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IF 4.066; International audience

Monocarboxylates have been implicated in the control of energy homeostasis. Among them, the putative role of ketone bodies produced notably during high-fat diet (HFD) has not been thoroughly explored. In this study, we aimed to determine the impact of a specific rise in cerebral ketone bodies on food intake and energy homeostasis regulation. A carotid infusion of ketone bodies was performed on mice to stimulate sensitive brain areas for 6 or 12 h. At each time point, food intake and different markers of energy homeostasis were analyzed to reveal the consequences of cerebral increase in ketone body level detection. First, an increase in food intake appeared over a 12-h period of brain ketone body perfusion. This stimulated food intake was associated with an increased expression of the hypothalamic neuropeptides NPY and AgRP as well as phosphorylated AMPK and is due to ketone bodies sensed by the brain, as blood ketone body levels did not change at that time. In parallel, gluconeogenesis and insulin sensitivity were transiently altered. Indeed, a dysregulation of glucose production and insulin secretion was observed after 6 h of ketone body perfusion, which reversed to normal at 12 h of perfusion. Altogether, these results suggest that an increase in brain ketone body concentration leads to hyperphagia and a transient perturbation of peripheral metabolic homeostasis.

Background and aim To accurately quantify the cost of physical activity and to evaluate the different components of energy expenditure in humans, it is necessary to evaluate external mechanical work ( W EXT ). Large platform systems surpass other currently used techniques. Here, we describe a calculation method for force-platforms to calculate long-term W EXT . Methods Each force-platform (2.46 × 1.60 m and 3.80 × 2.48 m) rests on 4 piezoelectric sensors. During long periods of recording, a drift in the speed of displacement of the center of mass (necessary to calculate W EXT ) is generated. To suppress this drift, wavelet decomposition is used to low-pass filter the source signal. By using wavelet decomposition coefficients, the source signal can be recovered. To check the validity of W EXT calculations after signal processing, an oscillating pendulum system was first used; then, 10 healthy subjects performed a standardized exercise (squatting exercise). A medical application is also reported in eight Parkinsonian patients during the timed “get-up and go” test and compared with the same test in ten healthy subjects. Results Values of W EXT with the oscillating pendulum showed that the system was accurate and reliable. During the squatting exercise, the average measured W EXT was 0.4% lower than theoretical work. W EXT and mechanical work efficiency during the “get-up and go” test in Parkinson's disease patients in comparison with that of healthy subjects were very coherent. Conclusions This method has numerous applications for studying physical activity and mechanical work efficiency in physiological and pathological conditions.

Introduction: Sweeteners are widespread in overweight or diabetic patients because of their high sweetening power and their light effect on glycaemia. Sweeteners effects on cortical activity, reflecting both cortical analysis and gustatory signalization pathway, are not well known. We observed previously that gustatory evoked potentials (GEPs) in response to sucrose varied in latency and/or in amplitude with sucrose concentration of the solution, the pleasantness of taste and the feeding status. The aim was to compare the gustatory cortical activity by recording GEPs in response to sucrose, aspartame and stevia solutions.Methods: Twenty healthy non-smoker adult volunteers were included (mean age 22 ± 2,47 years). GEPs were obtained after stimulation of taste receptors with sucrose and sweeteners solutions. Three randomly assigned sessions (during 3 different days) were performed for each subject: one with sucrose (10 g/100 ml of Evian water), one with aspartame (0.05 g/100 ml) and one with stevia (0.033 g/100 ml). Concentrations have been chosen in order to induce the same sweetening power for each molecule. Results: GEPs latency from primary gustatory cortex (Cz electrode) for sucrose, aspartame and stevia stimuli were respectively 149 ± 37 ms, 178 ± 42 ms and 203 ± 49ms, showing a significant shorter GEPs latency in response to the sucrose solution (p

International audience; The THESEUS project (Towards Human Exploration of Space: aEUropean Strategy) was initiated within the seventh FrameworkProgramme by the European Commission. This project aimed toprovide a cross-cutting, life science-based roadmap for Europe’sstrategy towards human exploration of space, especially for deepspace missions and its relevance to applications on Earth. Toaddress these challenges, relevance of space research on thecardiovascular system, the lungs and kidneys, was discussed in anexpert group and its principal conclusions will be presented in thisarticle.

Acute myocardial infarction (MI), a leading cause of morbidity and mortality worldwide, is the irreversible death of heart muscle secondary to ischemia. This ischemia, i.e. oxygen and nutrients deprivation, triggers a reticular stress disrupting the Ca2+ balance of the cardiac cell. Several Ca2+ pumps and channels located at the sarcolemma or at the reticulum membrane are key players in this maintenance of Ca2+ homeostasis. Among them, we find passive leak channels, such as TRPs and little is known about their precise role in MI.TRPV1 represents a non-selective cation channel that is activated by capsaicin, pH and noxious heat. In skeletal muscle, we previously demonstrated that TRPV1 is located in the longitudinal part of the SR and respond to pharmacological and physiological activations (Lotteau et al., 2013). We questioned here whether TRPV1 might have a similar role in heart physiology. Biochemical analysis and intracellular Ca2+ measurements were performed on cardiomyocytes from wild-type and TRPV1-KO mice. Our in vitro results show that: (i) TRPV1 is expressed in cardiac cells; (ii) an increase in intracellular calcium concentration ([Ca2+]i) is elicited under TRPV1 activation; (iii) TRPV1 could be a direct target of isoflurane. In parallel, our in vivo results indicate that a pharmacological preconditioning by isoflurane decrease the infarct size, probably though activation of TRPV1. According to the fact that TRPV1 activity can be modulated by a lot of pharmacological molecules, TRPV1 may serve as therapeutic target to reduce the infarct size. Most of published data have already evidenced this TRPV1 cardioprotective role in the peripheral heart system. The aim of the present work is to describe how TRPV1 channels behave in adult cardiomyocytes, L'infarctus du myocarde, une des causes majeures de mortalité à travers le monde, engendre une mort irréversible du muscle cardiaque suite à une ischémie. Cette ischémie, c'est-à-dire une privation de dioxygène et de nutriments, déclencher un stress réticulaire qui perturbe l'équilibre calcique de la cellule cardiaque. Plusieurs pompes et canaux calciques situés à la membrane plasmique ou réticulaire sont des intervenants clés dans le maintien de l'homéostasie calcique. Parmi eux, il existe des canaux de fuites calciques passives, comme les TRPs, et peu d'informations sont actuellement connus à propos de leur rôle précis au cours de l'infarctus du myocarde.TRPV1 est un canal cationique non sélectif qui est activé par la capsaïcine, le pH et la chaleur nocive (>42°C). Dans le muscle squelettique, nous avions démontré que TRPV1 est situé dans la partie longitudinale du réticulum sarcoplasmique et qu'il répond à différentes stimulations physiologiques et pharmacologiques (Lotteau et al., 2013). Ici, nous nous interrogeons sur un éventuel rôle similaire de TRPV1 dans la physiologie cardiaque. Des analyses biochimiques et des mesures de calcium intracellulaire furent réalisées sur des cardiomyocytes issus de souris WT et KO TRPV1. Nos résultats in vitro montrent que: (a) TRPV1 est exprimé dans les cellules cardiaques; (b) une activation de TRPV1 engendre une réduction de la concentration calcique réticulaire et que (c) TRPV1 pourrait être une cible directe de l'isoflurane.Dans la mesure où TRPV1 peut être modulé par de nombreuses molécules pharmacologiques, il pourrait constituer une cible thérapeutique pour réduire la taille d'infarctus. De nombreuses études antérieures ont déjà mis en évidence un rôle cardioprotecteur de TRPV1 dans le système nerveux entourant le cœur. Le but de cette étude est de décrire le fonctionnement des canaux TRPV1 dans des cardiomyocytes adultes

International audience; The fluoranthene (Fluo) is one of the most abundant polycyclic aromatic hydrocarbons (PAHs) in human food and in marine compartments. However, the existing data on its genotoxicity is poor and controversial. The aim of this study was to assess in vitro the potential genotoxicity of Fluo in sole and its possible effect on CYP450 modulation. Freshly isolated hepatocytes were exposed for 24 h to a range of Fluo concentrations from 0.5 to 50 μM in both culture flasks and microplate wells. The ethoxyresorufin-O-deethylase (EROD) activity was measured as an indicator of the activity of the cytochrome P450 1A1 (CYP1A1). The genotoxic effects were evaluated by measuring both DNA strand breaks and DNA adducts by the alkaline comet assay and the postlabeling technique respectively. Calf thymus DNA was also exposed to Fluo in the presence of sole liver microsomes in order to check for Fluo DNA adduct formation. In sole hepatocytes, Fluo was shown to induce a decrease in the EROD activity in a concentration-dependent manner. A significant genotoxic effect was observed in terms of DNA strand breakage from an exposure concentration of 5 μM: despite a concentration-dependent effect was observed, it did not follow a linear dose-response. The response was similar whatever the way of exposure in flasks or in wells. One reproducible adduct was detected in the hepatocytes exposed to the highest concentrations of Fluo. The formation of Fluo adducts was confirmed by the detection of one reproducible adduct following in vitro exposure of calf thymus DNA to 100 and 200 μM of Fluo in the presence of sole microsomes. These results demonstrate the potential of sole hepatocytes to metabolize Fluo in 24 h into reactive species, able to induce genotoxicity by DNA strand breakage and DNA adduct formation. Moreover, a miniaturized cell exposure system was validated for further experiments using fewer amounts of hepatocytes and contaminants, and allowing exposure to PAH metabolites.

