Thermal denaturation and aggregation of Myosin subfragment 1 isoforms with different essential light chains

We compared thermally induced denaturation and aggregation of two isoforms of the isolated myosin head (myosin subfragment 1, S1) containing different "essential" (or "alkali") light chains, A1 or A2. We applied differential scanning calorimetry (DSC) to investigate the domain structure of these two S1 isoforms. For this purpose, a special calorimetric approach was developed to analyze the DSC profiles of irreversibly denaturing multidomain proteins. Using this approach, we revealed two calorimetric domains in the S1 molecule, the more thermostable domain denaturing in two steps. Comparing the DSC data with temperature dependences of intrinsic fluorescence parameters and S1 ATPase inactivation, we have identified these two calorimetric domains as motor domain and regulatory domain of the myosin head, the motor domain being more thermostable. Some difference between the two S1 isoforms was only revealed by DSC in thermal denaturation of the regulatory domain. We also applied dynamic light scattering (DLS) to analyze the aggregation of S1 isoforms induced by their thermal denaturation. We have found no appreciable difference between these S1 isoforms in their aggregation properties under ionic strength conditions close to those in the muscle fiber (in the presence of 100 mM KCl). Under these conditions kinetics of this process was independent of protein concentration, and the aggregation rate was limited by irreversible denaturation of the S1 motor domain.     

From relative to absolute quantification of tryptic peptides with tandem mass tags: application to cerebrospinal fluid

Quantification is a major task in proteomics. Among the different analytical strategies to enable peptide and protein quantification, tagging with isotopic labels has emerged as a practical, versatile, and efficient alternative. In particular, isobaric labels, such as TMT or iTRAQ, are now widely employed to make relative comparison of the protein amounts in separate biological samples with tandem mass spectrometry (MS/MS). We used herein a shotgun proteomic approach based on labelling with tandem mass tags (TMTs) for the relative quantification of proteins, and the absolute quantification of their tryptic peptides in human cerebrospinal fluid (CSF). First, the comparison of ante- and post-mortem CSF samples was carried out for the discovery of protein marker candidates of brain-damage disorders. Second, tryptic peptides representative of these candidates were measured in CSF using reporter-ion calibration curves. These works highlighted the advantages and limitations of such strategies for quantification purposes in proteomics.     

Acyltransferase activities in the yolk sac membrane of the chick embryo

The activities of some enzymes of glycerolipid synthesis and fatty acid oxidation were measured in subcellular fractions of the yolk sac membrane (YSM), an extra-embryonic tissue that mediates the transfer of lipid from the yolk to the circulation of the chick embryo. The activities of monoacylglycerol acyltransferase and carnitine palmitoyl transferase-1 in the YSM (respectively, 284.8±13.2 nmol/min/mg microsomal protein and 145.6±9.1 nmol/min/mg mitochondrial protein; mean ±SE; n=4) at day 12 of development appear to be the highest yet reported for any animal tissue. Also, the carnitine palmitoyl transferase-1 of the YSM was very insensitive to inhibition by malonyl CoA. The maximal activities of glycerol-3-phosphate acyltransferase and diacylglycerol acyltransferase in the YSM (respectively, 26.7±2.2 and 36.1±2.1 nmol/min/mg microsomal protein) were also high compared with the reported values for various animal tissues. The very high enzymic capacity for glycerolipid synthesis supports the hypothesis that the yolk-derived lipids are subjected to hydrolysis followed by reesterification during transit across the YSM. The monoacylglycerol pathway appears to be the main route for glycerolipid resynthesis in the YSM. The results also suggest that the YSM has the capacity to perform simultaneously β-oxidation at a high rate in order to provide energy for the lipid transfer process.     

Ascorbic Acid-Assisted Eco-friendly Synthesis of NiCo<Subscript>2</Subscript>O<Subscript>4</Subscript> Nanoparticles as an Anode Material for High-Performance Lithium-Ion Batteries

We have synthesized NiCo₂O₄ nanoparticles (NCO NPs) using an ascorbic acid-assisted co-precipitation method for the first time. When NCO NPs are used as an anode material for lithium-ion batteries, the cell exhibits superior lithium storage properties, such as high capacity (700 mA h g^?1 after 300 cycles at 200 mA g^?1), excellent rate capabilities (applied current density range 100–1200 mA g^?1), and impressive cycling stability (at 1200 mA g^?1 up to 650 cycles). The enhanced electrochemical properties of NCO NPs are due to the nanometer dimensions which not only offers a smooth charge-transport pathway and short diffusion paths of the lithium ions but also adequate spaces for volume expansion during Li storage. Hence, this eco-friendly synthesis approach will provide a new strategy for the synthesis of various nanostructured metal oxide compounds, for energy conversion and storage systems applications.     

