Establishment of evaluation method to determine effects of veterinary medicinal products on manure fermentation using small-scale composting apparatus

To evaluate on a laboratory scale the influence of veterinary medicinal products (VMPs) excreted into feces on manure fermentation, we have developed an evaluation method that uses a small-scale composting apparatus. Each run is of approximately 3 kg scale and the operation can be conducted in an environmentally controlled laboratory. The main evaluation parameter is calorific value generated by aerobic fermentation. At the sulfadimethoxine (SDM) trial, the volume of CO(2) generated during fermentation and the disappearance of the inhibitory effect of immature manure on sprouting (using Komatsuna (Brassica rapa var. perviridis)) were measured. In addition, DNA of 16S rRNA was extracted from a manure sample and subjected to denaturing gradient gel electrophoresis (DGGE). The results suggest that the presence of such VMPs in feces affected the microbial community in manure fermentation, and indicate that the evaluation method may be used as a standard method to evaluate the effect of VMPs on the microbial community. Using the method, we obtained data of the influence of five VMPs approved for stockbreeding in Japan on swine manure fermentation. Erythromycin (EM) affected the calorific value even at a relatively low concentration (105 mg/3 kg manure). In contrast, oxytetracycline hydrochloride (OTC), norfloxacin (NFLX), and tylosin tartrate (TS) had no effect at that concentration. These VMPs also affected the increase of fermentation temperature when added at high concentrations.     

Comparison of efficiency of terminal differentiation of oligodendrocytes from induced pluripotent stem cells versus embryonic stem cells in vitro

Oligodendrocytes are the myelinating cells of the central nervous system (CNS), and defects in these cells can result in the loss of CNS functions. Although oligodendrocyte progenitor cells transplantation therapy is an effective cure for such symptoms, there is no readily available source of these cells. Recent studies have described the generation of induced pluripotent stem cells (iPS cells) from somatic cells, leading to anticipation of this technique as a novel therapeutic tool in regenerative medicine. In this study, we evaluated the ability of iPS cells derived from mouse embryonic fibroblasts to differentiate into oligodendrocytes and compared this with the differential ability of mouse embryonic stem cells (ES cells). Experiments using an in vitro oligodendrocyte differentiation protocol that was optimized to ES cells demonstrated that 2.3% of iPS cells differentiated into O4⁺ oligodendrocytes compared with 24.0% of ES cells. However, the rate of induction of A2B5⁺ oligodendrocyte precursor cell (OPC) was similar for both iPS-derived cells and ES-derived cells (14.1% and 12.6%, respectively). These findings suggest that some intracellular factors in iPS cells inhibit the terminal differentiation of oligodendrocytes from the OPC stage.     

Inter-laboratory evaluation studies for establishment of notified ELISA methods for allergic substances (peanuts)

Inter-laboratory evaluation studies were conducted for ELISA methods for allergic substances (peanuts). Extracts of biscuit, sauce, chocolate and butter spiked with peanut standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of the ELISA methods using a Peanut Protein ELISA Kit (Peanut kit) and a FASTKIT Peanut ELISA kit (Peanut ELISA kit) were mostly below 10%. Mean recoveries of the peanut standard protein from the food extracts were over 40% in the two ELISA methods. Repeatability relative standard deviations of peanut standard protein in four food extracts were in the ranges of 15.2-49.7% and 3.0-28.3% for the Peanut kit and the Peanut ELISA kit, respectively. Reproducibility relative standard deviations of peanut standard protein in four food extracts were 23.5-44.4%, 9.6-28.4% for the Peanut kit and the Peanut ELISA kit, respectively. The detection limits of both ELISA methods were 2-2.5 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of peanut protein levels in extracts of biscuit, sauce, chocolate and butter.     

Formation of protein adducts of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine in cooked foods

Heterocyclic amines (HCAs) are mutagenic and carcinogenic compounds found in cooked meat and fish. Although HCAs are known to form adducts with protein after metabolic activation, adduct formation during cooking has not been elucidated. In this study, we showed that 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP) is released from high molecular weight compounds by acid or enzymatic hydrolysis of cooked foods. Formation of free and protein adduct forms of PhIP was dependent on cooking temperature and time, and PhIP–protein adducts were estimated to form after formation of free PhIP. We also demonstrated that PhIP–protein adduct is formed by heating of PhIP and albumin as a model protein. A new adduct peak including [M+H]⁺ (m/z=225) of PhIP as a fragment ion was detected in the high molecular weight fraction of heat‐treated protein by LC–MS analysis. From model experiments by heating of PhIP and amino acids, the adduct was estimated to be produced by condensation of the amino group of PhIP and the carboxyl group of protein. PhIP–protein adducts were detected in several cooked meat and fish at ng/g food level as PhIP content. These results suggest that food‐borne protein adducts of HCAs may influence human HCA exposure and carcinogenic risk.     

Overexpression of 1-Deoxy-D-xylulose-5-phosphate reductoisomerase gene in chloroplast contributes to increment of isoprenoid production

Plants synthesize a large number of isoprenoid compounds that are of industrial, nutritional and medicinal importance. 1-Deoxy-D-xylulose reductoisomerase (DXR) catalyzes the first committed step of plastidial isoprenoid-precursor biosynthesis. In the present study, we generated transplastomic tobacco plants that overproduced DXR from Synechosystis sp. strain PCC6803. The transformants showed increase in the content of various isoprenoids such as chlorophyll a, beta-carotene, lutein, antheraxanthin, solanesol and beta-sitosterol, indicating that the DXR reaction is one of the key steps controlling isoprenoid level in tobacco leaves. A qualitative change in isoprenoid composition was also observed. The growth phenotype of the transplastomic plants was similar to that of wild-type plants. These results showed that plastid metabolic engineering is useful in manipulating the yield of isoprenoids in plants.     

A link selection method for web‐browser using eye‐gaze input

Methods for browsing Web pages using eye‐gaze input have been proposed for severely physically handicapped people who cannot handle a computer mouse to utilize convenient services on the Internet. There are two important functions for Web‐browsers, namely scroll and link selection. Since it is difficult to introduce existing link selection methods using eye‐gaze input for home usage because of high costs or complexity, we propose in this paper a novel link selection method using eye‐gaze input. The proposed method analyzes the HTML source in a Web page and makes a group of links by the content of each. In evaluation experiments, we compare the proposed method with a comparative method using eye‐gaze input with respect to the average response time in selecting the links. The results of the experiments show that the average response is nearly twice as fast as that of the compared method. © 2014 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.