Optimisation of recombinant production of active human cardiac SERCA2a ATPase

Methods for recombinant production of eukaryotic membrane proteins, yielding sufficient quantity and quality of protein for structural biology, remain a challenge. We describe here, expression and purification optimisation of the human SERCA2a cardiac isoform of Ca2+ translocating ATPase, using Saccharomyces cerevisiae as the heterologous expression system of choice. Two different expression vectors were utilised, allowing expression of C-terminal fusion proteins with a biotinylation domain or a GFP- His8 tag. Solubilised membrane fractions containing the protein of interest were purified onto Streptavidin-Sepharose, Ni-NTA or Talon resin, depending on the fusion tag present. Biotinylated protein was detected using specific antibody directed against SERCA2 and, advantageously, GFP-His8 fusion protein was easily traced during the purification steps using in-gel fluorescence. Importantly, talon resin affinity purification proved more specific than Ni-NTA resin for the GFP-His8 tagged protein, providing better separation of oligomers present, during size exclusion chromatography. The optimised method for expression and purification of human cardiac SERCA2a reported herein, yields purified protein (> 90%) that displays a calcium-dependent thapsigargin-sensitive activity and is suitable for further biophysical, structural and physiological studies. This work provides support for the use of Saccharomyces cerevisiae as a suitable expression system for recombinant production of multi-domain eukaryotic membrane proteins

The SPHERE data center: a reference for high contrast imaging processing

The objective of the SPHERE Data Center is to optimize the scientific return of SPHERE at the VLT, by providing optimized reduction procedures, services to users and publicly available reduced data. This paper describes our motivation, the implementation of the service (partners, infrastructure and developments), services, description of the on-line data, and future developments. The SPHERE Data Center is operational and has already provided reduced data with a good reactivity to many observers. The first public reduced data have been made available in 2017. The SPHERE Data Center is gathering a strong expertise on SPHERE data and is in a very good position to propose new reduced data in the future, as well as improved reduction procedures.Comment: SF2A proceeding

Modelling the hydrological behaviour of a coffee agroforestry basin in Costa Rica

UMR LISAH, Equipe Eau et Polluants en Bassins VersantsThe profitability of hydropower in Costa Rica is affected by soil erosion and sedimentation in dam reservoirs, which are in turn influenced by land use, infiltration and aquifer interactions with surface water. In order to foster the provision and payment for Hydrological Environmental Services (HES), a quantitative assessment of the impact of specific land uses on the functioning of drainage-basins is required. The present paper aims to study the water balance partitioning in a volcanic coffee agroforestry microbasin (1 km(2), steep slopes) in Costa Rica, as a first step towards evaluating sediment or contaminant loads. The main hydrological processes were monitored during one year, using flume, eddy-covariance flux tower, soil water profiles and piezometers. A new Hydro-SVAT lumped model is proposed, that balances SVAT (Soil Vegetation Atmosphere Transfer) and basin-reservoir routines. The purpose of such a coupling was to achieve a trade-off between the expected performance of ecophysiological and hydrological models, which are often employed separately and at different spatial scales, either the plot or the basin. The calibration of the model to perform streamflow yielded a Nash-Sutcliffe (NS) coefficient equal to 0.89 for the year 2009, while the validation of the water balance partitioning was consistent with the independent measurements of actual evapotranspiration (R-2 = 0.79, energy balance closed independently), soil water content (R-2 = 0.35) and water table level (R-2 = 0.84). Eight months of data from 2010 were used to validate modelled streamflow, resulting in a NS = 0.75. An uncertainty analysis showed that the streamflow modelling was precise for nearly every time step, while a sensitivity analysis revealed which parameters mostly affected model precision, depending on the season. It was observed that 64% of the incident rainfall R flowed out of the basin as streamflow and 25% as evapotranspiration, while the remaining 11% is probably explained by deep percolation, measurement errors and/or inter-annual changes in soil and aquifer water stocks. The model indicated an interception loss equal to 4% of R, a surface runoff of 4% and an infiltration component of 92%. The modelled streamflow was constituted by 87% of baseflow originating from the aquifer, 7% of subsurface non-saturated runoff and 6% of surface runoff. Given the low surface runoff observed under the current physical conditions (andisol) and management practices (no tillage, planted trees, bare soil kept by weeding), this agroforestry system on a volcanic soil demonstrated potential to provide valuable HES, such as a reduced superficial displacement- capacity for fertilizers, pesticides and sediments, as well as a streamflow regulation function provided by the highly efficient mechanisms of aquifer recharge and discharge. The proposed combination of experimentation and modelling across ecophysiological and hydrological approaches proved to be useful to account for the behaviour of a given basin, so that it can be applied to compare HES provision for different regions or management alternatives

A narrow, edge-on disk resolved around HD 106906 with SPHERE

HD~106906AB is so far the only young binary system around which a planet has been imaged and a debris disk evidenced thanks to a strong IR excess. As such, it represents a unique opportunity to study the dynamics of young planetary systems. We aim at further investigating the close (tens of au scales) environment of the HD~106906AB system. We used the extreme AO fed, high contrast imager SPHERE recently installed on the VLT to observe HD~106906. Both the IRDIS imager and the Integral Field Spectrometer were used. We discovered a very inclined, ring-like disk at a distance of 65~au from the star. The disk shows a strong brightness asymmetry with respect to its semi-major axis. It shows a smooth outer edge, compatible with ejection of small grains by the stellar radiation pressure. We show furthermore that the planet's projected position is significantly above the disk's PA. Given the determined disk inclination, it is not excluded though that the planet could still orbit within the disk plane if at a large separation (2000--3000 au). We identified several additional point sources in the SPHERE/IRDIS field-of-view, that appear to be background objects. We compare this system with other debris disks sharing similarities, and we briefly discuss the present results in the framework of dynamical evolution.Comment: 7 pages, 6 figures, accepted by Astronomy & Astrophysic

Ensemble Modeling of the Likely Public Health Impact of a Pre-Erythrocytic Malaria Vaccine

Using an ensemble modeling approach, Thomas Smith and colleagues find that targeted mass vaccination with a pre-erythrocytic malaria vaccine RTS,S in low-transmission settings might have better health effects than vaccination through national EPI programs