Immunohistochemical detection of porcine teschovirus antigen in the formalin-fixed paraffin-embedded specimens from pigs experimentally infected with porcine teschovirus

Porcine teschovirus (PTV) antigens were detected by a streptavidin-biotin complex method in formalin-fixed paraffin-embedded tissues of 3-week-old pigs that had been inoculated intravenously with PTV Talfan strain. PTV antigens were detected in cytoplasm of nerve cells, glial cells and endothelial cells in the cerebellar nuclei, the grey matter of the midbrain, pons and medulla oblongata and the ventral horn of the spinal cord and of ganglion cells in the spinal ganglion corresponding to those lesions characterized as non-suppurative encephalomyelitis and ganglionitis. The results of this study suggest that nerve cells of the brain stem and spinal cord and ganglion cells of the spinal ganglion permit PTV replication and represent the main target cell population of PTV. This is the first study to demonstrate PTV antigen by immunohistochemistry in formalin-fixed paraffin-embedded tissue specimens from pigs infected with PTV.     

Clinical and molecular analysis of GM2 gangliosidosis in two apparent littermate kittens of the Japanese domestic cat

This case report documents clinical and molecular findings in two littermate kittens of the Japanese domestic cat with GM2 gangliosidosis variant 0. Analysis included detailed physical, magnetic resonance imaging, biochemical, pathological and genetic examinations. At first, these littermate kittens showed typical cerebellar signs at approximately 2 months of age. About 2 months later, they progressively showed other neurological signs and subsequently died at about 7 months of age. Magnetic resonance imaging just before the death showed an enlarged ventricular system, T1 hyperintensity in the internal capsule, and T2 hyperintensity in the white matter of the whole brain. Histological findings suggested a type of lysosomal storage disease. Biochemical studies demonstrated that the kittens were affected with GM2 gangliosidosis variant 0, and a DNA assay finally demonstrated that these animals were homozygous for the mutation, which the authors had identified in a different family of the Japanese domestic cat. The findings in the present cases provide useful information about GM2 gangliosidosis variant 0 in Japanese domestic cats.     

Contrasting effect of perlecan on adipogenic and osteogenic differentiation of mesenchymal stem cells in vitro

Perlecan, a basement membrane component, shows diverse functions in different organs and tissues. However, the role of perlecan in differentiation of mesenchymal stem cells (MSCs) has been barely investigated. In this study, we examined the effect of perlecan on adipogenic and osteogenic differentiation of MSCs in vitro by adding extrinsic perlecan to culture media or blocking the function of intrinsic perlecan expressed into culture media by differentiating MSCs. Extrinsic perlecan suppressed adipogenic differentiation; however, it promoted osteogenic differentiation. These functions were further confirmed by a study of blocking intrinsic perlecan. Perlecan treated with heparitinase‐I also showed the suppressive effect on adipogenic differentiation. In contrast, the promotive effect on osteogenic differentiation was found to be heparan sulfate‐dependent. Intrinsic perlecan was suggested to be effective at the late stage of adipogenic differentiation by a study of perlecan‐blocking performed at distinct periods, but was suggested to be effective at the early stage of osteogenic differentiation. Our results showed perlecan has contrasting effect on adipogenic and osteogenic differentiation of MSCs due to its diverse actions. Based on these outcomes, we recognized that employing extrinsic perlecan or blocking intrinsic perlecan is effective for regulating adipogenic and osteogenic differentiation of MSCs by restricting its direction.     

The role estrogens play in sex differentiation and sex changes of fish

Using genetically controlled all-female and all-male tilapia (Oreochromis niloticus), the role steroid hormones play in the sex differentiation was analyzed histologically, ultrastructurally, immunohistichemically and experimenntally. The results strongly suggest that endogenous estrogen acts as an ovarian inducer, and that the lack of steroid hormone including androgen is important for testicular differentiation. Moreover, the roles of steroid hormones in protogynous sex change of three-spotted wrasse (Halichoeres trimaculatus) and saddleback wrasse (Tharassoma duperrey) were examined. The results strongly support the hypothesis that the endogenous estrogen plays an important role in protogynous sex change.     

Regulation of steriodogenic enzyme mRNAs in rainbow trout (Oncorhynchus mykiss) ovarian follicles in vitro

The effect of gonadotropins, forskolin and IGF-I on steroidogenic enzyme gene expression and E2 production by the rainbow trout follicle were measured. The results indicate that 3β-HSD gene expression is regulated by gonadotropins partially through the cAMP/PKA mechanism. The effect of IGF-I on P450arom mRNA levels suggests that IGF-I is a major regulator of P450arom gene expression.     

