Molecular cloning, expression, and characterization of a phi-type glutathione S-transferase from Oryza sativa

The structural gene for glutathione S-transferase in Oryza sativa was successfully cloned from a cDNA library by the polymerase chain reaction method. The deduced amino acid sequence of this gene showed 44-66% similarity to the sequences of the class phi GSTs from Arabidopsis thaliana and Zea mays. This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTF3-3 was a homo-dimer composed of 24 kDa subunit and its pI value was approximately 7.3. The OsGSTF3-3 was retained on GSH affinity column and its K_m value for GSH was 0.28 mM. The OsGSTF3-3 displayed high activity toward 1-chloro-2,4-dinitrobenzene, a general GST substrate and also had high activities towards acetanilide herbicides, alachlor, and metolachlor. The OsGSTF3-3 was highly sensitive to inhibition by benastatin A and S-hexyl-GSH. From these results, the expressed OsGSTF3-3 is a phi class GST and seems to play an important role in the conjugation of the chloroacetanilide herbicides.     

Canine adenovirus type 1 infection of a Eurasian river otter (Lutra lutra)

A 10-year-old female Eurasian river otter (Lutra lutra) died after prolonged anorexia and weight loss in the Seoul Grand Park Zoo, Seoul, Republic of Korea. On necropsy, the liver was found to be swollen and friable with 1 lobe enlarged and necrotic. The other organs showed no significant alterations except for mild atrophy of the right kidney. Microscopically, there was multifocal hepatic necrosis. The hepatocytes around the necrotic areas were swollen and contained large basophilic intranuclear inclusions. Periportal infiltration by plasma cells and lymphocytes was also evident. Transmission electron microscopy revealed characteristic hexagonal virus particles sized approximately 70 nm in diameter in the nuclei of the hepatocytes, which were consistent with an adenovirus. Polymerase chain reaction of the formalin-fixed, paraffin-embedded liver sections was used to determine whether the virus was either the canine adenovirus type 1 (CAV-1), canine adenovirus type 2 (CAV-2), or some other viral agent. The results of these tests showed that the virus was CAV-1. To our knowledge, this is the first report on a CAV-1 infection in an otter.     

Chitosan nanoparticles enhance developmental competence of in vitro-matured porcine oocytes

Oxidative stress is inevitable as it is derived from the handling, culturing, inherent metabolic activities and medium supplementation of embryos. This study was performed to investigate the protective effect of chitosan nanoparticles (CNPs) on oxidative damage in porcine oocytes. For this purpose, cumulus–oocyte complexes (COCs) derived from porcine slaughterhouse ovaries were exposed to different concentrations of CNPs (0, 10, 25 and 50 µg/ml) during in vitro maturation (IVM). Oocytes treated with 25 µg/ml CNPs showed significantly higher levels of GSH, along with a significant reduction in ROS levels compared to control, CNPs10 and CNPs50 groups. In parthenogenetic embryo production, the maturation rate was significantly higher in the CNPs25 group than that in the control and all other treated groups. In addition, when compared to the CNPs50 and control groups, CNPs25-treated oocytes showed significantly higher cleavage and blastocyst development rates. The highest concentration of CNPs reduced the total cell number and ratio of ICM: TE cells in parthenogenetic embryos, suggesting that there is a threshold where benefits are lost if exceeded. In cloned embryos, the CNPs25 group, as compared to all other treated groups, showed significantly higher maturation and cleavage rates. Furthermore, the blastocyst development rate in the CNPs25-treated group was significantly higher than that in the CNPs50-treated group, as was the total cell number. Moreover, we found that cloned embryos derived from the CNPs25-treated group showed significantly higher expression levels of Pou5f1, Dppa2, and Ndp52il genes, compared with those of the control and other treated groups. Our results demonstrated that 25 µg/ml CNPs treatment during IVM improves the developmental competence of porcine oocytes by reducing oxidative stress.     

Compensatory growth and body composition of juvenile black rockfish <Emphasis Type="Italic">Sebastes schlegeli</Emphasis> following feed deprivation

ABSTRACT:   Compensatory growth, feeding rate, feed efficiency and chemical composition of juvenile black rockfish (mean weight 1.43 g) were investigated for 35 days after a 14-day feed deprivation treatment under four feeding conditions: one group continuously fed (control) and the other three groups fasted for 5 days (F5), 10 days (F10) and 14 days (F14). All fasted fish were re-fed from day 15. Only F5 achieved the same body weight as the control, indicating that complete compensation occurred in F5. The specific growth rate (SGR) of F5 was the highest at day 21 and then decreased thereafter, showing higher values than the control at days 21, 28 and 42. In contrast, although SGRs of F10 and F14 were higher than that of the control during the whole refeeding period except day 21, they did not catch up the control in body mass, indicating that only partial compensation occurred in F10 and F14. The feeding rate (FR) of all groups except F14 changed in a pattern similar to SGR (Spearman's rank correlation, r _s > 0.9), suggesting that SGR varied depending on FR. Similar feeding efficiencies (FEs) were found in the four groups and they did not vary significantly during the whole refeeding period, suggesting that FE was not the factor affecting SGR. At day 14, the ratios of lipid to lean body mass in F10 and F14 were lower than those in the control and F5, and there was no difference between the control and F5. At day 49, however, only F14 showed a lower value than the other three groups, and there was no difference among the three groups. These results indicate that juvenile black rockfish fasted for 5–14 days can exhibit compensatory growth after refeeding, but timing and degree vary depending on the duration of feed deprivation.     

