Functional insights from the crystal structure of the N-terminal domain of the prototypical toll receptor.

Drosophila melanogaster Toll is the founding member of an important family of pathogen-recognition receptors in humans, the Toll-like receptor (TLR) family. In contrast, the prototypical receptor is a cytokine-like receptor for Spätzle (Spz) protein and plays a dual role in both development and immunity. Here, we present the crystal structure of the N-terminal domain of the receptor that encompasses the first 201 amino acids at 2.4 Å resolution. To our knowledge, the cysteine-rich cap adopts a novel fold unique to Toll-1 orthologs in insects and that is not critical for ligand binding. However, we observed that an antibody directed against the first ten LRRs blocks Spz signaling in a Drosophila cell-based assay. Supplemented by point mutagenesis and deletion analysis, our data suggests that the region up to LRR 14 is involved in Spz binding. Comparison with mammalian TLRs reconciles previous contradictory findings about the mechanism of Toll activation.This work is financed by the Wellcome Trust Award (RG47206). We thank Dr. Martin Moncrieffe for helpful discussions and Ms. Irina Ogay from the Baculovirus Facility, Department of Biochemistry, Cambridge, for protein expression. We are grateful to Dr. Katherine Stott, from the Biophysics Facility, for her aid with analytical ultracentrifugation. Thanks to Dr. Dimitri Chirdgaze, from the Crystallographic X-ray Facility, for his assistance. We are grateful to Prof. Abel Moreno for help with capillary crystallization. We thank the staff at beamlines IO3 at Diamond Light Source, England, and ID23EH1 at the ESRF, Grenoble, France, for help with the data collection.This is the final version of the article. It first appeared from Elsevier (Cell Press) via http://dx.doi.org/10.1016/j.str.2012.11.00

Northeast IPM Center Participation by the NYS IPM Program, 2006

NYS IPM Program staff were involved with several key activities of theNortheast IPM Center in 2006. Included were participation and leadership in the Center’s Working Groups and meeting with Natural Resource Conservation Service representatives

Présence d’un pergélisol würmien sur le piémont des Pyrénées Atlantiques

Des cryoturbations se sont produites sur le sommet de terrasses anciennes sur le piémont nord des Pyrénées. Elles déforment le contact entre une couche de loess wùrmien et les cailloutis altérés sous-jacents. Deux fentes de gel fossiles, sous la zone cryoturbée, indiquent la présence, sous la couche active, d'un pergélisol à plus de 15 km au nord des moraines externes de Lourdes.Cryoturbations developed on top of old terraces of the northern piedmont of the Pyrénées. They deformed the contact between a Wurmian loess and weathered gravels underneath. Two fossil ice wedge casts observed under the cryoturbated layer are indicative of a permafrost of Wurmian age, extending more than 15 km north of the external moraines of Lourdes.Kryturbationen haben an der Oberfläche der alten Terrassen der Bergfussebene der Pyrenaen stattgefunden. Sie haben den Kontakt zwischen einer Lôss-Schicht aus der Wurmeiszeit und dem darunter liegenden verwitterten Gerôll verformt. Zwei fossile Frost-Klùfte unter der kryoturbaten Zone weisen auf das Vorhandensein eines Permafrostbodens unter der aktiven Schicht, mehr als 15 km nôrdlich der externen Morànen von Lourdes

Note sur un site à macrorestes végétaux tardiglaciaires au sud-ouest de Montréal, Québec

À une cinquantaine de kilomètres au sud-ouest de Montréal, dans la région de Huntingdon, des dépôts littoraux attribués à la mer de Champlain renferment, vers 110 m d'altitude, deux minces lits organiques fossiles contenant des macrorestes de type arctique-alpin. La datation au radiocarbone de l'un des lits organiques a donné un âge de 10 480 ± 140 BP (Beta-8270). Le contexte géomorphologique, l'identification de la flore macrofossile et l'analyse pollinique indiquent que les lits organiques résultent du mélange, sur un estran tardiglaciaire, de macrorestes provenant de deux habitats d'une toundra sise à proximité du site.Two thin organic layers containing plant macrofossils of arctic-alpine taxa were found in shore deposits, attributed to the Champlain Sea, at an elevation of 110 m above present sea level, near Huntingdon (Québec) about fifty kilometres southwest of Montréal. A radiocarbon date of 10 480 ± 140 BP (Beta-8270) was obtained on one of the organic layers. Geomorphic context, macrofossil flora and pollen analysis suggest that the organic layers were the result of a mix of plant remains from two distinct tundra habitats on a lateglacial strand

LPS ligand and culture additives improve production of monomeric MD-1 and 2 in Pichia pastoris by decreasing aggregation and intermolecular disulfide bonding.