Lunar-related spawning in honeycomb grouper, Epinephelus merra

Lunar-related spawning in honeycomb grouper was investigated by histological and behavioral analyses. Ovarian development and spawning of the fish were related to lunar periodicity. And also the fish migrated to out side of the reef area and spawned for a few days after the full moon.     

Surface disinfection of Pacific threadfin, Polydactylus sexfilis, and amberjack, Seriola rivoliana, eggs

The effects of exposing the eggs of Pacific threadfin and amberjack eggs (AEs) to different concentrations of hydrogen peroxide for 5 min on hatch rate and survival were assessed in a series of experiments using a petri dish model rearing system. Despite significant inter‐batch variation in hatch rate, it was shown that eggs of both species could be safely exposed to up to 11 340 mg L⁻¹ H₂O₂ for 5 min. Exposure to 34 230 mg L⁻¹ H₂O₂ for 5 min was shown to be lethal to AEs at a late stage of development. In two further experiments, it was demonstrated that Pacific threadfin eggs were resistant to all tested concentrations of a range of polyvinylpyrrolidone iodine (PVP‐I) concentrations and contact times (up to 1000 mg L⁻¹ PVP‐I for 10 min). The level of bacteria adhering to the eggs of both species was highly variable. Where eggs were heavily colonized (>10⁴ cfu egg⁻¹), hydrogen peroxide concentrations of at least 11 340 mg L⁻¹, or PVP‐I concentrations higher than 500 mg L⁻¹ for 10 min, were required for effective sterilization. In less colonized batches, rinsing in sterile seawater or exposure to lower (550 mg L⁻¹) concentrations of H₂O₂ was sufficient to result in high apparent levels of surface sterility (<1 cfu egg⁻¹).     

Activity patterns of abalone under experimental conditions

Locomotion behaviour of the abalone, Nordotis discus (Reeve), N. gigantea (Gmelin) and Sulculus aquatilis (Reeve), was observed for 5‐ to 10‐day periods during the early summer and autumn from 1997 to 1999. Before sunset, the animals were individually put beneath each shelter situated on the flat concrete floor of indoor and outdoor tanks. For tracing of the locomotion path, a luminous rod was fixed on the animal's shell. Except for non‐appearance, the locomotion behaviour showed leaving or homing; these ratios against each total observation according to each species were 70.0% and 16.7% in N. discus, 47.1 and 23.5% in N. gigantea and 57.6 and 24.2% in S. aquatilis respectively. Irrespective of species, recurrences to the home were frequently recognized during the locomotion. For the homing behaviour, its critical distance was approximately 1 m from the home. Moonlight did not completely restrain the abalone appearance from the home, though the appearance frequency during the period around the full moon was lowered and in some cases the time of initial appearance was retarded.     

Elevation of lipid peroxide level and production of hydroxy lipids in cultured Hepa-T1 cells by oxidative stressors

ABSTRACT:   In order to investigate the relationships between levels of lipid peroxides (LPO), hydroxy lipids (L-OH) and of antioxidative enzymes in live fish, cultured cells from tilapia liver (Hepa-T1) were treated with oxidative stressors, 2,2'-azobios(2-amidino-propane) dihydrochloride (AAPH) and H₂O₂. By treating with 5 mM AAPH, LPO and L-OH of the cells significantly increased time-dependently. In particular, L-OH increased from 7.6 ± 0.7 to 23.6 ± 1.8 nmol/mg protein after 2 h of treatment, and synchronously the glutathione peroxidase (GPx) activity of the cells increased from 259 ± 106 to 1970 ± 135 mU/mg protein. In the case of H₂O₂, however, no elevation of L-OH was observed. In this case, catalase (CAT) and superoxide dismutase (SOD) activities increased time-dependently. Especially, after 2 h exposure, the CAT activity increased remarkably from 217 ± 21 to 2510 ± 120 mU/mg protein, though the GPx activity did not change significantly. These results suggest that the elevation of the L-OH level is closely associated with the elevation of GPx activity, but not with that of CAT or SOD. However, when vitamin E-enriched Hepa-T1 cells were treated with AAPH and H₂O₂, the production of LPO and L-OH was suppressed significantly. The findings suggest that vitamin E also plays an important role in the levels of the oxidation products in vivo .