Hyperglycemia is detected and integrated by the mediobasal hypothalamus (MBH) which, in turn, inhibits food intake and triggers insulin secretion. The MBH houses specialized glucose-sensitive (GS) neurons, which directly or indirectly modulate their electrical activity in response to changes in glucose level. In a first study, we hypothesized that indirect detection of glucose by MBH GS neurons involves the secretion of endozepine by astrocytes, a gliotransmitter known to inhibit food intake in response to hyperglycemia. The present work shows that endozepines selectively activate anorexigenic MBH pro-opiomelanotortine (POMC) neurons. In the second study, we show that the direct detection of increased glucose level involves hypothalamic glucose-excited (HGE) neurons. Using pharmacological and genetic approaches, we demonstrate that the redox-sensitive Transient Receptor Potential Canonical 3 et 4 (TRPC3/4) channels are involved in MBH HGE response to glucose in vitro and increased insulin secretion and decreased food intake in response to cerebral hyperglycemia in vivo. We also obtained evidences that MBH TRPC3 channel is a critical new player for energy homeostasis. This thesis work identifies two new mechanisms involved in hypothalamic detection of hyperglycemia: the first based on the involvement of TRPC3/4 channels in HGE neurons and the second highlighting the astroglial endozepines as a relay of the “glucose” signal to POMC neurons.; L’hyperglycémie est détectée et intégrée au niveau de l’hypothalamus médio-basal (MBH) qui inhibe la prise alimentaire et déclenche la sécrétion d’insuline. Le MBH renferme des neurones spécialisés gluco-sensibles (GS) qui détectent directement ou indirectement des variations de la concentration extracellulaire en glucose.Dans une première étude, nous suggérons que la détection indirecte du glucose par les neurones GS hypothalamiques repose sur la libération d’endozépines par les astrocytes, un gliotransmetteur connu pour inhiber la prise alimentaire en réponse à l’hyperglycémie. Nous travaux montrent que les endozépines activent spécifiquement les neurones à pro-opiomélanocortine (POMC) du MBH pour générer leur effet anorexigène.Dans une seconde étude, nous montrons que la détection directe de l’hyperglycémie implique les neurones hypothalamiques dits « high gluco-excited » (HGE). Grâce à des approches pharmacologiques et génétiques, nous mettons en évidence que les canaux redox sensibles Transient Receptor Potential Canonical 3 et 4 (TRPC3/4) sont fondamentaux pour la détection du glucose par les neurones HGE in vitro, la stimulation de la sécrétion d’insuline et la diminution de la prise alimentaire en réponse à l’hyperglycémie cérébrale in vivo. De plus, nos travaux démontrent que les canaux TRPC3 du MBH jouent un rôle clef dans le contrôle de l’homéostasie énergétique.Les travaux de cette thèse permettent de mettre en évidence deux nouveaux mécanismes de détection hypothalamique de l’hyperglycémie : l’un reposant sur l’implication des canaux TRPC3/4 dans les neurones HGE et l’autre proposant les endozépines astrocytaires comme relai du signal « glucose » aux neurones POMC.

International audience; During the early period of running after cycling, EMG patterns of the leg are modified in only some highly trained triathletes. The majority of studies have analysed muscle EMG patterns at arbitrary, predetermined time points. The purpose of this study was to examine changes to EMG patterns of the lower limb at physiologically determined times during the cycle-run transition period to better investigate neuromuscular adaptations. Six highly trained triathletes completed a 10 min isolated run (IR), 30 min of rest, then a 20 min cycling procedure, before a 10 min transition run (C-R). Surface EMG activity of eight lower limb muscles was recorded, normalised and quantified at four time points. Oxygen uptake and heart rate values were also collected. Across all muscles, mean (+/- SD) EMG patterns, demonstrated significant levels of reproducibility for each participant at all four time points (alpha < 0.05; r = 0.52-0.97). Mean EMG patterns during C-R correlated highly with the IR patterns (alpha < 0.05). These results show that EMG patterns during subsequent running are not significantly affected by prior cycling. However, variability of muscle recruitment activity does appear to increase during C-R transition when compared to IR.

International audience; This study aimed to compare the response of salivary hormones of track and field athletes induced by preparation and pre-competitive training periods in an attempt to comment on the physiological effects consistent with the responses of each of the proteins measured. Salivary testosterone, cortisol, alpha-amylase, immunoglobulin A (IgA), chromogranin A, blood creatine kinase activity, and profile of mood state were assessed at rest in 24 world-class level athletes during preparation (3 times in 3 months) and pre-competitive (5 times in 5 weeks) training periods. Total mood disturbance and fatigue perception were reduced, while IgA (+61%) and creatine kinase activity (+43%) increased, and chromogranin A decreased (−27%) during pre-competitive compared to preparation period. A significant increase in salivary testosterone (+9 to +15%) and a decrease in testosterone/cortisol ratio were associated with a progressive reduction in training load during pre-competitive period (P < 0.05). None of the psycho-physiological parameters were significantly correlated to training load during the pre-competitive period. Results showed a lower adrenocortical response and autonomic activity, and an improvement of immunity status, in response to the reduction in training load and fatigue, without significant correlations of salivary hormones with training load. Our findings suggest that saliva composition is sensitive to training contents (season period) but could not be related to workload resulting from track and field athletics training.

International audience; Membrane conductance (Dm) and capillary lung volume (Vc) derived from NO and CO lung transfer measurements in humans depend on the blood conductance (θ) values of both gases. Many θ values have been proposed in the literature. In the present study, measurements of CO and NO transfer while breathing 15% or 21% O2 allowed the estimation of θNO and the calculation of the optimal equation relating 1/θCO to pulmonary capillary oxygen pressure (PcapO2). In 10 healthy subjects, the mean calculated θNO value was similar to the θNO value previously reported in the literature (4.5mmHgmin(-1)) provided that one among three θCO equations from the literature was chosen. Setting 1/θCO=a·PcapO2+b, optimal values of a and b could be chosen using two methods: 1) by minimizing the difference between Dm/Vc ratios for any PcapO2, 2) by establishing a linear equation relating a and b. Using these methods, we are proposing the equation 1/θCO=0.0062·PcapO2+1.16, which is similar to two equations previously reported in the literature. With this set of θ values, DmCO reached the morphometric range.

A biomarker is a molecule (or a cluster of molecule) which will reflect the occurrence of a pathological state, giving us the ability to detect a disease, to predict its severity or to assess drug efficiency. Biological fluids are the golden standards for biomarker research in human as they are routinely collected for patients’ follow-up and are less invasive than biopsies. During my PhD, I focused on exosomes that can be found in these biological fluids. Exosomes are nanovesicles with a diameter ranging between 30 and 100 nanometers. Exosomes are secreted by all cell types and harbor cytoplasmic and membranous proteins specific of their cells of origins. One of the major interest of exosomes enriched from biological fluids is that they represent a valuable source of biomarkers. They can be considered as a « liquid biopsy ». Their analysis could complete classical diagnosis and follow-up tools. In this project, we applied high resolution, high throughput proteomic techniques for exosomes analysis. We firstly focused on protein profiles in urinary exosomes in the context of two urinary tract diseases: cystinuria and kidney cancer. Cystinuria is an inherited autosomal recessive disease that is characterized by the formation of cystine stones in the kidneys. To date, there are no markers to predict the evolution toward end stage renal disease. We developed a method to prepare exosomes in order to reproducibly analyze their protein profiles. We applied this method to eight cystinuria patients and compared their profiles to those of ten healthy subjects. A panel of 38 differentially expressed proteins in patients were found and validated by western blots. We also applied this method to patients with clear cell renal cell carcinoma, for which invasive biopsies are necessary for clear diagnosis. We analyzed urinary exosomes form eight patients before and after nephrectomy. We were able to highlight 25 overexpressed proteins in patients’ exosomes. Eventually, the last part of my thesis was dedicated to the analysis of exosomes enriched from bronchoalveolar lavage fluid collected in cystic fibrosis patients, a disease that affects mostly the lungs. Bronchoalveolar lavage fluid exosomes analysis could give a new insight on the mechanisms of this disease. We compared protein profiles in exosomes from four cystic fibrosis patients and six asthmatic patients. The whole point of this work is to show that proteomic analysis of exosomes isolated from biological fluids could become a golden standard for the discovery of diagnosis or prognosis biomarkers.; Un biomarqueur est une molécule (ou un ensemble de molécules) présente dans l’organisme qui témoigne de l’apparition d’un processus pathologique. Il permet ainsi de dépister une maladie, d’en prédire sa gravité ou encore d’évaluer l’efficacité d’un traitement. Les liquides biologiques représentent des milieux de choix pour la recherche de biomarqueurs en pathologie humaine car leur collection est habituelle dans la prise en charge des patients et moins invasive comparée aux biopsies d’organes ou de tissus. Dans cette thèse, nous nous sommes intéressés plus particulièrement aux exosomes présents dans ces liquides biologiques. Les exosomes sont des nanovésicules dont le diamètre est compris entre 30 et 100 nanomètres. Ils sont sécrétés par tous les types cellulaires et contiennent des protéines cytoplasmiques et membranaires spécifiques de leur cellule d’origine. L’intérêt majeur des exosomes isolés à partir des liquides biologiques, est qu’ils constituent une source de biomarqueurs. Ils peuvent donc être assimilés à une « biopsie liquide ». L’analyse des exosomes pourrait compléter utilement des examens classiques de dépistage, de diagnostic et de suivi d’une pathologie. Dans le cadre de projet de cette thèse, nous avons appliqué des techniques de protéomique à haut débit pour l’analyse des exosomes. Nous nous sommes tout d’abord intéressés à l’analyse du profil protéique des exosomes urinaires dans le contexte de deux pathologies du tractus urinaire : la cystinurie et le cancer du rein. La cystinurie est une néphropathie lithiasique d’origine génétique pour laquelle il y a peu de marqueurs biologiques pouvant prédire son évolution vers l’insuffisance rénale terminale. Nous avons développé une méthode de préparation des exosomes urinaire permettant d’analyser de façon reproductible leurs profils protéiques. Nous avons appliqué cette méthode à huit patients cystinuriques et comparé les résultats aux profils obtenus chez dix sujets sains. Un panel de 38 protéines différentiellement exprimé dans les exosomes des patients a été identifié et en partie validé par Western blot. Concernant le cancer du rein à cellules claires pour lequel le diagnostic nécessite des prélèvements invasifs par biopsie, nous avons analysé les exosomes urinaires de huit patients avant et après néphrectomie. Nous avons ainsi pu mettre en évidence un panel de 25 protéines surexprimées dans les exosomes des patients. Enfin, le dernier volet de cette thèse a été consacré à l’analyse des exosomes du lavage broncho-alvéolaire provenant de patients MV, maladie d’origine génétique qui atteint principalement les poumons. L’analyse des exosomes de lavage broncho-alvéolaire pourrait permettre de donner un éclairage nouveau sur la physiopathologie de la maladie. Nous avons réalisé la comparaison des profils protéiques des exosomes de quatre patients MV, et six patients asthmatiques. L’ensemble des résultats obtenus au cours de cette thèse montre que l’analyse protéomique des exosomes issus de fluides biologiques peut aider la recherche de biomarqueurs diagnostics ou pronostics de maladies.

