Poly[(R)-3-hydroxybutyrate] formation in Escherichia coli from glucose through an enoyl-CoA hydratase-mediated pathway

In this study, a new metabolic pathway for the synthesis of poly[(R)-3-hydroxybutyrate] [P(3HB)] was constructed in a recombinant Escherichia coli strain that utilized forward and reverse reactions catalyzed by two substrate-specific enoyl-CoA hydratases, R-hydratase (PhaJ) and S-hydratase (FadB), to epimerize (S)-3HB-CoA to (R)-3HB-CoA via a crotonyl-CoA intermediate. The R-hydratase gene (phaJ(Ac)) from Aeromonas caviae was coexpressed with the PHA synthase gene (phaC(Re)) and 3-ketothiolase gene (phaA(Re)) from Ralstonia eutropha in fadR mutant E. coli strains (CAG18497 and LS5218), which had constitutive levels of the beta-oxidation multienzyme FadB(Ec). When grown on glucose as the sole carbon source, the cells accumulated P(3HB) up to an amount 6.5 wt% of the dry cell weight, whereas the control cells without phaJ(Ac) or fadR mutation accumulated significantly smaller amounts of P(3HB). These results suggest that PhaJ(Ac) and FadB(Ec) played an important role in supplying monomers for P(3HB) synthesis in the pathway. Furthermore, by using this pathway, a P(3HB)-concentration-dependent fluorescent staining screening technique was developed to rapidly identify cells that possess active R-hydratase.     

Dependence of property, cathode characteristics, thermodynamic stability, and average and local structures on heat-treatment condition for LiNi0.5Mn0.5O2 as a cathode active material for Li-ion battery

LiNi_0.5Mn_0.5O₂ was heat-treated under high oxygen-pressure and Ar-reducing conditions, and then the cathode properties, thermodynamic stability and average and local structures were investigated. From X-ray diffraction and ICP measurements, it was found that the pristine LiNi_0.5Mn_0.5O₂ had a single phase of the layered rock-salt structure although the Ni content was slightly rich compared with the nominal one. These characteristics were kept even after the heat-treatments. Charge–discharge cycle tests clarified that the cycle performance of LiNi_0.5Mn_0.5O₂ was improved by both the reducing and oxidizing treatments. From neutron diffraction and X-ray absorption fine structure measurements, the local distortion around the transition metal, especially Ni, was supposed to be one of the important factors to determine the cathode properties. It was also found that the sample with higher thermodynamic stability exhibited better capacity retention in the discharge–charge cycle tests.     

Protective effects of galacturonic acid-rich vinegar brewed from Japanese pear in a dextran sodium sulfate-induced acute colitis model

The effects of pear vinegar (PV), which was specially brewed for enhanced galacturonic acid content, on the DSS-induced ulcerative colitis (UC) mouse model were evaluated. PV improved clinical symptoms, colon inflammation, and histological tissue injury in the DSS-induced acute UC mouse model. Moreover, PV suppressed inflammation due to acute UC by suppressing the myeloperoxidase (MPO)-mediated activation of inflammatory cells such as leukocytes and decreasing the serum concentration of IL-6. Our results demonstrated the protective action of PV in the DSS-induced acute UC mouse model. On the other hand, commercial apple vinegar did not show a protective effect in the DSS-induced acute UC mouse model. Our findings indicate that PV may act as a new functional food for inflammatory bowel disease patients.     

Inhibitory effects of proadrenomedullin N-terminal 20 peptide on antidiuresis and norepinephrine overflow induced by stimulation of renal nerves in anesthetized dogs

In an earlier study, we demonstrated that chronic ethanol (EtOH) exposure down-regulated the cannabinoid receptors (CB1) in mouse brain synaptic plasma membrane. In the present study, we investigated the effect of chronic EtOH on the formation of anandamide (AnNH), an endogenous cannabimimetic compound, and its precursor N-arachidonoylphosphatidylethanolamine (N-ArPE) in SK-N-SH cells that were prelabeled with [3H]arachidonic acid. The results indicate that exposure of SK-N-SH cells to EtOH (100 mM) for 72 h significantly increased levels of [3H]AnNH and [3H]N-ArPE (p < 0.05) (1.43-fold for [3H]AnNH and 1.65-fold for [3H]N-ArPE). Exposure of SK-N-SH cells to EtOH (100 mM, 24 h) inhibited initially the formation of [3H]AnNH at 24 h, followed by a progressive increase, reaching a statistical significance level at 72 h (p < 0.05). [3H]N-ArPE increased gradually to a statistically significant level after 48 and 72 h (p < 0.05). Incubation with exogenous ethanolamine (7 mM) and EtOH (100 mM, 72 h) did not result in an additive increase in the formation of [3H]AnNH. The formation of [3H]AnNH and [3H]N-ArPE by EtOH was enhanced by the Ca2+ ionophore A23187 or by the depolarizing agent veratridine and the K+ channel blocker 4-aminopyridine. Further, the EtOH-induced formation of [3H]AnNH and [3H]N-ArPE was inhibited by exogenous AnNH, whereas only [3H]AnNH formation was inhibited by the CB1 receptor antagonist SR141716A and pertussis toxin, suggesting that the CB1 receptor and G(i/o) protein mediated the regulation of AnNH levels. The observed increase in the levels of these lipids in SK-N-SH cells may be a mechanism for neuronal adaptation and may serve as a compensatory mechanism to counteract the continuous presence of EtOH. The present observation taken together with our previous results indicate the involvement of the endocannabinoid system in mediating some of the pharmacological actions of EtOH and may constitute part of a common brain pathway mediating reinforcement of drugs of abuse including EtOH.     