Myeloid differentiation proteins MD-1 and MD-2 have both been shown to form a heterogeneous collection of oligomers when expressed in absence of their respective receptor, RP105 and TLR4. The biological relevance of these oligomers is not clear. Only monomeric proteins have been found to be active and able to trigger an immune response to endotoxin by modulating the TLR4 pathway. In this study, we produced variants of MD-1 and MD-2 in Pichia pastoris. To minimize the time and expense of initial expression tests, small-scale cultures have been set up to allow the rapid identification of the highest expressing clone and the optimal expression conditions. The expression vectors used, the site of linearization and the locus of integration affected the yield of transformation. Next we screened culture additives and found that they significantly increased the fraction of monomeric proteins secreted in the culture medium (up to 15% of the total MD protein produced). We confirmed their presence by size-exclusion chromatography. Optimal anti-aggregation agents were protein-dependent except for LPS that presented stabilizing effects for all MD proteins. Contrary to previous reports, this study suggests that MD-1 can bind to LPS.WELLCOME TRUST; ward Number RG47206.This is the final version of the article. It first appeared from Elsevier at http://dx.doi.org/10.1016/j.pep.2010.11.018

Planetary Science Virtual Observatory architecture

In the framework of the Europlanet-RI program, a prototype of Virtual Observatory dedicated to Planetary Science was defined. Most of the activity was dedicated to the elaboration of standards to retrieve and visualize data in this field, and to provide light procedures to teams who wish to contribute with on-line data services. The architecture of this VO system and selected solutions are presented here, together with existing demonstrators

Chronologie de la déglaciation en Gaspésie : nouvelles données et implications

De nouvelles datations à l'accélérateur de particules des premiers macrorestes végétaux terricoles accumulés dans les sédiments postglaciaires d'une douzaine de lacs répartis en Gaspésie et au Bas-Saint-Laurent bouleversent entièrement la chronologie générale de la déglaciation de ces régions. Ces données indiquent que la Gaspésie est restée sous l'emprise des glaces jusque vers 10 500-10 000 ans BP sauf en bordure de la péninsule, notamment le long de la côte septentrionale où la déglaciation de l'arrière-pays date d'au moins 12 500 ans BP par endroits. La déglaciation finale se serait produite vers 9500-9000 ans BP dans la région de Murdochville. La chronologie de déglaciation qui se dégage de ces données s'accorde maintenant avec celle des provinces maritimes, notamment pour ce qui est de l'existence de masses glaciaires actives durant l'épisode du Dryas récent. Les conséquences sont importantes, tant pour la modélisation du retrait des glaces, la colonisation végétale et l'accessibilité du territoire aux populations humaines que pour les reconstitutions climatiques.The general chronology of ice retreat in the Lower-St. Lawrence and Gaspé Peninsula regions is drastically changed by new AMS dates on the first terrestrial plant macrofossils recovered from postglacial sediments from a dozen lakes throughout the area. The dates indicate that most of the Gaspé Peninsula was ice-covered until 10 500-10 000 year BP except along coasts, particularly the northern coast, where ice retreat on land dates back over 12 500 year BP in places. Final deglaciation occurred in the Murdochville area at around 9500-9000 year BP. The implied chronology of ice retreat is now in accordance with that from the Maritimes, especially with respect to active ice masses during the Younger Dryas episode. This finding has important implications for the modelling of ice retreat and for climatic reconstructions, as well as for accessibility of land to plant, animal and human populations.Neue Datierungen mit dem Teilchenbeschleuniger der ersten pflanzlichen Erd-Makroreste aus den postglazialen Sedimenten eines Dutzends von Seen der Gaspé-Halbinsel und des unteren Sankt-Lorenz-Gebiets haben die allgemeine Chronologie der Enteisung dieser Regionen drastisch veràndert. Diese Daten zeigen, daB die Gaspé-Halbinsel bis gegen 10 500 - 10 000 Jahre v.u.Z. unter dem EinfluB des Eises stand, auBer am Rand der Halbinsel, besonders entlang der nôrdlichen Kiiste, wo die Enteisung des Hinterlandes stellenweise mindestens vor 12 500 Jahren v.u.Z. stattfand. Die endgultige Enteisung soil gegen 9500 - 9000 Jahre v.u.Z. in der Gegend von Murdochville stattgefunden haben. Die so erhaltene Chronologie der Enteisung stimmt jetzt mit der der Atlantikprovinzen ùberein, vor allem was die Existenz der aktiven Eismassen der jungeren Dryas-Episode betrifft. Die Konsequenzen sind wichtig, sowohl fur das Modellieren des Eisrùckzugs, die Pflanzen-Ansiedlung und die Zuganglichkeit des Gebiets fur menschliche Populationen wie auch fur die klimatischen Rekonstruktionen

From performance curves to performance surfaces: Interactive effects of temperature and oxygen availability on aerobic and anaerobic performance in the common wall lizard

