Accumulation of pyrraline-modified albumin in phagocytes due to reduced degradation by lysosomal enzymes

Previous studies suggested that the interaction between proteins modified by advanced glycation end products (AGEs) and cells, such as macrophages, may be involved in diabetic angiopathy. Pyrraline is one of the AGEs and known to be elevated in plasma of diabetic rats and humans, and is present in vascular lesions of diabetic and elderly subjects. We examined whether modification of albumin by pyrraline influences its degradation by macrophage-like cell line, P388D1 cells. Degradation of pyrraline-modified albumin by these cells was diminished, causing accumulation of the albumin in these cells. The susceptibility of pyrraline-modified albumin to lysosomal proteolytic enzymes was reduced by approximately 40% in vitro, while lysosomal activity in the cells per se was not affected. This phenomenon was also observed when human monocytes were used instead of P388D1 cells. Our results suggest that accumulation of pyrraline-modified albumin in P388D1 cells is due to the reduced susceptibility of the protein to lysosomal enzymatic degradation. Such alterations in the interaction between AGEs-modified protein and phagocytes may contribute to angiopathy in elderly subjects and patients with diabetes.     

Structure and properties of 6061 + 26 mass% Si aluminum alloy produced via coupled rapid solidification and KOBO-extrusion of powder

Experiments on mechanical consolidation of rapidly solidified (RS) powder of 6061 + 26 mass% Si alloy were performed using the oscillating-die extrusion method. The RS powder was wrapped in thin-wall 6061-alloy cup 35 mm in diameter and vacuum-compressed by means of 100 ton press. Bars 8 mm in diameter were extruded with cross-section reduction of λ = 19 without any preheating of the charge. Tubes with a diameter/wall thickness of 14 mm/1 mm and cross-section reduction of λ = 33 were also manufactured with success. TEM/STEM observations revealed a very fine structure of as-extruded material and bimodal distribution of quasi-spherical silicon particles. Statistical analysis revealed a silicon fine fraction of 0.1–0.7 μm and a coarse fraction 2.1–2.5 μm in diameter. Examination by means of TEM did not reveal any significant changes in the morphology of the silicon particles, even when a high extrusion ratio and the material annealing after deformation were used. Hot compression tests on as-extruded rods (λ = 19) and preliminary annealed samples were performed at a constant true strain rate of 5 × 10^?3 s^?1 within the temperature range of 293–823 K. High strength of the material and relatively high ductility of samples deformed by compression up to ?_t ? 0.4 were observed. The maximum flow stress value for as-extruded material was reduced with deformation temperature from ～390 to ～3.5 MPa for 293 and 823 K, respectively. Annealing of the samples at 773 K/30 min was found to reduce the maximum flow stress by 30–40%. Tensile strengths of similar as-cast alloys and materials manufactured by means of other powder metallurgy methods were shown for the purpose of comparison.     

X-ray Photoemission Spectroscopy (XPS) and Thermally Stimulated Current (TSC) Studies on the Resistance Degradation of Iron-Doped Titania Ceramics

An anomalous increase of current with time has been found in iron-doped titania ceramics that have been subjected to a constant field. The result of thermally stimulated current analysis shows a peak at a temperature of 170°–180°C, which increases because of the depolarization of oxygen vacancies. X-ray photoemission spectroscopy indicates that an electrical stress results in an electrochemical reduction inside the specimen, particularly on the cathodic side. These results reinforce the viewpoint that the space charge that is responsible for the anomalous current results from a blockage of the O₂( g )/O²⁻( s ) ion transfer at the cathode.     

STD‐NMR‐Based Protein Engineering of the Unique Arylpropionate‐Racemase AMDase G74C

Structure‐guided protein engineering achieved a variant of the unique racemase AMDase G74C, with 40‐fold increased activity in the racemisation of several arylaliphatic carboxylic acids. Substrate binding during catalysis was investigated by saturation‐transfer‐difference NMR (STD‐NMR) spectroscopy. All atoms of the substrate showed interactions with the enzyme. STD‐NMR measurements revealed distinct nuclear Overhauser effects in experiments with and without molecular conversion. The spectroscopic analysis led to the identification of several amino acid residues whose substitutions increased the activity of G74C. Single amino acid exchanges increased the activity moderately; structure‐guided saturation mutagenesis yielded a quadruple mutant with a 40 times higher reaction rate. This study presents STD‐NMR as versatile tool for the analysis of enzyme–substrate interactions in catalytically competent systems and for the guidance of protein engineering.     

