Antibacterial action on pathogenic bacterial spore by green tea catechins

Antibacterial effects of catechins, the major green tea polyphenols, were studied using Clostridium and Bacillus spores. Incubation with crude catechins decreased the number of C botulinum and C butyricum spores but not B cereus spores. Furthermore, the effects of six catechin derivatives on spores were investigated. (−)‐Epicatechin gallate (ECg), (−)‐epigallocatechin (EGC), (−)‐epigallocatechin gallate (EGCg) and (+)‐gallocatechin gallate (GCg) were more effective in decreasing C botulinum and C butyricum spore numbers than (+)‐catechin (C) and (−)‐epicatechin (EC). The vegetative growth of C botulinum and B cereus was inhibited by crude extracts of the catechins. Specifically, purified GCg and EGCg inhibited the vegetative growth of C botulinum and B cereus. The inhibitory effect of ECg on B cereus was similar to that of GCg. However, toxin‐production by B cereus was not inhibited by catechin. Damage to the membrane of C butyricum spores by catechin derivatives was shown using fluorescent microscopy. This study shows that low concentrations of catechins, although requiring a long exposure time, inhibited the growth of bacterial spores. However, the effects of the purified derivatives of the catechins were not the same and GCg and EGCg were found to be the most potent. Spores that are generally resistant to many disinfectants were sensitive to catechins. Copyright © 2005 Society of Chemical Industry     

Isolation and characterization of the ATF2 gene encoding alcohol acetyltransferase II in the bottom fermenting yeast Saccharomyces pastorianus

The ATF2 gene encodes alcohol acetyltransferase II, which catalyses the synthesis of isoamyl acetate from acetyl coenzyme A and isoamyl alcohol. To characterize the ATF2 gene from the bottom fermenting yeast Saccharomyces pastorianus, the S. pastorianus ATF2 gene was cloned by colony hybridization using the S. cerevisiae ATF2 gene as a probe. When an atf1 null mutant strain was transformed with a multi-copy plasmid carrying the S. pastorianus ATF2 gene, the AATase activity of this strain was increased by 2.5-fold compared to the control. The S. pastorianus ATF2 gene has 99% nucleic acid homology in the coding region and 100% amino acid homology with the S. cerevisiae ATF2 gene. Southern blot analysis of chromosomes separated by pulse-field gel electrophoresis indicated that the ATF2 gene probe hybridized to chromosome VII in S. cerevisiae and to the 1100 kb chromosome in S. pastorianus. As S. pastorianus is thought to be a hybrid of S. cerevisiae and S. bayanus, the S. bayanus-type gene, which has a relatively low level of homology with the S. cerevisiae-type gene, is also usually detected. Interestingly, an S. bayanus-type ATF2 gene could not be detected. These results suggested that the cloned ATF2 gene was derived from S. cerevisiae. Analysis using an ATF2-lacZ fusion gene in S. pastorianus showed that expression of the ATF2 gene was relatively lower than that of the ATF1 gene and that it is repressed by aeration but activated by the addition of unsaturated fatty acids. The S. pastorianus ATF1, Lg-ATF1 and ATF2 Accession Numbers in the DDBJ Nucleotide Sequence Database are D63449, D63450 and D86480, respectively.     

Hydrogen embrittlement behavior induced by dynamic martensite transformation of Ni–Ti superelastic alloy

The hydrogen embrittlement behavior induced by the martensite transformation of Ni–Ti superelastic alloy subjected to a dynamic cyclic tensile test with hydrogen cathodic charging has been investigated by hydrogen thermal desorption analysis. The critical stress for the martensite transformation steeply decreases with increasing number of deformation cycles, whereas the critical stress for the reverse transformation only slightly changes. The dynamic stress-induced martensite transformation markedly enhances hydrogen absorption, compared with that of the martensite phase itself. The hydrogen concentration at the surface layer of the specimen is evaluated to be above 3500 mass ppm; nevertheless, no fracture associated with the stress-induced martensite transformation occurs. In addition, no hardening is observed at the surface layer of the specimen despite the formation of the hydride and hydrogen enrichment. The hydrogen thermally desorbed at a low temperature markedly increases, indicating that the hydrogen states are changed by the dynamic martensite transformation. Note that interactions between hydrogen and the phase transformation are probably irreversible, although the phase transformation is reversible. The present study shows, for the first time, that the hydrogen embrittlement behavior of the alloy strongly depends on the dynamic change of the hydrogen states accompanied by the martensite transformation.     

A hybrid model for estimating global solar radiation

In view of the site-dependence of Ångström correlation, this study developed a hybrid model to estimate global radiation H. Unlike Ångström correlation H=(α+βS/S₀)H₀, this model suggested that H=(a+b S/S₀)H_b+(c+d S/S₀)H_d, H_b and H_d are effective beam radiation and effective diffuse radiation, which imply latitude, elevation and seasonal effect on radiation. H_b and H_d are calculated by an arithmetic model derived from spectral model. The hybrid model was designed for estimating monthly mean daily global radiation with hourly-recorded bright sunshine time, and its applicability was verified at observatories in Japan.     

