Scanning electron microscopy of food-poisoning bacterium Bacillus cereus using a variable-pressure SEM

A variable-pressure scanning electron microscopy (VP-SEM) with a cooling stage permits long hours of observation of water-containing specimens in their natural or close to natural state, without the conventional specimen preparations of fixation, dehydration, drying and metal coating. It reduces water vaporization and beam damage by keeping the specimens at a low temperature. We observed Bacillus cereus colonies on nutrient agar, which would shrink significantly if any conventional specimen preparation technique were used. We also studied the growing process of the bacteria on raw and steamed rice using the VP-SEM without conventional preparation techniques. Original specimens were directly mounted onto specimen holders and their backscattered electron images observed under the following conditions: specimen stage temperature, -10 degrees C; specimen chamber vacuum level, 30-70 Pa; and accelerating voltage, 15-20 kV. We recognized that the VP-SEM minimized deformation of the colonies due to shrinkage of the nutrient agar, and successfully imaged the morphology of the colonies and bacteria without a decline in bacteria number, which is apt to occur during fixation and dehydration. Also, the growth process of the bacteria on raw or steamed rice could be observed promptly, since there is no specimen preparation step.     

Influence of network chain orientation on the mechanical property of epoxy resin filled with silica particles

When tensile stress was applied to epoxy resin filled with silica particles, we expected that the stress concentration would occur in the epoxy matrix near the interface between the matrix and the silica particles. We investigated the plastic deformation of the network chains near the interface, which was quantitatively evaluated using a polarized microscope FTIR technique. A biphenol‐type epoxy resin, which has a mesogenic group in the backbone moiety, was used as the matrix resin. As a result, reorientation of the network chains along the direction of the tensile stress near the interface with the silica particles was observed. Especially when the silica/matrix interface has good bonding properties, the reorientation of the network chains was observed at a larger area around the silica particles. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 787–794, 2003     

Mechanical properties of the Ni3(Si,Ti) alloys doped with carbon and beryllium

The Ni₃(Si, Ti) alloys doped with small amounts of carbon and beryllium were tensile tested in two environments, vacuum and air, over a wide range of test temperatures. The yield stresses of the carbon-doped alloys were almost identical to the undoped alloys while those of the beryllium-doped alloys were slightly higher than the undoped Ni₃(Si, Ti) alloys. The doping with carbon enhanced the elongation and ultimate tensile strength (UTS) whereas doping with beryllium reduced the elongation over the entire temperature range tested. The fracture patterns were primarily associated with the ductility behaviour. As the elongation (or UTS) increased, the fracture pattern changed from the intergranular to the transgranular fracture patterns. No environmental embrittlement of the ductility of the carbon-doped alloys was found at ambient temperatures but it was evident at elevated temperatures. Ductilities were reduced at high temperatures when the carbon-doped alloys were tensile tested in air. At high temperatures the environmental embrittlement observed is suggested to be due to the penetration of (free) oxygen into the grain boundaries causing the ductility loss in the carbondoped alloys.     

A study of living plant specimens by low-temperature scanning electron microscopy

Young fresh Tradescantia reflexa stamen hair cells were used to clarify the optimal conditions for direct viewing and taking photographs with a scanning electron microscope (SEM) equipped with a cryo-system. The rate of protoplasmic streaming in the cells was measured under an optical microscope after examining and photographing them in the SEM over a period of a few minutes. Almost the same rate of streaming (5.5 μm/second, 20°C) was observed in nonirradiated control cells and irradiated cells photographed in the SEM using an accelerating voltage of 10 kV with the cryo-stage at a temperature of – 15°C. (The specimen holder and specimen were not at this temperature, but, rather, probably somewhat higher.) Fresh plant organs, tissues, and cells were also tested under the same conditions. The fine structure was well preserved in detail. The procedures were as follows: (1) prompt attachment of fresh plant materials on an aluminum specimen holder with double-faced adhesive Scotch tape or a small amount of plastic adhesive for woodcraft; (2) setting the holder on the cryo-stage cooled to –15°C in advance and rapid evacuation; and (3) quick SEM examination and photography (within several minutes). The advantages of this method are summarized as follows: (1) high possibility of viewing living materials; (2) minimal artifacts: freedom from chemical fixation and additional procedures utilized in ordinary SEM specimen preparation; and (3) simplicity, speediness, and economy in preparation for viewing. Since the specimens were not likely to be frozen during quick examination and photography, this method might well be called “low-temperature SEM” (LT-SEM) as distinguished from “cryo-SEM”.     

