High hydrostatic pressure treatment impairs AcrAB-TolC pump resulting in differential loss of deoxycholate tolerance in Escherichia coli

We reported previously that high hydrostatic pressure-injured stationary phase cells of Escherichia coli K-12 lost their intrinsic deoxycholate tolerance. The AcrAB-TolC multi-drug resistance pump driven by proton motive force has been argued to be responsible for the tolerance to deoxycholate. In this report, we tested the sensitivity of the AcrAB-TolC (three components) pump to high hydrostatic pressure treatment (HPT). E. coli K-12 treated with HPT became sensitive to AcrAB-TolC-specific drugs such as ethidium bromide, but not to tetracycline which is pumped out by a one-component transporter, Tet. Only E. coli K-12 overproducing both AcrAB and TolC exhibited restored tolerance to deoxycholate after HPT but not E. coli overproducing either TolC or AcrAB. These observations strongly suggest that three-component pumps such as AcrAB-TolC are more susceptible to HPT than one-component pumps such as Tet, resulting in the differential loss of deoxycholate tolerance in high hydrostatic pressure-injured E. coli cells.     

Effect of coexisting metals on determination of lead and cadmium in the plastic materials test of the Japanese Food Sanitation Law

The effect of coexisting metals in a sample on the determination of lead and cadmium in plastics used for food contact materials was investigated. In the official method specified in the Japanese Food Sanitation Law, contents of lead and cadmium are determined by a dry incineration method using sulfuric acid. It was assumed that sometimes, coexisting metals in a sample may form insoluble sulfate and that lead sulfate might be adsorbed into the insoluble sulfate. Therefore, hydrochloric acid was added to the ash, to turn formed insoluble sulfate into soluble compounds (HCl addition method). We found that recoveries of cadmium were not affected in the presence of other metals except when calcium exceeded 20 mg/g in both methods. Recoveries of lead decreased in the presence of barium exceeding 0.1 mg/g or calcium exceeding 10 mg/g in the official method. However, improvement of recoveries was achieved with the HCl addition method and by reducing the sample amount to one-tenth (0.1 g) of that specified in the official method.     

Interlaboratory validation of quantitative duplex real-time PCR method for screening analysis of genetically modified maize

To reduce the cost and time required to routinely perform the genetically modified organism (GMO) test, we developed a duplex quantitative real-time PCR method for a screening analysis simultaneously targeting an event-specific segment for GA21 and Cauliflower Mosaic Virus 35S promoter (P35S) segment [Oguchi et al., J. Food Hyg. Soc. Japan, 50, 117-125 (2009)]. To confirm the validity of the method, an interlaboratory collaborative study was conducted. In the collaborative study, conversion factors (Cfs), which are required to calculate the GMO amount (%), were first determined for two real-time PCR instruments, the ABI PRISM 7900HT and the ABI PRISM 7500. A blind test was then conducted. The limit of quantitation for both GA21 and P35S was estimated to be 0.5% or less. The trueness and precision were evaluated as the bias and reproducibility of the relative standard deviation (RSD(R)). The determined bias and RSD(R) were each less than 25%. We believe the developed method would be useful for the practical screening analysis of GM maize.     

Improvement of carbon monoxide analysis in fish meat

Carbon monoxide (CO) treatment of fish meat of tuna, yellowtail, tilapia etc. is not allowed in Japan, since it can maintain the red color for a longer period than the microbiological shelf life of fish meat. The official method for quantification of CO has a problem, in that a part of the CO is lost during the preparation of the fish sample. To solve this problem, we modified the official method in this study. We also applied this modified method to survey the contents of CO in tuna, yellowtail, young yellowtail, and tilapia. As a result, the modified method was found to be more suitable for CO quantification than the official method. An inter-laboratory study by 4 laboratories confirmed that the CO content of many samples of tilapia exceeded the regulation value, apparently due to the higher recovery of CO, compared to the official method. Therefore, it was suggested that the regulation value in the case of tilapia should be changed if this method is introduced as an official method.     

