Quantitative analysis of canine plasma lipoproteins

A combined ultracentrifugationl/precipitation method for the measurement of lipoprotein cholesterol concentrations was developed and validated for use with canine plasma. Very low density lipoproteins (VLDL) were isolated by flotation ultracentrifugation and low density lipoproteins (LDL) separated from high density lipoproteins (HDL) by precipitation with heparin-manganese chloride. Effective separation of these classes was confirmed by agarose gel electrophoresis of native lipoproteins and by sodium dodecyl sulphate polyacrylamide gel electrophoresis of their apolipoprotein distributions. There was trace contamination of the LDL precipitate with HDL, but this represented less than 4 and 9 per cent of the total plasma HDL in normo- and hypercholesterolaemic dogs, respectively. The intra-assay and interassay coefficients of variation for LDL- and HDL-cholesterol concentrations were between 3·3 and 6·9 per cent, and 7·2 and 9·0 per cent, respectively, for plasma cholesterol concentrations between 2·67 and 8·14 mmoll/litre. The intra-assay coefficient of variation for VLDL-cholesterol was 53·8 and 18·4 per cent at plasma cholesterol concentrations of 2·67 and 8·14 mmol/litre, respectively. The interassay coefficient of variation for VLDL was 22·5 per cent. Storage of plasma at -20°C for between two and eight weeks did not affect VLDL-cholesterol concentrations, but led to an increase in LDL-cholesterol and a decrease in HDL-cholesterol concentrations of approximately 10 per cent. The method described is appropriate for the measurement of lipoprotein concentrations in plasma from normo- and hypercholesterolaemic dogs, but samples should not be subjected to prolonged storage before analysis.     

Radiographic interpretation of normal skeletal variations and pseudolesions in the equine foot.

Effective radiographic interpretation requires a veterinarian who is knowledgeable of equine limb anatomy and the various principles that affect the resulting image. The normal and its variations must be recognized and understood before the abnormal can be confidently identified as pathologic. Proper patient positioning and sound radiographic technique are mandatory if reliable diagnostic radiographs are to be produced. This review emphasizes equine foot radiographic variations of normal and pseudolesions that occur with commonly used radiographic views performed in equine practice.     

Nitrogen uptake by ryegrass from organic wastes applied to a sandy loam soil

Sandy‐textured Mediterranean soils are invariably depleted in organic matter and supply only small amounts of N to crops. To compensate for these deficiencies, we tested the N supply from six organic wastes applied to a Cambic Arenosol in pots growing ryegrass. The results showed that the behaviour of the wastes in supplying N to a ryegrass crop grown in this soil can be predicted by observing their performance in laboratory aerobic incubations. The N made available during these incubations fitted well to a one‐pool kinetic model.     

A novel approach to studying the effects of temperature on soil biogeochemistry using a thermal gradient bar

The temperature dependence of chemical reaction rates and microbial metabolism mean that temperature is a key factor regulating soil trace gas emissions and hydrochemistry. Here we evaluated a novel approach for studying the thermal response of soils, by examining the effects of temperature on gas emissions and hydrochemistry in (a) peat and (b) soil from a Sitka spruce plantation. A thermal gradient was applied along an aluminium bar, allowing soil to be incubated contemporaneously from 2 to 18 °C. The approach demonstrated clear differences in the biogeochemical responses of the two soil types to warming. The peat showed no significant emission of CH₄ at temperatures below 6 °C, while above 6 °C, a marked increase in the rate of release was apparent up to 15 °C (Q₁₀ = 2.5) with emissions being similar between 15 and 18 °C. Conversely, CH₄ emissions from the forest soil did not respond to warming. Nitrate availability in the peat decreased by 90% between 2 and 18 °C (P < 0.01), whereas concentrations in the forest soil did not respond. Sulphate availability in the peat decreased significantly with warming (60%, P < 0.01), while the forest soil showed the opposite response (a 30% increase, P < 0.01). Conventionally, thermal responses are studied by incubating individual soil samples at different temperatures, involving lengthy preparation and facilities to incubate samples at different temperatures simultaneously. Data collected on a given thermal response is usually limited and thus interpolated or extrapolated. The thermal gradient method overcomes these problems, is simple and flexible, and can be adapted for a wide range of sample types (not confined to soil). Such apparatus may prove useful in the optimization of management practices to mitigate the effects of climate change, as thermal responses will differ depending on land use and soil type.     

Characteristics of cytotoxic cells induced by Toxoplasma lysate antigen in mouse spleen.

Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL male-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement inhibited cytotoxic activities of effector cells. These results suggested that spleen cells sensitized with TLA both in vivo and in vitro were asialo GM1 positive and Thy-1 positive, and the majority of cytotoxic cells induced by TLA were similar to lymphokine-activated killer (LAK) cells induced by IL-2.     

Oxidative damage to the membrane of canine erythrocytes with inherited high Na, K-ATPase activity.

Oxidative damage to the membrane in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) was compared with that in normal canine cells (LK cells). When 30 mM beta-acetylphenylhydrazine (APH) was applied to HK and LK cells, lipid peroxidation and hemoglobin denaturation occurred. Lipid peroxidation determined from malondialdehyde (MDA) formation was significantly lower in HK than in LK cells so far as endogenous glutathione (GSH) concentration was maintained at appropriate levels. With the depletion of GSH, MDA formation was accelerated and difference between HK and LK cells was not significant. Denatured hemoglobin bound to the membrane protein was less in HK than in LK cells. During incubation with APH, osmotic fragility increased markedly in LK cells, while HK cells showed very little change. The amounts of total lipid, total and free cholesterol, glycolipid, phospholipid and fatty acids were essentially the same in both cell types. Fatty acid compositions showed very small differences. The membrane of HK cells thus appear to have greater protection against oxidative damage induced by APH, owing to the presence of excess GSH in HK cells. The capability of HK cells to withstand oxidative damage would not be due to differences in membrane lipid compositions.     

Prevention of estrus in the queen with chlormadinone acetate administered orally.

The possibility of estrus prevention in the queen by the oral administration of chlormadinone acetate was examined. The animals used were 29 mature and 15 immature queens. For 16 mature females, 4-12.5 mg was given daily by mouth for 7 days every 3 months. Ten of the 16 queens given this treatment came into estrus within 4 months of the first treatment. For 28 females including the immature, 2-12.5 mg was given once a week throughout the experiment. This treatment prevented estrous activity for at least 1 year. In the queens in this study, the side effects were not observed excepting an increase in body weight during treatment. Our results showed that oral administration of this drug weekly is safe and reliable for long-range prevention of estrus in queens.