Associations of visual forms with colors: The minor role of emotion as the mediator

Associations of colors with a wide variety of visual forms were examined. Fifty visual forms were presented to Japanese participants, who were asked to report a color suitable to each visual form out of 15 colors. Correspondence analysis was applied to the responses, and four dimensions of configurations were obtained. The first and second dimensions were related to chromaticity or hue. In these dimensions, red and orange were associated with round symmetric shapes, yellow with shapes with sharp corners, violet and purple with irregular shapes with smooth curves, and cold colors such as blue and green with forms that consisted of simple geometrical elements such as triangles, squares, and circles. The third dimension was related to L* and b* in the CIELAB color space and to the stability of visual forms. Dimension 4 was not significantly related to any of L*, a*, and b*. In another experiment, participants rated the affective impressions of the colors and visual forms on semantic differential (SD) scales. The analyses of the SD data combined with the results of the color‐form associations suggest that the role of emotional meanings of colors and visual forms in mediating the associations was rather minor, and that world‐knowledge contributed considerably to the observed color‐form associations. Kandinsky's color‐form assignment was also examined, and except for the yellow‐triangle association, the assignment was not supported.     

The COX-2 Selective Blocker Etodolac Inhibits TNFα-Induced Apoptosis in Isolated Rabbit Articular Chondrocytes

Chondrocyte apoptosis contributes to the disruption of cartilage integrity in osteoarthritis (OA). Recently, we reported that activation of volume-sensitive Cl⁻ current (I_Cl,vol) mediates cell shrinkage, triggering apoptosis in rabbit articular chondrocytes. A cyclooxygenase (COX) blocker is frequently used for the treatment of OA. In the present study, we examined in vitro effects of selective blockers of COX on the TNFα-induced activation of I_Cl,vol in rabbit chondrocytes using the patch-clamp technique. Exposure of isolated chondrocytes to TNFα resulted in an obvious increase in membrane Cl⁻ conductance. The TNFα-evoked Cl⁻ current exhibited electrophysiological and pharmacological properties similar to those of I_Cl,vol. Pretreatment of cells with selective COX-2 blocker etodolac markedly inhibited I_Cl,vol activation by TNFα as well as subsequent apoptotic events such as apoptotic cell volume decrease (AVD) and elevation of caspase-3/7 activity. In contrast, a COX-1 blocker had no effect on the decrease in cell volume or the increase in caspase-3/7 activity induced by TNFα. Thus, the COX-2-selective blocker had an inhibitory effect on TNFα-induced apoptotic events, which suggests that this drug would have efficacy for the treatment of OA.     

Tensile Fracture Behavior and Strength of Surface-Modified SiTiCO Fiber SiC-Matrix Minicomposites Fabricated by the PIP Process

The effect of surface-modified SiTiCO (Tyranno®) fiber on the tensile behavior of SiC-matrix minicomposites has been examined. Standard SiTiCO fiber and two kinds of surface-modified fibers were used and the composites were fabricated by a polymer infiltration pyrolysis process. The strengths of the surface-modified-fiber-reinforced minicomposites were significantly improved compared to that of the standard fiber minicomposite. The scattering of tensile strength also decreased with the use of surface-modified fibers. The results demonstrate the achievement of SiTiCO-fiber-reinforced SiC-matrix composite by modifying the surface chemistry of the fiber.     