Epub 2015 Jan 31; The objective of this study was to characterize the mechanics of maximal running sprint acceleration in high-level athletes. Four elite (100-m best time 9.95-10.29 s) and five sub-elite (10.40-10.60 s) sprinters performed seven sprints in overground conditions. A single virtual 40-m sprint was reconstructed and kinetics parameters were calculated for each step using a force platform system and video analyses. Anteroposterior force (FY), power (PY), and the ratio of the horizontal force component to the resultant (total) force (RF, which reflects the orientation of the resultant ground reaction force for each support phase) were computed as a function of velocity (V). FY-V, RF-V, and PY-V relationships were well described by significant linear (mean R(2) of 0.892 ± 0.049 and 0.950 ± 0.023) and quadratic (mean R(2) = 0.732 ± 0.114) models, respectively. The current study allows a better understanding of the mechanics of the sprint acceleration notably by modeling the relationships between the forward velocity and the main mechanical key variables of the sprint. As these findings partly concern world-class sprinters tested in overground conditions, they give new insights into some aspects of the biomechanical limits of human locomotion.

Import JabRef | WosArea Physiology; Sport Sciences; International audience; The use of electrical stimulation (ES) can contribute to our knowledge of how our neuromuscular system can adapt to physical stress or unloading. Although it has been recently challenged, the standard technique used to explore central modifications is the twitch interpolated method which consists in superimposing single twitches or high-frequency doublets on a maximal voluntary contraction (MVC) and to compare the superimposed response to the potentiated response obtained from the relaxed muscle. Alternative methods consist in (1) superimposing a train of stimuli (central activation ratio), (2) comparing the MVC response to the force evoked by a high-frequency tetanus or (3) examining the change in maximal EMG response during voluntary contractions, if this variable is normalized to the maximal M wave, i.e. EMG response to a single stimulus. ES is less used to examine supraspinal factors but it is useful for investigating changes at the spinal level, either by using H reflexes, F waves or cervicomedullary motor-evoked potentials. Peripheral changes can be examined with ES, usually by stimulating the muscle in the relaxed state. Neuromuscular propagation of action potentials on the sarcolemma (M wave, high-frequency fatigue), excitation-contraction coupling (e.g. low-frequency fatigue) and intrinsic force (high-frequency stimulation at supramaximal intensity) can all be used to non-invasively explore muscular function with ES. As for all indirect methods, there are limitations and these are discussed in this review. Finally, (1) ES as a method to measure respiratory muscle function and (2) the comparison between electrical and magnetic stimulation will also be considered.

International audience; Aim: Voluntary apnoea induces several physiological adaptations, including bradycardia, arterial hypertension and redistribution of regional blood flows. Elite breath-hold divers (BHDs) are able to maintain very long apnoea, inducing severe hypoxaemia without brain injury or black-out. It has thus been hypothesized that they develop protection mechanisms against hypoxia, as well as a decrease in overall oxygen uptake. Methods: To test this hypothesis, the apnoea response was studied in BHDs and non-divers (NDs) during static and dynamic apnoeas (SA, DA). Heart arterial oxygen saturation (SaO(2)), and popliteal artery blood flow were rate, recorded to investigate the oxygen-conserving effect of apnoea response, and the internal carotid artery blood flow was used to examine the mechanisms of cerebral protection. Results: The bradycardia and peripheral vasoconstriction were accentuated in BHDs compared with NDs (P < 0.01), in association with a smaller SaO(2) decrease (-2.7% vs. -4.9% during SA, P < 0.01 and -6% vs. -11.3% during DA, P < 0.01). Greater increase in carotid artery blood flow was also measured during apnoea in BHDs than in controls. Conclusion: These results confirm that elite divers present a potentiation of the well-known apnoea response in both SA and DA conditions. This response is associated with higher brain perfusion which may partly explain the high levels of world apnoea records.

International audience; BACKGROUND: Parecoxib, a selective cyclooxygenase-2 inhibitor, may reduce postoperative pain without increasing bleeding when administered before surgery. METHODS: We randomly assigned 62 patients scheduled for total hip arthroplasty to the following IV dosing schedule: 1) placebo at induction, at wound closure, and 12 h after induction (control); 2) parecoxib 40 mg at induction, placebo at wound closure, and parecoxib 40 mg 12 h after induction (pre); or, 3) placebo at induction, parecoxib 40 mg at wound closure, and parecoxib 40 mg 12 h after induction (post). Pain scores at rest and with movement were recorded every 4 h for 24 h using a visual analog scale. Treatment side effects were recorded every 4 h. Red cell loss for 5 days after surgery was calculated. RESULTS: Postoperative pain scores were less in the pre and post groups than in the control group. Postoperative bleeding was similar in the three groups. There were no significant differences between the pre and post groups, nor was their any trend suggesting a preemptive analgesic efficacy from preincision administration of parecoxib. Morphine use in the Postanesthesia Care Unit was reduced in the pre and post groups compared with the control group (14.2 +/- 2.0, and 15.7 +/- 2.0, vs 20.4 +/- 2.3 mg), although the trend was only significant (P < 0.05) in the pre group. The first pain score was also reduced in the pre and post groups compared to the control group (56.1 +/- 7.5 and 64.2 +/- 7.0 vs 78.3 +/- 5), but this was also only significant for the pre group (P = 0.001). The delay for first analgesic demand was increased for both the pre and post group compared to the control group (38 +/- 9 and 28.2 +/- 6.6 vs 18 +/- 6 min) but, again, this was only significant for the pre group (P = 0.05). Twenty-four hour consumption of morphine was similar in the pre (26 +/- 12 mg) and post groups (25 +/- 13 mg); both were significantly less than in the control group (47 +/- 27 mg, P < 0.001). CONCLUSIONS: Administration of parecoxib before hip arthroplasty did not provide preemptive analgesia. There was a trend towards improved analgesia immediately after surgery with preincision administration, consistent with the expected time course of nonsteroidal antiinflammatory drug's effect. Perioperative parecoxib administration, consisting of two injections spaced 12 h apart, improved postoperative analgesia over the first 24 h without increasing bleeding.

The kallikrein-kinin(s) system (KKS) is a peptide system with various pathophysiological effects. Kinins exert their actions through activation of two different receptor subtypes: B1 receptor (B1R) and B2 receptor (B2R). The aim of my thesis was to study the role of this system in diabetic complications in mice. We used pharmacological approaches using new specific agonists of B1R or B2R, or a specific antagonist of B2R. In the first part, we showed that selective pharmacological activation of B1R or B2R overcomes the effect of diabetes on post-ischemic neovascularization and restores tissue perfusion through inflammation. In a second model, we showed that B2R agonist impairs wound repair in mice, inducing skin disorganization and epidermis thickening. Interestingly, B2R blockade improves skin wound healing in two mouse models of diabetes. In a last part, we showed that B2R activation increases mortality after transient cerebral ischemia. In diabetic mice, B1R activation has neuroprotective effects. Indeed, B1R agonist treatment decreases infarct size and improves neurological deficit at day 2 after transient cerebral ischemia. To conclude, KKS activation has contradictory effects depending on the organs studied. The study of new B1 or B2 kinin receptor agonists opens new therapeutic options in diabetic complications.; Le système kallicréine-kinine(s) (SKK) est un système peptidique vasomoteur avec des effets multiples et complexes qui dépendent principalement du tissu et des pathologies étudiés. Les kinines activent deux types de récepteurs : le récepteur B1 (RB1) et le récepteur B2 (RB2). L’objectif de mon travail de thèse a été d’étudier les effets de nouveaux agonistes spécifiques des RB1 ou des RB2 dans certaines complications associées au diabète chez la souris. Dans une première partie, nous avons pu montrer que l’administration chronique d’un agoniste spécifique des RB1 ou des RB2 corrige entièrement le défaut de néovascularisation post-ischémique observé chez la souris diabétique notamment en augmentant l’inflammation. Dans un deuxième modèle, nous avons pu montrer que l’activation des RB2 entraîne une désorganisation des couches cutanées et un retard important de cicatrisation chez la souris. En revanche, l’administration d’un antagoniste des RB2 accélère de façon significative la cicatrisation dans deux modèles de diabète. La dernière partie de ce travail a permis de montrer que l’administration chronique des RB2 augmente la mortalité des animaux ayant subit une ischémie cérébrale. Chez la souris diabétique, l’activation des RB1 a des effets neuroprotecteurs puisqu’elle réduit la taille de l'infarctus et les déficits neurologiques après une ischémie cérébrale transitoire. Pour conclure, nous avons pu montrer au cours de ces travaux que l’activation du SKK a des effets contradictoires en fonction des organes étudiés. Ainsi, l’étude de ces nouveaux agonistes des récepteurs des kinines ouvre des nouvelles voies thérapeutiques dans les complications du diabète.

Epub 2015 Jul 17; Overground sprint studies have shown the importance of net horizontal ground reaction force impulse (IMPH) for acceleration performance, but only investigated one or two steps over the acceleration phase, and not in elite sprinters. The main aim of this study was to distinguish between propulsive (IMPH+) and braking (IMPH−) components of the IMPH and seek whether, for an expected higher IMPH, faster elite sprinters produce greater IMPH+, smaller IMPH−, or both.Nine high-level sprinters (100-m best times range: 9.95–10.60 s) performed 7 sprints (2×10 m, 2×15 m, 20 m, 30 m and 40 m) during which ground reaction force was measured by a 6.60 m force platform system. By placing the starting-blocks further from the force plates at each trial, and pooling the data, we could assess the mechanics of an entire “virtual” 40-m acceleration.IMPH and IMPH+ were significantly correlated with 40-m mean speed (r=0.868 and 0.802, respectively; P

The Neural networks underlying the liking and wanting responses to food odors are modified in anorexia nervosa. 25. Congress of the european chemoreception research organisation (ECRO)

International audience; Mouse cortical astrocytes express VAMP3 but not VAMP2. VAMP3 vesicles undergo Ca(2+) -independent exo- and endocytotic cycling at the plasma membrane. VAMP3 vesicle traffic regulates the recycling of plasma membrane glutamate transporters. cAMP modulates VAMP3 vesicle cycling and glutamate uptake. Previous studies suggest that small synaptic-like vesicles in astrocytes carry vesicle-associated vSNARE proteins, VAMP3 (cellubrevin) and VAMP2 (synaptobrevin 2), both contributing to the Ca(2+) -regulated exocytosis of gliotransmitters, thereby modulating brain information processing. Here, using cortical astrocytes taken from VAMP2 and VAMP3 knock-out mice, we find that astrocytes express only VAMP3. The morphology and function of VAMP3 vesicles were studied in cultured astrocytes at single vesicle level with stimulated emission depletion (STED) and total internal reflection fluorescence (TIRF) microscopies. We show that VAMP3 antibodies label small diameter (∼80 nm) vesicles and that VAMP3 vesicles undergo Ca(2+) -independent exo-endocytosis. We also show that this pathway modulates the surface expression of plasma membrane glutamate transporters and the glutamate uptake by astrocytes. Finally, using pharmacological and optogenetic tools, we provide evidence suggesting that the cytosolic cAMP level influences astrocytic VAMP3 vesicle trafficking and glutamate transport. Our results suggest a new role for VAMP3 vesicles in astrocytes.

