An order-disorder twin crystal of L-2-haloacid dehalogenase from Sulfolobus tokodaii.

Journal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov'tThe L-2-haloacid dehalogenase enzymes catalyse the hydrolytic cleavage of a halogen from the C2 position of short-chain haloacids. The recombinant dehalogenase from the thermophilic archaeon Sulfolobus tokodaii has been cloned, overexpressed and purified to homogeneity. The 24 kDa enzyme was crystallized using the microbatch method in the monoclinic space group C2, with unit-cell parameters a = 127.6, b = 58.1, c = 51.2 A, beta = 97.2 degrees . Data were collected to 1.9 A resolution using synchrotron radiation and the structure was solved by molecular replacement. Analysis of the data and the preliminary refined model showed that the crystal was an order-disorder twin by reticular merohedry with a twin index of 10. It was possible to de-twin the experimental data utilizing the symmetry of the molecular layers from which the crystal is built.Engineering and Physical Sciences Research CouncilNIHBBSRCNWDA project awar

Inferring the thermal resistance and effective thermal mass of a wall using frequent temperature and heat flux measurements

Evaluating how much heat is lost through external walls is a key requirement for building energy simulators and is necessary for quality assurance and successful decision making in policy making and building design, construction and refurbishment. Heat loss can be estimated using the temperature differences between the inside and outside air and an estimate of the thermal transmittance (U-value) of the wall. Unfortunately the actual U-value may be different from those values obtained using assumptions about the materials, their properties and the structure of the wall after a cursory visual inspection. In-situ monitoring using thermometers and heat flux plates enables more accurate characterisation of the thermal properties of walls in their context. However, standard practices require that the measurements are carried out in winter over a two-week period to significantly reduce the dynamic effects of the wall's thermal mass from the data. A novel combination of a lumped thermal mass model, together with Bayesian statistical analysis is presented to derive estimates of the U-value and effective thermal mass. The method needs only a few days of measurements, provides an estimate of the effective thermal mass and could potentially be used in summer

Biochemical and structural studies of a L-haloacid dehalogenase from the thermophilic archaeon Sulfolobus tokodaii

addresses: Henry Wellcome Building for Biocatalysis, School of Biosciences, University of Exeter, Stocker Road, Exeter EX4 4QD, UK.types: Journal Article; Research Support, Non-U.S. Gov'tThis a post-print, author-produced version of an article accepted for publication in Extremophiles. Copyright © 2009 Springer Verlag. The definitive version is available at http://link.springer.com/article/10.1007%2Fs00792-008-0208-0Haloacid dehalogenases have potential applications in the pharmaceutical and fine chemical industry as well as in the remediation of contaminated land. The L: -2-haloacid dehalogenase from the thermophilic archaeon Sulfolobus tokodaii has been cloned and over-expressed in Escherichia coli and successfully purified to homogeneity. Here we report the structure of the recombinant dehalogenase solved by molecular replacement in two different crystal forms. The enzyme is a homodimer with each monomer being composed of a core-domain of a beta-sheet bundle surrounded by alpha-helices and an alpha-helical sub-domain. This fold is similar to previously solved mesophilic L: -haloacid dehalogenase structures. The monoclinic crystal form contains a putative inhibitor L: -lactate in the active site. The enzyme displays haloacid dehalogenase activity towards carboxylic acids with the halide attached at the C2 position with the highest activity towards chloropropionic acid. The enzyme is thermostable with maximum activity at 60 degrees C and a half-life of over 1 h at 70 degrees C. The enzyme is relatively stable to solvents with 25% activity lost when incubated for 1 h in 20% v/v DMSO

The Role of the Substantia Nigra Pars Compacta in Regulating Sleep Patterns in Rats

