Research data on pavement texture changes for D6.5

<p>In-situ surface texture measurements</p><p> </p><p>Three different tests were conducted to monitor the evolution in the NEMO micro-texture of the carousel pavements (S1-S4): British pendulum (SRT), Dynamic Friction Tester (DFT) and T2GO. Measurements were taken at three different locations per section in the initial state and at increasing numbers of cycles : 0, 20,000, 50,000, 100,000, 200,000, 500,000, 750,000 and 1 million  load cycles. </p><p> </p><p>The macrotexture of NEMO pavements was evaluated using two complementary techniques. The first is a classical volumetric techniques called the sand patch test. According to this techniques, the macrotexture is approached by mean texture depth (MTD). The second technique relies on the profilometry according to ISO EN 13473-1 standard. Two devices named Elatextur and LTS9500 measure the depth of surface roughness using a laser beam.  The Elatextur laser scan has a width 400 mm diameter and a 0.2 mm of path while the LTS9500 scans a 4″ x 4″ area with a lateral resolution of less than 0.5 mm</p><p> </p&gt

Discrimination of dog palatant solutions using human olfactory receptors

International audienc

Capacity of myeloid and plasmacytoid dendritic cells especially at mature stage to express and secrete HLA-G molecules

International audienc

Discrimination of dog palatant solutions using human olfactory receptors platform

International audienc

Proteomics of cypress pollen allergens using double and triple one-dimensional electrophoresis.

International audienceItalian cypress (Cupressus sempervirens, Cups) pollen causes allergic diseases in inhabitants of many of the cities surrounding the Mediterranean basin. However, allergens of Cups pollen are still poorly known. We introduce here a novel proteomic approach based on double one-dimensional gel electrophoresis (D1-DE) as an alternative to the 2-DE immunoblot, for the specific IgE screening of allergenic proteins from pollen extracts. The sequential one-dimensional combination of IEF and SDS-PAGE associated with IgE immunoblotting allows a versatile multiplexed immunochemical analysis of selected groups of allergens by converting a single protein spot into an extended protein band. Moreover, the method appears to be valuable for MS/MS identification, without protein purification, of a new Cups pollen allergen at 43 kDa. D1-DE immunoblotting revealed that the prevalence of IgE sensitization to this allergen belonging to the polygalacturonase (PG) family was 70% in tested French allergic patients. In subsequent triple one-dimensional gel electrophoresis, the Cups pollen PG was shown to promote lectin-based protein-protein interactions. Therefore, D1-DE could be used in routine work as a convenient alternative to 2-DE immunoblotting for the simultaneous screening of allergenic components under identical experimental conditions, thereby saving considerable amounts of sera and allergen extracts

Analyse immunoprotéomique des allergènes non-hydrosolubles de farines de quatre légumineuses : arachide, soja, sésame et lentille

International audiencePeanut, soybean, sesame and lentil are members of legumes worldwide consumed by human that can induce food allergy in genetically predisposed individuals. Several protein allergens, mainly water-soluble, have been described. We studied the non water-soluble fraction from these 4 food sources using immunoproteomics tools and techniques. Flour extracts were solubilized in detergent and chaotropes and analysed in 1 and 2 dimensional gel electrophoresis (2D). Results showed numerous proteins exhibiting wide ranges of isoelectric points and relative molecular masses. When IgE immunoreactivities of 18 food allergy patients were individually tested in 1 and 2D western-blots, a very diversified IgE repertoire was observed, reflecting extensive cross-reactivities but also co-sensitizations. Besides already well known and characterized allergens, mass spectrometry analysis allowed the identification of 22 allergens undescribed until now: 10 in peanut, 2 in soybean, 3 in sesame and 7 in lentil. Three allergens are legume storage proteins and the others belong to transport proteins, nucleotide binding proteins and proteins involved in the regulation of metabolism. Seven proteins are potentially similar to allergens described in plants and fungi and 11 are not related to any known allergen. Our results contribute to increase the repertoire of legume allergens that may improve the diagnosis, categorize patients and thus provide a better treatment of patients.L’arachide, le soja, le sésame et la lentille sont des légumineuses consommées dans l’alimentation humaine. Elles peuvent provoquer des allergies alimentaires chez les individus génétiquement prédisposés. Plusieurs allergènes protéiques, principalement hydrosolubles, ont été décrits. Nous avons étudié la fraction non-hydrosoluble de ces 4 sources alimentaires par les techniques et les outils de l’analyse immunoprotéomique. Les extraits obtenus en détergent et agents chaotropiques ont révélé par électrophorèse en 1 et 2 dimensions (2D) de nombreuses protéines de points isoélectriques et de masses moléculaires relatives très variés. L’immunoréactivité des IgE de 18 patients souffrant d’allergie alimentaire testés par immunoempreinte 1 et 2D a montré une très grande diversité de répertoire des IgE anti-allergène compatible avec des réactivités croisées mais aussi des co-sensibilisations. L’analyse en spectrométrie de masse des protéines reconnues par les IgE a permis d’identifier, outre des allergènes déjà caractérisés et répertoriés dans les banques de données, 22 allergènes non décrits jusqu’à présent : 10 dans l’arachide, 2 dans le soja, 3 dans le sésame et 7 dans la lentille. Trois allergènes correspondent à des protéines de réserve de légumineuses et les autres incluent des protéines de transport, des protéines liant des nucléotides et des protéines impliquées dans la régulation du métabolisme. Sept protéines sont potentiellement homologues à des allergènes décrits dans des plantes et champignons et 11 n’ont aucune relation avec des allergènes connus. Nos résultats complexifient le répertoire des allergènes des légumineuses et permettront, à terme, par un diagnostic plus précis, de catégoriser les patients pour une prise en charge thérapeutique adaptée

