Cyclotrivératrylènes fluorescents pour la détection d'ammoniums d'intérêt biologique

Le développement de sondes moléculaires fluorescentes pour le suivi ex vivo de phénomènes biologiques (communication nerveuse, par exemple) est en plein essor. Dans cette optique, des dérivés fluorescents de cyclotrivératrylènes (CTV), cavitands propices à la complexation de petites molécules organiques, ont été synthétisés. La stratégie utilisée consiste à introduire des groupements électro-donneurs et électro-attracteurs conjugués sur chaque unité aromatique du CTV afin d avoir des systèmes de type push-pull . L extension de la conjugaison entre les groupements électro-attracteurs et électro-donneurs a été envisagée afin d améliorer les propriétés spectroscopiques des sondes. Des réactions de couplage organométallique de Sonogashira, ainsi que des cycloadditions alcynes-azotures catalysées par le cuivre (CuAAc), ont été conduites sur le squelette CTV dans ce but. La caractérisation spectroscopique des différentes sondes fluorescentes développées a été réalisée en milieu organique et en conditions physiologiques, pour celles dont la solubilité le permettait. L étude des propriétés de reconnaissance de ces dernières, pour des ammoniums d intérêt biologique (tels que les neurotransmetteurs, acétylcholine, dopamine et dérivés) en conditions physiologiques, a également été menée par spectrofluorimétrie, ainsi que par d autres techniques (RMN, calorimétrie). La formation d assemblages moléculaires en milieu aqueux a également été mise en évidence (expériences de diffusion dynamique de la lumière, microscopie électronique en transmission), pour certaines sondes lors de ce travail.The development of fluorescent probes for the ex vivo detection of biological phenomena (neuronal communication, for example) presents a growing interest. In this context, fluorescent cyclotriveratrylene (CTV) derivatives have been developed, as these cavitands are known to complex small organic molecules. The strategy used to get interesting spectroscopic properties, was to introduce conjugated electron-donating and electron-withdrawing groups on each aromatic unit of the CTV (leading to push-pull systems). To improve the spectroscopic properties of the probes, the conjugation between the electron-donating and the electron-withdrawing groups was extended. Hence, the Sonogashira organometalic coupling reaction and the copper catalyzed cycloaddition directly on CTV skeleton were used. The spectroscopic characterization of the new fluorescent probes synthesized was done, in organic solvent but also in physiological conditions when the solubility permits it. The recognition studies for biological ammoniums, such as neurotransmitters (acetylcholine, dopamine, and derivatives) were performed in physiological conditions by spectrofluorimetry but also other methods (like MNR or micro-calorimetry). The formation of molecular assemblies was also observed (by dynamic light scattering and transmission electron microscopy) during this work with some of the probes.BORDEAUX1-Bib.electronique (335229901) / SudocSudocFranceF

TLR1/TLR2 Heterodimers Play an Important Role in the Recognition of Borrelia Spirochetes

After infection with Borrelia species, the risk for developing Lyme disease varies significantly between individuals. Recognition of Borrelia by the immune system is mediated by pattern recognition receptors (PRRs), such as TLRs. While TLR2 is the main recognition receptor for Borrelia spp., little is known about the role of TLR1 and TLR6, which both can form functionally active heterodimers with TLR2. Here we investigated the recognition of Borrelia by both murine and human TLR1 and TLR6. Peritoneal macrophages from TLR1- and TLR6- gene deficient mice were isolated and exposed to Borrelia. Human PBMCs were stimulated with Borrelia with or without specific TLR1 and TLR6 blocking using specific antibodies. Finally, the functional consequences of TLR polymorphisms on Borrelia-induced cytokine production were assessed. Splenocytes isolated from both TLR1−/− and TLR6−/− mice displayed a distorted Th1/Th2 cytokine balance after stimulation with B.burgdorferi, while no differences in pro-inflammatory cytokine production were observed. In contrast, blockade of TLR1 with specific neutralizing antibodies led to decreased cytokine production by human PBMCs after exposure to B.burgdorferi. Blockade of human TLR6 did not lead to suppression of cytokine production. When PBMCs from healthy individuals bearing polymorphisms in TLR1 were exposed to B.burgdorferi, a remarkably decreased in vitro cytokine production was observed in comparison to wild-type controls. TLR6 polymorphisms lead to a minor modified cytokine production. This study indicates a dominant role for TLR1/TLR2 heterodimers in the induction of the early inflammatory response by Borrelia spirochetes in humans

Airborne laser scanning reveals uniform responses of forest structure to moose (Alces alces) across the boreal forest biome

1. The moose Alces alces is the largest herbivore in the boreal forest biome, where it can have dramatic impacts on ecosystem structure and dynamics. Despite the importance of the boreal forest biome in global carbon cycling, the impacts of moose have only been studied in disparate regional exclosure experiments, leading to calls for common analyses across a biome-wide network of moose exclosures. 2. In this study, we use airborne laser scanning (ALS) to analyse forest canopy re-sponses to moose across 100 paired exclosure-control experimental plots dis-tributed across the boreal biome, including sites in the United States (Isle Royale), Canada (Quebec, Newfoundland), Norway, Sweden and Finland. 3. We test the hypotheses that canopy height, vertical complexity and above- ground biomass (AGB) are all reduced by moose and that the impacts vary with moose density, productivity, temperature and pulse disturbances such as logging and insect outbreaks. 4. We find a surprising convergence in forest canopy response to moose. Moose had negative impacts on canopy height, complexity and AGB as expected. The responses of canopy complexity and AGB were consistent across regions and did not vary along environmental gradients. The difference in canopy height be-tween exclosures and open plots was on average 6 cm per year since the start of exclosure treatment (±2.1 SD). This rate increased with temperature, but only when moose density was high. 5. The difference in AGB between moose exclosures and open plots was 0.306 Mg ha−1 year−1 (±0.079). In browsed plots, stand AGB was 32% of that in the exclosures, a difference of 2.09 Mg ha−1. The uniform response allows scaling of the estimate to a biome-wide impact of moose of the loss of 448 (±115) Tg per year, or 224 Tg of carbon. 6. Synthesis: Analysis of ALS data from distributed exclosure experiments identified a largely uniform response of forest canopies to moose across regions, facilitat-ing scaling of moose impacts across the whole biome. This is an important step towards incorporating the effect of the largest boreal herbivore on the carbon cycling of one of the world's largest terrestrial biomes.publishedVersio