Thermodynamic assessment of systems of actinide or rare earth with Cd

A pyrometallurgical process is being developed for recycling nuclear reactor fuels. Thermodynamic information on multicomponent systems of actinides and rare earths (REs) with liquid Cd is very useful in the design of a process in which a liquid Cd electrode is used for the selective recovery of Pu and U, and a reductive extraction process using a molten salt/liquid Cd system for the recovery of minor actinides, such as Np, Pu, etc. A key issue in the design of these processes is a variation in solubility or activity of actinides or REs in multielement systems. In the present study, phase diagrams of U-Cd, Pu-Cd, Np-Cd, Y-Cd, La-Cd, Ce-Cd, Pr-Nd, Nd-Cd, and Gd-Cd were optimized by the CALPHAD method. For these systems, thermodynamic data, such as the activity of solutes in liquid Cd and the Gibbs energies of formation of the intermetallic compounds as well as the phase diagram data were available for the optimization. For optimization, the calculated primary results were entered into a database. Then, some ternary systems were preliminarily assessed through the use of the optimized data for the binary systems. Two extreme conditions were assumed: one condition was complete miscibility between the compounds that have the same mole ratio between solutes and Cd; the other condition was no solid solubility between the compounds. The results indicated the tendencies toward solubility and activity of actinides and REs in multielement systems.     

Polymerization of β-lactoglobulin and bovine serum albumin at oil-water interfaces in emulsions by transglutaminase

Polymerization of β-lactoglobulin and bovine serum albumin at the oil—water interfaces in n-tetradecane-in-water emulsions induced by the transglutaminase reaction was studied. The emulsions were incubated with transglutaminase for various times, and adsorbed and unadsorbed protein fractions at the oil—water interfaces were analyzed by sodium dodecyl sulfate—polyacrylamide gel electrophoresis. While only monomers were detected in the unadsorbed fractions, polymers were observed in the adsorbed fractions of the both proteins. The sizes and amounts of the polymers increased with incubation time. The incubation with transglutaminase caused much flocculation of the emulsion stabilized by β-lactoglobulin. An increase in viscosity was also observed with the flocculation. The flocculation was probably initiated by the formation of ε-(γ-glutamyl)-lysyl isopeptide bonds between β-lactoglobulin molecules adsorbed on different oil droplets. In the case of the emulsion stabilized by bovine serum albumin, however, the flocculation and the increase in viscosity occurred to only limited extents by the transglutaminase reaction. This suggests that ε-(γ-glutamyl)-lysyl isopeptide bonds induced by the transglutaminase reaction were formed only between neighboring molecules of bovine serum albumin on the same droplet.     

Expression of mRNA encoding IFN alpha in macrophages stimulated with Lactobacillus gasseri

The N7 of guanine is thought to be the primary target for adduct and crosslink formation between cisplatin and DNA. However, reactive sites in DNA other than the N7 of guanine may also participate in the formation of adducts with cisplatin. The possibility that these interactions arise and form DNA polymerase blocking lesions was investigated by primer extension reactions with Taq DNA polymerase. To differentiate between damage produced at relatively weak sites from those formed at the N7 of guanine, a modified DNA template was synthesised with the N7 of guanine replaced with a carbon atom. This was achieved in a PCR designed to incorporate 7-deazaguanine instead of normal guanine. The sequence specificity of cisplatin damage in the modified and unmodified DNA substrates was compared (after linear amplification) by DNA sequencing gel analysis. For concentrations of cisplatin (1 to 5 microM) that induce blocking lesions in normal DNA, no significant damage was observed in the modified DNA. This confirmed that the N7 of guanine is the major site of adduct formation in normal DNA. At higher concentrations of cisplatin (50 microM and 100 microM), lesions were found at AA dinucleotides and other novel sites in the modified DNA. These results indicate that the N7 of guanine is not required in the formation of some cisplatin adducts.     

Effect of silver content on thermal fatigue life of Sn-xAg-0.5Cu flip-chip interconnects

The thermal fatigue properties of Sn-xAg-0.5Cu (x=1, 2, 3, and 4 in mass%) flip-chip interconnects were investigated to study the effect of silver content on thermal fatigue endurance. The solder joints with lower silver context (x=1 and 2) had a greater failure rate compared to those with higher silver content (x=3 and 4) in thermal fatigue testing. Cracks developed in the solders near the solder/chip interface for all joints tested. This crack propagation may be mainly governed by the nature of the solders themselves because the strain-concentrated area was similar for tested alloys independent of the silver content. From the microstructural observation, the fracture was a mixed mode, transgranular and intergranular, independent of the silver content. Higher silver content alloys (x=3 and 4) had finer Sn grains before thermal cycling according to the dispersion of the Ag₃Sn intermetallic compound, and even after the cycling, they suppressed microstructural coarsening, which degrades the fatigue resistance. The fatigue endurance of the solder joints was strongly correlated to the silver content, and solder joints with higher silver content had better fatigue resistance.     

Study for improvement of solar cell efficiency by impurity photovoltaic effect

This paper is to study for efficiency improvement of solar cells by utilizing impurity traps introduced in the band gap of semiconductor, that is, impurity photovoltaic (IPV) effect. It is revealed theoretically that there is a certain energy range where impurity-traps act as stepping stones in two-step excitation of electrons from the valence band to the conduction band under suppression of carrier recombination through such traps. Indium is selected as one of proper impurities that satisfy this condition in crystalline silicon, and theoretical prediction is experimentally verified. A good agreement between theory and experiment is obtained concerned with photoconductive properties. It is concluded that the IPV effect is useful to improve the cell efficiency.