Large-scale in vivo synthesis of the carbohydrate moieties of gangliosides GM1 and GM2 by metabolically engineered Escherichia coli

Two metabolically engineered Escherichia coli strains have been constructed to produce the carbohydrate moieties of gangliosides GM2 (GalNAcbeta-4(NeuAcalpha-3)Galbeta-4Glc; Gal = galactose, Glc = glucose, Ac = acetyl) and GM1 (Galbeta-3GalNAcbeta-4(NeuAcalpha-3)Galbeta-4Glc. The GM2 oligosaccharide-producing strain TA02 was devoid of both beta-galactosidase and sialic acid aldolase activities and overexpressed the genes for CMP-NeuAc synthase (CMP = cytidine monophosphate), alpha-2,3-sialyltransferase, UDP-GlcNAc (UDP = uridine diphosphate) C4 epimerase, and beta-1,4-GalNAc transferase. When this strain was cultivated on glycerol, exogenously added lactose and sialic acid were shown to be actively internalized into the cytoplasm and converted into GM2 oligosaccharide. The in vivo synthesis of GM1 oligosaccharide was achieved by taking a similar approach but using strain TA05, which additionally overexpressed the gene for beta-1,3-galactosyltransferase. In high-cell-density cultures, the production yields for the GM2 and GM1 oligosaccharides were 1.25 g L(-1) and 0.89 g L(-1), respectively.     

Aortic aneurysms. Technique--indications--results. 1958]

Cardiomyocytes subjected to brief episode of hypoxia possess a resistance to serious damaging effect exerted by a subsequent long-time hypoxia on these cells, which is called hypoxic preconditioning (PC). The pathway of intracellular signal transduction during hypoxia PC has not yet been validated. On a model of hypoxia/reoxygenation (H/R) of cultured neonatal rabbit cardiomyocytes, the present study is taken to investigate the changes of mitogen-activated protein kinase (MAPK) and ribosomal S6 kinase (S6K) activity. It was found that intracellular total MAPK and nuclear MAPK, after a 15-min period of reoxygenation preceded by a single 60-min period of hypoxia, were increased by 95% and 230%, respectively. Intracellular S6K activity increased by 142% at 30 min of H/R vs the control group (P < 0.01). Phosphatase 1 (PPase 1) inhibitor (ocadaic acid, OA 1 mumol/L) augmented the increase of MAPK and S6K activity induced by H/R. However, tyrosine kinase (Tyr K) inhibitor (genistein), protein kinase C (PKC) inhibitor (H7) and preincubation of cardiomyocytes with PKC activator PMA all reduced MAPK activation by H/R. Protein kinase A (PKA) inhibitor (H89), Ca2+/Calmodulin-dependent protein kinase (PKM) inhibitor (W7) or PPase 2a inhibitor (OA 10 nmol/L) had no significant effect on MAPK and S6K activity. The above results suggested that activation of MAPK and S6K activity during hypoxia/reoxygenation there might require participation of PKC, Tyr K and PPase 1, while PKA, PKM and PPase 2a were not involved.     

Theoretical and numerical study of strain rate influence on AA5083 formability

With the application of new forming techniques (hydroforming, incremental forming), it is necessary to improve the characterization of the formability of materials and in particular the influence of strain rate. This paper begins with the characterization of material behavior of an aluminum alloy 5083 at high temperatures. To describe its visco-plastic behavior, Swift’s hardening law is used and the corresponding parameter values are identified. Then, two different approaches are introduced to construct FLDs (forming limit diagrams) of this alloy sheet and evaluate the effect of the rate sensitivity index on its formability. The first one is theoretical (the M-K model), and an algorithm is developed to calculate the limit strains by this model. In the second approach, the Marciniak test is simulated with the commercially available finite-element program ABAQUS. Based on FEM results, different failure criteria are discussed and an appropriate one is chosen to determine the onset of localized necking. With the material behavior data corresponding to AA5083 at 150 °C, parametric studies are carried out to evaluate the effect of the strain rate sensitivity index. The comparison of results by these two approaches shows the same tendency that an improvement of the formability with increasing strain rate sensitivity is observed. Finally, by consideration of the compensating effects of the strain hardening and rate sensitivity indices, the FLDs of this sheet at 150, 240 and 300 °C are determined and compared. Results show that the formability of AA5083 seems not to be improved up to a certain temperature (between 240 and 300 °C), above this temperature, the formability is greatly enhanced.     

Path‐following algorithms and experiments for an unmanned surface vehicle

This paper addresses the problem of path following in two‐dimensional space for underactuated unmanned surface vehicles (USVs), defining a set of guidance laws at the kinematic level. The proposed nonlinear Lyapunov‐based control law yields convergence of the path‐following error coordinates to zero. Furthermore, the introduction of a virtual controlled degree of freedom for the target to be followed on the path removes singularity behaviors present in other guidance algorithms proposed in the literature. Some heuristic approaches are then proposed to face the problem of speed of advance adaptation based on path curvature measurement and steering action prediction. Finally a set of experimental results of all the proposed guidance laws, carried out with the Charlie USV, demonstrates the feasibility of the proposed approach and the performance improvements, in terms of precision in following the reference path and transient reduction, obtained by introducing speed adaptation heuristics. © 2009 Wiley Periodicals, Inc.     