Negative Pore Air Pressure Generation in Backfill of Retaining Walls During Earthquakes and its Effect on Seismic Earth Pressure

In order to investigate the seismic behavior of conventional type and geosynthetic-reinforced soil retaining walls, 1-g model shaking tests were conducted. Model walls having a height of about 50 cm were placed on a subsoil layer and backfilled with a layer of dense dry Toyoura sand. They were subjected to several steps of horizontal irregular excitations. As a result, generation of negative pore air pressure in the backfill was observed. The maximum amplitude of the negative pore air pressure during each shaking step increased with the base acceleration. Based on analyses of the measured data, it was inferred that such negative pore air pressure was caused by outward wall displacement relative to the backfill and not by dilative behavior of the backfill. It would cause a reduction in the seismic earth pressures exerted from the backfill. This feature suggests an advantage of a rigid full-height facing for reinforced soil walls over the segmental types of facing. A simplified numerical procedure to evaluate earth pressure was applied while considering the effects of the negative pore air pressure, and it could qualitatively simulate the measured behavior in terms of the seismic earth pressure and the angle of failure plane in the backfill.     

Ca-α SiAlON hollow spheres prepared by carbothermal reduction–nitridation from different SiO2 powders

The formation process of hollow spheres composed of nanosized Ca-α SiAlON particles was investigated using SiO₂ starting powders with different characteristics in particle size, shape and crystalline state. TEM observations showed Ca-α SiAlON hollow spheres composed of a large number of nanosized particles in the products prepared at 1450 °C for 120 min in nitrogen. In all systems, the Ca-α SiAlON hollow spheres were always produced through an intermediate Si–Al–Ca–O liquid phase in the same mechanism, regardless of the characteristics of SiO₂ starting powders used. Spherical solid particles consisted of amorphous phase containing Si, Al, Ca, O and a small amount of N were generated at the initial stage of carbothermal reduction–nitridation. These spherical solid particles changed into hollow particles with the progression of the reaction from the liquid phase to the crystalline Ca-α SiAlON with increasing temperature.     

砂土地基地震液化沉降的两种简易计算方法的对比分析

利用两种简易方法对日本某核电站的砂性土地基在地震液化后的大变形沉降进行了计算和对比分析。这两种简易的实用方法只需要利用标准贯入试验数据就可以操作。计算结果表明,该核电站的砂性土地基在L2级地震作用下会发生较大的地面沉降,并可能对地下管线造成损坏。由于两种方法的计算原理相同,因此计算所得到的地基分层沉降趋势基本一致,但是在数值上存在着较大的差异,这主要是由于两种方法建立的基础试验数据不同以及具体计算方法上存在着差异所造成的。     

Removal of a high load of ammonia gas by a marine bacterium, Vibrio alginolyticus

A newly isolated marine bacterium, Vibrio alginolyticus was used to remove a high load of ammonia gas. By a stepwise increase in ammonia supply over the concentration range of 120-2000 ppm (v/v), complete removal of ammonia was observed from the start of the experiment in a suspended culture of the bacterium in basal medium containing 3% NaCl. When cells were inoculated onto an inorganic packing material in a biofilter, and a high load of ammonia was introduced continuously under nonsterile conditions, the average percentage of gas removed exceeded 85% for a 61-d operation. The maximum removal capacity and the complete removal capacity were 22.8 g-N/kg-dry packing material/d and 18.6 g-N/kg-dry packing material/d, respectively, which were about four times larger than those obtained in nitrifying sludge inoculated onto the same packing material. During this operation, the nonsterile air supply had no adverse effect on the removability of ammonia by V. alginolyticus.     

Study on the determination of pesticide residues in crops by ion-trap GC/MS/MS

Ion-trap GC/MS/MS was evaluated for the multi-residue determination of pesticides in agricultural products. Matrices were extracted from samples (spinach, carrot, onion and brown rice) with acetone and submitted to gel permeation chromatography, followed by a clean-up step through a graphite carbon cartridge. Thirty-five pesticides were added to either matrix, and analyzed by GC/MS/MS. Detection limits of pesticides by GC/MS/MS was almost the same as those by GC/MS (SIM). Coefficients of variation of peak area in 5 measurements of each pesticide at 0.1 microgram/mL or 0.05 microgram/mL with or without matrices were mostly acceptable, though those of 20 pesticides out of 35 were higher than 10% at a concentration of 0.02 microgram/mL. It was indicated that matrix artifacts, which interfere with GC/MS-Scan analysis, could be eliminated in some cases by using GC/MS/MS.