Purification, bacteriolytic activity, and specificity of beta-lytic protease from Lysobacter sp. IB-9374

Lysobacter sp. IB-9374, which was isolated from soil as a high lysyl endopeptidase-producing strain (Chohnanet al., FEMS Microbiol. Lett., 213, 13-20, 2002), was found to produce a beta-lytic protease capable of lysing gram-positive bacteria such as Staphylococcus aureus, Microccocuseus, and Bacillus subtilis. The Lysobacter strain secreted the beta-lytic protease into the culture medium at a 2.4-fold higher level than Achromobacter lyticus. The enzyme was highly purified through a series of six steps with a high yield. The enzyme was strongly inhibited by tetraethylene-pentamine and 1,10-phenanthroline. The purified enzyme lysed more efficiently almost all the gram-positive bacteria tested than lysozyme, lysostaphin, and mutanolysin. The enzyme was very similar to Achromobacter beta-lytic protease containing one zinc atom in terms of amino acid composition and N-terminal sequence. The nucleotide sequence revealed that the mature enzyme was composed of 179 amino acid residues with additional 198 amino acids at the amino-terminal end of the enzyme. The deduced amino acid sequence of the mature enzyme coincided with that of the Achromobacter enzyme, although the prepro-region showed a 41% sequence identity with the counterpart. These results indicate that Lysobacter sp. is a useful strain for an efficient large-scale preparation of beta-lytic protease capable of lysing bacteria.     

A Dynamic Motion Control Technique for Human-like Articulated Figures

This paper presents a dynamic motion control technique for human-like articulated figures in a physically based character animation system. This method controls a figure such that the figure tracks input motion specified by a user. When environmental physical input such as an external force or a collision impulse are applied to the figure, this method generates dynamically changing motion in response to the physical input. We have introduced comfort and balance control to compute the angular acceleration of the figure's joints. Our algorithm controls the several parts of a human-like articulated figure separetely through the minimum number of degrees-of-freedom. Using this approach, our algorithm simulates realistic human motions at efficient computational cost. Unlike existing dynamic simulation systems, our method assumes that input motion is already realistic, and is aimed at dynamically changing the input motion in real-time only when unexpected physical input is applied to the figure. As such, our method works efficiently in the framework of current computer games.     

Magnetic field analysis of a 70‐MW‐class superconducting generator by the 3D finite element method

The three‐dimensional magnetic field and eddy current distribution of the 70‐MW‐class superconducting generator was calculated by a finite element method. The condition for the exciting armature coil was calculated by the jω‐method. In the magnetic flux distribution of the armature winding bore, the value obtained by analysis had good agreement with the measurement. An analysis that let the rotor move by the synchronizing speed was also done. The armature voltage for the nonload condition and the armature current for the three‐phase short‐circuit condition obtained by calculations had good agreement with the measurements. The validity of the analytical model for three‐dimensional magnetic field analysis of superconducting generators was confirmed from these results. In addition, the synchronous reactances were calculated using these results with eddy current in the facing. It was found that the facing had the effect of decreasing synchronous reactance by about 5%. © 2000 Scripta Technica, Electr Eng Jpn, 134(2): 53–60, 2001     

Versatile gas/particle ion chromatograph

A new, compact gas/particle ion chromatograph has been developed for measuring ionic constituents in PM2.5 (particulate matter of aerodynamic diameter < or = 2.5 microm) and water-soluble ionogenic gases. The instrument has separate sampling channels for gases and particles. In one, a membrane denuder collects soluble gases for preconcentration and analysis. In the other, a cyclone removes larger particles, a membrane denuder removes soluble gases, and a continuously wetted hydrophilic filter collects particles. A single, multiport, syringe pump handles liquid transport, and one conductivity detector measures anions and ammonium for both channels. Electrodialytically generated gradient hydroxide eluent permits 20 min chromatographic runs. Gas/particle samples are each collected for 40 min, butthe sampling intervals are staggered by 20 min. Liquid samples from the gas denuder and particle collector are aspirated and preconcentrated on sequential cation and anion concentrators and transferred respectively to an ammonia transfer device and an anion separation column. The flow configuration results in an ammonium peak before anion peaks in the chromatogram. The system measures ammonia, organic acids (such as acetic, formic, and oxalic acids), HCl, HONO, SO2, HNO3, and the corresponding ions in the aerosol phase. Low ng/m3 to sub-ng/m3 limits of detection (LODs) are attained for most common gases and particulate constituents, the LODs for gaseous SO2 to NH3 range, for example, from sub parts per trillion by volume (sub-pptv) to approximately 5 pptv.     