Piezoelectric properties and phase transition temperatures of the solid solution of (1 − x)(Bi0.5Na0.5)TiO3–xSrTiO3

The phase diagram of (1 ? x)(Bi_0.5Na_0.5)TiO₃–xSrTiO₃ was completed and investigations on polarization and strain in this system were carried out. (1 ? x)(Bi_0.5Na_0.5)TiO₃–xSrTiO₃-ceramics were prepared by conventional mixed oxide processing. The depolarization temperature (T_d), the temperature of the rhombohedral–tetragonal phase transition (T_r–t) and the Curie temperature (T_m) were determined by measuring the temperature dependence of the relative permittivity. All solid solutions of (1 ? x)(Bi_0.5Na_0.5)TiO₃–xSrTiO₃ show relaxor behavior (A-site relaxor). From XRD-measurements a broad maximum of the lattice parameter can be observed around x = 0.5 but no structural evidence for a morphotropic phase boundary was found. SEM-analysis revealed a decrease of the grain size for increasing SrTiO₃-content. At room temperature a maximum of strain of about 0.29% was found at x = 0.25 which coincides with a transition from a ferroelectric to an antiferroelectric phase. The temperature dependence of the displacement indicates an additional contribution from a structural transition (rhombohedral–tetragonal), which would be of certain relevance for the existence of a morphotropic phase boundary.     

Squeezing-to-dripping transition for bubble formation in a microfluidic T-junction

The aim of this paper is to investigate the squeezing-to-dripping transition for bubble formation in a microfluidic T-junction by cross-flowing rupture technique using a high-speed digital camera. Experiments were conducted in a glass microfluidic T-junction with the cross-section of the microchannel of 120 μm wide and 40 μm deep. N₂ bubbles were generated in glycerol–water mixtures with several concentrations of surfactant sodium dodecyl sulfate (SDS). Three different regimes were identified for generating different kinds of bubbles: squeezing, dripping and transition regimes. Various forces exerted on the gaseous thread in different regimes were analyzed. Long slug bubbles were formed in the squeezing regime, while dispersed bubbles in the dripping regime. The transition regime formed short slug bubbles. The bubble sizes in various regimes could be correlated with several dimensionless numbers such as the ratio of gas/liquid flow rates and capillary number. The two-step model for droplets (Steegmans et al., 2009) was extended to describe the bubble formation.     

Sunflower oil coating on the nanoparticles of iron(III) oxides

High-temperature solution phase reaction of iron(III)-furoate in sunflower oil, in the presence of trichloroacetic acid and 1,2-hexadecylamine leads to iron(III) oxide (amorphous and γ-Fe2O3) nanoparticles with sizes about 1.3 nm in diameter (method “A”), and about 5.5 nm (amorphous Fe2O3) from the “sunflower oil/oxadiazol” system (method “B”).     

Influence of various process conditions on surface finishes induced by the direct metal deposition laser technique on a Ti–6Al–4V alloy

The direct metal deposition (DMD) with laser is a free-form metal deposition process for manufacturing dense pieces, which allows generating a prototype or small series of near net-shape structures. One of the most critical issues is that produced pieces have a deleterious surface finish which systematically requires post machining steps. This problem has never been fully addressed before.The present work describes investigations on the DMD process, using an Yb-YAG disk laser, and a widely used titanium alloy (Ti–6Al–4V) to understand the influence of the main process parameters on the surface finish quality. The focus of our work was: (1) to understand the physical mechanisms responsible for deleterious surface finishes, (2) to propose different experimental solutions for improving surface finish.In order to understand the physical mechanisms responsible for deleterious surface finishes, we have carried out: (1) a precise characterization of the laser beam and the powder stream; (2) a large number of multi-layered walls using different process parameters (P(W), V(m/min), D_m (g/min), Gaussian or uniform beam distribution); (3) a real time fast camera analysis of melt pool dynamics and melt-pool – powder stream coupling; (4) a characterization of wall morphologies versus process parameters using 2D and 3D profilometry.The results confirm that surface degradation depends on two distinct aspects: the sticking of non-melted or partially melted particles on the free surfaces, and the formation of menisci with more or less pronounced curvature radii. Among other aspects, a reduction of layer thickness and an increase of melt-pool volumes to favor re-melting processes are shown to have a beneficial effect on roughness parameters. Last, a simple analytical model was proposed to correlate melt-pool geometries to resulting surface finishes.     

Improved evidence for the existence of an intermediate phase during hydration of tricalcium silicate

Tricalcium silicate (Ca₃SiO₅) with a very small particle size of approximately 50 nm has been prepared and hydrated for a very short time (5 min) by two different modes in a paste experiment, using a water/solid-ratio of 1.20, and by hydration as a suspension employing a water/solid-ratio of 4000. A phase containing uncondensed silicate monomers close to hydrogen atoms (either hydroxyl groups or water molecules) was formed in both experiments. This phase is distinct from anhydrous tricalcium silicate and from the calcium-silicate-hydrate (C-S-H) phase, commonly identified as the hydration product of tricalcium silicate. In the paste experiment, approximately 79% of silicon atoms were present in the hydrated phase containing silicate monomers as determined from ²⁹Si{¹H} CP/MAS NMR. This result is used to show that the hydrated silicate monomers are part of a separate phase and that they cannot be attributed to a hydroxylated surface of tricalcium silicate after contact with water. The phase containing hydrated silicate monomers is metastable with respect to the C-S-H phase since it transforms into the latter in a half saturated calcium hydroxide solution. These data is used to emphasize that the hydration of tricalcium silicate proceeds in two consecutive steps. In the first reaction, an intermediate phase containing hydrated silicate monomers is formed which is subsequently transformed into C-S-H as the final hydration product in the second step. The introduction of an intermediate phase in calculations of the early hydration of tricalcium silicate can explain the presence of the induction period. It is shown that heterogeneous nucleation on appropriate crystal surfaces is able to reduce the length of the induction period and thus to accelerate the reaction of tricalcium silicate with water.