National audience

Introduction: Early nutrition through maternal diet during pregnancy and breastfeeding was shown to have an impact on obesity in childhood. Experiments on animal models have also indicated alterations regarding taste preferences. In addition, salivary physiology has a documented role in taste preferences and food intake in rodents. The objective of this study was therefore to study the impact of maternal diet on salivary glands’ protein profiles of the offspring.Materials and methods: Female rats received during gestation and lactation either a control (C) or a high fat / high sugar diet (WD for “Western Diet”). 18 offspring rats (9C/9WD) were sacrificed at 25 days of age, i.e. shortly after weaning. Protein expression in their submandibular glands was studied by 2D electrophoresis, and proteins differentially expressed (p

International audience; Introduction: Gustatory evoked potentials (GEP), detected in response to an intermittent stimulation of the gustatory receptors by a primary flavor, are a reliable and safe record of the human gustatory function. We previously highlighted the modifications of GEP in response to saccharose stimulation by the concentration of the saccharose solution and its hedonic sensation. Fatty taste might be considered as the sixth primary flavor. We aimed to demonstrate that GEP in response to fatty taste stimulation do exist, and to compare these GEP with the GEP in response to saccharose and salty taste.Methods: Healthy and voluntary male subjects were included. Each subject underwent six GEP recording with different taste solutions: 5g and 20g of saccharose per 100mL of water, 0,5g and 2g of NaCl per 100mL of water, 0.25% and 1% linoleic acid solution. GEP were recorded from 9 cortical sites with EEG sensors: Cz, Fz, Pz, C3, C4, F3, F4, Fp1 and Fp2 of the 10/20 system.Results:We already included 12 subjects. We recorded GEP in response to the two concentrations of linoleic acid, like as saccharose and salt stimuli. GEP in response to saccharose stimulation had shorter latency and higher amplitude than fatty or salty taste. Contrary to salty and fatty stimulation, saccharose solution strongly activated the bilateral orbito-frontal cortex.Discussion/Conclusion:Recording GEP in response to linoleic acid stimulation shows that it exists gustative receptors which response to fatty taste. So, it is a strong argument to say that fatty taste can be considered as the sixth primary flavor.

International audience; Introduction: Unstable footwear has been shown to increase lower extremity muscle activity, but the reflex response to perturbations induced by this intervention is unknown. Methods: Twenty healthy subjects stood in stable and unstable footwear conditions (presented randomly) while H-reflex amplitude and background muscle activity were measured in the soleus and lateral gastrocnemius (LG) muscles. Results: Wearing unstable footwear resulted in larger H-reflexes (normalized to the maximal M-wave) for the LG (+12%; P=0.025), but not for the soleus (+4%; P>0.05). Background activity of both muscles was significantly higher in the unstable condition. Conclusions: The H-reflex facilitation observed with unstable footwear was unexpected, as challenging postural conditions usually result in reflex depression. Increased muscle activity, decreased presynaptic inhibition, and/or more forward postural position may have (over-)compensated the expected reflex depression. Differences between LG and soleus H-reflex modulation may be due to diverging motor unit recruitment thresholds.

International audience; The purpose of this cross-sectional study was to investigate the change in 100-km running performance and in the age of peak performance for 100-km ultramarathoners. Age and running speed of the annual fastest women and men in all 100-km ultramarathons held worldwide between 1960 and 2012 were analyzed in 148,017 finishes with 18,998 women and 129,019 men using single, multivariate, and nonlinear regressions. Running speed of the annual fastest men increased from 8.67 to 15.65 km.h(-1) and from 8.06 to 13.22 km.h(-1) for the annual fastest women. For the annual 10 fastest men, running speed increased from 10.23 ± 1.22 to 15.05 ± 0.29 km.h(-1) (p < 0.0001) and for the annual 10 fastest women from 7.18 ± 1.54 to 13.03 ± 0.18 km.h(-1) (p < 0.0001). The sex difference decreased from 56.1 to 16.3% for the annual fastest finishers (p < 0.0001) and from 46.7 ± 8.7% to 14.0 ± 1.2% for the annual 10 fastest finishers (p < 0.0001). The age of the annual fastest men increased from 29 to 40 years (p = 0.025). For the annual fastest women, the age remained unchanged at 35.0 ± 9.7 years (p = 0.469). For the annual 10 fastest women and men, the age remained unchanged at 34.9 ± 3.2 (p = 0.902) and 34.5 ± 2.5 years (p = 0.064), respectively. To summarize, 100-km ultramarathoners became faster, the sex difference in performance decreased but the age of the fastest finishers remained unchanged at ∼ 35 years. For athletes and coaches to plan a career as 100-km ultramarathoner, the age of the fastest female and male 100-km ultramarathoners remained unchanged at ∼ 35 years between 1960 and 2012 although the runners improved their performance over time.

Epub 2015 Oct.; Previous studies have reported that reaction time in gait initiation was affected by emotion eliciting pictures. This study examined the effect of a change in the delay between image onset and the imperative “go” on reaction time. From a standing posture, 19 young adults had to walk (several steps) toward pleasant or unpleasant images in two conditions. In the short condition, the word “go” appeared 500 ms after image onset and participants were instructed to initiate gait as soon as possible after the word go appeared. In the long condition, the same procedure was used but the word “go” appeared 3000 ms after image onset. Results demonstrated that motor responses were faster for pleasant pictures than unpleasant ones in the short condition. In contrast, no significant difference was found between both categories of pictures in the long condition. Moreover, we found that self ratings of valence of unpleasant pictures were less unpleasant in the long condition than in the short one whereas there was no difference for pleasant pictures between both conditions. This result reflected a change in the affective significance of unpleasant pictures in the long condition. We also found in the long condition, that the body was inclined forward and to the stance limb during the standing posture and importantly with a similar extent for pleasant and unpleasant pictures. This change clearly reflected a facilitation of the gait initiation process. Overall, results suggested that this gait facilitation when confronted to unpleasant pictures resulted from emotional regulation processes enabling to reappraise these pictures and to override the initial avoidance tendency that they caused.

Obesity is now considered a chronic low-grade inflammatory disease with increased levels of inflammatory mediators both in circulation and adipose tissue. Among adipose tissue cell types, adipocytes and macrophages are known to play key roles in the progression of inflammation by establishing and maintaining it. In this PhD project, we particularly focus on the transcriptional mechanisms behind the chronic low-grade inflammation by deciphering the action of GPS2 in adipocytes and adipose tissue macrophages. We initially studied the gene regulation and the actions of GPS2 and its partners in adipose tissue and adipocytes of human obese subjects compared to lean subjects. In this first study we identified a novel regulatory pathway that participates in the transcriptional control of inflammation associated with obesity, both in adipose tissue and adipocytes. We have shown that GPS2 and SMRT were differentially expressed and regulated in obese adipocytes. In addition, this dysregulation contributes to inflammation of the adipose tissue by allowing the derepression of specific inflammatory genes. In a second study, in order to go further in the characterisation of the in vivo function of GPS2, we generated a mouse model were GPS2 was specifically invalidated in macrophages. Models of diet-induced obesity were applied in these experiments. Interestingly, GPS2-MacKO mice showed an increased expression of inflammatory genes both in adipose tissue and isolated ATMs (F4/80+ cells) associated with a significant increase of macrophages infiltration in the adipose tissue. Finally, we observed that GPS2-MacKO mice had impaired glucose metabolism as they presented high glucose intolerance as well as an important insulin resistance.; L'obésité est aujourd’hui considérée comme une maladie inflammatoire chronique dite de « bas grade » principalement caractérisée par une augmentation de l’inflammation du tissu adipeux. Les adipocytes et les macrophages sont connus pour jouer un rôle clé dans l’établissement, la progression et le maintien de l'inflammation. Dans mon projet de thèse, nous nous sommes particulièrement intéressés aux mécanismes transcriptionnels impliqués dans l'inflammation chronique en décodant l'action du corégulateur transcriptionnel GPS2 (G protein pathway suppressor 2) dans les adipocytes et les macrophages du tissu adipeux. Dans un premiers temps, nous avons étudié la régulation et les actions de GPS2 (et ses partenaires SMRT et NCOR) dans le tissu adipeux humains de sujets obèses par rapport à des sujets minces. Dans cette première étude, nous avons identifié un mécanisme épigénomique qui participe à la régulation de la transcription des gènes inflammatoires dans les adipocytes lors de l’obésité. Nous avons démontré que la dérégulation de GPS2 contribuait à l'inflammation du tissu adipeux en permettant à la dérépression de certains gènes inflammatoires dont l’interleukine 6. Dans la deuxième étude, nous avons caractérisé les conséquences de l’invalidation de GPS2 dans le phénotype inflammatoire des macrophages ainsi que les conséquences in vivo sur la progression de l’insulino-résistance. Pour ceci, nous avons généré un modèle de souris où GPS2 a été spécifiquement invalidé dans les macrophages (GPS2-MacKO). De manière intéressante, les souris GPS2-MacKO, présentent une expression accrue des gènes impliqués dans la voie de signalisation des TLR et des chimiokines dans les macrophages isolés. Par conséquent, une augmentation significative de l'infiltration des macrophages dans le tissu adipeux est observée dans un contexte d’obésité induisant une altération de l’homéostasie glucidique. Par nos approches génomiques, transcriptomiques et épigénomiques, nous avons pu révéler les voies de signalisations spécifiquement contrôlées par GPS2. Ces travaux démontrent également l’importance des régulations épigénomiques dans l'inflammation métabolique du tissu adipeux durant l'obésité.

