Purification of tubulin from limited volumes of cultured cells.

International audienceA method was designed to purify tubulin from limited volumes of cultured cells, which can be performed in less than 4 h. The method is based on the preservation of intact microtubule arrays during cell lysis in a large volume of buffer, followed by disassembly of microtubules in a small volume of cold buffer. This allows a good enrichment in tubulin, which is then purified by one cycle of polymerisation/depolymerisation and a cation exchange chromatography. Such a procedure has been employed successfully on suspension-cultured and on adherent HeLa cells. Tubulin obtained was 90% pure, assembly-competent and composed of alpha/beta I and alpha/beta IV isotypes. Microtubules made with this tubulin displayed specific properties such as resistance to dilution, maybe related to their specific dynamic behaviour

Separation and determination of some carboxylic acids by capillary electrophoresis

RADIOCHSeparation and determination of some organic acids, mono-carboxylic (formic and acetic), dicarboxylic (oxalic and tartaric), tricarboxylic (citric) acids and aromatic acids (phtalic, benzoic, mellitic and trimellitic), by capillary electrophoresis are reviewed. The method development parameters, such as separation and injection mode, are discussed. Special attention is paid to the comparison of different detection types (spectroscopic and electrochemical). The optimisation of the carrier electrolyte composition (choice of carrier electrolyte, effect of pH, ionic strength, electro-osmotic flow modifier) is treated. Different additives (alkali-earth and transition metal ions, cyclodextrins and alcohol), which are often used for improving organic acid separation, are also considered

Comparative approach of the solubility of protactinium oxy/hydroxides

RADIOCHThe hydrolysis constants and the solubility products of the oxide/hydroxides of Pa(IV) and Pa(V) have been first reviewed and re-evaluated through the application of three comparative methods: the simple Hard Sphere Electrostatic model (HSE), the Brown and Wanner Theory (BWT) and the Correlation between Solubility and Complex stability constants of a metal ion (CSC). The latter method is the most empirical one, but it gives coherent results, in agreement with the few published experimental values, for all the possible aqua species of protactinium (Pa4+, Pa5+, PaO2+ and PaO3+). The selected thermodynamic data have then been used to predict the protactinium solubility in non complexing media of various pH and redox potential

Development and validation of a composite material law for crash simulations

International audienceMore and more structural parts are building using organic matrix composite materials in order to reduce the weight of aircraft structures. In order to reduce the number of tests required at the different scale of the building block approach, an increase of the knowledge regarding strain rate dependency of such materials is still a topic of interest. Particularly most of the strain rate dependent material laws are based on or validated with tests performed on split Hopkinson bars [1,2]. Few results are available in the literature concerning the intermediate strain rate, typically ranging between 10-3 s-1 and 100 s-1. Consequently, this study is first dedicated to the experimental characterisation of the rate effect for this specific range of strain rate. For that purpose, tensile tests have been performed on a servo-hydraulic jack on off-axis specimens. Various off-axis angles have been considered: 15°, 30°, 45°, 60°, 75° and 90°. The geometry of the coupon for each angle has been defined thanks to a numerical study in order to reduce stress concentration near to the tabs and maximise the stress field homogeneity within the specimens. Three different loading speeds with at least three specimens were tested in order to evaluate discrepancy. A Photron SA-X high speed camera has been used to evaluate strain using a Digital Image Correlation software (Aramis GOM). Moreover, as multiple failure locations are observed for high loading rate tests, high speed imaging allow the localisation of the first failure.These results have been used in order to include rate dependency in an advanced material law initially proposed by ONERA for quasi-static loadings. This model written at the ply scale is based on a visco-elastic behaviour law until a ply failure criterion is reached. A Hashin like failure criterion is used and two failure modes are considered: fibre failure and matrix failure. The degradation of the ply properties is obtained with 2 damage variables: d1 for fibre failure and d2 for matrix failure. Improvements regarding rate dependency of this material law have been studied for its three main aspects: the visco-elastic behaviour, the failure criterion and the damage evolution law. Regarding the visco-elastic behaviour, previous works [3] have shown that a bi-spectral visco-elastic model is required in order to take into account the short relaxation times required to accurately described rate dependency for high strain rate tests and long relaxation times for low strain rate tests. The main drawback of this model is its computational cost due to the important number of viscous mechanisms that have to be considered. As full-scale crash simulation is the targeted application case, a modified version of this model has been proposed. It still has an accurate prediction of the different modulus evolutions with the loading rate but it has a reduce computational cost. Concerning the loading speed dependency of the failure criterion, dynamic off-axis tensile tests will be used as input data to analyse and improve the failure criterion. Finally, improvements of the different damage evolution laws will be based on results available in the literature [4]. Finally, this material law has been implemented in an explicit finite element code in order to validate it on more complex coupons

