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.     

Hydrothermal Assessment of Temporal Variability in Seedbed Microclimate

The microclimatic requirements for successful seedling establishment are much more restrictive than those required for adult plant survival. The purpose of the current study was to use hydrothermal germination models and a soil energy and water flux model to evaluate intra- and interannual variability in seedbed microclimate relative to potential germination response of six perennial grasses and cheatgrass. We used a 44-yr weather record to parameterize a seedbed microclimate model for estimation of hourly temperature and moisture at seeding depth for a sandy loam soil type at the Orchard Field Test Site in southwestern Ada County, Idaho. Hydrothermal germination response was measured in the laboratory for two seed lots of cheatgrass (Bromus tectorum L.), four seed lots of bluebunch wheatgrass (Pseudoroegneria spicata [Pursh] Löve), three seed lots of bottlebrush squirreltail (Elymus elymoides [Raf] Swezey), and one seed lot each of Sandberg bluegrass (Poa secunda J. Presl.), big squirreltail (Elymus multisetus [J.G. Smith] M.E. Jones), thickspike wheatgrass (Elymus lanceolatus [Scribn. And J.G. Smith] Gould) and Idaho fescue (Festuca idahoensis Elmer). Germination response models were developed to estimate potential germination rate for 13 subpopulations of each seed lot for every hour of the 44-yr simulation. Seedbed microclimate was assessed seasonally and for each day, month, and year, and germination rate-sum estimates integrated for a numerical index of relative site favorability for germination for each time period. The rate-sum favorability index showed a consistent pattern among seed lots for different years, and provides a relatively sensitive indicator of annual and seasonal variability in seedbed microclimate. This index could be used with field data to define minimum weather thresholds for successful establishment of alternative plant materials, in conjunction with weather forecast models for making restoration and fire-rehabilitation management decisions in the fall season, for evaluation of potential climate-change impacts on plant community trajectories, and in optimization schemes for selecting among alternative restoration/rehabilitation management scenarios.     

Endometrial polyp in an African wild dog (Lycaon pictus)

An 8-year-old female African wild dog (Lycaon pictus) from a zoo in Gyeonggi province, Republic of Korea presented with a 3.0 x 2.0 x 2.5 cm in size, smooth-surfaced, solitary pedunculated mass protruding into the uterine lumen. Microscopically, the mass was covered with epithelium, contained endometrial gland tissue, and was dilated in the vascularised stroma. Within the mass, there was extensive diffuse haemorrhage with several blood vessels apparently plugged with fibrin. At the base of the mass, the spaces lined with epithelium near the attachment of the stalk were interpreted to be glandular structures. There were segments of cuboidal epithelium found on the surface of the mass, which was similar to the lining the uterus. A diagnosis of an endometrial polyp was made based on the gross and histology findings. This is the first case report of a spontaneous endometrial polyp in an African wild dog.     

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.     

Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

Interleukin (IL)-10 exerts potent anti-inflammatory effects by suppression of both T-help (Th) 1 and Th2 cells. Previous studies have reported that IL-10 can ameliorate various inflammatory disorders. The present study was performed to examine whether IL-10 plasmid DNA could suppress development of atopic dermatitis (AD)-like skin lesions in NC/Nga mice, as an initial step towards the development of an appliance for use in dogs with AD. Intradermal injection of IL-10 plasmid DNA markedly inhibited the development of AD-like skin lesions, as evidenced by a marked decrease in skin symptoms and reduced inflammation within the skin lesions. Efficacy was confirmed by significant decreases in eosinophil ratio and serum IgE concentration, and a reduction in the number of Staphylococcus aureus recovered from the ear. Moreover, relative mRNA expression levels of IL-4 and interferon-γ in the skin lesions of mice injected with IL-10 plasmid DNA were also decreased compared with those of control mice. Of note, higher serum IL-10 levels in mice injected with IL-10 plasmid DNA were maintained compared with those in control mice. Taken together, the results indicate that IL-10 plasmid DNA can suppress the development of AD-like skin lesions by suppressing both Th1 and Th2 cell responses. Beneficial effects of IL-10 plasmid DNA may be expected in dogs with AD.     

Comparison of serological techniques to measure antibody to Pasteurella haemolytica A1.

Analysis of 45 sera was performed employing five techniques which are currently in use in three laboratories to measure anti-Pasteurella haemolytica antibodies. The enzyme linked immunosorbent assay, passive hemagglutination, complement fixation and direct and indirect bacterial agglutination assays were employed and a relationship between tests in the measurement of anti-P. haemolytica antibodies was demonstrated. Regression analysis together with prediction and confidence intervals were tabulated also. The conclusion drawn from statistical analysis was that all five tests are similar in their ability to detect immune responses (antibody and antigen(s) interactions) to Pasteurella haemolytica.