The mineralocorticoid hormone aldosterone binding its receptor, the mineralocorticoid receptor (MR),regulates the renal reabsorption of sodium. Several studies showed the involvement of the aldosterone/MR complex in cardiovascular diseases, even if the activated signalling pathways are still unclear. This thesis work has been established to increase the knowledge on the mechanisms of thecardiac signalling of MR using two main purposes i) the identification of new molecular targets of the MR in the heart and ii) the understanding of the pathophysiological effects of its activation. Apharmacological approach showed that the diuretic torasemide cannot block the mineralocorticoid signalling in the cell line cardiomyocytes transfected with MR, H9C2-MR+. The study of the MR’s cardiac transactivation activity formed the most important part of this thesis work. We demonstrated with a candidate gene approach that the MR increases the expression of the gene coding the connective tissue growth factor (CTGF) and the aldosterone increases even more this effect in vivo. We foundCTGF specifically expressed in cardiomyocytes and we identified in vitro that the MR binds tohormonal responsive elements on the promoter of the gene coding CTGF. In order to investigate the whole genes differentially expressed by the two ligands of MR in the heart, we treated mice with cardiomyocyte-targeted human MR over expression and their controls with aldosterone or corticosterone. The cardiac transcriptomic analyses show that the majority of aldosterone-regulatedgenes is involved in cell division as Cyclin B1 or Cyclin-dependent kinase 1 (Cdk1). Also, we identified that aldosterone promotes cardiac endothelial cells proliferation.; L’hormone minéralocorticoïde aldostérone en se fixant à son récepteur, le récepteur minéralocorticoïde (RM), module la réabsorption de sodium au niveau du rein. De nombreuses étude sont rapporté l’implication du complexe aldostérone/RM dans les pathologies cardiovasculaires, sans que les voies de signalisation activées soient encore entièrement élucidées à ce jour. Ce travail de thèse se propose d’approfondir les connaissances sur les mécanismes de la signalisation cardiaque du RM à travers deux objectifs principaux i) l’identification de nouvelles cibles moléculaires du RM dans lecoeur et ii) la compréhension des effets physiopathologiques de son activation. Par une approche pharmacologique, nous avons montré que le diurétique torasémide n’est pas capable de bloquer la voie de signalisation minéralocorticoïde dans la lignée cellulaire de cardiomyocyte transfectée avec le RM,H9C2-RM+. L’étude de l’activité transcriptionnelle du RM cardiaque a concerné la majeure partie de ce travail de thèse. Par une approche gène candidat, nous avons mis en évidence que l’expression dugène codant pour le facteur de croissance du tissu conjonctif (CTGF, pour connective tissue growthfactor) est augmentée par le RM et que l’aldostérone potentialise cet effet in vivo. Nous avons pu localiser CTGF spécifiquement dans les cardiomyocytes, et une étude in vitro nous a permis d’identifier que le RM se lie au niveau d’éléments de réponse hormonale dans le promoteur du gène codant pour CTGF. Des souris surexprimant le RM humain spécifiquement dans les cardiomyocytes et traitées avec de l’aldostérone ou de la corticostérone ont permis une exploration plus large des gènesdifférentiellement exprimés par les deux ligands du RM dans le coeur. L’analyse des transcriptomes cardiaques de ces souris et de leurs contrôles montre qu’une augmentation modeste de la concentration plasmatique en aldostérone induit dans le coeur l’expression de gènes impliqués dans le cycle cellulaire comme la Cycline B1 ou sa kinase associée Cdk1 (pour Cyclin-dependent kinase 1). Nous avons montré également que l’aldostérone active la prolifération des cellules endothéliales cardiaques.

Cyclosporine A (cyclo) is a widely used drug in kidney transplantation: its anticalcineurin actioninhibits T lymphocytes activation and prevents allograft rejection. Despite a huge benefit on graftsurvival, cyclo exerts a side effect that limits its use: nephrotoxicity. Vasculotoxicity appears to becentral: cyclo enhances renal vasoconstriction by altering vasoactive factors and vascular smoothmuscle cells (VSMC) response to vasoactive factors. Beyond its effects on sodium reabsorption,Mineralocorticoid Receptor (MR) acts on vascular tone by modulating both endothelial and VSMCresponses to vasoactive factors. Our working hypothesis was that MR could participate to cycloinducedvasoconstriction and that MR inactivating (pharmacologically or genetically) could alleviatecyclo-induced nephrotoxicity. Two genetically MR-knock out (MR-KO) were generated: inendothelial or VSMC. Only VSMC MR-KO mice were protected from cyclo-induced nephrotoxicity.We also show that such an effect was mediated by vascular tone modulation. This prevention was alsoconferred by the systemic pharmacological antagonism of MR (by canrenoate) in mice but not duringnephrotoxicity induced by tacrolimus (another anticalcineurine drug used in kidney transplantation).Then, we proposed to use MR pharmacological antagonism in humans (by eplerenone) during kidneytransplantation. We first had to prove its safety in such a population. Among 31 cyclo-treated patients,only 9 developed hyperkalemia (>5mmol/L) and none presented serious side effect. We propose akalemia higher than 4.35mmol/L at baseline to be the marker of a higher risk of developinghyperkalemia under treatment. The efficiency of eplerenone to prevent/alleviate cyclo-inducednephrotoxicity during kidney transplantation should be tested during a randomized controlled trial.; La ciclosporine est un traitement immunosuppresseur très utilisé : elle inhibe l'activation des lymphocytes T via la calcineurine. Sa néphrotoxicité limite son utilisation : la ciclosporine induit une augmentation de la vasoconstriction ainsi qu'une augmentation de la réponse des cellules musculaires lisses vasculaires (CMLV) aux agents vasoactifs. Le récepteur minéralocorticoïde (RM), au-delà de ses effets sur la réabsorption sodée, agit sur le tonus vasculaire en modulant la réponse des cellules (endothéliales et musculaires lisses) vasculaires aux agents vasoactifs. Notre hypothèse était que le RM pouvait participer à l'action vasoconstrictrice de la ciclosporine ; son inactivation pourrait limiter la néphrotoxicité de la ciclosporine. Deux modèles de souris ont été invalidés génétiquement pour le RM : dans les cellules endothéliales et les CMLV (KO-RM CMLV). Seules les souris KO-RM CMLV étaient protégées contre la néphrotoxicité de la ciclosporine. Ces effets impliquent une action sur le tonus vasculaire rénal. L'antagonisme pharmacologique du RM (par le canrénoate) administré per os confère la même protection. En revanche, la néphrotoxicité induite par le tacrolimus (une autre anticalcineurine) n'est pas prévenue par l'antagonisme du RM. Utiliser un antagoniste sélectif du RM (l'éplérénone) pourrait prévenir la néphrotoxicité de la ciclosporine. Nous avons prouvé sa bonne tolérance en association à la ciclosporine chez les patients transplantés et insuffisants rénaux chroniques. Une kaliémie supérieure à 4,35mmol/L à l'initiation indique un sur-risque de développer une hyperkaliémie. L'efficacité reste à démontrer au cours d'un essai prospectif et randomisé.

Obesity and type 2 diabetes are associated with a sedentary lifestyle and a diet rich in fat. Indeed, saturated fatty acids accumulate in non-adipose tissue such as skeletal muscle to generate lipids called ceramides (CER). My thesis project was divided into two parts with the objective to prevent CER to act. We have shown that, depending on the structure of the plasma membrane, CER alter the insulin signaling pathway by targeting PKB, a key insulin signalling protein, via a PKCζ pathway in L6 myotubes and a PP2A pathway in C2C12 myotubes. We also demonstrated that CER affect insulin sensitivity via the PKCζ pathway in human muscle cells. Once CER generated in the endoplasmic reticulum (ER), they are transported to the Golgi by a carrier called CERT to be metabolized into sphingomyelin (SM). Studies have shown that the transformation of CER into SM could be a crucial step to prevent CER to act. In several muscle insulin resistance models, expression of CERT is decreased and we demonstrated the importance of the transport of ceramide from the ER to the Golgi by inhibiting artificially the activity or the expression of CERT. In contrast, overexpression of CERT enhances insulin sensitivity in muscle cells in lipotoxiques conditions. Our results show that CERT plays a crucial role in mechanisms leading to the development of muscle insulin resistance since its presence is essential for maintaining normal traffic of CER between the ER and the Golgi., L'obésité et le diabète de type 2 sont associés à la sédentarité et à une alimentation riche en graisses. En effet, les acides gras saturés s'accumulent dans les tissus non adipeux, comme les muscles squelettiques pour générer des lipides appelés céramides (CER). Mon projet de thèse s'est articulé en deux parties dont l'objectif est d'empêcher les CER d'agir. Nous avons montré que, selon la structure de la membrane plasmique, les CER altèrent la voie de signalisation insulinique en ciblant la PKB, protéine clef de la voie insulinique, via la voie PKC? dans les myotubes L6 et la voie PP2A dans les myotubes C2C12. Nous avons aussi mis en évidence que les CER altèrent la sensibilité à l'insuline via la voie PKC? dans les cellules musculaires humaines. Une fois les CER produits au niveau du réticulum endoplasmique (RE), ils sont transportés au Golgi par un transporteur CERT pour y être métabolisés en sphingomyéline (SM) et des études ont montré que la transformation des CER en SM pouvait être une étape cruciale pour empêcher les CER d'agir. Dans plusieurs modèles d'insulino-résistance musculaire, l'expression de CERT est diminuée et nous avons démontré l'importance du transport des céramides du RE vers le Golgi en inhibant artificiellement l'activité ou l'expression de CERT. A l'opposé, la surexpression de CERT améliore la sensibilité à l'insuline dans les cellules musculaires dans des conditions lipotoxiques. Nos résultats montrent que CERT joue un rôle crucial dans les mécanismes conduisant au développement de l'insulinorésistance musculaire puisque sa présence est essentielle pour le maintien d'un trafic normal des CER entre le RE et le golgi.

