A novel HPLC method for glutathione-insulin transhydrogenase assay using fluorescence labeled insulin

The activity of rat liver glutathione-insulin transhydrogenase (GIT) was measured by HPLC. The degradation of fluorescein isothiocyanate-I (FITC-I)-labeled insulin is separated into several peaks, which are bound different amount of FITC-I. We selected mono-fluorescein-thiocarbamylated insulin to estimate the decrease of insulin content and it became possible to assay GIT activity. This novel method was time-saving and simple, and this system could utilize instead of previous method.     

Postovulatory Effect of Intravenous Administration of Lipopolysaccharide (Escherichia coli, O55:B5) on the Endocrine Status of Recently Ovulated Sows

The effects of lipopolysaccharide ( Escherichia coli , O55:B5), administered 18 h after ovulation in the second oestrus after weaning on the hormonal profiles in 14 Swedish cross-bred (Landrace × Yorkshire) multiparous sows were studied. The endotoxin group (E-group) sows were administered with 300 ng/kg of lipopolysaccharide (LPS) whereas the control group (C-group) sows were administered 5 ml of saline intravenously via an indwelling jugular cannula. Blood samples for hormonal analyses were collected from all sows until slaughter. In the E-group, progesterone, cortisol and prostaglandin F_2α metabolite levels increased significantly (p < 0.05) following LPS compared with the C-group. It can be concluded from this study that apart from elevating cortisol and prostaglandin F_2α metabolite, LPS also elevates progesterone levels.     

Fluid accumulation in mouse ligated intestine inoculated with the vascular permeability factor produced by Bacillus cereus.

Partially purified vascular permeability (VP) factor (VPF) of Bacillus cereus induced fluid accumulation in the ligated intestinal loops of mouse (MIL) and rabbit (RIL), suggesting that the VP activity may correlate with fluid accumulation in ligated intestinal loops of these animals. Fluid accumulation was observed at 6-8 hr in 55-67% of mouse intestinal loops inoculated with 40-50 immunodiffusion units (IDU) of partially purified VPF, whereas it was found at 2 hr in all loops with 400-600 IDU of partially purified VPF. In rabbit intestinal loops with 120-190 IDU of partially purified VPF, fluid accumulation was observed at 6 hr. From these findings, VPF produced by B. cereus can be easily detected in both MIL and RIL. The intestinal tissue of mouse intestinal loops was histopathologically damaged at different concentrations of the VPF to induce fluid accumulation. With 50 IDU of partially purified VPF, severe edema was found in the laminia proprial layer and submucosa. With 600 IDU of partially purified VPF, on the other hand, severe necrosis in the surface epithelium of villus and laminia proprial layer, and hyperemia in the submucosa were observed, suggesting that partially purified VPF may be cytotoxic and/or intestinecrotic.     

Effect of orally administered furazolidone on volume and sperm concentration of dwarf and non-dwarf cock semen

2 groups of 20 cocks each were selected at random from non-dwarf White Leghorn (28 weeks post-hatch) and dwarf Krishna-J (38 weeks post-hatch) genotypes. The treated groups comprised 10 White Leghorn and 10 Krishna-J cocks. The remaining birds served as controls. 8 weeks prior to furazolidone treatment, semen was collected from both control groups at regular 4-day intervals, for 4 weeks. Cocks of the treated groups of both genotypes were administered furazolidone (0.14 g/bird/day) for 7 consecutive days. Semen was collected from all cocks at regular 4-day intervals for 4 weeks. Semen from the cocks of the same group was pooled. The pooled ejaculate volume and sperm density did not differ significantly in the 2 genotypes. The semen output as well as sperm density increased along with progressive attainment of sexual maturity. Furazolidone treatment caused significant reduction in semen volume as well as sperm concentration in either genotype.     

Evaluation of heparin for prophylaxis of equine laminitis: 71 cases (1980-1986)

The records of 71 horses with small intestinal disorders requiring surgical correction were disorders requiring surgical correction were reviewed to compare the prevalence of laminitis in those horses treated prophylactically with heparin and the prevalence of horses not treated with heparin. The prevalence of laminitis was 13% (9/71), and there was no significant difference (P less than 0.05) in the prevalence of laminitis between the 2 groups. The lack of significant benefit after treatment with heparin indicates that further work needs to be done on the equine coagulation system before heparin can be advocated for prevention of laminitis.     

Polyarteritis in a beagle

A 13-month-old Beagle became anorectic and had fever, stiff gait, and tenderness in the inguinal region. Clinical signs of disease were associated with neutrophilia and a decrease in the albumin-to-globulin ratio. The dog became clinically normal for 5 days after 3 days of treatment with penicillin G and dihydrostreptomycin. Clinical signs of disease recurred, and the dog was euthanatized after failing to respond to administration of a trimethoprim-sulfamethoxazole combination for 9 days. Disseminated arteritis was seen in the testes, epididymides, mesentery, coronary arteries, aorta, and thyroid gland. Lesions were seen in large and medium-sized arteries and varied from acute necrotizing arteries to a chronic lesion with organization and recanalization of thrombi. The clinical signs of disease resembled those of Beagle pain syndrome, described in laboratory Beagles.     

Cellular injury and lipid peroxidation induced by hexavalent chromium in isolated rat hepatocytes

In order to elucidate the relationship between cellular injury and lipid peroxidation induced by hexavalent chromium (CrVI), isolated rat hepatocytes treated with any one of scavengers of active oxygen species, antioxidants or antichromium agent were incubated with K2Cr2O7 as CrVI (1 mM Cr). After the incubation, the development of lipid peroxidation was determined as thiobarbituric acid (TBA)-reacting materials in total lipid extracts from the incubated hepatocytes. Cellular injury was observed as a leakage of lactate dehydrogenase (LDH) from hepatocytes into incubation medium. The contents of reduced glutathione (GSH) in hepatocytes were also assessed. Results obtained were as follows: (1) CrVI facilitated lipid peroxidation in isolated hepatocytes after 20 min of incubation. On the other hand, the cellular injury induced by CrVI was barely observed even after 60 min of incubation. (2) The CrVI-induced lipid peroxidation was inhibited by catalase and mannitol as scavengers of active oxygen species, or N,N'-diphenyl-p-phenylenediamine and alpha-tocopherol as antioxidants. However the cytotoxicity of CrVI could not be prevented by these chemicals. (3) CrVI depleted the contents of intracellular GSH and diminished the activities of glutathione reductase (GR) and glutathione-S-transferase (GST) except glutathione peroxidase. (4) The scavengers of active oxygen species and the antioxidants could not prevent the depletion of intracellular GSH induced by CrVI. (6) Ascorbic acid, antichromium agent, prevented all of the lipid peroxidation, the cellular injury and intracellular GSH depletion induced by CrVI.(ABSTRACT TRUNCATED AT 250 WORDS)