Radiosensitivity of canine osteosarcoma cells transfected with wild‐type p53 in vitro*

The p53 gene is one of the important tumour suppressor genes that are involved with the cell survival signal pathway. One of the major functions of the p53 protein is to organize cell cycle regulation and induction of apoptosis for cellular genetic stability. It has been documented that more than 50% of all human cancers include a p53 mutation. We evaluated the difference in radiosensitivity between upregulating the expression of canine wild‐type p53 (cp53) in cultured osteosarcoma (D17) cells and naive D17 cells in vitro. We found that upregulating transfected cp53 D17 cells increased their radiation sensitivity in vitro, and there was a significant decrease (P < 0.009) in survival between cp53‐transfected D17 cells and naive D17 cells. In this experiment, a p53 enhancement ratio (p53ER) reached approximately 3.0 at high doses. The transfected cp53 D17 cells were significantly more radiosensitive at all doses evaluated than naive D17 cells, except at 1 Gy where too few data points were available. The p53ER increased rapidly at doses less than 4 Gy, achieving a maximum of about 3.0 for doses of 4 Gy and above. This study shows the enhanced radiosensitivity of the transfected p53 at clinically relevant doses.     

Effects of exercise on the morphology of the myenteric neurons of the duodenum of Wistar rats during the ageing process

This study aimed to evaluate the effects of regular physical activity on the morphology of the myenteric plexus of the duodenum in rats during the ageing process. To this end, 45 Wistar rats were divided into three groups: C (sedentary – 6 months old), S (sedentary – 12 months old) and T (trained – 12 months old). The animals of group S were given with a physical activity programme consisting of a 10-min-treadmill workout once a week. The animals of group T were submitted to the physical activity programme five times a week. Their duodenums were collected and submitted to the techniques of nicotinamide adenine dinucleotide (NADH)-diaphorase enzyme histochemistry for whole-mount preparations and transmission electron microscopy. No differences in the constitution of the myenteric plexuses were found when the sedentary and trained groups were compared with the control group. The ultrastructural features were similar for the three groups. However, it was verified that the physical activity of the trained animals resulted in a similar myenteric neuron morphology to that of the adult animals (6 months old), thereby confirming its beneficial effect, as the sedentary animals had larger alterations in the collagen fibrils and the basal membrane that occur through ageing. The quantitative analysis showed that the NADH-diaphorase positive neurons decreased with ageing and increased with physical activity ( P  > 0.05). No significant alteration ( P  > 0.05) in the neuronal profile area of the NADH-diaphorase positive neurons has been observed with ageing.     

Differential response of cull cow muscles to the hypertrophic actions of ractopamine-hydrogen chloride

Ractopamine-HCl (RAC) is a beta-adrenergic agonist with variable effects on cattle performance and carcass variables. Cull cows fed RAC (200 mg . head(-1) . d(-1)) demonstrate an increased size of type I and II muscle fibers that does not translate into a larger ribeye area. The objective of this study was to examine the dose-dependent effects of RAC on cull cow muscle morphometrics. Eighty-eight cull beef cows representing 2 breed types (n = 44 each) were fed 0, 100, 200, and 300 mg . head(-1) . d(-1) of RAC for the last 28 d of a 54-d feeding period. On d 54, cows were slaughtered, and samples of the LM and semimembranosus muscle (SM) from 16 randomly selected carcasses (n = 4 per treatment) were taken for measurement of beta (2)-adrenergic receptors and type I, IIA, and IIX myosin heavy chain (MyHC) gene expression. Twenty-four hours postmortem, LM, SM, infraspinatus (INF), and vastus lateralis samples from 40 randomly selected carcasses (n = 10 per treatment) were obtained and frozen for immunohistochemical analysis. Muscle fiber cross-sectional area and diameter, MyHC isoform expression, and fiber-associated nuclei numbers were measured. Ractopamine dosage exhibited differential effects on muscle morphometrics and MyHC gene expression. Muscle fiber cross-sectional area and diameter were increased (P < 0.05) by RAC in INF type I and IIA fibers and SM type IIA fibers. Ractopamine increased (P < 0.05) MyHC type IIX mRNA and tended to increase (P < 0.10) beta(2)-adrenergic receptors in the SM; a change in mRNA abundance was not detected for either gene in the LM. Treatment with RAC decreased (P < 0.05) fiber-associated nuclei numbers in the INF, vastus lateralis, and LM but did not affect (P > 0.05) MyHC or beta-adrenergic receptor expression. These results indicate that cull cow feeding programs may consider supplementing RAC as a means of adding value to cuts within the chuck, such as the INF.     