International audience; We analysed (i) the gender difference in cycling speed and (ii) the age of winning performers in the 508-mile "Furnace Creek 508". Changes in cycling speeds and gender differences from 1983 to 2012 were analysed using linear, non-linear and hierarchical multi-level regression analyses for the annual three fastest women and men. Cycling speed increased non-linearly in men from 14.6 (s=0.3) km center dot h(-1) (1983) to 27.1 (s=0.7) km center dot h(-1) (2012) and non-linearly in women from 11.0 (s=0.3) km center dot h(-1) (1984) to 24.2 (s=0.2) km center dot h(-1) (2012). The gender difference in cycling speed decreased linearly from 26.2 (s=0.5)% (1984) to 10.7 (s=1.9)% (2012). The age of winning performers increased from 26 (s=2) years (1984) to 43 (s=11) years (2012) in women and from 33 (s=6) years (1983) to 50 (s=5) years (2012) in men. To summarise, these results suggest that (i) women will be able to narrow the gender gap in cycling speed in the near future in an ultra-endurance cycling race such as the "Furnace Creek 508" due to the linear decrease in gender difference and (ii) the maturity of these athletes has changed during the last three decades where winning performers become older and faster across years.

Pantethine, a natural low-molecular-weight thiol, shows a broad activity in a large range of essential cellular pathways. It has been long known as a hypolipidemic and hypocholesterolemic agent. We have recently shown that it exerts a neuroprotective action in mouse models of cerebral malaria and Parkinson's disease through multiple mechanisms. In the present study, we looked at its effects on membrane lipid rafts that serve as platforms for molecules engaged in cell activity, therefore providing a target against inappropriate cell response leading to a chronic inflammation. We found that pantethine-treated cells showed a significant change in raft fatty acid composition and cholesterol content, with ultimate downregulation of cell adhesion, CXCL12-driven chemotaxis, and transendothelial migration of various T cell types, including human Jurkat cell line and circulating effector T cells. The mechanisms involved include the alteration of the following: (i) CXCL12 binding to its target cells; (ii) membrane dynamics of CXCR4 and CXCR7, the two CXCL12 receptors; and (iii) cell redox status, a crucial determinant in the regulation of the chemokine system. In addition, we considered the linker for activation of T cells molecule to show that pantethine effects were associated with the displacement from the rafts of the acylated signaling molecules which had their palmitoylation level reduced.. In conclusion, the results presented here, together with previously published findings, indicate that due to its pleiotropic action, pantethine can downregulate the multifaceted process leading to pathogenic T cell activation and migration.

Introduction :Saliva plays numerous roles in the oral cavity, especially in digestion and taste perception. It is well admitted that tastants are effective stimuli of salivation and that the salivary response (flow and composition) depends on the nature of the tastant. However, the impact of odours on salivation is poorly known and not consensual. Odours are the first chemical signal informing about food palatability before ingestion. In this context, it is necessary to understand if the salivary secretions are modulated in function of this sensory signal. The aim of this study is to compare the salivary flow response after stimulation by different odours having different hedonic values.Materials and Methods :Thirty men subjects participated to 3 sessions of total saliva sampling after olfactory stimulation. Four odours were used: strawberry, lemon, bacon, thyme and a control (no odour). Then, five subjects for whom an increase of salivary flow was observed, participated to 3 similar sessions during which parotid salivary flow was measured using a Lashley cup coupled to a flow meter.Results :Results show a significant effect of the odours on the total and parotid salivary flow, the most effective stimuli being lemon and bacon. Correlation between total and parotid saliva flow were observed with an important variability between subjects.Discussion/Conclusion :These results open interesting perspectives about the role of salivary response on food intake anticipation in relation with sensory properties.

Current lifestyle changes, notably sedentary behavior, are associated with chronic diseases, while regular activity improves metabolic functions The purpose of my work is to examine the effects of different level of physical activity, until extreme inactivity on cross-talk mediators, such as adiponectine and microRNAs, and on metabolism, notably in muscle or liver. We demonstrate a significant increase of hepatic markers under severe physical inactivity (60d bed-rest), showing the onset of a development of NASH. Our data support that regular exercise can limit these physical inactivity-induced metabolic alterations. We demonstrate that independently of effects of physical activity and inactivity on fat mass, adiponectin plasma concentrations were negatively related to physical activity level. Our data highlight that variations in total plasma adiponectin with physical activity level are mainly due to changes in HMW adiponectin plasma concentrations, and are unlikely explained by variations in expression of adiponectin in adipose tissue and muscle. We also show an inverse relation between fasting insulin plasma concentrations with both adiponectin receptors, and one of its downstream effector (APPL1) in the muscle, suggesting an improvement of adiponectin muscular sensitivity. During a contrasted and moderate physical activity intervention in human and mice, we show that transition from activity toward inactivity results in muscle miR-148b content increase, leading to muscle metabolism alteration. Our study on miR-148b expression modulation in vitro in human myotubes, show its involvement in insulin signaling pathway in muscle, suggesting that miR-148b accumulation in muscle could participate in the whole body insulin sensitivity degradation, which is a feature of persistent sedentary behavior. Altogether, these results demonstrate that the study of cross-talk actors is crucial to understand the mechanisms involved in active and inactive behaviors, and their effects on health, Les travaux de thèse présentés dans ce manuscrit ont pour but d’étudier les effets de différents niveaux d’activité physique, jusqu’à l’inactivité extrême sur des acteurs du dialogue inter-organe comme l’adiponectine et les microARNs, et sur le métabolisme, notamment dans le muscle ou le foie. Nous démontrons que l’inactivité physique extrême, dans le cadre d’un bedrest de 60 jours, augmente les marqueurs de l’inflammation hépatique, prémices du développement d’une stéato-hépatite non alcoolique. Nos données montrent que des exercices réguliers peuvent limiter les altérations métaboliques induites par l’inactivité physique. Nous montrons, ensuite qu’indépendamment des effets de l’activité et l’inactivité physique sur la masse grasse, les concentrations circulantes d’adiponectine sont inversement associées avec le niveau d’activité physique. Nous montrons notamment que les variations des taux circulants d’adiponectine totale avec le niveau d’activité physique sont principalement dû aux variations de la forme de haut poids moléculaire, et ne sont pas expliquées par des variations d’expression de l’adiponectine dans le tissu adipeux ou musculaire. Nous montrons également une relation inverse entre l’insulinémie plasmatique à jeun et les récepteurs de l’adiponectine, et un de ses effecteurs (APPL1) dans le muscle supportant l’amélioration de la sensibilité à l’adiponectine musculaire. Au cours d’une intervention contrastée et modérée sur le niveau d’activité physique chez l’homme et la souris, nous avons pu montrer que la transition d’un statut actif vers inactif augmentait le niveau d’expression de miR-148b dans le muscle, participant à la dégradation du métabolisme. Notre étude de modulation de l’expression de miR-148b in vitro dans les myotubes humains, montre son implication dans la voie de signalisation de l’insuline, et suggère que l’accumulation de miR-148b dans le tissu musculaire peut participer à l’altération de la sensibilité à l’insuline qui est caractéristique des comportements sédentaires persistants. L’ensemble de ces résultats démontrent que l’étude des acteurs de la signalisation inter-organe est cruciale pour comprendre les mécanismes mis en jeu par les comportements actifs et inactifs, et leurs effets sur la santé

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Lettre à l'éditeur ("Journal of Applied physiology" vol.118, no.10). Fait partie de : "Commentaries on Viewpoint: The two-hour marathon: what's the equivalent for women?". http://jap.physiology.org/content/118/10/1324#ref-list-2

Newborn mice are attracted to mammary odor cues carried in murine milk and nipple secretions. However, murine milk odor is not equally attractive along lactation. The present study focuses on the differential response of 2day-old mouse pups of C57Bl/6 (C) and Balb/C (B) strains to the odor of milk (Experiment 1) and nipples (Experiment 2) that are matched/unmatched in terms of pup's age or strain. In Experiment 1, C and B pups were tested in a series of tests simultaneously opposing either murine milk and a blank (water), or two milks collected in early and late lactation (lactation days 2 and 15, respectively) from females belonging to their own or the other strain. Results showed that C and B pups were attracted to the odor of the different milks regardless of the lactation age and the strain of the donor female. Nevertheless, C and B pups preferred the odor conveyed by early- than late-lactation milk of either strain. Moreover, early-lactation milk from C females was more attractive than early-lactation milk from B females for pups of either strain. In Experiment 2, differential nipple grasping response of C and B pups was measured when they were exposed to nipples of females in early or late lactation. The proportion of C pups that grasped a nipple was greater when they were exposed to a nipple in early lactation regardless of the strain of the donor females, whereas the proportion of B pups that grasped a nipple was greater when they were exposed to a nipple in early lactation, but only from own strain. Thus, newborn mice prefer the odor of milk and nipples from females that are matched in lactation age. This result is discussed in terms of reciprocally adaptive mechanisms between lactating females and their newborn offspring.

International audience; The purpose of this study was to assess the prognostic value of a large panel of cytokines in aggressive non-Hodgkin's lymphoma (NHL) and to confront it to parameters of the International Prognostic Index (IPI). It investigated the concomitant determination of interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), interleukin-4 (IL-4), interleukin-6 (IL-6) and interleukin-10 (IL-10) on a uniform population of 116 previously untreated patients. Commercially available enzyme-linked immunoassay kits were used for cytokines measurements. Results were correlated with complete remission (CR), overall survival (OS) and failure free survival (FFS). In univariate analysis, sIL-2R and IL-6 demonstrated prognostic significance for CR (p = 0.016 and p = 0.048), OS (p = 0.0011 and p = 0.0387) and FFS (p = 0.0001 and p = 0.0363), but multi-variate analysis failed to demonstrate an independent prognostic significance. In the intermediate group risk defined by IPI, patients presenting high level of sIL-2R or IL-6 demonstrated lower CR rate and survival than those with low level. In conclusion, sIL-2R and IL-6 serum levels are elevated in high grade NHL and are correlated to CR, OS and FFS, but this study did not support their independent prognostic value. However, sIL-2R and IL-6 measurements may improve risk assignment by IPI and allow a better prognostic evaluation of patients with intermediate prognosis NHL.