Alloy design for Al addition on microstructure and mechanical properties of Ni3(Si,Ti) alloy

The effect of Al addition on the microstructure and tensile properties of Ni₃(Si,Ti) alloys with an L1₂ ordered structure, which were fabricated through thermomechanical processing from arc-melted ingots, was investigated. Al was added to a Ni₃(Si,Ti) alloy by using two methods such that Al substituted for (1) only Ti and (2) both Ni and Ti along a Ni₃(Si,Ti)-Ni₃Al pseudo-binary line. In the case of the alloys prepared by the former method, the addition of more than 4 at.% Al resulted in a two-phase microstructure consisting of disordered fcc Ni solid solution dispersions in the L1₂ matrix, while in the case of the alloys prepared by the latter method, the addition of 4 at.% Al retained the L1₂ single-phase microstructure. In the case of the 4 at.% Al-added alloys, the room-temperature tensile properties were similar and independent of the alloying methods, whereas the high-temperature yield stress was higher in the alloys prepared by the latter method than in the case of the alloys prepared by the former method. These results suggest that a single-phase microstructure consisting of an entire L1₂ structure is favorable for obtaining high-temperature tensile properties.     

Evaluation of Potential Anti-Hepatitis A Virus 3C Protease Inhibitors Using Molecular Docking

Hepatitis A virus (HAV) infection is a major cause of acute hepatitis worldwide and occasionally causes acute liver failure and can lead to death in the absence of liver transplantation. Although HAV vaccination is available, the prevalence of HAV vaccination is not adequate in some countries. Additionally, the improvements in public health reduced our immunity to HAV infection. These situations motivated us to develop potentially new anti-HAV therapeutic options. We carried out the in silico screening of anti-HAV compounds targeting the 3C protease enzyme using the Schrodinger Modeling software from the antiviral library of 25,000 compounds to evaluate anti-HAV 3C protease inhibitors. Additionally, in vitro studies were introduced to examine the inhibitory effects of HAV subgenomic replicon replication and HAV HA11-1299 genotype IIIA replication in hepatoma cell lines using luciferase assays and real-time RT-PCR. In silico studies enabled us to identify five lead candidates with optimal binding interactions in the active site of the target HAV 3C protease using the Schrodinger Glide program. In vitro studies substantiated our hypothesis from in silico findings. One of our lead compounds, Z10325150, showed 47% inhibitory effects on HAV genotype IB subgenomic replicon replication and 36% inhibitory effects on HAV genotype IIIA HA11-1299 replication in human hepatoma cell lines, with no cytotoxic effects at concentrations of 100 μg/mL. The effects of the combination therapy of Z10325150 and RNA-dependent RNA polymerase inhibitor, favipiravir on HAV genotype IB HM175 subgenomic replicon replication and HAV genotype IIIA HA11-1299 replication showed 64% and 48% inhibitory effects of HAV subgenomic replicon and HAV replication, respectively. We identified the HAV 3C protease inhibitor Z10325150 through in silico screening and confirmed the HAV replication inhibitory activity in human hepatocytes. Z10325150 may offer the potential for a useful HAV inhibitor in severe hepatitis A.     

Mechanical properties of Ni3(Si,Ti) polycrystals alloyed with substitutional additions

The mechanical properties of the L1₂-type Ni₃(Si, Ti) polycrystals, which were alloyed with 1–2 at% of various transition metals and also doped with boron, were investigated over a wide range of temperatures. The addition of Hf enhanced the levels of yield stress whereas the addition of Cr, Mn and Fe reduced the levels of the yield stress over a wide range of temperatures. Ni₃(Si, Ti) alloyed with Cr, Mn and Fe showed a shallow minimum in the yield stress-temperature curves. This result was correlated with the modification of the micro-cross-slip process by the additives. At low temperatures, the addition of Hf and Nb slightly reduced the elongation, while the addition of Cr, Mn and Fe improved elongation. This elongation behaviour was interpreted as the alloying effect on the intergranular cohesive strength of L1₂ ordered alloys. At high temperatures, the elongation of Ni₃(Si, Ti) alloyed with Hf showed a particularly high value. This elongation behaviour is discussed based on the alloying effect on the competition between dynamic recrystallization and cavitation at grain boundaries. The fracture surfaces exhibited a variety of fracture patterns, depending on temperature and the alloy, and were primarily well correlated with the elongation behaviour. The ductilities of most of the alloys at high temperatures were reduced by the tests in air.