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 dobutamine on cardiac index and arterial blood pressure in isoflurane-anaesthetized horses under clinical conditions

Volatile agent-induced hypotension may contribute to anaesthetic-related morbidity and mortality in horses. Dobutamine is commonly used to support arterial blood pressure (ABP) but little is known about its cardiovascular effects under clinical conditions. The aim of this clinical study was to elucidate the relationship between cardiovascular function and dobutamine infusion in isoflurane-anaesthetized horses. Forty-four horses anaesthetized for a variety of surgical procedures were studied. Premedication with acepromazine, methadone and detomidine was followed by induction of anaesthesia with ketamine and midazolam. Anaesthesia was maintained with isoflurane vaporized in oxygen. Routine anaesthetic monitoring was applied and cardiac output was measured by lithium dilution. Dobutamine was infused to maintain mean ABP above 70 mmHg. The relationship between dobutamine infusion rate, heart rate (HR), ABP and cardiac index was investigated immediately prior to ( T ₀) and 15 min ( T ₁) after dobutamine infusion started, followed at 30 min intervals ( T ₂, etc.). Arterial blood pressure increased significantly after dobutamine infusion started, HR and cardiac index increased significantly only with dobutamine infusion in combination with surgical stimulus. Although isoflurane decreases blood pressure mainly by vasodilation, dobutamine is an effective treatment for hypotension under clinical conditions in isoflurane-anaesthetized horses. The effect of dobutamine is not directly proportional to dose and surgical stimulus probably contributes to the cardiovascular improvement.     

Validation of the periodicity of growth zone formation in the otoliths of four fish species from the Upper Zambezi ecoregion,southern Africa

In the Upper Zambezi and Okavango ecoregions, Brycinus lateralis, Hepsetus cuvieri, Schilbe intermedius and Serranochromis macrocephalus are important in subsistence fisheries, while S. intermedius and S. macrocephalus are often caught in commercial catches. Despite their importance, there is little information on their age and growth and age validation. Growth zone deposition rate in otoliths of B. lateralis, H. cuvieri, S. intermedius and S. macrocephalus was validated as annual using edge analysis in this study. Annual deposition of growth zones was corroborated by a fluorochrome marking experiment. Both methods demonstrated that one growth zone was deposited annually. Edge analysis demonstrated that the highest proportion of opaque zones was detected between August and December, most likely as a result of slow growth during lower temperature water during the preceding winter. With growth zone deposition validated as annual, future research should focus on determining the growth, maturity and mortality rates of these species, parameters which are important for their management.     

Salivary testosterone measurements in growing pigs: validation of an automated chemiluminescent immunoassay and its possible use as an acute stress marker

This study aimed to validate the use of an automated chemiluminescent immunoassay analyser for salivary testosterone measurements in growing pigs and study how circadian pattern during daytime and stress can influence its values. The test method had intra- and inter-assay coefficient of variation lower than 10%. The method showed good linearity and recovery, and detection limits were low enough to detect salivary testosterone levels. No significant differences were observed in testosterone concentrations at different sampling time, and age and gender did not influence circadian pattern. In addition, this assay was used to quantify testosterone in two models of acute stress and, in both cases, significant increases (P < 0.01) in salivary testosterone were detected. Therefore, the automated assay system tested for porcine testosterone determinations would be suitable for its use in saliva samples and, furthermore, salivary testosterone levels could be used as a possible marker of acute stress in pigs.     

High infection rate of Trichophyton verrucosum in calves from Central Italy

Recent epidemiological features of the zoonotic dermatophyte Trichophyton verrucosum were investigated in 294 calves from 20 farms located in Central Italy. By the combination of microscopic examinations and fungal cultures, the total prevalence rate of T. verrucosum infection was found to be high, with 87.7% of samples yielding positive results and 100% of farms being infected. Farm to farm prevalence of infection varied from 25% to 100% of the examined calves. Prevalence rates were higher in females than males (91.6% versus 84%), in >3-< or =6 than >6-< or =9 month old calves (89.8% versus 85.7%), in purebred than in crossbred (88.5% versus 81.8%), and in symptomatic than asymptomatic ones (95.7% versus 80.4%). T. verrucosum was as prevalent in semi-intensive as in intensive breeding system (88.2% versus 87.6%) but more prevalent in calves raised for meat rather than milk production (91.1% versus 84.9%), and reared under conventional than organic production management system (89.3% versus 82.6%). Prevalence rates were also higher when the cattle present in the farm were of mixed origin, rather than when the whole stock was purchased or was born in the farm (92% versus 88.2% and 85.7%), when other cattle in the farm were showing skin signs (90% versus 84.2%), and at least one case of human ringworm was recorded in the farm (90.9% versus 86.1%). No statistically significant association was found. Therefore, T. verrucosum infection may be more widespread in calves from Central Italy than it was previously thought to be. Calves of any sex, breed, age, and of any of the farm condition considered, are highly exposed to the risk of infection. As calves may play a considerable role as reservoirs of the infection, our results highlight the need to determine the incidence and the importance of T. verrucosum ringworm in personnel involved with cattle care in Italy and elsewhere.     

