84 research outputs found
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Hiking and digital innovation: Analysis strategies for tourist destinations management
Hiking is a recreational and outdoor activity that could experience significant growth in France with an increasing role in destination attractiveness and development. Having become a fully-fledged territorial marketing tool, hiking comes becomes integral to many destination management strategies. Hiking benefits from a growing social enthusiasm nurtured by the adoption of digital tools. The research question is the following: How does hiking fit into territorial development strategies for tourist destinations that are investing in digital tools? This paper is aims at presenting the latest developments of the research project, the conceptual approach (actor network theory), and the methodology. The results of an exploratory study will be presented, as well as the perspectives they present for the second phase of the research project
Diplomatique et les arts numériques (La)
Mémoire du Master Archives numériques portant sur les oeuvres d\u27art numérique et la diplomatique comme méthode possible d\u27authentification suite aux recherches du groupe InterPARES
Sensitive proton-detected solid-state NMR spectroscopy of large proteins with selective CH3 labelling: application to the 50S ribosome subunit
International audienceSolid-state NMR spectroscopy allows the characterization of structure, interactions and dynamics of insoluble and/or very large proteins. Sensitivity and resolution are often major challenges for obtaining atomic-resolution information, in particular for very large protein complexes. Here we show that the use of deuterated, specifically CH3-labelled proteins result in significant sensitivity gains compared to previously employed CHD2 labelling, while line widths only marginally increase. We apply this labelling strategy to a 468 kDa-large dodecameric aminopeptidase, TET2, and the 1.6 MDa-large 50S ribosome subunit of Thermus thermophilus
Specific isotopic labelling and reverse labelling for protein NMR spectroscopy : using metabolic precursors in sample preparation
The study of protein structure, dynamics and function by NMR spectroscopy commonly requires samples that have been enriched ('labelled') with the stable isotopes 13C and/or 15N. The standard approach is to uniformly label a protein with one or both of these nuclei such that all C and/or N sites are in principle 'NMR-visible'. NMR spectra of uniformly labelled proteins can be highly complicated and suffer from signal overlap. Moreover, as molecular size increases the linewidths of NMR signals broaden, which decreases sensitivity and causes further spectral congestion. Both effects can limit the type and quality of information available from NMR data. Problems associated with signal overlap and signal broadening can often be alleviated though the use of alternative, non-uniform isotopic labelling patterns. Specific isotopic labelling 'turns on' signals at selected sites while the rest of the protein is NMR-invisible. Conversely, specific isotopic unlabelling (also called 'reverse' labelling) 'turns off' selected signals while the rest of the protein remains NMR-visible. Both approaches can simplify NMR spectra, improve sensitivity, facilitate resonance assignment and permit a range of different NMR strategies when combined with other labelling tools and NMR experiments. Here, we review methods for producing proteins with enrichment of stable NMR-visible isotopes, with particular focus on residue-specific labelling and reverse labelling using Escherichia coli expression systems. We also explore how these approaches can aid NMR studies of proteins
Lignosulfonic Acid Exhibits Broadly Anti-HIV-1 Activity – Potential as a Microbicide Candidate for the Prevention of HIV-1 Sexual Transmission
Some secondary metabolites from plants show to have potent inhibitory activities against microbial pathogens, such as human immunodeficiency virus (HIV), herpes simplex virus (HSV), Treponema pallidum, Neisseria gonorrhoeae, etc. Here we report that lignosulfonic acid (LSA), a polymeric lignin derivative, exhibits potent and broad activity against HIV-1 isolates of diverse subtypes including two North America strains and a number of Chinese clinical isolates values ranging from 21.4 to 633 nM. Distinct from other polyanions, LSA functions as an entry inhibitor with multiple targets on viral gp120 as well as on host receptor CD4 and co-receptors CCR5/CXCR4. LSA blocks viral entry as determined by time-of-drug addiction and cell-cell fusion assays. Moreover, LSA inhibits CD4-gp120 interaction by blocking the binding of antibodies specific for CD4-binding sites (CD4bs) and for the V3 loop of gp120. Similarly, LSA interacts with CCR5 and CXCR4 via its inhibition of specific anti-CCR5 and anti-CXCR4 antibodies, respectively. Interestingly, the combination of LSA with AZT and Nevirapine exhibits synergism in viral inhibition. For the purpose of microbicide development, LSA displays low in vitro cytotoxicity to human genital tract epithelial cells, does not stimulate NF-κB activation and has no significant up-regulation of IL-1α/β and IL-8 as compared with N-9. Lastly, LSA shows no adverse effect on the epithelial integrity and the junctional protein expression. Taken together, our findings suggest that LSA can be a potential candidate for tropical microbicide
