Purification and crystallization of complexes modeling the active state of the fragile histidine triad protein

Fragile histidine triad protein (Fhit) is a diadenosine triphosphate (ApppA) hydrolase encoded at the human chromosome 3 fragile site which is frequently disrupted in tumors. Reintroduction of FHIT coding sequences to cancer cell lines with FHIT deletions suppressed the ability of these cell lines to form tumors in nude mice even when the reintroduced FHIT gene had been mutated to allow ApppA binding but not hydrolysis. Because this suggested that the tumor suppressor activity of Fhit protein depends on substrate-dependent signaling rather than ApppA catabolism, we prepared two crystalline forms of Fhit protein that are expected to model its biologically active, substrate-bound state. Wild-type and the His96Asn forms of Fhit were overexpressed in Escherichia coli, purified to homogeneity and crystallized in the presence and absence of ApppA and an ApppA analog. Single crystals obtained by vapor diffusion against ammonium sulfate diffracted X-rays to beyond 2.75 A resolution. High quality native synchrotron X-ray data were collected for an orthorhombic and a hexagonal crystal form.      

Influence of moisture content on the specific methanogenic activity of dry mesophilic municipal solid waste digestate

The objective of this study was to evaluate the influence of moisture content on the specific methanogenic activity (SMA) of a fresh dry mesophilic digestate from a municipal solid waste digester plant. For this purpose, SMA tests were performed under mesophilic conditions in 500 mL glass bottles of volume used as batch reactors, during a period of 20–25 days. Cellulose, propionate and acetate were used as substrates (5 g_COD kg^?1 digestate) at four different moisture contents, ranging from 65 to 82%. The moisture content strongly influenced the specific methanogenic activity. The highest SMA values were observed at a moisture content of 82% (11.1, 7.8 and 6.0 mg_COD g_VS^?1 d^?1 for cellulose, propionate and acetate spikes, respectively). SMA and moisture content were found to be linearly linked. Dry digestion at low water content is thus detrimental to the biological activity, probably due to physical limitations. Copyright © 2011 Society of Chemical Industry     

Chemical modification of hemp fibers by silane coupling agents

Natural hemp fibers were chemically modified using silane coupling agents to reduce their hydrophilic character. The existence of a chemical bond between coupling agents and hemp fibers was confirmed by ATR‐FTIR spectroscopy, ²⁹Si Nuclear Magnetic Resonance (NMR), thermogravimetric analysis (TGA), energy dispersive spectroscopy (EDS), and BET surface area measurements. It was shown that the initial concentration and the chemical structure of the organosilane coupling agent have an effect on the grafted quantity on the hemp fiber surfaces. The grafted quantity increased proportionally to the initial concentration of silane molecules. The presence of polar amino end group (NH₂) in silane structure can cause an increase in the grafted quantity, compared with results obtained in the case of silane molecules containing methacryloxy groups. This effect is attributed to the formation of hydrogen bonds between NH₂ and unreacted hydroxyl groups of hemp fibers. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Evaluation of <Emphasis Type="Italic">Rhizopus oryzae</Emphasis> Lipase for the Determination of Regiodistribution in Triacylglycerols with Medium Chain Fatty Acids

The nutritional profile and rheological behaviors of lipids is both due to fatty acid composition and regiodistribution on external and internal positions of triacylglycerol. Actual methods for regiodistribution analysis having some restrictions, there is still a need for investigating a safe, simple and environmentally friendly method for the sn-2 position analysis that could especially be used for the analysis of fats containing medium and short chain fatty acids. The objective of this study was to evaluate the 1,3-selectivity and typoselectivity of Rhizopus oryzae lipase in the presence of short/medium chain fatty acids in partial hydrolysis conditions used for regiodistribution analysis. Structured triacylglycerols containing eight-carbon-chain length fatty acids in the sn -2 position were chemically synthesized using DCC/DMAP coupling agent and purification steps by flash-chromatography. The final product showed very high purity and was used as the substrate for 1,3-selectivity evaluation. Typoselectivity was assessed by investigating partial hydrolysis of equimolar blends of homogeneous TAG. This study confirmed the 1,3-selectivity of Rhizopus oryzae lipase in the hydrolysis conditions used, and revealed that this lipase was less influenced by fatty acids chain length than pancreatic lipase. Considering this, Rhizopus oryzae lipase appeared to be a good candidate for regiodistribution analysis of fats containing medium and short chain fatty acids.     

Plastic deformation of dense nanocrystalline yttrium oxide at elevated temperatures

Nanocrystalline yttrium oxide, Y₂O₃ with 110 nm average grain size was plastically deformed between 800 °C and 1100 °C by compression at different strain rates and by creep at different stresses. The onset temperature for plasticity was at 1000 °C. Yield stress was strongly temperature dependent and the strain hardening disappeared at 1100 °C. The polyhedral and equiaxed grain morphology were preserved in the deformed specimens. The experimentally measured and theoretically calculated stress exponent n = 2 was consistent with the plastic deformation by grain boundary sliding. Decrease in the grain size was consistent with decrease in the brittle to ductile transition temperature.     

