Purification and characterisation of chicken liver monomeric glutathione peroxidase

1. A novel glutathione peroxidase, which is distinct from tetrameric glutathione peroxidase, was purified to homogeneity from a broiler chick liver cytosolic fraction using 5 different column chromatographic methods.

2. The enzyme in cytosol was separated from ‘classic’ tetrameric glutathione peroxidase and glutathione S‐transferases by DEAE‐Sephacel and Sephadex G‐100 chromatographies and further purified by Mono Q, hydroxylapatite and sulphobro‐mophthalein‐S‐glutathione‐agarose chromatographies.

3. The molecular weight of the purified enzyme determined by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis was 19,500 and that found by gel filtration chromatography was comparable. This indicates that the enzyme protein is a single polypeptide. The isoelectric point of the enzyme was determined as 7.0 by polyacrylamide gel isoelectric focusing.

4. The purified enzyme catalysed the reduction of hydrogen peroxide, cumene hydroperoxide, tert‐butyl hydroperoxide and linoleic acid hydroperoxide. Furthermore, it reduced phosphatidylcholine hydroperoxide in the absence of phospholi‐pase A2. The optimum pH for the enzyme reaction was 7.0. The antiserum against the purified enzyme reacted with the 19.5 kDa polypeptide in the liver cytosol of duck and quail.     

Oral administration of L-citrulline changes the concentrations of plasma hormones and biochemical profile in heat-exposed broilers

We examined the effects of oral administration of L-citrulline (L-Cit) on plasma metabolic hormones and biochemical profile in broilers. Food intake, water intake, and body temperature were also analyzed. After dual oral administration (20 mmol/head/administration) of L-Cit, broilers were exposed to a high ambient temperature (HT; 30 ± 1°C) chamber for 120 min. Oral administration of L-Cit reduced (p < .001) rectal temperature in broilers. Food intake was increased (p < .05) by heat stress, but it was reduced (p < .05) by L-Cit. Plasma levels of 3,5,3′-triiodothyronine, which initially increased (p < .0001) due to heat stress, were reduced (p < .01) by oral administration of L-Cit. Plasma insulin levels were increased by heat exposure (p < .01) and oral L-Cit (p < .05). Heat stress caused a decline (p < .05) in plasma thyroxine. Plasma lactic acid (p < .05) and non-esterified fatty acids (p < .01) were increased in L-Cit-treated heat-exposed broilers. In conclusion, our results suggest that oral L-Cit can modulate plasma concentrations of major metabolic hormones and reduces food intake in broilers.     

Molecular characterization of the equine herpesvirus 1 strains RacL11 and Kentucky D

The pathogenicities of RacL11 and Kentucky D strains of equine herpesvirus 1 in the hamster infection model are different from those of Ab4p and the Japanese isolates. Virus genome restriction fragment length polymorphism analysis and sequence comparison of an intergenic region, glycoproteins and tegument genes showed higher conservation but with some strain-specific differences. These results indicate that point nucleotide differences in RacL11 and Kentucky D might be responsible for their pathogenicity in rodent models.     

GM1 gangliosidosis in a Japanese domestic cat: a new variant identified in Hokkaido,Japan

A male Japanese domestic cat with retarded growth in Hokkaido, Japan, showed progressive motor dysfunction, such as ataxia starting at 3 months of age and tremors, visual disorder and seizure after 4 months of age. Finally, the cat died of neurological deterioration at 9 months of age. Approximately half of the peripheral blood lymphocytes had multiple abnormal vacuoles. Magnetic resonance imaging showed bisymmetrical hyperintensity in the white matter of the parietal and occipital lobes in the forebrain on T2-weighted and fluid-attenuated inversion recovery images, and mild encephalatrophy of the olfactory bulbs and temporal lobes. The activity of lysosomal acid β-galactosidase in leukocytes was negligible, resulting in the biochemical diagnosis of GM1 gangliosidosis. Histologically, swollen neurons characterized by accumulation of pale, slightly granular cytoplasmic materials were observed throughout the central nervous system. Dysmyelination or demyelination and gemistocytic astrocytosis were observed in the white matter. Ultrastructually, membranous cytoplasmic bodies were detected in the lysosomes of neurons. However, genetic analysis did not identify the c.1448G>C mutation, which is the single known mutation of feline GM1 gangliosidosis, suggesting that the cat was affected with a new variant of the feline disease.     

Supplemental effect of cholyltaurine and soybean lecithin to a soybean meal-based fish meal-free diet on hepatic and intestinal morphology of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

The supplemental effects of cholyltaurine and soybean lecithin to a defatted soybean meal (SBM)-based fish meal-free diet on the hepatic and intestinal morphology of rainbow trout Oncorhynchus mykiss were investigated. A fish meal-based diet (FM), three fish meal-free diets supplemented either with soybean lecithin (SL), bovine gall powder (BG) or cholyltaurine (C-tau) and an unsupplemented fish meal-free diet (NFM), were given to fish (−11 g) for 10 weeks. The growth was the lowest in fish fed diet NFM, intermediate in fish fed diet SL and the highest in fish fed diet FM, BG and C-tau. Hepatocytes of fish fed diet NFM were atrophied, and the distal intestine of these fish showed abnormal features: disintegrated microvilli and fatty degeneration in epithelial cells and increased amount of connective tissue in the submucosa. Hepatic and intestinal histological features in fish fed diets SL, BG and C-tau were similar to those in fish fed diet FM. These results indicate that soybean lecithin, bovine bile salts and cholyltaurine have similar effects on normalizing the hepatic and intestinal morphologies of rainbow trout fed the SBM-based diet, although growth promotion effect was limited in the soybean lecithin.     