Background. As of late, dopaminergic neurotransmission has been recognized to be involved in the generation of sleep disturbances. Increasing evidence shows that sleep disturbances in Parkinson's disease (PD) patients are mostly related to the disease itself, rather than being a secondary phenomenon. Evidence contained in the literature lends support to the hypothesis that the dopaminergic nigrostriatal pathway is closely involved in the regulation of sleep patterns. Methodology/Principal Findings. To test this hypothesis we examined the electrophysiological activity along the sleep-wake cycle of rats submitted to a surgically induced lesion of the SNpc by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). We demonstrated that a 50% lesion of the substantia nigra pars compacta (SNpc) suffices to produce disruptions of several parameters in the sleep-wake pattern of rats. A robust and constant decrease in the latency to the onset of slow wave sleep (SWS) was detected throughout the five days of recording in both light [F((22.16)) = 72.46, p<0.0001] and dark [F((22.16)) = 75.0, p<0.0001] periods. Also found was a pronounced increase in the percentage of sleep efficiency during the first four days of recording [F((21.15)) = 21.48, p<0.0001], in comparison to the sham group. Additionally, the reduction in the SNpc dopaminergic neurons provoked an ablation in the percentage of rapid eye movement sleep (REM) during three days of the sleep-wake recording period with a strong correlation (r = 0.91; p<0.0001) between the number of dopaminergic neurons lost and the percentage decrease of REM sleep on the first day of recording. On day 4, the percentage of REM sleep during the light and dark periods was increased, [F((22.16)) = 2.46, p<0.0007], a phenomenon consistent with REM rebound. Conclusions/Significance. We propose that dopaminergic neurons present in the SNpc possess a fundamental function in the regulation of sleep processes, particularly in promoting REM sleep.AFIPCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Psicobiol, São Paulo, BrazilUniv Fed Parana, Dept Farmacol, BR-80060000 Curitiba, Parana, BrazilUniversidade Federal de São Paulo, Dept Psicobiol, São Paulo, BrazilFAPESP: 98/14.303-3Web of Scienc

Search for new phenomena in final states with an energetic jet and large missing transverse momentum in pp collisions at √ s = 8 TeV with the ATLAS detector

Results of a search for new phenomena in final states with an energetic jet and large missing transverse momentum are reported. The search uses 20.3 fb−1 of √ s = 8 TeV data collected in 2012 with the ATLAS detector at the LHC. Events are required to have at least one jet with pT > 120 GeV and no leptons. Nine signal regions are considered with increasing missing transverse momentum requirements between Emiss T > 150 GeV and Emiss T > 700 GeV. Good agreement is observed between the number of events in data and Standard Model expectations. The results are translated into exclusion limits on models with either large extra spatial dimensions, pair production of weakly interacting dark matter candidates, or production of very light gravitinos in a gauge-mediated supersymmetric model. In addition, limits on the production of an invisibly decaying Higgs-like boson leading to similar topologies in the final state are presente

Isolation, Characterization, and Stability of Discretely-Sized Nanolipoprotein Particles Assembled with Apolipophorin-III

Background: Nanolipoprotein particles (NLPs) are discoidal, nanometer-sized particles comprised of self-assembled phospholipid membranes and apolipoproteins. NLPs assembled with human apolipoproteins have been used for myriad biotechnology applications, including membrane protein solubilization, drug delivery, and diagnostic imaging. To expand the repertoire of lipoproteins for these applications, insect apolipophorin-III (apoLp-III) was evaluated for the ability to form discretely-sized, homogeneous, and stable NLPs. Methodology: Four NLP populations distinct with regards to particle diameters (ranging in size from 10 nm to.25 nm) and lipid-to-apoLp-III ratios were readily isolated to high purity by size exclusion chromatography. Remodeling of the purified NLP species over time at 4uC was monitored by native gel electrophoresis, size exclusion chromatography, and atomic force microscopy. Purified 20 nm NLPs displayed no remodeling and remained stable for over 1 year. Purified NLPs with 10 nm and 15 nm diameters ultimately remodeled into 20 nm NLPs over a period of months. Intra-particle chemical cross-linking of apoLp-III stabilized NLPs of all sizes. Conclusions: ApoLp-III-based NLPs can be readily prepared, purified, characterized, and stabilized, suggesting their utilit