Effects of potential, temperature and pH on hydrogen absorption and thermal desorption behaviors of Ni-Ti superelastic alloy in 0.9% NaCl solution

The effects of electrochemical potential, solution temperature and pH on the hydrogen absorption and desorption behaviors of Ni-Ti superelastic alloy immersed in 0.9% NaCl solution for 2 h have been investigated systematically by hydrogen thermal desorption analysis. For hydrogen cathodic charging under constant applied potential, upon increasing solution temperature, the critical potential for the hydrogen absorption very slightly shifts to a noble direction. As solution pH decreases, the critical potential for the hydrogen absorption shifts markedly to a noble direction and approaches to the corrosion potential; the critical current density for the hydrogen absorption slightly decreases. At a less noble potential than the critical potential for the hydrogen absorption, the amount of absorbed hydrogen increases markedly with decreasing applied potential. For hydrogen cathodic charging under constant current density, the amount of absorbed hydrogen increases with increasing solution temperature and decreasing solution pH. The basic hydrogen desorption behavior only slightly depends on solution temperature or pH. Nevertheless, hydrogen desorption at low temperatures for specimens subjected to cathodic charging under constant current density is observed distinctly as compared with that under constant applied potential.     

Ghosts of B‐type Wheat Starch Granules in Concentrated KI/I2 Solution

When B‐type (2.0–8.0 µm) wheat starch granules containing various amounts (2.1–25%) of amylose were treated with 25% KI/10% I₂ solution, low‐amylose (below 10% amylose) B‐type wheat starch granules changed to the ghost form. It is known that A‐type (25–35 µm) wheat starch granules change to the ghost form and show a typical double structure (a black‐brown central portion and red‐brown surrounding portion), however, the B‐type wheat starch ghosts did not show the same double structure but rather a simple (red‐brown portion) sack form. The relative ghost areas in the B‐type wheat starch granules were highly correlated to the amylose content (%), which was similar to the results of A‐type starch granules. This suggests that the amylose molecule in B‐type starch contributes to the structural stability of the starch granule.     

Multiresidue method for pesticides and veterinary drugs in bovine milk using GC/MS and LC/MS/MS

A simple, sensitive and selective method with gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed to detect 342 pesticides and veterinary drugs contaminating bovine milk at the maximum residue limits (MRLs) defined in the "positive list system". Sample preparation was performed by extracting the analytes with acetonitrile, followed by salting-out with sodium chloride. For some pesticides, the extract was further cleaned up by n-hexane partitioning and PSA cartridge column chromatography. GC/MS-EI or -NCI was used to determine pesticide residues, while LC/MS/MS-ESI was applicable to the determination of pesticide and veterinary drug residues. The variation of the recoveries of these drugs at MRL was relatively wide; however the relative standard deviations of the recovery of each drug were within 28%, suggesting that the present method is good enough for use as a screening test for contaminants at the MRLs. These results show that this method is useful for multiresidue analysis of numerous pesticides and veterinary drugs in bovine milk.     

Dietary exposure to acrylamide in a group of Japanese adults based on 24-hour duplicate diet samples

Acrylamide is a probable human carcinogen and known human neurotoxin that can be generated in food through heating. Using a mathematical modelling approach, our previous study estimated long-term average dietary exposure to acrylamide in the Japanese people; however, the validity of these estimates remained unknown. Here, we aimed to obtain a more accurate estimate of acrylamide exposure that would reflect the usual practice of heat processing and consumption of foods in the population. We collected duplicate diet samples and dietary records during 24 h from a group of Japanese adults. A total of 110 duplicate diet samples were analysed for acrylamide by LC-MS/MS. Data from individual dietary records were used to examine the association between dietary acrylamide exposure and consumption of selected food groups (e.g., coffee, tea, confectioneries, and vegetables prepared at high temperature [deep-frying, stir-frying, sautéing, and baking]). Of the 110 homogenised diet samples, 108 contained detectable levels of acrylamide. Dietary exposure to acrylamide ranged from 8 to 1582 ng/kg body weight (bw)/day, with the mean value of 215 ng/kg-bw/day and median value of 143 ng/kg-bw/day. This mean value was higher than the value we previously estimated for Japanese adults using a mathematical approach. Multiple linear regression analysis showed log dietary acrylamide exposure was significantly associated with consumption of coffee and vegetables prepared at high temperature during 24-hr of sampling (adj. R² = 0.250, p < 0.001). We revealed significant difference in dietary acrylamide exposure between participants who had coffee and vegetables prepared at high temperature (median, 169 ng/kg-bw/day; range, 35–1224 ng/kg-bw/day, n = 42) and those who had none of them (median, 75 ng/kg-bw/day; range, 8–311 ng/kg-bw/day, n = 15) (Steel-Dwass test, p < 0.05).     

Association of Ligamentum Flavum Hypertrophy with Adolescent Idiopathic Scoliosis Progression—Comparative Microarray Gene Expression Analysis

The role of the ligamentum flavum (LF) in the pathogenesis of adolescent idiopathic scoliosis (AIS) is not well understood. Using magnetic resonance imaging (MRI), we investigated the degrees of LF hypertrophy in 18 patients without scoliosis and on the convex and concave sides of the apex of the curvature in 22 patients with AIS. Next, gene expression was compared among neutral vertebral LF and LF on the convex and concave sides of the apex of the curvature in patients with AIS. Histological and microarray analyses of the LF were compared among neutral vertebrae (control) and the LF on the apex of the curvatures. The mean area of LF in the without scoliosis, apical concave, and convex with scoliosis groups was 10.5, 13.5, and 20.3 mm², respectively. There were significant differences among the three groups (p < 0.05). Histological analysis showed that the ratio of fibers (Collagen/Elastic) was significantly increased on the convex side compared to the concave side (p < 0.05). Microarray analysis showed that ERC2 and MAFB showed significantly increased gene expression on the convex side compared with those of the concave side and the neutral vertebral LF cells. These genes were significantly associated with increased expression of collagen by LF cells (p < 0.05). LF hypertrophy was identified in scoliosis patients, and the convex side was significantly more hypertrophic than that of the concave side. ERC2 and MAFB genes were associated with LF hypertrophy in patients with AIS. These phenomena are likely to be associated with the progression of scoliosis.