Structure-related behavior of hybrid organic–inorganic materials prepared in different synthesis conditions from Zr-based NBBs and 3-methacryloxypropyl trimethoxysilane

The copolymerization of zirconium oxo-clusters (Zr12) with 3-methacryloxypropyl (trimethoxy)silane (MPTMS), using a Si/Zr molar ratio of 4, was investigated. The hybrid samples were prepared both with and without organosilane prehydrolysis. Differential scanning calorimetry (DSC), multinuclear liquid, and solid state NMR analyses and Fourier transform infrared (FTIR) spectroscopy were used to characterize the hybrid materials and study the influence of synthesis conditions on condensation and polymerization. The degree of condensation (DOC) of the silsesquioxane network and the polymerization yield are generally high. However, the organosilane prehydrolysis step leads to the reduction of the extent of phase interaction, thus favoring the phase separation between silica-zirconia-based domains. Dynamic mechanical spectroscopy (DMS) analyses were performed on the hybrid polymers obtained by means of the two synthetic pathways. The sample prepared without the organosilane prehydrolysis step presents a higher glass transition temperature (T_g) than the one with silane prehydrolysis. By heating above the T_g, the samples retain shape and size, due to the lack of viscous flow. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

Numerical and experimental investigations of mixing in T-shaped and cross-shaped micromixers

Mass transfer within the T-shaped and cross-shaped micromixers has been studied using CFD and confocal laser scanning microscopy methods. The concentration profiles, based on flow regimes, were used to compare the T- and cross-geometries. The cross-shaped micromixer tends to intensify the mixing and this is occurring for lower flow rates in comparison to the T shape. The improvement made by the cross geometry is attributed to the stronger vortex stretching and high shear rate, which reduces the liquid transfer length. The presence of a single outlet in the T-shaped micromixer induces a smaller degree of freedom for the fluid. A higher pressure drop is calculated in T-shaped micromixer than in cross-shaped micromixer.     

Design and optimization of electrochemical microreactors for continuous electrosynthesis

The study focuses on the design and construction, as well as the theoretical and experimental optimization of electrochemical filter press microreactors for the electrosynthesis of molecules with a high added value. The main characteristics of these devices are firstly a high-specific electrochemical area to increase conversion and selectivity, and secondly the shape and size of the microchannels designed for a uniform residence time distribution of the fluid. A heat exchanger is integrated into the microstructured electrode to rapidly remove (or supply) the heat required in exo- or endothermic reactions. The microreactors designed are used to perform-specific electrosynthesis reactions such as thermodynamically unfavorable reactions (continuous NADH regeneration), or reactions with high enthalpy changes.     

Production of 2,3-butanediol from diverse saccharides via fermentation

The present study aimed to evaluate the potential use of whey to produce 2,3-BD via the fermentation of lactose and its monosaccharides, glucose and galactose, in a synthetic culture medium (medium 9, M9) using a modified strain of Escherichia coli K12 MG1655 (E. coli JFR12) at a 0.1 L/L (10 vol%) inoculum ratio, 37 °C, atmospheric pressure, an initial pH 7.4, and 100 rpm for 72 h varying the saccharide concentration from 12.5, 25, and 50 g/L. The 2,3-BD yield was ∼80 % of the theoretical yield using 25 g/L of glucose and lactose, corresponding to 0.38 g/g saccharides at a fermentation time of 48 h (glucose) and 72 h (lactose). However, the 2,3-BD yield was halved (0.19 g/g galactose), fermenting 25 g/L of galactose at 48 h. Taking into account these results, two important conclusions were determined: i) E. coli JFR12 could transform galactose into 2,3-BD although its yield was half of the yield observed with glucose at 48 h; and ii) E. coli JFR12 was as efficient as other natural 2,3-BD producers such as Klebsiella species fermenting lactose. However, the E. coli strain has the advantage of being an innocuous strain. To the best of our knowledge, there is no other study presenting the production of 2,3-BD from galactose and lactose with a genetically modified E. coli strain.     

Synthesis and Anticancer Properties of Oxazepines Related to Azaisoerianin and IsoCoQuines

In this article, we report the synthesis and biological properties of a series of novel oxazepines related to isoCA-4 having significant antitumor properties. Among them, three oxazepin-9-ol derivatives display a nanomolar or a sub-nanomolar cytotoxicity level against five human cancer cell lines (HCT116, U87, A549, MCF7, and K562). It was demonstrated that the lead compound in this series inhibits tubulin assembly with an IC₅₀ value of 1 μM and totally arrests the cellular cycle in the G2/M phase at the low concentration of 5 nM in HCT116 and K562 cells. Molecular modeling studies perfectly corroborates these promising results.