Mechanistic studies of the electropolymerization of 2,2′-bithiophene and of pyrrole to form conducting polymers

The electropolymerizations of 2,2′-bithiophene and of pyrrole to form conducting polymers were investigated by several electrochemical methods. Cyclic voltammetry indicates that a peak at 1.1 V is associated with the polymer and that the same product is formed from thiophene as from 2,2′-bithiophene. The rate of polymer formation has a zero kinetic order dependence on monomer concentration for polymerization of pyrrole or 2,2′-bithiophene. A dependence on concentration of supporting electrolyte is interpreted as a consequence of anion diffusion and oxidative doping in the polymer. The kinetic results are consistent with an oxidative coupling mechanism for polymer formation. The electrochromic switching of the polymer film was investigated by spectroelectrochemical methods and the kinetics were examined over a range of film thicknesses.     

Carbon–polymer composites with extreme electrical conductivity

The aim of this study was to develop Carbon–polymer composites with extreme electrical conductivity (100 S/cm) combined with good flexural strength. Despite the many optimization methods described in the literature, no comprehensive optimization procedure was to be found because the formulation did not control by itself the final properties. This study showed the major influence of the processing conditions with these peculiar materials. A detailed study of the influence of the processing conditions on the microstructural and macroscopic properties was performed. We, thereby, proposed a comprehensive way to optimize the properties of the final product. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42274.     

On the Pre-oxidation Treatments of Four Commercial Ni-Based Superalloys in Air and in Ar–H<Subscript>2</Subscript>O at 950 °C

The short-term oxidation behaviour of RA 602CA, Inconel 693, Manaurite 40XO and Sumitomo 696, which are four alloys recommended for hydrocarbon processing, was studied both in air and in Ar–H₂O to determine the conditions of pre-oxidation treatments. Regardless of the considered material, the oxidation rate at 950 °C was systematically higher in Ar–H₂O than in dry air. Surface examination of the Al-containing alloys indicated that they were not uniformly oxidized all over the surface. All Al-containing alloys (from 1.6 to 3.2% wt) formed an external protective alumina scale and behaved as alumina-forming alloys in dry air at 950 °C. In contrast, these alloys developed a rate-controlling chromia scale and severe internal oxidation in a H₂O-containing atmosphere. Compared with their oxidation behaviour in air + H₂O, the phenomenon was significantly enhanced in the atmosphere that coupled water vapour with a low oxygen pressure. Consequently, the Al-containing alloys should not be pre-oxidized in a water vapour atmosphere prior to long exposure to a corrosive atmosphere. In contrast, the chromia-forming Al-free 696 alloy exhibited identical oxidation behaviour in both atmospheres, demonstrating that the degradation of this alloy was not significantly affected by water vapour.     

Autonomic Self‐Repairing Glassy Materials

A new process that enables glassy materials to self‐repair from mechanical damage is presented in this paper. Contrary to intrinsic self‐healing, which involves overheating to enable crack healing by glass softening, this process is based on an extrinsic effect produced by vanadium boride (VB) particles dispersed within the glass matrix. Self‐repair is obtained through the oxidation of VB particles, and thus without the need to increase the operating temperature. The VB healing agent is selected for its capacity to oxidize at a lower temperature than the softening point of the glass. Thermogravimetric analyses indeed show that VB oxidation is rapid and occurs below the glass transition temperature. Solid‐state nuclear magnetic resonance spectroscopy indicates that VB is oxidized into V₂O₅ and B₂O₃, which enable the local formation of glass. The autonomic self‐healing effect is demonstrated by an in situ experiment visualized using an environmental scanning electron microscope. It is shown that a crack could be healed by the VB oxidation products.     

Additional vectors for PCR-based gene tagging in Saccharomyces cerevisiae and Schizosaccharomyces pombe using nourseothricin resistance

The one-step PCR-mediated technique used for modification of chromosomal loci is a powerful tool for functional analysis in yeast. Both Saccharomyces cerevisiae and Schizosaccharomyces pombe are amenable to this technique. However, the scarce availability of selectable markers for Sz. pombe hampers the easy use of this technique in this species. Here, we describe the construction of new vectors deriving from the pFA6a family, which are suitable for tagging in both yeasts owing to the presence of a nourseothricin-resistance cassette. These plasmids allow various gene manipulations at chromosomal loci, viz. N- and C-terminal tagging with 3HA (haemagglutinin) or 13Myc epitopes, GST (glutathione S-transferase), 4TAP (tandem affinity purification) and several GFP (green fluorescent protein) isoforms. For N-terminal modifications, the use of different promoters allows constitutive (PADH1) or regulatable (PGAL1) promoters for S. cerevisiae and derivatives of Pnmt1 for Sz. pombe expression.