Sinonasal cysts causing dyspnoea in two cattle - case report.

Two cattle were referred to the University Clinic for Ruminants of the University of Veterinary Medicine in Vienna. The main clinical sign in both cattle was dyspnoea with nasal stridor. Clinical examination of the upper respiratory tract was conducted, supplemented by ultrasonography, endoscopy and radiography. In addition, histological, bacteriological, and cytological examinations of different specimen materials were performed. The cow of Case 1 suffered from cystic nasal conchae, which was treated successfully by a laser technique. The cow of Case 2 also suffered from cystic nasal conchae. No surgery was performed in this case because the cyst opened spontaneously the day after diagnostic endoscopic procedures had been performed and the animal did not show any respiratory signs anymore. Pathological changes in the upper respiratory tract, such as nasal obstructions, should be included in the list of differential diagnoses in cattle showing respiratory distress.

Cytokine-induced interleukin-1 receptor antagonist protein expression in genetically engineered equine mesenchymal stem cells for osteoarthritis treatment.

BACKGROUND: A combination of tissue engineering methods employing mesenchymal stem cells (MSCs) together with gene transfer takes advantage of innovative strategies and highlights a new approach for targeting osteoarthritis (OA) and other cartilage defects. Furthermore, the development of systems allowing tunable transgene expression as regulated by natural disease-induced substances is highly desirable. METHODS: Bone marrow-derived equine MSCs were transduced with a lentiviral vector expressing interleukin-1 receptor antagonist (IL-1Ra) gene under the control of an inducible nuclear factor-kappa B-responsive promoter and IL-1Ra production upon pro-inflammatory cytokine stimulation [tumor necrosis factor (TNF)α, interleukin (IL)-1ß] was analysed. To assess the biological activity of the IL-1Ra protein that was produced and the therapeutic effect of IL-1Ra-expressing MSCs (MSC/IL-1Ra), cytokine-based two- and three-dimensional in vitro models of osteoarthritis using equine chondrocytes were established and quantitative real-time polymerase chain reaction (PCR) analysis was used to measure the gene expression of aggrecan, collagen IIA1, interleukin-1ß, interleukin-6, interleukin-8, matrix metalloproteinase-1 and matrix metalloproteinase-13. RESULTS: A dose-dependent increase in IL-1Ra expression was found in MSC/IL-1Ra cells upon TNFα administration, whereas stimulation using IL-1ß did not lead to IL-1Ra production above the basal level observed in nonstimulated cells as a result of the existing feedback loop. Repeated cycles of induction allowed on/off modulation of transgene expression. In vitro analyses revealed that IL-1Ra protein present in the conditioned medium from MSC/IL-1Ra cells blocks OA onset in cytokine-treated equine chondrocytes and co-cultivation of MSC/IL-1Ra cells with osteoarthritic spheroids alleviates the severity of the osteoarthritic changes. CONCLUSIONS: Thus, pro-inflammatory cytokine induced IL-1Ra protein expression from genetically modified MSCs might represent a promising strategy for osteoarthritis treatment.

Inflammation-induced transgene expression in genetically engineered equine mesenchymal stem cells.

BACKGROUND: Osteoarthritis, a chronic and progressive degenerative joint disorder, ranks amongst the top five causes of disability. Given the high incidence, associated socioeconomic costs and the absence of effective disease-modifying therapies of osteoarthritis, cell-based treatments offer a promising new approach. Owing to their paracrine, differentiation and self-renewal abilities, mesenchymal stem cells (MSCs) have great potential for regenerative medicine, which might be further enhanced by targeted gene therapy. Hence, the development of systems allowing transgene expression, particularly when regulated by natural disease-dependent occuring substances, is of high interest. METHODS: Bone marrow-isolated equine MSCs were stably transduced with an HIV-1 based lentiviral vector expressing the luciferase gene under control of an inducible nuclear factor κB (NFκB)-responsive promoter. Marker gene expression was analysed by determining luciferase activity in transduced cells stimulated with different concentrations of interleukin (IL)-1ß or tumour necrosis factor (TNF)α. RESULTS: A dose-dependent increase in luciferase expression was observed in transduced MSCs upon cytokine stimulation. The induction effect was more potent in cells treated with TNFα compared to those treated with IL-1ß. Maximum transgene expression was obtained after 48 h of stimulation and the same time was necessary to return to baseline luciferase expression levels after withdrawal of the stimulus. Repeated cycles of induction allowed on-off modulation of transgene expression without becoming refractory to induction. The NFκB-responsive promoter retained its inducibility also in chondrogenically differentiated MSC/Luc cells. CONCLUSIONS: The results of the present study demonstrate that on demand transgene expression from the NFκB-responsive promoter using naturally occurring inflammatory cytokines can be induced in undifferentiated and chondrogenically differentiated equine MSCs. Copyright © 2016 John Wiley & Sons, Ltd.