Effects of Atmospheric Composition on the Molecular Structure of Synthesized Silicon Oxycarbides

The dependence of silicon oxycarbides' chemical composition and molecular structure on their reaction conditions was tested by varying the atmosphere under which pyrolysis was performed. To obtain the silicon oxycarbides, densely cross‐linked silicone resin particles with an averaged diameter of 2 μm were pyrolyzed in various atmospheres of H₂, Ar, and CO₂, in the temperature range 700°C–1100°C. The residual mass of resin after pyrolysis was almost constant at 700°C, although their apparent colors varied distinctly. The sample obtained from the H₂ atmosphere was white, whereas that obtained from the CO₂ atmosphere was dark brown. Fourier‐transform infrared (FT‐IR) spectra of the residues suggested that the Si–O–Si network evolution was accelerated in the CO₂ atmosphere. Beyond 800°C, the chemical compositions of the compounds obtained from a H₂ atmosphere increasingly approached near‐stoichiometric SiO₂–xSiC composition with increasing the pyrolysis temperature. Compounds from a CO₂ atmosphere approached a composition of SiO₂–xC with no free SiC as the pyrolysis temperature increased. In the products from an Ar atmosphere, SiO₂–xSiC–yC compositions were typically obtained. The observed effects of the pyrolysis atmosphere on the resulting chemical compositions were analyzed in terms of thermodynamic calculations. Electron spin resonance (ESR) spectra revealed broad and intense signals from products obtained from either Ar or CO₂. Estimating from the signal intensity, the residual spin concentrations were in the range 10¹⁸–10¹⁹ g^?1. Meanwhile, the spectra from the samples obtained in H₂ showed weak and sharp signals with estimated spin concentrations ranging from 10¹⁶–10¹⁷ g^?1. This signal attenuation may have been due to the hydrogen capping of dangling bond formed during pyrolysis.     

Tensile behavior of polypropylene blended with bimodal distribution of styrene‐ethylene‐butadiene‐styrene particle size

The objective is to characterize the effects of the bimodal distribution of rubber particles and its blend ratio on the mechanical properties of the thermoplastic polypropylene blended with two different styrene‐ethylene‐butadiene‐styrene triblock copolymer at the intermediate and high strain rates. Tensile tests are conducted at the nominal strain rates from 3 × 10^?1 to 10² (1/s). Phase morphology is investigated to estimate the bimodal rubber particle size distribution. In addition, the in situ observation is conducted during uniaxially stretching within transmission electron microscopy step by step to investigate the deformation events depending on the elongation of samples. The elastic modulus increased gradually as the blend ratio of large rubber particle increased. An increase in the rupture strain and the strain energy up to failure was found for the bimodal rubber particle distributed blend system where the blend ratios of small rubber particle and large rubber particle were same. This is because the smaller particles dominant blend systems show the bandlike craze deformation while the localized plastic deformation is taken place in the larger particles dominated blend systems. The synergistic effect of these rubber particles gives rise to a strong increase in the ductility of these bimodal rubber particle distributed polypropylene systems. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

A rechargeable lithium metal battery operating at intermediate temperatures using molten alkali bis(trifluoromethylsulfonyl)amide mixture as an electrolyte