Earth: Atmospheric Evolution of a Habitable Planet

Our present-day atmosphere is often used as an analog for potentially habitable exoplanets, but Earth's atmosphere has changed dramatically throughout its 4.5 billion year history. For example, molecular oxygen is abundant in the atmosphere today but was absent on the early Earth. Meanwhile, the physical and chemical evolution of Earth's atmosphere has also resulted in major swings in surface temperature, at times resulting in extreme glaciation or warm greenhouse climates. Despite this dynamic and occasionally dramatic history, the Earth has been persistently habitable--and, in fact, inhabited--for roughly 4 billion years. Understanding Earth's momentous changes and its enduring habitability is essential as a guide to the diversity of habitable planetary environments that may exist beyond our solar system and for ultimately recognizing spectroscopic fingerprints of life elsewhere in the Universe. Here, we review long-term trends in the composition of Earth's atmosphere as it relates to both planetary habitability and inhabitation. We focus on gases that may serve as habitability markers (CO2, N2) or biosignatures (CH4, O2), especially as related to the redox evolution of the atmosphere and the coupled evolution of Earth's climate system. We emphasize that in the search for Earth-like planets we must be mindful that the example provided by the modern atmosphere merely represents a single snapshot of Earth's long-term evolution. In exploring the many former states of our own planet, we emphasize Earth's atmospheric evolution during the Archean, Proterozoic, and Phanerozoic eons, but we conclude with a brief discussion of potential atmospheric trajectories into the distant future, many millions to billions of years from now. All of these 'Alternative Earth' scenarios provide insight to the potential diversity of Earth-like, habitable, and inhabited worlds.Comment: 34 pages, 4 figures, 4 tables. Review chapter to appear in Handbook of Exoplanet

Performance of the upgraded PreProcessor of the ATLAS Level-1 Calorimeter Trigger

The PreProcessor of the ATLAS Level-1 Calorimeter Trigger prepares the analogue trigger signals sent from the ATLAS calorimeters by digitising, synchronising, and calibrating them to reconstruct transverse energy deposits, which are then used in further processing to identify event features. During the first long shutdown of the LHC from 2013 to 2014, the central components of the PreProcessor, the Multichip Modules, were replaced by upgraded versions that feature modern ADC and FPGA technology to ensure optimal performance in the high pile-up environment of LHC Run 2. This paper describes the features of the newMultichip Modules along with the improvements to the signal processing achieved.ANPCyTYerPhI, ArmeniaAustralian Research CouncilBMWFW, AustriaAustrian Science Fund (FWF)Azerbaijan National Academy of Sciences (ANAS)SSTC, BelarusNational Council for Scientific and Technological Development (CNPq)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Natural Sciences and Engineering Research Council of CanadaCanada Foundation for InnovationNational Natural Science Foundation of China (NSFC)Departamento Administrativo de Ciencia, Tecnología e Innovación ColcienciasMinistry of Education, Youth & Sports - Czech Republic Czech Republic GovernmentCzech Republic GovernmentDNRF, DenmarkDanish Natural Science Research CouncilCentre National de la Recherche Scientifique (CNRS)CEA-DRF/IRFU, FranceFederal Ministry of Education & Research (BMBF)Max Planck SocietyGreek Ministry of Development-GSRTRGC and Hong Kong SAR, ChinaIsrael Science FoundationBenoziyo Center, IsraelIstituto Nazionale di Fisica Nucleare (INFN)Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT)Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT) Japan Society for the Promotion of ScienceCNRST, MoroccoRCN, NorwayPortuguese Foundation for Science and TechnologyMNE/IFA, RomaniaMES of RussiaMESTD, SerbiaMSSR, SlovakiaSlovenian Research Agency - SloveniaMIZS, SloveniaSpanish GovernmentSRC, SwedenWallenberg Foundation, SwedenSNSF Geneva, SwitzerlandMinistry of Science and Technology, TaiwanMinistry of Energy & Natural Resources - TurkeyScience & Technology Facilities Council (STFC)United States Department of Energy (DOE)National Science Foundation (NSF)BCKDF, CanadaCANARIE, CanadaCRC, CanadaEuropean Research Council (ERC)European Union (EU)French National Research Agency (ANR)German Research Foundation (DFG)Alexander von Humboldt FoundationGreek NSRF, GreeceBSF-NSF, IsraelGerman-Israeli Foundation for Scientific Research and DevelopmentLa Caixa Banking Foundation, SpainCERCA Programme Generalitat de Catalunya, SpainPROMETEO, SpainGenT Programmes Generalitat Valenciana, SpainGoran Gustafssons Stiftelse, SwedenRoyal Society of LondonLeverhulme TrustNRC, CanadaCERNANID, ChileChinese Academy of SciencesMinistry of Science and Technology, ChinaSRNSFG, GeorgiaHGF, GermanyNetherlands Organization for Scientific Research (NWO) Netherlands GovernmentMinistry of Science and Higher Education, PolandNCN, PolandNRCKI, Russia FederationJINRDST/NRF, South AfricaSERI, Geneva, SwitzerlandCantons of Bern and Geneva, SwitzerlandCompute Canada, CanadaHorizon 2020Marie Sklodowska-Curie ActionsEuropean Cooperation in Science and Technology (COST)EU-ESF, Greec