Accurately predicting the responses of organisms to novel or changing environments requires the development of ecologically-appropriate experimental methodology and process-based models. For ectotherms, thermal performance curves (TPCs) have provided a useful framework to describe how organismal performance is dependent on temperature. However, this approach often lacks a mechanistic underpinning, which limits our ability to use TPCs predictively. Furthermore, thermal dependence varies across traits, and performance is also limited by additional abiotic factors, such as oxygen availability. We test a central prediction of our recent Hierarchical Mechanisms of Thermal Limitation (HMTL) Hypothesis which proposes that natural hypoxia exposure will reduce maximal performance and cause the TPC for whole-organism performance to become more symmetrical. We quantified TPCs for two traits often used as fitness proxies, sprint speed and aerobic scope, in lizards under conditions of normoxia and high-elevation hypoxia. In line with the predictions of HMTL, anaerobically fuelled sprint speed was unaffected by acute hypoxia while the TPC for aerobic scope became shorter and more symmetrical. This change in TPC shape resulted from both the maximum aerobic scope and the optimal temperature for aerobic scope being reduced in hypoxia as predicted. Following these results, we present a mathematical framework, which we call Temperature–Oxygen Performance Surfaces, to quantify the interactive effects of temperature and oxygen on whole-organism performance in line with the HMTL hypothesis. This framework is transferrable across traits and levels of organization to allow predictions for how ectotherms will respond to novel combinations of temperature and other abiotic factors, providing a useful tool in a time of rapidly changing environmental conditions.info:eu-repo/semantics/publishedVersio

Identification of key residues that confer Rhodobacter sphaeroides LPS activity at horse TLR4/MD-2.

The molecular determinants underpinning how hexaacylated lipid A and tetraacylated precursor lipid IVa activate Toll-like receptor 4 (TLR4) are well understood, but how activation is induced by other lipid A species is less clear. Species specificity studies have clarified how TLR4/MD-2 recognises different lipid A structures, for example tetraacylated lipid IVa requires direct electrostatic interactions for agonism. In this study, we examine how pentaacylated lipopolysaccharide from Rhodobacter sphaeroides (RSLPS) antagonises human TLR4/MD-2 and activates the horse receptor complex using a computational approach and cross-species mutagenesis. At a functional level, we show that RSLPS is a partial agonist at horse TLR4/MD-2 with greater efficacy than lipid IVa. These data suggest the importance of the additional acyl chain in RSLPS signalling. Based on docking analysis, we propose a model for positioning of the RSLPS lipid A moiety (RSLA) within the MD-2 cavity at the TLR4 dimer interface, which allows activity at the horse receptor complex. As for lipid IVa, RSLPS agonism requires species-specific contacts with MD-2 and TLR4, but the R2 chain of RSLA protrudes from the MD-2 pocket to contact the TLR4 dimer in the vicinity of proline 442. Our model explains why RSLPS is only partially dependent on horse TLR4 residue R385, unlike lipid IVa. Mutagenesis of proline 442 into a serine residue, as found in human TLR4, uncovers the importance of this site in RSLPS signalling; horse TLR4 R385G/P442S double mutation completely abolishes RSLPS activity without its counterpart, human TLR4 G384R/S441P, being able to restore it. Our data highlight the importance of subtle changes in ligand positioning, and suggest that TLR4 and MD-2 residues that may not participate directly in ligand binding can determine the signalling outcome of a given ligand. This indicates a cooperative binding mechanism within the receptor complex, which is becoming increasingly important in TLR signalling.This work was supported by a project grant from the Horserace Betting Levy Board to CEB and a Horserace Betting Levy Board Veterinary Research Training Scholarship to KLI. This work was also supported by a Wellcome Trust program grant to NJG and CEB. CEB is a BBSRC Research Development Fellow.This is the final version of the article. It first appeared from PLOS at http://dx.doi.org/10.1371/journal.pone.0098776

Localized CVD growth of oriented and individual carbon nanotubes from nanoscaled dots prepared by lithographic sequences

International audienceUsing a combination of top-down lithographic techniques, isolated, individual and oriented multi-wall carbon nanotubes (MWNTs) were grown on nickel or iron nanoscaled dots. In the first step of the process, micron-sized catalytic metallic dots (either iron or nickel) were prepared using UV lithography. MWNTs were then synthesized from these catalysts using a direct current plasma-assistance and hot-filament-enhanced chemical vapor deposition (CVD) reactor. Samples were characterized by means of scanning electron microscopy. It turns out that the splitting up of the micron-sized dot is favored in the iron case and that the surface diffusion of the metal is enhanced using ammonia in the gaseous mixture during the CVD process. The results are discussed giving arguments for the understanding of the MWNT growth mechanism. In a second step, a focused ion beam (FIB) procedure is carried out in order to reduce the initial dot size down to submicronic scale and subsequently to grow one single MWNT per dot. It is found that nickel is most appropriate to control the size of the dot. Dots of size 200 nm ± 40 nm are then required to grow individual MWNTs