Immunolocalization of steroidogenic enzymes (P450scc, P450c17, P450arom) in gonad of Japanese eel

Antibodies against P450scc, P450c17 and P450arom were generated using recombinant proteins. In eel testis, P450scc and P450c17 were immunolocalized as clusters in Leydig cells. In vitellogenic eel ovary, P450scc and P450c17 immunoreactive cells were localized as clusters in the outer layer of the ovarian follicle. In contrast, P450arom seemed to be immunolocalized in the innermost follicle layer.     

Photosynthetic response and nitrogen use efficiency of sugarcane under drought stress conditions with different nitrogen application levels

Drought stress which often occurs during early growth stage is one constraint in sugarcane production. In this study, the response of sugarcane to drought and nitrogen application for physiological and agronomical characteristics was investigated. Two water regimes (well-watered and drought stress from 60 to 120 day after transplanting) and four nitrogen levels (0, 4.4, 8.8 and 13.2 g pot^?1 equivalent to 0, 90, 180 and 270 kg ha^?1, respectively) were assigned in a Split-plot design with three replications. The results showed that photosynthetic responses to light intensity and intercellular CO₂ concentrations of sugarcane were different between fertilized and non-fertilized treatments. Photosynthetic rates of 180 and 270 N treatments, normally, were significantly higher than that of 90 N, but not significant at drought conditions. Photosynthetic rates of 0 N treatment were the lowest under both conditions. Higher nitrogen application supported higher photosynthetic rate, stomatal conductance, and chlorophyll content because of higher nitrogen concentration accumulated into the leaf. Drought significantly reduced the potential photosynthetic rate, stomatal conductance, SPAD, leaf area, and biomass production. Higher nitrogen applications with larger root system could support higher photosynthetic activities to accumulate more dry mass. Strong positive coefficient between photosynthetic and biomass nitrogen use efficiency and drought tolerance index may suggest that higher nitrogen use efficiency could help plants have higher ability to tolerate drought stress.     

Availability of Micro-Tom mutant library combined with TILLING in molecular breeding of tomato fruit shelf-life

Novel mutant alleles of an ethylene receptor Solanum lycopersicum ETHYLENE RESPONSE1 (SlETR1) gene, Sletr1-1 and Sletr1-2, were isolated from the Micro-Tom mutant library by TILLING in our previous study. They displayed different levels of impaired fruit ripening phenotype, suggesting that these alleles could be a valuable breeding material for improving shelf life of tomato fruit. To conduct practical use of the Sletr1 alleles in tomato breeding, genetic complementation analysis by transformation of genes carrying each allele is required. In this study, we generated and characterized transgenic lines over-expressing Sletr1-1 and Sletr1-2. All transgenic lines displayed ethylene insensitive phenotype and ripening inhibition, indicating that Sletr1-1 and Sletr1-2 associate with the ethylene insensitive phenotype. The level of ethylene sensitivity in the seedling was different between Sletr1-1 and Sletr1-2 transgenic lines, whereas no apparent difference was observed in fruit ripening phenotype. These results suggested that it is difficult to fine-tune the extent of ripening by transgenic approach even if the weaker allele (Sletr1-2) was used. Our present and previous studies indicate that the Micro-Tom mutant library combined with TILLING could be an efficient tool for exploring genetic variations of important agronomic traits in tomato breeding.     

Antimicrobial resistance of Staphylococcus pseudintermedius isolates from healthy dogs and dogs affected with pyoderma in Japan

Staphylococcus pseudintermedius strains were isolated from healthy dogs and dogs with pyoderma in 2000-2002 and 2009. All the isolates from dogs with pyoderma in 1999-2000 and from healthy dogs in 2000-2002 and 2009 were susceptible to cefalexin and/or other cephalosporins and oxacillin. However, 7.1-12.5 and 11.4% of S. pseudintermedius isolates from dogs with pyoderma in 2009 were resistant to cephalosporins and oxacillin, respectively. All S. pseudintermedius isolates from dogs with pyoderma in 1999-2000 and those from healthy dogs in 2000-2002 were susceptible to fluoroquinolones; however, 50% of the S. pseudintermedius strains isolated from dogs with pyoderma in 2009 and 30% of the S. pseudintermedius strains isolated from healthy dogs in 2009 were resistant to fluoroquinolones. Of the 21 oxacillin-resistant S. pseudintermedius (MRSP) isolates, 11 carried SCCmec type V and 10 carried hybrid SCCmec types II-III. Staphylococcus pseudintermedius strains that were resistant to only one of three fluoroquinolones had a mutation in the quinolone resistance determination region of grlA, whereas S. pseudintermedius strains that were resistant to two or more fluoroquinolones had mutations in the quinolone resistance determination regions of both grlA and gyrA.     

