Accumulation of gamma-aminobutyrate (GABA) caused by heat-drying and expression of related genes in immature vegetable soybean (edamame)

We studied the effects of drying of immature seeds of vegetable soybean (Glycine max L. Merrill) on the accumulation of gamma-aminobutyrate (GABA) in the seeds. GABA accumulated after heat-drying, with the maximum at 40°C. The GABA content (447.5 mg/100 g DW) increased to more than 5 times the value in untreated seeds (79.6 mg/100 g DW). In contrast, the glutamate content decreased rapidly to 1/3 the level in the untreated seeds. The GABA content increased early in the heat-drying treatment: after 30 min, it had increased to 1.5 times the value in the untreated seeds. GABA did not accumulate in the vacuum-drying treatment. Among genes related to the GABA shunt, the gene for glutamate decarboxylase (EC 4.1.1.15), which catalyzes the decarboxylation of glutamate to produce GABA, showed relatively high expression, decreasing to only 70% of the value in untreated seeds even after 4 h of treatment. In contrast, expression of the genes for two catabolic mitochondrial enzymes, GABA transaminase (GABA-T; EC 2.6.1.19) and succinate semialdehyde dehydrogenase (SSADH; EC 1.2.1.16), decreased rapidly during heat-drying. These results suggest that the accumulated GABA was not metabolized rapidly by GABA-T and SSADH and therefore remained at high levels.     

Overexpression of the LASAP2 gene for secretory acid phosphatase in white lupin improves the phosphorus uptake and growth of tobacco plants

Secretion of acid phosphatase (APase) from the roots to take up phosphorus (P) is a well-known strategy of plants under P-deficient conditions. White lupin, which shows vigorous growth in low-P soils, is noted for its ability to secrete APase under P-deficient conditions. The APase secreted by white lupin roots is stable in soil solution and shows low substrate specificity, suggesting that genetic modification of plants using the APase gene LASAP2 might improve their ability to use organic P. The objective of the present study was to evaluate the potential of LASAP2 transgenic plants to increase organic P utilization. Dry matter production and P accumulation were higher in LASAP2 transgenic tobacco plants grown in gel media containing soluble phytate as the sole P source than in wild-type tobacco plants. Phosphorus uptake by the transgenic plants also increased in soil culture conditions. LASAP2 was apparently more effective in the liberation of organic P, including phytate, in the soil than the native tobacco APase. Thus, the enzymatic stability of LASAP2 in the soil appears to be an important factor for P acquisition.     

Biological nitrification inhibition by Brachiaria humidicola roots varies with soil type and inhibits nitrifying bacteria, but not other major soil microorganisms

The tropical pasture grass Brachiaria humidiola (Rendle) Schweick releases nitrification inhibitory compounds from its roots, a phenomenon termed 'biological nitrification inhibition' (BNI). We investigated the influence of root exudates of B. humidicola on nitrification, major soil microorganisms and plant growth promoting microorganisms using two contrasting soil types, Andosol and Cambisol. The addition of root exudates (containing BNI activity that is expressed in Allylthiourea unit (ATU) was standardized in a bioassay against a synthetic inhibitor of nitrification, allylthiourea, and their function in soil was compared to inhibition caused by the synthetic nitrification inhibitor dicyandiamide. At 30 and 40 ATU g⁻¹soil, root exudates inhibited nitrification by 95% in fresh Cambisol after 60 days. Nitrification was also similarly inhibited in rhizosphere soils of Cambisol where B. humidicola was grown for 6 months. Root exudates did not inhibit other soil microorganisms, including gram-negative bacteria, total cultivable bacteria and fluorescent pseudomonads. Root exudates, when added to pure cultures of Nitrosomonas europaea , inhibited their growth, but did not inhibit the growth of several plant growth promoting microorganisms, Azospirillum lipoferum , Rhizobium leguminosarum and Azotobacter chroococcum. Our results indicate that the nitrification inhibitors released by B. humidicola roots inhibited nitrifying bacteria, but did not negatively affect other major soil microorganisms and the effectiveness of the inhibitory effect varied with soil type.     

Pathogenicity of Bordetella bronchiseptica isolated from apparently healthy rabbits in guinea pig,rat, and mouse

Bordetella bronchiseptica (B. bronchiseptica) is associated with respiratory tract infections in laboratory animals. In our laboratory animal facility, B. bronchiseptica was isolated from 21 of 27 apparently healthy rabbits obtained from a breeding farm contaminated with B. bronchiseptica. Restriction fragment length polymorphism (RFLP) analysis showed that the flagellin genotype of isolates from the laboratory animal facility and breeding farm was type A, which is seen relatively frequently in rabbits in Europe. To examine its pathogenicity, guinea pigs, rats, and mice were inoculated intranasally with a representative strain isolated in the laboratory animal facility. Following inoculation of 10⁷ colony forming unit (cfu), severe inflammation was observed in the lungs of guinea pig and mice, although the inflammation was less severe in rats. The strain was recovered from the trachea and lungs of these species after inoculation with lower dose such as 10³ or 10⁴ cfu. These results suggest that the isolated strain causes respiratory tract infection in guinea pigs, rats, and mice, and that its pathogenicity higher in mice than in rats. This study extends our knowledge of interpreting the microbiologic status of laboratory animals, which will contribute to the development of reliable and reproducible animal experiments.     