The physicochemical properties of molten alkali bis(trifluoromethylsulfonyl)amide, MTFSI (M = Li, K, Cs), mixture (x_LiTFSI = 0.20, x_KTFSI = 0.10, x_CsTFSI = 0.70) were studied to develop a new rechargeable lithium battery operating at intermediate temperature (100–180 °C). The viscosity and ionic conductivity of this melt at 150 °C are 87.2 cP and 14.2 mS cm⁻¹, respectively. The cyclic voltammetry revealed that the electrochemical window at 150 °C is as wide as 5.0 V, and that the electrochemical deposition/dissolution of lithium metal occurs at the cathode limit. A Li/MTFSI (M = Li, K, Cs)/LiFePO₄ cell showed an excellent cycle performance at a constant current rate of C/10 at 150 °C; 95% of the initial discharge capacity was maintained after 50 cycles. Except for the initial few cycles, the coulombic efficiencies were approximately 100% for all the cycles, indicating the stabilities of the molten MTFSI mixture and all the electrode materials.     

A pre-heating vaporization technology of coal-water-slurry for the gasification process

A pre-heating vaporization technology of coal water slurry, CWS, was developed to increase the efficiency of coal gasification with the reduction of oxygen consumption. A CWS pre-heating experimental unit having a capacity of 2.0 tons-coal/day was manufactured. After the CWS was heated to 573 K at about 10 MPa, water in CWS was continuously vaporized in a tube of which the diameter increased gradually (2–10 mm I.D.), and dry coal particles with steam were atomized and fed to a vessel. No aggregation of coal particles was observed in the atomized samples. The temperature and the pressure distributions of the fluid were calculated with empirical heat transfer and pressure drop correlations. The calculated results well corresponded to the experimental data measured in some test points. Using the calculation model, a large scale apparatus of 100 tons-coal/day was designed using KNO₃ + NaNO₂ + NaNO₃ molten salt as a heating medium.     

Main R&D issues for fast reactor structural design standard

For realization of economical and reliable fast reactor (FR) plants, the Japan Atomic Energy Agency (JAEA) and the Japan Atomic Power Company (JAPC) are cooperating on the “Feasibility Study on Commercialized FR Cycle Systems”. To certify the design concepts through evaluation of the structural integrity of FR plants, the research and development of the “Elevated Temperature Structural Design Guide for Commercialized Fast Reactor (FDS)” is recognized as an essential theme. The FDS focuses on particular failure modes of FRs such as ratchet deformation and creep-fatigue damage due to cyclic thermal loads. For precise evaluation of these modes, the research and development for three main issues is in progress. First, the “Refinement of Failure Criteria” needs to be addressed for particular failure modes of FRs. Secondly, the development of “Guidelines for Inelastic Design Analysis” is conducted to predict elastic plastic and creep deformation under elevated temperature conditions. Lastly, efforts are being made toward preparing “Guidelines for Thermal Load Modeling” for the design of FR components where thermal loads are dominant.     

Investigation of pathways of advanced glycation end-products accumulation in macrophages

Advanced glycation end-products (AGE) play a role in the pathogenesis of several diseases, including diabetic complications and atherosclerosis. In atherosclerotic lesions of human aortas, AGE are localized in the extracellular matrix and intracellularly in foam cells. Two interpretations are possible for AGE accumulation inside macrophages, one is endocytic uptake of extracellular AGE-proteins by scavenger receptors; the other is intracellular AGE formation inside the macrophages. In the present study, we determined the pathways involved in AGE accumulation inside macrophages. RAW 264.7 cells, a murine macrophage cell line, incubated with BSA and 1600 mM glucose for 40 weeks, recognized heavily modified AGE- BSA. In contrast, the cells showed no ligand activity for mildly modified AGE-BSA, prepared by incubating BSA with 50 mM glucose for 24 weeks. Nepsilon-(carboxymethyl)lysine (CML)-modified proteins of about 65 kDa were detected in human monocyte-derived macrophages incubated for 7 days with 30 mM glucose and phorbol myristate acetate. Furthermore, CML was generated when glycated protein was incubated with hypochloric acid. Taken together, our results indicate that AGE detected inside foam cells in atherosclerotic lesions are generated intracellularly rather than representing endocytic uptake of extracellular AGE-proteins by scavenger receptors.