An Asymptotic-Preserving all-speed scheme for the Euler and Navier-Stokes equations

We present an Asymptotic-Preserving 'all-speed' scheme for the simulation of compressible flows valid at all Mach-numbers ranging from very small to order unity. The scheme is based on a semi-implicit discretization which treats the acoustic part implicitly and the convective and diffusive parts explicitly. This discretization, which is the key to the Asymptotic-Preserving property, provides a consistent approximation of both the hyperbolic compressible regime and the elliptic incompressible regime. The divergence-free condition on the velocity in the incompressible regime is respected, and an the pressure is computed via an elliptic equation resulting from a suitable combination of the momentum and energy equations. The implicit treatment of the acoustic part allows the time-step to be independent of the Mach number. The scheme is conservative and applies to steady or unsteady flows and to general equations of state. One and Two-dimensional numerical results provide a validation of the Asymptotic-Preserving 'all-speed' properties

A Human Minor Histocompatibility Antigen Specific for B Cell Acute Lymphoblastic Leukemia

Human minor histocompatibility antigens (mHags) play an important role in the induction of cytotoxic T lymphocyte (CTL) reactivity against leukemia after human histocompatibility leukocyte antigen (HLA)-identical allogeneic bone marrow transplantation (BMT). As most mHags are not leukemia specific but are also expressed by normal tissues, antileukemia reactivity is often associated with life-threatening graft-versus-host disease (GVHD). Here, we describe a novel mHag, HB-1, that elicits donor-derived CTL reactivity in a B cell acute lymphoblastic leukemia (B-ALL) patient treated by HLA-matched BMT. We identified the gene encoding the antigenic peptide recognized by HB-1–specific CTLs. Interestingly, expression of the HB-1 gene was only observed in B-ALL cells and Epstein-Barr virus–transformed B cells. The HB-1 gene–encoded peptide EEKRGSLHVW is recognized by the CTL in association with HLA-B44. Further analysis reveals that a polymorphism in the HB-1 gene generates a single amino acid exchange from His to Tyr at position 8 within this peptide. This amino acid substitution is critical for recognition by HB-1–specific CTLs. The restricted expression of the polymorphic HB-1 Ag by B-ALL cells and the ability to generate HB-1–specific CTLs in vitro using peptide-loaded dendritic cells offer novel opportunities to specifically target the immune system against B-ALL without the risk of evoking GVHD

Selective cancer-germline gene expression in pediatric brain tumors

Cancer-germline genes (CGGs) code for immunogenic antigens that are present in various human tumors and can be targeted by immunotherapy. Their expression has been studied in a wide range of human tumors in adults. We measured the expression of 12 CGGs in pediatric brain tumors, to identify targets for therapeutic cancer vaccines. Real Time PCR was used to quantify the expression of genes MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A10, MAGE-A12, MAGE-C2, NY-ESO-1 and GAGE-1,2,8 in 50 pediatric brain tumors of different histological subtypes. Protein expression was examined with immunohistochemistry. Fifty-five percent of the medulloblastomas (n = 11), 86% of the ependymomas (n = 7), 40% of the choroid plexus tumors (n = 5) and 67% of astrocytic tumors (n = 27) expressed one or more CGGs. Immunohistochemical analysis confirmed qPCR results. With exception of a minority of tumors, the overall level of CGG expression in pediatric brain tumors was low. We observed a high expression of at least one CGG in 32% of the samples. CGG-encoded antigens are therefore suitable targets in a very selected group of pediatric patients with a brain tumor. Interestingly, glioblastomas from adult patients expressed CGGs more often and at significantly higher levels compared to pediatric glioblastomas. This observation is in line with the notion that pediatric and adult glioblastomas develop along different genetic pathways

Denudation outpaced by crustal thickening in the eastern Tianshan

The modern high topography of the Tianshan resulted from the reactivation of a Paleozoic orogenic belt by the India/Asia collision. Today, the range exhibits tectonically active forelands and intermontane basins. Based on quantitative morphotectonic observations and age constraints derived from cosmogenic 10Be dating, single-grain post-infrared infrared stimulated luminescence (p-IR IRSL) dating and modeling of fault scarp degradation, we quantify the deformation in the Nalati and Bayanbulak intermontane basins in the central Eastern Tianshan. Our results indicate that at least 1.4 mm/yr of horizontal crustal shortening is accommodated within these two basins. This shortening represents over 15% of the 8.5 ± 0.5 mm/yr total shortening rate across the entire range at this longitude. This shortening rate implies that the Eastern Central Tianshan is thickening at a mean rate of ∼1.4 mm/yr, a rate that is significantly higher than the average denudation rate of 0.14 mm/yr derived from our cosmogenic analysis. This discrepancy suggests that the Tianshan range has not yet reached a steady-state topography and remains in a transient state of topographic growth, most likely due to limited denudation rates driven by the arid climate of Central Asia