Acute lead intoxication in a pregnant mare

Lead (Pb) intoxication in horses is usually a chronic phenomenon with clinical signs associated with central nervous dysfunction. This report gives details of a case of acute Pb intoxication in a 9-year-old American Saddlebred mare with severe, progressive and ultimately fatal neurological deterioration. During the 4 days of hospitalisation, clinical signs progressed from intermittent headshaking and depression to severe, continuous, uncontrollable manic behaviour. At autopsy, three grey-coloured, hard metal particles were present in the gastrointestinal tract and subsequently found to contain 2614 ppm Pb. Lead concentrations in the brain, liver, stomach and kidney were 29, 4, 6 and 2 ppm wet weight, respectively.     

Pharmacokinetics of morphine and its metabolites in freshwater rainbow trout (Oncorhynchus mykiss)

In this study, we injected morphine sulfate IP into rainbow trout and measured the concentration of morphine and all potential metabolites in plasma using LC-MS/MS at a series of times after the injection. The pharmacokinetics of morphine were similar to those previously reported for seawater-acclimated rainbow trout, i.e. they were about one order of magnitude slower than in similarly sized mammals. The only metabolite of morphine present in the plasma was morphine-3-β- d -glucuronide (M3G); morphine-6-β- d -glucuronide (M6G) was not detected. M3G gradually increased after the morphine injection, peaked about 2 days later, then gradually decreased. In mammals, M3G plasma levels exceed morphine levels extremely rapidly, i.e. in less than an hour, regardless of dose, route of administration, or species. In trout, it took 2 days for M3G levels to exceed morphine levels. This is the first study of the metabolites of morphine in any ectotherm. We conclude that trout can metabolize morphine, but at a rate much slower than in mammals.     

The effect of dietary supplementation with 9-cis:12-trans and 10-trans:12-cis conjugated linoleic acid (CLA) for nine months on serum cholesterol, lymphocyte proliferation and polymorphonuclear cells function in Beagle dogs

Conjugated linoleic acids (CLA), 9-cis:11-trans and 10-trans:12-cis, have been shown to be able to modify some immune cells parameters and plasma lipids in a variety of experiment models. Since lymphocytes and polymorphonuclear cells (PMNC) have a large spectrum functions in the immune response, the knowledge in this field has to be expanded. Beagle dogs were fed a control diet or a CLA supplemented diet for nine months. Blood was collected for biochemical analysis and lymphocyte and PMNC isolation. PMNC were assayed for lysosome content, phagocytic activity and superoxide anion production. A lymphocyte proliferation capacity assay was done. The CLA fed dogs had a 34% reduction in total cholesterol (P < 0.05), 28% in LDL (P < 0.05) and 28% non-HDL-cholesterol (P < 0.05). Neither of the PMNC parameters evaluated demonstrated significant alteration. Lymphocytes from CLA group increased by 45% their mitotic capacity (P < 0.05). Our study demonstrates that CLA can successfully modify the lipid profile of dogs (monogastrics) when fed at reasonable levels, but did not significantly alter inflammatory function as would generally predicted. Further, we had some indication that CLA modulated T cell responsiveness.     

Epiphyseal cartilage canal blood supply to the tarsus of foals and relationship to osteochondrosis

REASON FOR PERFORMING STUDY: Pathological changes in the blood supply to growth cartilage have been implicated in the pathogenesis of osteochondrosis (OC) in horses, but have not been reported using vascular perfusion techniques. OBJECTIVE: To describe the developmental pattern of cartilage canal vessels in the distal tibial epiphysis and talar growth cartilage of foals. METHODS: Nine foals bred from parents with OC were sacrificed between the ages of 0 and 7 weeks to undergo a barium perfusion procedure. The distal end of the tibia and the entire talus were cleared in methyl salicylate and perfused vessels studied in the intact bones. Slabs with a thickness of 4-5 mm from 3 predilection sites for OC were examined in the stereomicroscope and with light microscopy. RESULTS: Cartilage canals were present for a limited period of growth. Perfused vessels initially entered canals from the perichondrium. Vessels in the proximal portion of canals retained their perichondrial arterial source throughout. With time, the ossification front advanced to incorporate the mid-portion of canals; and anastomoses formed between canal vessels and subchondral vessels. A shift occurred and vessels in the distal terminus of canals came to use subchondral vessels as their arterial source. Twelve histological lesions were found in 7 foals. All contained necrotic vessels surrounded by necrotic growth cartilage and 3 caused macroscopically visible delay in endochondral ossification. Lesions were located where vessels traversed the ossification front to enter the distal terminus of canals. CONCLUSION: Cartilage canal vessels are particularly susceptible to failure at the point where they cross the ossification front, with consequences for the viability of those chondrocytes that depend on them. POTENTIAL RELEVANCE: A better understanding of how lesions of OC arise may improve the ability to identify, monitor, prevent and treat this disorder. Involvement of cartilage canals in the pathogenesis of equine tarsal OC plausibly explains several clinical features of this disease.     