Our objective was to test the hypothesis that the beneficial effect of the administration of the recombinant form of tissue plasminogen activator (rt-PA) in ischemic stroke patient not only results from its fibrinolytic activity but also from its ability to increase brain-derived neurotrophic factor (BDNF) in the brain. To this end, we conducted an initial study to evaluate the effect of rt-PA on brain BDNF levels in healthy animals. In a second study, we investigated the effect of rt-PA on serum BDNF levels in ischemic stroke patients and in animals subjected to permanent focal cerebral ischemia. Blood samples were obtained from patient on admission (D0), D1, D7 and D90 after stroke and in rats before and after (1h, 4h and 24h) ischemia. BDNF was measured in the brain by Western blot and in the blood by ELISA. In both studies, the rt-PA (Actilyse®) was administered as a bolus followed by an infusion of one hour. The first study evidences that 1) rt-PA increases the BDNF levels in the hippocampus, 2) treatment with MK801 (a NMDA receptor antagonist) but not with tranexamic acid (a plasmin inhibitor) canceled the effect of rt-PA on BDNF levels. The second study exhibits that 1) neurological recovery was higher in the patients receiving rt-PA, 2) treatment with rt-PA increases serum BDNF at D1 and D7 in patients, but does not change the blood BDNF levels in animals, 3) BDNF levels are not correlated with neurological recovery but are inversely correlated to the patient cardiovascular score. In conclusion, our results suggest that rt-PA may have a protective extra-fibrinolytic effect by increasing in BDNF levels through a potentiation of glutamatergic pathway. Although rt-PA induces a better neurological recovery and increases circulating BDNF levels in stroke patients, the lack of correlation between these two parameters is not in favor of using circulating BDNF as a predictive marker of neurological recovery, but could be a reflect of the endothelium ability to synthesize BDNF., L’objectif de ce travail a été de tester l’hypothèse selon laquelle l’effet bénéfique de l’administration de la forme recombinante de l’activateur tissulaire du plasminogène (rt-PA) chez le patient victime d’un accident vasculaire cérébral (AVC) ischémique ne résulte pas uniquement de son action fibrinolytique mais aussi de sa capacité à augmenter le brain-derived neurotrophic factor (BDNF) dans le cerveau. Dans ce but, nous avons conduit une première étude visant à évaluer chez l’animal sain, l’effet du rt-PA sur les taux cérébraux de BDNF. Dans une seconde approche, nous avons étudié l’effet du rt-PA sur les taux sériques de BDNF chez le patient victime d’un AVC ischémique et chez l’animal soumis à une ischémie cérébrale focale. Le sang a été prélevé chez l’Homme à l’admission (J0), J1, J7 et J90 après l’AVC, et chez le Rat avant et après (1h, 4h et 24h) l’ischémie. Le BDNF a été mesuré dans le cerveau par technique de Western blot et dans le sang par technique ELISA. Dans les deux études le rt-PA (Actilyse®) a été administré sous forme d’un bolus suivi d’une perfusion d’une heure. La première étude montre que 1) le rt-PA augmente les taux de BDNF dans l’hippocampe, 2) le traitement par MK801 (un antagoniste des récepteurs NMDA) mais pas par l’acide tranexamique (un inhibiteur de la plasmine) annule l’effet du rt-PA sur les taux de BDNF. La deuxième étude met en évidence que 1) la récupération neurologique est meilleure chez les patients recevant le rt-PA, 2) le traitement par rt-PA augmente les taux sériques de BDNF à J1 et J7 chez l’Homme, mais ne modifie pas les taux sanguins de BDNF chez l’animal, 3) les taux de BDNF ne sont pas corrélés à la récupération neurologique mais sont inversement corrélés au score cardiovasculaire du patient. En conclusion, nos résultats suggèrent que le rt-PA peut exercer un effet protecteur extra-fibrinolytique en augmentant les taux de BDNFm par une potentialisation de l’activité glutamatergique. Même si le rt-PA induit une meilleure récupération neurologique et augmente les taux circulants de BDNF chez les patients victimes d’un AVC, l’absence de corrélation entre ces deux paramètres n’est pas en faveur de l’utilisation du BDNF circulant comme un marqueur prédictif de récupération neurologique, mais pourrait être un reflet de la capacité de l’endothélium à sécréter le BDNF.

International audience; Torque time integral (TTI) is the combination of intensity and duration of a contraction. The aim of this study was to compare neuromuscular alterations following different isometric sub-maximal contractions of the knee extensor muscles but with similar TTI. Sixteen participants performed 3 sustained contractions at different intensities (25 %, 50 %, and 75 % of Maximal Voluntary Contraction (MVC) torque) with different durations (68.5 +/- 33.4 s, 35.1 +/- 16.8 s and 24.8 +/- 12.9 s, respectively) but similar TTI value. MVC torque, maximal voluntary activation level (VAL), M-wave characteristics and potentiated doublet amplitude were assessed before and immediately after the sustained contractions. EMG activity of the vastus lateralis (VL) and rectus femoris (RF) muscles was recorded during the sustained contractions. MVC torque reduction was similar in the 3 conditions after the exercise (-23.4 +/- 2.7 %). VAL decreased significantly in a similar extent (-3.1 +/- 1.3 %) after the 3 sustained contractions. Potentiated doublet amplitude was similarly reduced in the 3 conditions (-19.7 +/- 1.5 %), but VL and RF M-wave amplitudes remained unchanged. EMG activity of VL and RF muscles increased in the same extent during the 3 contractions (VL: 54.5 +/- 40.4 %; RF: 53.1 +/- 48.7 %). These results suggest that central and peripheral alterations accounting for muscle fatigue are similar following isometric contractions with similar TTI. TTI should be considered in the exploration of muscle fatigue during sustained isometric contractions.

The link between prolactin (PRL) and mammary tumorigenesis is supported by many experimental, clinical and epidemiologic data. However, mutations on the gene coding for PRL or its receptor (PRLR) have never been identified in the context of breast cancer. In 2008, our Laboratory identified the first gain-of-function polymorphism of the PRLR in patients presenting with a rare form of benign mammary tumors (mammary polyadenomatosis). Indeed, the one-residue substitution of isoleucine 146 by a leucine in the extracellular domain of the PRLR conferred a higher basal activity than that of the wild-type PRLR. Within the framework of my thesis, we have tried to determine the consequences of expressing a constitutively active PRLR in a healthy or cancerous mammary cell. With this aim, we have used two complementary approaches. On one hand, we generated two models of knock-in mice carrying only one copy of the mutated (I146L) or wild-type (as control) human PRLR. We then analyzed the mammary phenotypes of these mice at various ages. We did not observe any tumor development, indicating that the PRLR-I146L is not an oncogene. Nevertheless, the mammary glands of old mice presented certain histological anomalies suggesting a possible interference of the mutated PRLR with normal mammary differentiation. The analysis of these phenotypes must continue to determine their extent and the potential underlying mechanisms. On the other hand, we carried out an in vitro study using human breast cancer cell lines. A parallel study in the Laboratory had showed that the I146D substitution led to a stronger constitutive activity of the PRLR than the natural I146L polymorphism. Thus, we generated stable clones of two human breast cancer cell lines expressing the PRLR-I146D. In spite of its constitutive activity, the expression of PRLR-I146D did not result in any selective advantage for tumor cells in terms of proliferation, nor did it modify their histological phenotype (Zhang, Cherifi et al., in revision). In conclusion, our work has shown that the expression of a gain-of-function mutation in the PRLR is insufficient to transform a healthy mammary cell, or to enhance the proliferation of mammary cancer cells.; Le lien entre la prolactine (PRL) et la tumorigenèse mammaire est soutenu par de nombreux arguments expérimentaux, cliniques et épidémiologiques. Cependant, aucune mutation des gènes de la PRL ou de son récepteur (PRLR) n'a jamais été identifiée dans le contexte du cancer du sein. En 2008, notre Laboratoire a identifié le premier polymorphisme gain-de-fonction du PRLR chez des patients présentant une forme rare de tumeur mammaire bénigne (polyadénomatose mammaire). En effet, la seule substitution de l'isoleucine 146 par une leucine dans le domaine extracellulaire du PRLR lui confère une activité de signalisation basale plus élevée que le PRLR sauvage. Dans le cadre de ma thèse, nous avons essayé de déterminer expérimentalement quelles pouvaient être les conséquences de l'expression d'un PRLR constitutivement activé dans une cellule mammaire saine ou tumorale. Pour ce faire, nous avons utilisé deux approches complémentaires. D'une part, nous avons généré deux modèles de souris knock-in ayant intégré une seule copie du PRLR humain, muté (I146L) ou sauvage (en contrôle). Nous avons ensuite analysé les phénotypes mammaires de ces souris à différents âges. Nous n'avons observé aucun développement tumoral, indiquant que le PRLR-I146L n'est pas un oncogène. Néanmoins, les glandes mammaires des souris âgées présentent certaines anomalies histologiques suggérant une interférence possible du PRLR muté avec la différenciation mammaire. L'analyse des ces phénotypes doit être poursuivie pour en déterminer l'ampleur exacte et les mécanismes potentiels. D'autre part, nous avons réalisé une étude in vitro en utilisant des cellules tumorales mammaires humaines. Une étude parallèle au Laboratoire ayant permis de montrer que la substitution I146D conduisait à une activité constitutive plus forte que le polymorphisme naturel I146L, nous avons généré des clones stables de deux lignées tumorales mammaires humaines exprimant le PRLR-I146D. Malgré une signalisation constitutive démontrée, l'expression de ce PRLR muté ne procure aucun avantage sélectif aux cellules tumorales en termes de prolifération, ni ne modifie leur phénotype histologique (Zhang, Cherifi et al, en révision). En conclusion, notre travail a permis de montrer que la seule expression d'un variant gain-de-fonction du PRLR est insuffisante pour transformer une cellule mammaire saine, ou pour favoriser la prolifération de cellules mammaires tumorales.