Equine Granulocytic Anaplasmosis: A Case Report and Review

A 15 year-old Thoroughbred mare was examined for lethargy, fever, and inappetence of 1-day duration. A hard-bodied tick was removed from the horse. A complete blood count (CBC) demonstrated leukopenia with lymphopenia and thrombocytopenia. Morulae were visualized in circulating granulocytes. A polymerase chain reaction (PCR) confirmed the identity of these organisms as Anaplasma phagocytophilum. The horse was treated symptomatically for fever and inappetence with flunixin meglumine (1.1 mg/kg [0.5 mg/lb]) and oral electrolyte paste. Oxytetracycline (6.6 mg/kg [3 mg/lb] intravenously every 24 hours) treatment was begun as soon as a definitive diagnosis was determined. The mare responded to treatment, but she was switched to oral doxycycline (10 mg/kg [4.5 mg/lb] every 12 hours) after 5 days because of perivascular swelling at the injection site. Complete resolution of clinical signs was seen. There was no evidence of recurrence 1 year later. No additional horses at the farm were affected. The horse in this report presented for lethargy, inappetence, and fever, with limited other abnormalities. This represents a classical presentation of a mild to moderate case of anaplasmosis, which had not previously been reported in Virginia. The disease may be more widespread than has been previously reported, and it should warrant inclusion on a complete differential diagnosis list in a case of fever of unknown origin.     

A retrospective analysis of radiation therapy for the treatment of feline vaccine‐associated sarcoma*

We retrospectively evaluated predictive prognostic factors in 73 cats with vaccine‐associated sarcoma given postsurgical curative (n = 46, most with clean margins) or coarse fractionated radiotherapy (n = 27, most with either macroscopic disease or dirty margins). The former animals displayed a median survival of 43 months and a median progression free interval (PFI) of 37 months, the latter reached a median survival of 24 months and a median PFI of 10 months. In cats undergoing coarse fractionated therapy, factors predictive of a better outcome included lack of visible mass (n = 10) as opposed to macroscopic disease (n = 17, survival: 30 versus 7 months, P = 0.025; PFI: 20 versus 4 months, P = 0.01), adjuvant chemotherapy for gross disease (n = 5/17, survival: 29 versus 5 months, P = 0.04) and a smaller number of surgeries preceding radiation therapy (coeff = 0.41, P = 0.03). The Ki67 index was not predictive for survival. We concluded that postsurgical curative and coarse fractionated radiotherapy are effective legitimate options for managing vaccine‐associated sarcomas.     

In vitro evaluation of metacarpophalangeal joint loading during simulated walk

Reasons for performing study: Insight into the loading pattern of the articular cartilage surface during the complete stride is important as biomechanical factors play a pivotal role in the pathogenesis of joint trauma and osteoarthritis (OA). Objectives: To determine the loading pattern in the equine MCP articulation in vitro during simulated walk. Methods: Eight cadaveric limbs from mature Dutch Warmblood horses were loaded in a pneumatic loading device in 6 different positions (A1‐A6). The pressure distribution on the articular surface of the proximal phalanx (P1) was measured at 7 sites (S1‐7) using intra‐articularly placed pressure sensitive films, which were analysed by scanning and densitometry. Results: Pressures recorded after mid‐stance (A4, 5, 6) were significantly (P<0.05) higher than those before (A1, 2, 3) and showed the biphasic loading pattern of the walk at all sites, except for the site halfway along the sagittal groove (S7). At S7, there was a linear increase in pressure during the progress of the stance phase of the stride in most horses. Medially (S4, 5, 6) the pressure was significantly higher than laterally (S1, 2, 3) (P<0.05). Conclusions and potential relevance: The heavier medial loading coincides with the location where articular cartilage degeneration in the process of OA in the equine MCP joint is known to start. The discrepancy between the loading of the central groove and the other parts of the joint may result in large stress differences at the end of the stance phase, which might be related to the pathogenesis of stress fractures in the first phalanx and distal third metacarpal bone.