Tocopherol and fluorescence levels in deep-frying oil and their measurement for oil assessment

This study examined how tocopherol retention is affected by the presence or absence of food coatings, and also tested the measurement of fluorescent substance levels in cooking oil to evaluate oil deterioration. Potato slices were tempura-fried (with a coating) or french-fried (without a coating). The three tocopherol isomers decreased with heating time, and better retention was found in the tempura process. The decomposition rates of tocopherol were in the order γ> δ ≥α for the three isomers for both processes over repeated fryings. The fluorescence of frying oil increased 15-and 17-fold after tempura-and french-frying, respectively, for 32 consecutive times. Changes in the amounts of tocopherol and the fluorescence correlated well with the changes found by the chemiluminescent intensity and five conventional methods of oil quality measurement. These results indicated that tocopherol retention is affected by the food coating, and that measurements of vitamin E loss and fluorescence increase in oil should be useful for assessing the progressive deterioration of frying oil with its repeated usage.     

Elucidation of Softening Mechanism in Rinse-Cycle Fabric Softeners. Part 2: Uneven Adsorption—The Key Phenomenon to the Effect of Fabric Softeners

We investigated the actual factor determining the softening effect of a fabric softener. The adsorption area of the softener on model cotton cloths and yarns was identified using bromophenol blue. There was almost no softener at the cross-points of the yarns in the cloth samples or in the inner part of the yarns. The softening performance was better when there was less softener at the cross-points of the yarns than when the yarns were evenly covered by the softener. Thus we conclude that the presence of softener at the cross-points of yarns is not a vital factor in the softening effect. In addition, more softener was found on the outer part of the yarn than the inner part, indicating gradation in the adsorption pattern of the softener. Thus, we propose that more softener is adsorbed on the exposed part of the yarn in a cloth, and the formation of a hydrogen-bonding network containing bound water is inhibited, thus softening the outer part of the yarn. However, the presence of a small amount of softener in the inner part of the yarn preserves the hydrogen-bonding network. Favorable elasticity, or bounce, of the yarns and cloth is realized when an appropriate amount of softener is used. Excess softener would reach the inner part of the yarn, reducing the diameter of the core part of the yarn, making the cloth appear wilted.     

Multi-parametric profiling network based on gene expression and phenotype data: a novel approach to developmental neurotoxicity testing

The establishment of more efficient approaches for developmental neurotoxicity testing (DNT) has been an emerging issue for children's environmental health. Here we describe a systematic approach for DNT using the neuronal differentiation of mouse embryonic stem cells (mESCs) as a model of fetal programming. During embryoid body (EB) formation, mESCs were exposed to 12 chemicals for 24 h and then global gene expression profiling was performed using whole genome microarray analysis. Gene expression signatures for seven kinds of gene sets related to neuronal development and neuronal diseases were selected for further analysis. At the later stages of neuronal cell differentiation from EBs, neuronal phenotypic parameters were determined using a high-content image analyzer. Bayesian network analysis was then performed based on global gene expression and neuronal phenotypic data to generate comprehensive networks with a linkage between early events and later effects. Furthermore, the probability distribution values for the strength of the linkage between parameters in each network was calculated and then used in principal component analysis. The characterization of chemicals according to their neurotoxic potential reveals that the multi-parametric analysis based on phenotype and gene expression profiling during neuronal differentiation of mESCs can provide a useful tool to monitor fetal programming and to predict developmentally neurotoxic compounds.     

Highly Fluorescent Nanodiamonds Protein‐Functionalized for Cell Labeling and Targeting

Fluorescent nanodiamond (FND) is attracting much attention as a bioimaging agent because of its inherent biocompatibility and superior optical properties (e.g., excellent photostability and far‐red emission). However, for practical use in life science research, some issues such as higher brightness and ease of bioconjugation have to be solved. Here, it is shown that the 100‐nm FND particles fabricated by using nitrogen‐rich type Ib diamonds and high‐energy proton irradiation are highly fluorescent and readily functionalizable with proteins for biological applications. In the first approach, acid‐treated FND is noncovalently coated with glycoproteins or neoglycoproteins (i.e., proteins chemically modified with multiple sugar residues) for targeting hepatocytes via carbohydrate receptors. In the second approach, FND is first PEGylated and then covalently conjugated with streptavidin, to which biotin‐labeled antibodies of interest are linked. High targeting specificity of the bioconjugated FND is demonstrated with the human hepatoma cell line, HepG2, and breast cancer cell lines, ASB145‐1R, MCF‐7, and MDA‐MB‐231 cells. These approaches should be widely applicable to a variety of situations for specific targeting and labeling of cells.