The possible peripheral and/or central origin in the mechanisms responsible for day-time fluctuation in maximal torque of the triceps surae muscle were investigated with a special emphasis on antagonist muscle coactivation. Eleven healthy male subjects (physical education students) took part in this investigation. The electromechanical properties of the plantar flexor muscles were recorded at two different times of day: between 06:00 h and 08:00 h in the morning and between 17:00 h and 19:00 h in the evening. To investigate peripheral mechanisms, the posterior tibial nerve was stimulated at rest, using percutaneous electrical stimuli, to evoke single twitch, double twitch, and maximal tetanic contraction (100 Hz). Maximal voluntary contraction of the plantar flexors was also assessed by means of the relative electromyographic activity of respective agonist and antagonist muscles (soleus, gastrocnemius medialis, gastrocnemius lateralis, and tibialis anterior). A double twitch was delivered during maximal voluntary plantar flexion to record muscle activation (i.e., interpolated twitch technique). The coactivation level of the tibialis anterior muscle during plantar flexion was calculated. The results indicated a significant decrease in maximal voluntary muscle torque of triceps surae in the evening as compared with the morning (-7.0 %; p < 0.05). Concerning the central command, when extrapolated by the twitch interpolation technique, the decrease in mean activation level of -6.8 % was consistent with the fluctuation in torque (-7.0 %). Soleus muscle electromyographic activity (normalized to the M-wave) showed a significant decline (21.6 %; p < 0.001). Moreover, individual changes in MVC percentage were significantly related to those of normalized electromyographic activity of the soleus muscle (r = 0.688; p < 0.01). Thus, it indicated that the subject's capacity to activate the soleus muscle was affected by the time of day. The coactivation level in the tibialis anterior muscle during plantar flexion did not change significantly in the evening. Concerning peripheral mechanisms, we observed a decrease in maximal M-wave amplitude for soleus and gastrocnemii, associated with unchanged single twitch and tetanus torque. To conclude, impairment in soleus muscle central command seemed to be the mechanism in the origin of torque failure. Such information would be of importance in the investigation of day-time fluctuations in complex motor task performances implicating the triceps surae muscle.

Cardiovascular diseases are the leading cause of death worldwide. Among the mechanisms involved in these pathologies, there is the activation of the mineralocorticoid signaling pathway. Our laboratory has previously identified Lipocalin 2 (Lcn2) as a direct target of the aldosterone/mineralocorticoid receptor (MR) complex in the cardiovascular system. Lcn2, also called Neutrophil Gelatinase Associated Lipocalin (NGAL), is a circulating protein, a member of the lipocalin family. It is described as being involved in inflammation or as regulating the activity and stability of matrix metallopeptidase 9. The aim of this work is to investigate the possible involvement of Lcn2 in pro-fibrotic and pro-inflammatory pathological effects of aldosterone/MR complex in the cardiovascular system. For this purpose, mice with constitutive and overall Lcn2 inactivation (Lcn2 KO) and their littermates were subjected to a treatment mimicking overactivation of the mineralocorticoid pathway (nephrectomy-aldosterone-salt treatment) during 4 weeks. This work has highlighted the role of Lcn2 in the development of perivascular fibrosis and inflammation induced by the complex aldo/MR and in the blood pressure increase. However, Lcn2 is not involved in interstitial fibrosis or vascular dysfunction. The action of Lipocalin 2 in these pathological phenomena mechanisms remains to be elucidated. In conclusion, the results show the direct involvement of Lcn2 in the pro-fibrotic effects of aldo/MR complex in the cardiovascular system, suggesting a potential therapeutic target in cardiovascular fibrosis., Les pathologies cardiovasculaires sont la première cause de mortalité dans le monde. Parmi les mécanismes participant à ces pathologies figurent l'activation de la voie de signalisation minéralocorticoïde. Notre laboratoire a précédemment identifié la Lipocaline 2 (Lcn2) comme cible directe du complexe aldostérone/récepteur minéralocorticoïde (RM) dans le système cardiovasculaire. Lcn2, aussi appelée Neutrophil Gelatinase Associated Lipocalin (NGAL), est une protéine circulante, membre de la famille des lipocalines. Elle est décrite comme participant à l'inflammation ou comme régulant l'activité et la stabilité de la métalloprotéinase matricielle 9. L'objectif de ce travail de thèse est d'étudier l'implication possible de Lcn2 dans les effets pathologiques pro-fibrosants et pro-inflammatoires du complexe aldostérone/RM dans le système cardiovasculaire. Pour ce faire, des souris présentant une inactivation globale et constitutive de Lcn2 (KO Lcn2), ainsi que leurs contrôles, ont été soumises à un traitement mimant une sur-activation de la voie minéralocorticoïde (traitement néphrectomie-aldostérone-sel) durant 4 semaines. Ce travail a mis en évidence le rôle de Lcn2 dans le développement de la fibrose périvasculaire et l'inflammation induite par le complexe aldo/RM ainsi que dans l'augmentation de pression artérielle. En revanche, Lcn2 n'intervient pas dans la fibrose interstitielle ni dans la dysfonction vasculaire. Les mécanismes d'action de la Lipocaline 2 dans ces différents phénomènes pathologiques restent à élucider. En conclusion, les résultats obtenus montrent l’implication directe de Lcn2 dans les effets pro-fibrosants du complexe aldo/RM au niveau cardiovasculaire, suggérant une potentielle cible thérapeutique dans la fibrose cardiovasculaire.

It is well established that vasopressin (AVP) level is high in both human and experimental diabetes. In humans, several recent studies have shown an association between copeptin (biomarker of AVP secretion) and the occurrence of diabetes mellitus or hyperglycemia, metabolic syndrome and obesity. Our team has shown a reverse association between water consumption (decrease AVP secretion) and the risk of hyperglycemia in the general population (D.E.S.I.R cohort). The aim of my thesis was to determine the role of AVP and fluid intake in glucose homeostasis in healthy rats and in a rat model of metabolic syndrome. AVP, administered acutely or chronically in healthy rats, increases glycaemia and this effect is reversed by a V1a receptor antagonist. V1b receptor activation does not influence insulin secretion but stimulates moderately basal glucagon production by the pancreas. These effects were observed in two different healthy strains of rats. In obese Zucker rats, a high AVP level worsens fasting hyperinsulinaemia and glucose intolerance whereas hydration does not affect glucose tolerance but drastically reduces hepatic steatosis, the content of cholesterol and triglycerides in liver and expression of genes involved in hepatic lipogenesis. In conclusion, these studies show for the first time, that AVP aggravates glucose tolerance whereas a highly hydrated diet is protective. These results, in agreement with our epidemiological data, demonstrate a causal link between vasopressin and/or hydration and glucose metabolism disorders.; Il est bien établi que la vasopressine (AVP) est élevée dans le diabète tant humain qu'expérimental. Chez l'homme, plusieurs études récentes ont montré une association entre la copeptine (biomarqueur de la sécrétion d'AVP), et la survenue d'un diabète ou d'une hyperglycémie, le syndrome métabolique et l'obésité. Dans l'équipe, nous avons montré une association inverse entre la consommation d'eau (diminue la sécrétion AVP) et le risque de survenue d'hyperglycémie dans la cohorte D.E.S.I.R. Le but de mon projet de thèse a été de déterminer le rôle de l'AVP et de la prise hydrique dans l'homéostasie glucidique chez le rat sain et dans un modèle de rat présentant un syndrome métabolique. L'administration aigüe ou chronique d'AVP augmente la glycémie et cet effet est réversé par un antagoniste des récepteurs V1a. L'activation des récepteurs V1b ne modifie pas l'insulino-sécrétion mais stimule en permanence et de façon modérée la glucagonémie. Ces effets ont été observés sur deux souches différentes de rats sains. Chez le rat Zucker obèse, l'AVP aggrave l'hyperinsulinémie à jeun et l'intolérance au glucose alors que le régime hydraté ne modifie pas la tolérance au glucose mais réduit très fortement la stéatose hépatique ainsi que le contenu hépatique en cholestérol et triglycérides et l'expression des gènes impliqués dans la lipogenèse. En conclusion, ces travaux montrent pour la première fois, que l'AVP dégrade à long terme la tolérance au glucose ; a contrario, un régime fortement hydraté est protecteur. Ces résultats, en accord avec nos données épidémiologiques, démontrent un lien de causalité entre vasopressine/hydratation et désordre du métabolisme glucidique.

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International audience; Abstract The capacity to predict the heart rate (HR) and speed at the first (VT1) and second (VT2) ventilatory thresholds was evaluated during an incremental ski-mountaineering test using heart rate variability (HRV). Nine skiers performed a field test to exhaustion on an alpine skiing track. VT1 and VT2 were individually determined by visual analysis from gas exchanges (VT1V and VT2V) and time-varying spectral HRV analysis (VT1fH, VT2fH and VT2H). VT1 could not be determined with the HRV methods used. On the contrary, the VT2 was determined in all skiers. No significant difference between HR and speed at VT2H and VT2V was observed (174.3 ± 5.6 vs. 174.3 ± 5.3 bpm, and 6.3 ± 0.9 and 6.3 ± 0.9 km h-1, respectively). Strong correlations were obtained for HR (r = 0.91) and speed (r = 0.92) at VT2H and VT2V with small limits of agreement (±3.6 bpm for HR). Our results indicated that HRV enables determination of HR and speed at VT2 during a specific ski-mountaineering incremental test. These findings provide practical applications for skiers in order to evaluate and control specific training loads, at least when referring to VT2.

International audience; Abstract The capacity to predict the heart rate (HR) and speed at the first (VT1) and second (VT2) ventilatory thresholds was evaluated during an incremental ski-mountaineering test using heart rate variability (HRV). Nine skiers performed a field test to exhaustion on an alpine skiing track. VT1 and VT2 were individually determined by visual analysis from gas exchanges (VT1V and VT2V) and time-varying spectral HRV analysis (VT1fH, VT2fH and VT2H). VT1 could not be determined with the HRV methods used. On the contrary, the VT2 was determined in all skiers. No significant difference between HR and speed at VT2H and VT2V was observed (174.3 ± 5.6 vs. 174.3 ± 5.3 bpm, and 6.3 ± 0.9 and 6.3 ± 0.9 km h-1, respectively). Strong correlations were obtained for HR (r = 0.91) and speed (r = 0.92) at VT2H and VT2V with small limits of agreement (±3.6 bpm for HR). Our results indicated that HRV enables determination of HR and speed at VT2 during a specific ski-mountaineering incremental test. These findings provide practical applications for skiers in order to evaluate and control specific training loads, at least when referring to VT2.