Radiation therapy communication: equine hemangioma.

A 13-month-old Standardbred Colt had a recurrent hemangioma at the level of the coronary band. Multiple excisions had led to a nonhealing skin and hoof defect. Using 14 MV electrons, a total dose of 36 Gy was administered, given as six fractions of 6 Gy twice a week. Wound healing by second intention was achieved over the next 4 months and the colt began race training 6 months after the end of therapy. Twenty months later the colt is sound and there is no evidence of tumor recurrence.

[Evaluation of a modified exogenous creatinine clearance as a suitable renal function test for the small animal practice]. / Bewertung einer modifizierten Plasma-Clearance mit exogenem Kreatinin als ein für die Kleintierpraxis geeignetes Verfahren der renalen Funktionsdiagnostik.

A suitable method in the routine veterinary practice for the quantitative determination of the glomerular filtration rate (GFR) in dogs and cats has not been available until to date. Therefore, we modified the known plasma clearance model (=P-CL). The resulting P-CLterminal was assessed concerning its diagnostic value. P-CL of exogenous creatinine (P-CLcrea) and of inulin were determined in dogs (n=12, Beagle, 6 months of age) and cats (n=11, Domestic Short Hair, 14 months of age). The marker substances were administered as a bolus injection. In fasted dogs, P-CLcrea was 84.3 +/- 14.85 ml/min/m2 after a creatinine dose of 2.4 g/m2. An electrolyte infusion during the clearance determination did not alter the resulting values (p>0.05). In fasted cats, P-CLcrea was 54.7 +/- 5.8 ml/min/m2 (creatinine dose 2.0 g/m2). The inulin clearance, determined at the same time, was 104.5 +/- 19.81 ml/min/m2. Feeding the cats just before and during the test increased P-CL of both markers significantly (p<0.05). In order to adapt the clearance method for diagnostic assessment of GFR in the small animal practice, we aimed at minimizing the number of required blood samples (3 instead of 7 or more) and introduced the modified exogenous creatinine clearance (P-CLterminal). These values determined were 108.4 +/- 20.81 ml/min/m2 in fasted dogs and 66.3 +/- 11.81 ml/min/m2 in fasted cats. An electrolyte infusion (dogs) and feeding (cats) had the same effect on P-CLterminal values as described above for P-CL. In conclusion,the modified exogenous creatinine clearance is a suitable renal function test for the early diagnosis of renal disease in dogs and cats presented in small animal practices.

Tracing axons of peripheral nerves in rats: a potential technique to study the equine recurrent laryngeal nerve.

To study the fascicular anatomy of peripheral nerves, three different groups of retrograde axonal tracers were evaluated: fluorophores, horseradish peroxidase conjugated to subunit B of cholera toxin (CT-HRP), and adeno-associated virus (AAV). The hindlimb nerves in rats served as a model to identify the most efficient tracer in regard to labeling axons within peripheral nerves. The rat's tibial and common peroneal nerves were injected with the different tracers and the sciatic nerve was subsequently examined for evidence of labeled axons. The CT-HRP clearly provided the best results in this rat model. Subsequently, CT-HRP was injected into the recurrent laryngeal nerve (RLN) of two horses in order to identify the location and distribution pattern of the RLN axons within the course of the cervical vagus nerve trunk. No labeling could be observed in either of the two horses.