Thermal stability of carp G-actin monitored by loss of polymerization activity using an extrinsic fluorescent probe

ABSTRACT:   The thermal stability of carp G-actin was investigated by monitoring loss of actin polymerization ability. To determine the amount of native actin remaining after heat treatment, actin was labeled with a fluorescence reagent, N-(1-pyrene)iodoacetamide. The loss of polymerization ability of carp actin during heat treatment, at between 45 and 55°C, occurred faster than that of chicken actin. The inactivation rate was influenced by concentrations of ATP and Ca²⁺ in solution. With the increase of Ca²⁺ concentration, the inactivation of carp actin was markedly suppressed. Furthermore, the activation energy of the inactivation of carp actin obtained from an Arrhenius plot was similar to that of chicken actin. These results indicated that the thermal instability of carp G-actin was due to the low affinites of ATP and Ca²⁺ for carp actin described in a previous report.     

Intrapancreatic accessory spleen mimicking pancreatic insulinoma with intrapancreatic metastasis in a cat

An 11-year-old neutered male Domestic Shorthair cat presented with a 3-month history of hypoglycemia, two episodes of seizure, and intermittent tick-like signs. Serum biochemistry revealed severe hypoglycemia associated with high insulin concentrations. Dynamic abdominal computed tomography (CT) indicated two pancreatic masses, which were enhanced most during the late arterial phase but had different degrees and variations of attenuation. Partial pancreatectomy was performed. Histopathology and immunohistochemistry confirmed that one mass was an insulinoma and the other was an ectopic splenic tissue, consistent with the differences in imaging findings. When an intrapancreatic lesion with hyper-attenuation on dynamic abdominal CT is detected, not only insulinoma or metastasis of malignancies but also intrapancreatic accessory spleen (IPAS) should be considered as differential diagnoses.     

Effects of contact with wood on blood pressure and subjective evaluation

This study examined the effects of contact with wood on the living human body using a physiological index and subjective evaluation. Consecutive blood pressure measurements were used as the physiological index, and sensory evaluation using the semantic differential (SD) method was used for subjective evaluation. Consideration was also given to cases in which materials were cooled and heated as well as kept at room temperature, to eliminate the effects of heat flux due to differences in thermal conductivity between wood and other materials. It was found that contact with wood produced coarse/natural sensations, with no associated increase in systolic blood pressure. Contact with cold wood created subjectively dangerous/uncomfortable but still coarse/natural sensations, also with no associated increase in blood pressure; therefore, there was no correspondence between subjective evaluation and physiological responses. Contact with aluminum kept at room temperature and cold acrylic plastic created flat/artificial and dangerous/uncomfortable sensations, with an associated significant increase in blood pressure; thus, there was a close correlation between subjective evaluation and physiological responses. It was therefore concluded that contact with wood, unlike artificial materials such as aluminum, induces no physiological stress even when kept at room temperature or cooled. Part of this report was presented at the 48th Annual Meeting of the Japan Wood Research Society in Shizuoka, April 1998     

Possible active transport mechanism in pharmacokinetics of flunixin-meglumin in rabbits

The plasma and urine kinetics of flunixin-meglumin (FNX, 2 mg/kg, i.v.) in rabbits were examined. Unusual pharmacokinetic profiles were obtained, including high binding percentage with plasma protein (> 99%), a short elimination half-life (< 4 hr) and a relatively large Vd-area (0.5 L/kg). These profiles indicate that some active transport mechanisms are involved in FNX disposition. The recovery of FNX from urine was approximately 9 % of the dose within 24 hr following the injection. The estimated renal clearance of the unbound drug nearly corresponded to the renal blood flow rates, indicating that active tubular secretion in the renal re-absorptive tract may be involved in the disposition. The effect of a concomitant administration of pravastatin (PV) on FNX disposition was also examined. PV is a representative substrate of a transporter in human liver cells (OATP-2). After the PV administrations, the Vd-area of FNX and total body clearance markedly decreased, indicating that FNX is actively taken up and metabolized in liver cells by an OATP-2 like transporter. In conclusion, there are at least 2 active transport pathways for FNX pharmacokinetics in rabbits, one is renal tubular secretion and the other is in the sinusoidal section of the liver.     

Silicon content in rice seedlings to protect rice blast fungus at the nursery stage

To control rice blast effectively at the nursery stage, the absolute SiO₂ content necessary for rice plants to resist blast disease was investigated using various rice cultivars and soils. Nine rice cultivars with different complete resistance genes and different degrees of partial resistance were grown on nursery soils amended with silica gel at different rates to change the SiO₂ content of rice plant. The rice seedlings were then inoculated 28 days after sowing with Pyricularia grisea to estimate their blast resistance. In all rice cultivars, the number of lesions was significantly reduced when SiO₂ content increased in the rice seedling; lesions were reduced to 5%–20% of the number on the seedlings grown in soil without silica gel when the seedling SiO₂ content reached 5%. Additionally, the susceptibility to blast disease of rice seedlings grown on eight soils collected from different districts, with varying amounts of silica gel, was compared. The number of lesions decreased significantly when the SiO₂ content in the seedlings reached 5%. These results suggest that SiO₂ content of at least 5% in the rice plant can control this disease at the nursery stage under any conditions.