Effect of a stepwise lighting method termed “stage reduced lighting” using LED and metal halide fishing lamps in the Japanese common squid jigging fishery

Lighting systems combining light-emitting diodes (LEDs) and metal halide lamps (MHs) are expected to be energy-saving tools in Japan??s squid jigging fishery. Previous research has shown the need for light stronger than LEDs (9?kW) and 36 MHs (108?kW) to catch the Japanese common squid Todarodes pacificus. We tested a stepwise lighting method termed ??stage reduced lighting?? in the Tsushima Strait in January and February 2010 using nine fishing boats. LEDs (9?kW) and 50 MHs (150?kW) were lit for 3.9?h on average, and then the number of MHs was reduced to either 30 or 36 until the end of fishing (7.3?h on average). This method reduced fuel consumption by 22?C25?% compared to the continuous use of all fishing lamps (159?kW). We carried out a catch analysis of nine experimental boats and 21 commercial boats during the experimental period. Generalized linear modeling analysis suggested that the squid catch can be explained by the illuminated fraction of the moon and monthly changes in squid abundance, and the lighting method. The stage reduced lighting using LEDs and MHs has the potential to reduce fuel consumption while maintaining the squid catch.     

Histopathology of acute colchicine intoxication: novel findings and their association with clinical manifestations

A 32-year-old woman attempted suicide by ingesting Gloriosa bulbs and died approximately 2 days later. Toxicological examination revealed a potentially fatal blood concentration of colchicine (0.096 mg/L). In addition to the increased mitotic figures in the gastrointestinal mucosa, a unique finding for acute colchicine intoxication, pathological examination showed microvesicular lipid droplets in the liver, kidney, heart, and conduction system. Furthermore, central chromatolysis of neurons was observed in the pontine nucleus, medial accessory olivary nucleus, nucleus of the solitary tract, and nucleus ambiguus. Grumose degeneration of the cerebellar dentate nucleus was also evident. These pathological findings may help identify colchicine intoxication, even in the absence of evidence suggesting ingestion during autopsy. Moreover, pathological changes in the heart and central nervous system may be associated with the development of serious complications of acute colchicine intoxication.     

Molecular sexing of Japanese cormorants used for traditional fishing on the Nagara River in Gifu City

The Japanese cormorants used in traditional fishing in Japan are wild derived and their sex cannot be determined from their appearance. Applicability of molecular sex determination based on the size difference between CHD1Z and CHD1W introns was confirmed in male and female Japanese cormorants whose sexes had been ascertained by pathological autopsy. All of 21 birds of unknown sex reared by a cormorant fishing master were identified as males. The molecular sexing method will provide valuable information on sex differences of wild Japanese cormorants, including tameness, trainability, behavior and fishing capability, as well as for future trials involving artificial reproduction.     

Detection of specific antibodies against deflagellated Salmonella Enteritidis and S. Enteritidis FliC-specific 9kDa polypeptide

Specific antibody levels of laying hens and young chickens experimentally infected with Salmonella Enteritidis and vaccinated farm flocks were evaluated by enzyme-linked immunosorbent assays (ELISAs) with two different antigens, deflagellated S. Enteritidis whole cell (DEWC) and S. Enteritidis FliC-specific 9kDa polypeptide (SEP9). Infected laying hens excreted S. Enteritidis throughout the experimental period, and the specific antibody titers in DEWC-ELISA, were significantly higher than the uninfected group. It suggests that this DEWC-specific antibody will serve as an effective indicator of S. Enteritidis infection, especially for non-vaccinated laying flocks. SEP9-specific antibodies were detected in spray-inoculated young chickens but not in oral-inoculated young chickens. Compared with greatly high SEP9-specific antibody levels of vaccinated farm flocks, no response was observed in orally infected hens. These results indicate that S. Enteritidis discontinues expressing SEP9 once it has crossed the intestinal barrier, and that SEP9-ELISA will serve as a valuable monitoring tool for the status of S. Enteritidis vaccination on a flockwide basis, independent of stable S. Enteritidis infections.     

Alcohol and dietary fibre intakes affect circulating sex hormones among premenopausal women

BACKGROUND: The association of alcohol and fibre intake with breast cancer may be mediated by circulating sex hormone levels, which are predictors of breast cancer risk. OBJECTIVE: To evaluate the relationship of alcohol and dietary fibre intake with circulating sex hormone levels among premenopausal women. METHODS: A total of 205 premenopausal women completed a validated food-frequency questionnaire at baseline and after 2 years; blood samples taken at the same time were analysed for circulating sex hormone concentrations, including oestrone (E1), oestradiol (E2), free E2, progesterone, androstenedione and sex hormone-binding globulin, by radioimmunoassay. We used mixed models to estimate least-square means of sex hormone concentrations for alcohol intake categories and quartiles of dietary intake. RESULTS: After adjustment for covariates, alcohol consumption was moderately associated with higher circulating oestrogen levels; those who consumed more than one drink per day had 20% higher E2 (Ptrend=0.07) levels than non-drinkers. In contrast, higher dietary fibre intake was associated with lower serum levels of androstenedione (-8% between the lowest and highest quartiles of intake, Ptrend=0.06), but not oestrogens. Similarly, consumption of fruits (-12%, Ptrend=0.03), vegetables (-9%, Ptrend=0.15) and whole grains (-7%, Ptrend=0.07) showed inverse associations with androstenedione levels. CONCLUSIONS: The consistency of the observed differences in sex hormone levels associated with alcohol and fibre-rich foods indicates that these nutritional factors may affect sex hormone concentrations and play a role in breast cancer aetiology and prevention.