Structural heterogeneity of the microtubule lattice

Abstract Microtubules are polymers assembled from αβ-tubulin heterodimers. They typically display lateral α-α and β-β homotypic interactions, except at one region, called the seam, where heterotypic α-β and β-α interactions occur. Here, we decorated microtubules assembled in vitro or in cytoplasmic Xenopus egg extracts with kinesin-motor domains, and analyzed their lattice organization using dual axis cryo-electron tomography followed by segmented sub-tomogram averaging. In both conditions, microtubules incorporated variable protofilament and/or tubulin subunit helix start numbers. While microtubules assembled in vitro displayed variable numbers of seams, those assembled in extracts displayed preferentially one seam. The seam location varied within individual microtubules implying the presence of lattice holes. Thus, the formation of discontinuous microtubule lattices is an intrinsic property of tubulin assembly, a process that is controlled in cells. One-Sentence Summary Microtubules assembled from purified tubulin form discontinuous lattices, an intrinsic property strictly controlled in cytoplasmic Xenopus egg extracts

Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts

International audienceMicrotubules are tubes of about 25 nm in diameter that are critically involved in a variety of cellular functions, including motility, compartmentalization, and division. They are considered as pseudo-helical polymers whose constituent αβ-tubulin heterodimers share lateral homotypic interactions, except at one unique region called the seam. Here, we used a segmented sub-tomogram averaging strategy to reassess this paradigm and analyze the organization of the αβ-tubulin heterodimers in microtubules assembled from purified porcine brain tubulin in the presence of GTP and GMPCPP, and in Xenopus egg cytoplasmic extracts. We find that in almost all conditions, microtubules incorporate variable protofilament and/or tubulin subunit helical-start numbers, as well as variable numbers of seams. Strikingly, the seam number and location vary along individual microtubules, generating holes of one to a few subunits in size within their lattices. Together, our results reveal that the formation of mixed and discontinuous microtubule lattices is an intrinsic property of tubulin that requires the formation of unique lateral interactions without longitudinal ones. They further suggest that microtubule assembly is tightly regulated in a cytoplasmic environment

Blockade of substance P (neurokinin 1) receptors enhances extracellular serotonin when combined with a selective serotonin reuptake inhibitor: An in vivo microdialysis study in mice

Substance P antagonists of the neurokinin-1 receptor type (NK1) are gaining growing interest as new antidepressant therapies. It has been postulated that these drugs exert this putative therapeutic effect without direct interactions with serotonin (5-HT) neurones. Our recent microdialysis experiment performed in NK1 receptor knockout mice suggested evidence of changes in 5-HT neuronal function (Froger et al. 2001). The aim of the present study was to evaluate the effects of coadministration of the selective 5-HT reuptake inhibitor (SSRI) paroxetine with a NK1 receptor antagonist (GR205171 or L733060), given either intraperitoneally (i.p.) or locally into the dorsal raphe nucleus, on extracellular levels of 5-HT ([5-HT]ext) in the frontal cortex and the dorsal raphe nucleus using in vivo microdialysis in awake, freely moving mice. The systemic or intraraphe administration of a NK1 receptor antagonist did not change basal cortical [5-HT]ext in mice. A single systemic dose of paroxetine (4 mg/kg; i.p.) resulted in a statistically significant increase in [5-HT]ext with a larger extent in the dorsal raphe nucleus (+ 138% over basal AUC values), than in the frontal cortex (+ 52% over basal AUC values). Co-administration of paroxetine (4 mg/kg; i.p.) with the NK1 receptor antagonists, GR205171 (30 mg/kg; i.p.) or L733060 (40 mg/kg; i.p.), potentiated the effects of paroxetine on cortical [5-HT]ext in wild-type mice, whereas GR205171 (30 mg/kg; i.p.) had no effect on paroxetine-induced increase in cortical [5-HT]ext in NK1 receptor knock-out mice. When GR205171 (300 µmol/L) was perfused by ‘reverse microdialysis’ into the dorsal raphe nucleus, it potentiated the effects of paroxetine on cortical [5-HT]ext, and inhibited paroxetine-induced increase in [5-HT]ext in the dorsal raphe nucleus. Finally, in mice whose 5-HT transporters were first blocked by a local perfusion of 1 µmol/L of citalopram into the frontal cortex, a single dose of paroxetine (4 mg/kg i.p.) decreased cortical 5-HT release, and GR205171 (30 mg/kg i.p.) reversed this effect. The present findings suggest that NK1 receptor antagonists, when combined with a SSRI, augment 5-HT release by modulating substance P/5–HT interactions in the dorsal raphe nucleus.Peer reviewe