Computational Reverse-Engineering of a Spider-Venom Derived Peptide Active Against Plasmodium falciparum SUB1
merozoites and invasion into erythrocytes. As PfSUB1 has emerged as an interesting drug target, we explored the hypothesis that PcFK1 targeted PfSUB1 enzymatic activity. culture in a range compatible with our bioinformatics analysis. Using contact analysis and free energy decomposition we propose that residues A14 and Q15 are important in the interaction with PfSUB1.Our computational reverse engineering supported the hypothesis that PcFK1 targeted PfSUB1, and this was confirmed by experimental evidence showing that PcFK1 inhibits PfSUB1 enzymatic activity. This outlines the usefulness of advanced bioinformatics tools to predict the function of a protein structure. The structural features of PcFK1 represent an interesting protein scaffold for future protein engineering
Spermatozoa capture HIV-1 through heparan sulfate and efficiently transmit the virus to dendritic cells
Semen is the main vector for HIV-1 dissemination worldwide. It contains three major sources of infectious virus: free virions, infected leukocytes, and spermatozoa-associated virions. We focused on the interaction of HIV-1 with human spermatozoa and dendritic cells (DCs). We report that heparan sulfate is expressed in spermatozoa and plays an important role in the capture of HIV-1. Spermatozoa-attached virus is efficiently transmitted to DCs, macrophages, and T cells. Interaction of spermatozoa with DCs not only leads to the transmission of HIV-1 and the internalization of the spermatozoa but also results in the phenotypic maturation of DCs and the production of IL-10 but not IL-12p70. At low values of extracellular pH (∼6.5 pH units), similar to those found in the vaginal mucosa after sexual intercourse, the binding of HIV-1 to the spermatozoa and the consequent transmission of HIV-1 to DCs were strongly enhanced. Our observations support the notion that far from being a passive carrier, spermatozoa acting in concert with DCs might affect the early course of sexual transmission of HIV-1 infection
Caractérisation de l'interaction entre la glycoprotéine d'enveloppe gp120 du VIH-1 et les héparanes sulfate : importance des changements conformationnels induits par la liaison à CD4
During viral entry, HIV surface glycoprotein gp120 binds to CD4, thereby anchoring the virus to the host cell surface. This interaction induces conformational changes within gp120 that expose CD4-induced (CD4i) epitope, the binding site for coreceptors (usually a member of the chemokine receptor family). In addition, HIV binds to heparan sulphate (HS), an abundant cell surface polysaccharide, through several domains of gp120, including CD4i. Thus, it may be possible to inhibit HIV binding to cells, using soluble molecules derived from HS. In this context, we proposed to characterise further the structural features of the CD4i/HS interaction. First, we produced and purified gp120 proteins mutated on four residues within CD4i, suspected to interact with HS and, then, we studied their ability to bind to heparin by SPR. A second aspect of the project concerned the development of a new strategy for defining critical residues involved in protein/heparin interactions. Results led to the identification of four heparin binding domains within gp120 and, in particular, three residues within CD4i (R419, K421 and K432) were reported to bind heparin. These studies should contribute to clarify the role of HS in the mechanism of HIV binding to the cell surface and provide precise structural information enabling the definition of HS-derived inhibitors of viral entry.Lors de l'attachement du VIH à la surface d'une cellule, la protéine d'enveloppe gp120 se fixe au récepteur cellulaire CD4, exposant alors son site CD4i qui est alors reconnu par des corécepteurs. Par ailleurs, Le VIH est capable de se fixer aux héparanes sulfate (HS), des polysaccharides présents en abondance à la surface cellulaire, notamment via le site CD4i de gp120 (site de fixation des corécepteurs). Il serait donc possible d'inhiber l'attachement du VIH sur les cellules par l'utilisation de molécules solubles dérivées des HS. Dans ce contexte, nos travaux se sont attachés à définir les aspects structuraux de l'interaction VIH/HS au niveau du site CD4i. Pour cela, des protéines gp120 mutées sur quatre résidus du site CD4i, potentiellement engagés dans la fixation des HS, ont été produites, purifiées et étudiées, par BIAcore, pour leur capacité à interagir avec l'héparine. En parallèle, nous avons développé une méthode simple, permettant d'identifier les régions de fixation à l'héparine d'une protéine donnée. L'ensemble de ces travaux nous a permis d'identifier, au sein de gp120, quatre domaines de liaison à l'héparine et de valider, en particulier, l'engagement de trois résidus du site CD4i (R419, K421 et K432) dans l'interaction avec le polysaccharide. Ces différentes approches ont pour but de clarifier le rôle des HS dans le processus d'attachement du virus à la surface cellulaire et de fournir des informations structurales précises permettant la définition de composés issus des HS capables d'inhiber le mécanisme de l'entrée virale
L’économisation des loisirs pédestres. Structuration et dynamique du marché sur le littoral
International audienc
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