The effect of bentonite concentration on the drug delivery efficacy of a pH-sensitive alginate/bentonite hydrogel

We produced a protein loaded, pH-sensitive alginate-bentonite hydrogel for wound dressings. Alginate is a nontoxic polysaccharide with favorable pH-sensitive properties that make it useful for the intestinal delivery of protein drugs. However, the use of alginate for drug delivery is limited by drug leaching and rapid dissolution of alginate at the higher pH, which may result in lower entrapment efficiency and a burst in the release of entrapped protein drugs. To overcome these problems, we created a novel cross-linked alginate-bentonite hydrogel by combining mineral-rich bentonite with the alginate matrix along with an additive to ensure controlled release. We analyzed the gel in the drug loading process in an aqueous environment by looking at the release profiles of a model protein drug (BSA) from the hydrogel at pH values of 4.5, 5.2 (skin area) and 7.4, 9.2 (wound area). The swelling ratio decreased with bentonite concentration, but did not fall below 6. The rate of drug release was slowest at a pH value of 4.5 and fastest at a pH value of 9.2. The rate of drug release decreased with bentonite concentration. The presence of bentonite prevents the rapid dissolution of alginate at the higher pH, ensuring the controlled release of the entrapped drug.     

Hydroxycinnamoylmalic acids and their methyl esters from pear (Pyrus pyrifolia Nakai) fruit peel

Two novel caffeoylmalic acid methyl esters, 2-O-(trans-caffeoyl)malic acid 1-methyl ester (6) and 2-O-(trans-caffeoyl)malic acid 4-methyl ester (7), were isolated from pear (Pyrus pyrifolia Nakai cv. Chuhwangbae) fruit peels. In addition, 5 known hydroxycinnamoylmalic acids and their methyl esters were identified: 2-O-(trans-coumaroyl)malic acid (1), 2-O-(cis-coumaroyl)malic acid (2), 2-O-(cis-coumaroyl)malic acid 1-methyl ester (3), 2-O-(trans-coumaroyl)malic acid 1-methyl ester (4), and 2-O-(trans-caffeoyl)malic acid (phaselic acid, 5). The chemical structures of these compounds were determined by spectroscopic data from ESI MS and NMR. Of all the isolated compounds, five hydroxycinnamoylmalic acids and their methyl esters (2-4, 6, 7) were identified in the pear for the first time.     

Variation and correlation analysis of flavonoids and carotenoids in Korean pigmented rice (Oryza sativa L.) cultivars

Flavonoids and carotenoids of pigmented rice ( Oryza sativa L.), including five black cultivars and two red cultivars, from Korea were characterized to determine the diversity among the phytochemicals and to analyze the relationships among their contents. Black cultivars were higher in flavonoids and carotenoids than the red and white cultivars. The profiles of eight phytochemicals identified from the rice grains were subjected to principal component analysis (PCA) to evaluate the differences among cultivars. PCA could fully distinguish between these cultivars. The Heugjinjubyeo (BR-1) and Heugseolbyeo (BR-2) cultivars were separated from the others based on flavonoid and carotenoid concentrations. Flavonoid contents had a positive correlation with carotenoid contents among all rice grains. The BR-1 and BR-2 cultivars appear to be good candidates for future breeding programs because they have simultaneously high flavonoid and carotenoid contents.     

Molecular epidemiology of rabies virus circulating in South Korea, 1998-2010

We analyzed the nucleotide sequences of the G-L (glycoprotein-large protein) intergenic non-coding region of 33 strains of the rabies virus (RABV) isolated in South Korea in 1998-2010 and compared the sequences with those of previously reported non-Korean strains. The similarities of the nucleotide sequences of the G-L region among all Korean RABV isolates ranged from 97.1 to 100%. Based on the phylogenetic analysis of the G-L region, the Korean RABV isolates were classified into three distinct subgroups with high similarity and were most closely related to the non-Korean NeiMeng1025C isolate, which was isolated from a rabid raccoon dog in eastern China, suggesting that the Korean RABV isolates originate from a rabid raccoon dog in northeastern Asia. Our results indicated that G-L region, as a useful phylogenetic indicator, is equivalent to the nucleoprotein (N) or glycoprotein (G) gene for study of RABV molecular epidemiology and that the Korean RABV isolates showing a few substitutions in the G-L region are continuously circulating in South Korea.     

Beef quality traits of heifer in comparison with steer,bull and cow at various feeding environments

The present review has been focused largely on the sex type differences in beef quality among heifers, cows, steers and bulls in various feeding environments. Genetic groups, feeding systems and gender are the major factors that change carcass characteristics and fatty acid profiles of cattle. Studies identified that heifer beef has super characteristics in eating quality and a better healthy composition in fatty acids than steer, cow and bull. Diet influences the variation of fatty acid profile; particularly the level of polyunsaturated fatty acids (PUFA) interacts with breed and sex. Animals finished in pasture systems were reported to show better ratios of PUFA/ saturated fatty acids and n‐6/n‐3. Carcasses of roughage‐fed beef are lighter and have less marbling and lower quality grades but have higher cutability than carcasses of grain‐fed bulls. Heifers and cows are reported to deposit more fat than steers and bulls. Among males, lower production of testosterone by steers favors more fat thickness compared with bulls. Marbling greatly varies among cattle belonging to different sexes, and particularly, females have genetic makeup that efficiently controls deposition. The current review identified that heifers can be a premium beef brand, while steer beef currently take a large part of market share across the world.