile of the dorsum) has shown to be an effective method of distinguishing populations that are known to differ in terms of welfare states. Here we investigated photographs of 85 riding school horses differing in terms of welfare state, in order to determine if a specific posture (modelled by GM) is associated with altered welfare. The welfare state was estimated with the prevalence of stereotypic or abnormal repetitive behaviours, depressed-like posture and the ear positions. ANOVA results show that horses with stereotypic or abnormal behaviour, and to a lesser degree horses with depressed-like postures, tend to have a flatter, or even hollow, dorsal profile, especially at the neck and croup levels. These altered profiles could represent an additional indicator of poor welfare, easy to use in the field or by owners

Effect of the serotonin 5-HT7 recepteur on cellular proliferation in Drosophila melanogaster

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Threshold dose for peanut: Risk characterization based upon diagnostic oral challenge of a series of 286 peanut-allergic individuals

Clinical records of 286 consecutive patients reacting positively with objective symptoms to double-blind, placebo-controlled oral peanut challenges at University Hospital, Nancy, France were examined for individual No Observed Adverse Effect Levels (NOAELs) and Lowest Observed Adverse Effect Levels (LOAELs). After fitting to a log-normal probability distribution model, the ED10 and ED05 were 14.4 and 7.3 mg (expressed as whole peanut), respectively, with 95% lower confidence intervals of 10.7 and 5.2 mg, respectively. Compared to results from a previous study where the ED10 was based upon individual peanut thresholds gleaned from 12 publications, a statistically significant difference was observed between the ED50’s, but not the ED10’s of the two probability distribution curves. The Nancy patient group contains more sensitive subjects than the group from the published literature thus contributing to the observed differences. Minimum eliciting dose-distributions for patients with histories of more severe reactions (grade 4 or 5; 40 subjects) did not differ significantly from those of patients with histories of less severe reactions (grades 1–3; 123 subjects). These data and this modeling approach could be used to establish population thresholds for peanut-allergic consumers and thereby provide a sound basis for allergen control measures in the food industry

Complementary Nuclear Magnetic Resonance-Based Metabolomics Approaches for Glioma Biomarker Identification in a Drosophila melanogaster Model

International audienceHuman malignant gliomas are the most common type of primary brain tumor. Composed of glial cells and their precursors, they are aggressive and highly invasive, leading to a poor prognosis. Due to the difficulty of surgically removing tumors and their resistance to treatments, novel therapeutic approaches are needed to improve patient life expectancy and comfort. Drosophila melanogaster is a compelling genetic model to better understanding human neurological diseases owing to its high conservation in signaling pathways and cellular content of the brain. Here, glioma has been induced in Drosophila by co-activating the epidermal growth factor receptor and the phosphatidyl-inositol-3 kinase signaling pathways. Complementary nuclear magnetic resonance (NMR) techniques were used to obtain metabolic profiles in the third instar larvae brains. Fresh organs were directly studied by 1H high resolution-magic angle spinning (HR-MAS) NMR, and brain extracts were analyzed by solution-state 1H-NMR. Statistical analyses revealed differential metabolic signatures, impacted metabolic pathways, and glioma biomarkers. Each method was efficient to determine biomarkers. The highlighted metabolites including glucose, myo-inositol, sarcosine, glycine, alanine, and pyruvate for solution-state NMR and proline, myo-inositol, acetate, and glucose for HR-MAS show very good performances in discriminating samples according to their nature with data mining based on receiver operating characteristic curves. Combining results allows for a more complete view of induced disturbances and opens the possibility of deciphering the biochemical mechanisms of these tumors. The identified biomarkers provide a means to rebalance specific pathways through targeted metabolic therapy and to study the effects of pharmacological treatments using Drosophila as a model organism