Effects of feeding dried distillers grains with solubles (DDGS) on meat quality at the late stage of the fattening period of Holstein steers

Feeding dried distillers grains with solubles (DDGS) during the late stage of the fattening period of Holstein steers was studied in regard to the influence on meat quality. Sixteen Holstein steers approximately 18 months old were used in this study. Eight animals were fed commercial concentrated feed for the entire fattening period. The other eight animals were fed 15% DDGS in the concentrated feed for 3 months before slaughtering. The moisture, ether extract and crude protein from both groups was approximately the same. The thiobarbituric acid reactive substance (TBA) value of storage for 7 days at 5°C from the animals fed DDGS showed a tendency to be lower (P = 0.059). The change in the TBA value during storage was also lower for the animals not fed DDGS (P < 0.05). There were no differences in the subcutaneous fat color between the two groups. The a*(measure of redness) and b*(measure of yellowness) of the M. longissimus from the animals fed DDGS showed a tendency to be lower (P = 0.051, 0.070). The fatty acid composition of the M. longissimus, subcutaneous and perirenal fat were not widely influenced by the feeding of DDGS. It is suggested that feeding 15% DDGS during the late stage of the fattening period for Holstein steers reduced the oxidation of the beef.     

Regulated upon activation normal T-cell expressed and secreted (RANTES) contributes to abortion caused by Brucella abortus infection in pregnant mice

Brucella abortus (B. abortus) is a facultative intracellular pathogen that can survive inside macrophages and trophoblast giant cells, and the causative agent of brucellosis. In the present study, we found that production of regulated upon activation normal T-cell expressed and secreted (RANTES) due to B. abortus infection contributes to abortion in pregnant mice. B. abortus infected pregnant interferon-gamma (IFN-gamma) knockout mice died within 15 days of infection, but non-pregnant IFN-gamma knockout mice were still alive. With infection by wild type B. abortus, a large amount of RANTES production was observed in pregnant IFN-gamma knockout mice, and induction of RANTES was also observed in normal pregnant mice infected with the wild type, but not in those infected with the intracellular replication-defective mutant. Production of RANTES and IFN-gamma were inhibited in mice inoculated with the respective RANTES or IFN-gamma antibody. Neutralization of RANTES, induced by B. abortus infection, served to prevent abortion. These results indicate that the production and function of RANTES are correlated with IFN-gamma in pregnant mice infected with B. abortus.     

Zinc uptake system (znuA locus) of Brucella abortus is essential for intracellular survival and virulence in mice

Brucella spp. are facultative intracellular pathogens that have the ability to survive and multiply in professional and nonprofessional phagocytes, and cause abortion in domestic animals and undulant fever in humans. The mechanism and factors of virulence are not fully understood. High-affinity zinc uptake system protein mutant (znuA mutant) showed reduced growth in zinc chelated medium, and failed to replicate in HeLa cells and mouse bone marrow-derived macrophages. Transformation of znuA mutant with a shuttle vector pBBR1MCS-4 containing znuA gene restored the growth in zinc chelated medium and intracellular replication in HeLa cells and macrophages to a level comparable to that of wild-type strain. Bacterial internalization into HeLa cells and macrophages and co-localization with either late endosomes or lysosomes of znuA mutant were not different from those of wild-type strain. These results suggest that znuA does not contribute to intracellular trafficking of B. abortus, but contributes to utilization of zinc required for intracellular growth.     

Multiple high-throughput analyses monitor the response of E. coli to perturbations

Analysis of cellular components at multiple levels of biological information can provide valuable functional insights. We performed multiple high-throughput measurements to study the response of Escherichia coli cells to genetic and environmental perturbations. Analysis of metabolic enzyme gene disruptants revealed unexpectedly small changes in messenger RNA and proteins for most disruptants. Overall, metabolite levels were also stable, reflecting the rerouting of fluxes in the metabolic network. In contrast, E. coli actively regulated enzyme levels to maintain a stable metabolic state in response to changes in growth rate. E. coli thus seems to use complementary strategies that result in a metabolic network robust against perturbations.