Economic effect of pneumonia and pleurisy in lambs in New Zealand

AIM: To estimate the cost of pneumonia and pleurisy in lambs to the sheep industry in New Zealand, in order to provide a reference for future cost-benefit calculations for control programmes to reduce the incidence of pneumonia. METHODS: An estimate of the economic cost of pneumonia and pleurisy in lambs was based on: cohort studies of the association between growth rate and the extent of pneumonic lesions at slaughter (n=14 flocks), the prevalence of moderate to severe (MS) pneumonia (> or =10% lung surface area affected) and pleurisy (n=1,719 flocks), the correlation between the prevalence of MS pneumonia and economic loss at the flock level, and data on annual slaughter statistics and carcass value in New Zealand. A stochastic spreadsheet model was developed and run with 1,000 iterations. Input variables represented by probability distributions were growth rate, average cost of loss according to the prevalence of pneumonia per month, carcass value, prevalence of pleurisy, and carcasses downgraded for pleurisy, and annual national slaughter statistics. The output was a posterior distribution of the annual cost of disease. RESULTS: The cost of pneumonia only included the loss associated with reduced growth rate, while mortality due to pneumonia was assumed to be low and was ignored. The cost of pleurisy included the loss associated with downgraded or condemned carcasses. Thus, the simulated annual average cost of pneumonia was NZ$28.1 (95% CI=NZ$15.0-42.0) million, and that of pleurisy NZ$25.1 (95% CI=NZ$10.2-48.1) million. The combined cost of pneumonia and pleurisy averaged NZ$53.2 (95% CI=NZ$32.4-78.9) million. The parameters with the greatest impact on the combined cost of pneumonia and pleurisy were prevalence of pleurisy between March and May, and cost of reduced growth due to pneumonia for lambs slaughtered in June. CONCLUSIONS: The average cost of pneumonia and pleurisy to the sheep industry in New Zealand due to reduced lamb growth and decreased carcass value is likely to be between NZ$32.4 and $78.9 million. This is a conservative estimate, omitting mortalities, indicating that pneumonia and pleurisy have substantial economic impact on sheep farming in New Zealand. Considering that 23,833,000 lambs were slaughtered in 2003/2004 (average value $65.56/lamb), this cost would equate to NZ$1.36-3.31 per lamb. CLINICAL RELEVANCE: Pneumonia and pleurisy are common diseases in lambs in New Zealand, leading to substantial economic loss to producers.     

Immunohistochemical studies on the splenic lobe of the pancreas in young Japanese quails (Coturnix c. japonica)

The splenic lobe (Lobus splenicus) of the pancreas of young meat-type quails (Coturnix c. japonica) was examined by immunohistochemical and light microscopic methods. The endocrine cells are mainly grouped as alpha, beta and mixed islets. A large region consisting of alpha cells is located in the central region of the splenic lobe whereas numerous beta islets are detected in the periphery of the splenic lobe. Alpha islets are in the majority composed of toluidine blue positive A cells and a few toluidine blue negative D and / or avian pancreatic polypeptide (APP) endocrine cells. Beta islets contain only a few toluidine blue negative B and a few D cells. Immunohistochemical staining of the splenic lobe reveal in the centre of beta islets numerous insulin immunoreactive cells and scarcely in alpha islets, exocrine tissue and / or among acinar cells. Somatostatin immune-reactive cells form a circular layer in the periphery of beta islets whereas these cells are uniformly distributed throughout the alpha islet parenchyma and exocrine tissue. In conclusion, the morphology but also the endo- and exocrine functions of the splenic lobe of quails are similar to observations in other avian species such as chicken, duck, goose and pigeon.