Lowering the applied potential during successive scratching/re-inoculation improves the performance of microbial anodes for microbial fuel cells

Microbial anodes were formed under polarisation at -0.2 V/SCE on smooth graphite plate electrodes with paper mill effluents. Primary, secondary and tertiary biofilms were formed by a successive scratching and re-inoculation procedure. The secondary and tertiary biofilms formed while decreasing the polarisation potential allowed the anodes to provide current density of 6 A/m² at -0.4 V/SCE. In contrast, applying -0.4 V/SCE initially to form the primary biofilms did not lead to the production of current. Consequently, the scratching/re-inoculation procedure combined with progressive lowering of the applied potential revealed an efficient new procedure that gave efficient microbial anodes able to work at low potential. The observed progressive pH drift to alkaline values above 9 explained the open circuit potentials as low as -0.6 V/SCE. The remarkable performance of the electrode at alkaline pH was attributed to the presence of Desulfuromonas acetexigens as the single dominant species in the tertiary microbial anodes

Lowering the applied potential during successive scratching/re-inoculation improves the performance of microbial anodes for microbial fuel cells

Microbial anodes were formed under polarisation at -0.2 V/SCE on smooth graphite plate electrodes with paper mill effluents. Primary, secondary and tertiary biofilms were formed by a successive scratching and re-inoculation procedure. The secondary and tertiary biofilms formed while decreasing the polarisation potential allowed the anodes to provide current density of 6 A/m² at -0.4 V/SCE. In contrast, applying -0.4 V/SCE initially to form the primary biofilms did not lead to the production of current. Consequently, the scratching/re-inoculation procedure combined with progressive lowering of the applied potential revealed an efficient new procedure that gave efficient microbial anodes able to work at low potential. The observed progressive pH drift to alkaline values above 9 explained the open circuit potentials as low as -0.6 V/SCE. The remarkable performance of the electrode at alkaline pH was attributed to the presence of Desulfuromonas acetexigens as the single dominant species in the tertiary microbial anodes

The Perils of Fundamentalism in France Interview with Caroline Fourest by Stephen Cowden

Caroline Fourest is a French feminist writer, film director, journalist, radio presenter at France Culture, and co-founder and editor of the magazine ProChoix. She has been a columnist with Charlie Hebdo and Le Monde and has written several influential essays on the political and religious right in France and the US. She is the author of a biography of the far-right politician Marine Le Pen and of a number of books including In Praise of Blasphemy: Why Charlie Hebdo is Not ‘Islamophobic’, Brother Tariq: the Doublespeak of Tariq Ramadan and most recently The Offended Generation

STOP Proteins are Responsible for the High Degree of Microtubule Stabilization Observed in Neuronal Cells

Neuronal differentiation and function require extensive stabilization of the microtubule cytoskeleton. Neurons contain a large proportion of microtubules that resist the cold and depolymerizing drugs and exhibit slow subunit turnover. The origin of this stabilization is unclear. Here we have examined the role of STOP, a calmodulin-regulated protein previously isolated from cold-stable brain microtubules. We find that neuronal cells express increasing levels of STOP and of STOP variants during differentiation. These STOP proteins are associated with a large proportion of microtubules in neuronal cells, and are concentrated on cold-stable, drug-resistant, and long-lived polymers. STOP inhibition abolishes microtubule cold and drug stability in established neurites and impairs neurite formation. Thus, STOP proteins are responsible for microtubule stabilization in neurons, and are apparently required for normal neurite formation

Experimental and theoretical characterization of microbial bioanodes formed in pulp and paper mill effluent in electrochemically controlled conditions

Microbial bioanodes were formed in pulp and paper effluent on graphite plate electrodes under constant polarization at -0.3 V/SCE, without any addition of nutriment or substrate. The bioanodes were characterized in 3-electrode set-ups, in continuous mode, with hydraulic retention times from 6 to 48 h and inlet COD from 500 to 5200 mg/L. Current densities around 4 A/m2 were obtained and voltammetry curves indicated that 6 A/m2 could be reached at +0.1 V/SCE. A theoretical model was designed, which allowed the effects of HRT and COD to be distinguished in the complex experimental data obtained with concomitant variations of the two parameters. COD removal due to the electrochemical process was proportional to the hydraulic retention time and obeyed a Michaelis–Menten law with respect to the COD of the outlet flow, with a Michaelis constant KCOD of 400 mg/L. An inhibition effect occurred above inlet COD of around 3000 mg/L