Novel diabetes mellitus treatment: mature canine insulin production by canine striated muscle through gene therapy

Muscle-targeted gene therapy using insulin genes has the potential to provide an inexpensive, low maintenance alternative or adjunctive treatment method for canine diabetes mellitus. A canine skeletal muscle cell line was established through primary culture, as well as through transdifferentiation of canine fibroblasts after infection with a myo-differentiation gene containing adenovirus vector. A novel mutant furin-cleavable canine preproinsulin gene insert (cppI4) was designed and created through de novo gene synthesis. Various cell lines, including the generated canine muscle cell line, were transfected with nonviral plasmids containing cppI4. Insulin and desmin immunostaining were used to prove insulin production by muscle cells and specific canine insulin ELISA to prove mature insulin secretion into the medium. The canine myoblast cultures proved positive on desmin immunostaining. All cells tolerated transfection with cppI4-containing plasmid, and double immunostaining for insulin and desmin proved present in the canine cells. Canine insulin ELISA assessment of medium of cppI4-transfected murine myoblasts and canine myoblast and fibroblast mixture proved presence of mature fully processed canine insulin, 24 and 48 h after transfection. The present study provides proof of principle that canine muscle cells can be induced to produce and secrete canine insulin on transfection with nonviral plasmid DNA containing a novel mutant canine preproinsulin gene that produces furin-cleavable canine preproinsulin. This technology could be developed to provide an alternative canine diabetes mellitus treatment option or to provide a constant source for background insulin, as well as C-peptide, alongside current treatment options.     

Observations on animal and human health during the outbreak of Mycobacterium bovis in game farm wapiti in Alberta

This report describes and discusses the history, clinical, pathologic, epidemiologic, and human health aspects of an outbreak of Mycobacterium bovis infection in domestic wapiti in Alberta between 1990 and 1993, shortly after legislative changes allowing game farming. The extent and seriousness of the outbreak of M. bovis in wapiti in Alberta was not fully known at its onset. The clinical findings in the first recognized infected wapiti are presented and the postmortem records for the herd in which the animal resided are summarized. Epidemiologic findings from the subsequent field investigation are reviewed, the results of recognition and investigation of human exposure are updated, and recommendations for reduction of human exposure are presented.     

The Current State of Solar Modeling

Data from the Global Oscillation Network Group (GONG) project and other helioseismic experiments provide a test for models of stellar interiors and for the thermodynamic and radiative properties, on which the models depend, of matter under the extreme conditions found in the sun. Current models are in agreement with the helioseismic inferences, which suggests, for example, that the disagreement between the predicted and observed fluxes of neutrinos from the sun is not caused by errors in the models. However, the GONG data reveal subtle errors in the models, such as an excess in sound speed just beneath the convection zone. These discrepancies indicate effects that have so far not been correctly accounted for; for example, it is plausible that the sound-speed differences reflect weak mixing in stellar interiors, of potential importance to the overall evolution of stars and ultimately to estimates of the age of the galaxy based on stellar evolution calculations.     

Comparative effects of proligestone and megestrol acetate on basal plasma glucose concentrations and cortisol responses to exogenous adrenocorticotrophic hormone in cats

Cats were given megestrol acetate (MA, 5 mg once daily for 14 days), subcutaneous proligestone (PRG, 100 mg on two occasions one week apart) or subcutaneous saline (1 ml as for PRG). In cats given saline (n = 6) basal cortical concentrations, cortisol concentrations after adrenocorticotrophic hormone (ACTH) administration and fasting blood glucose concentrations did not change significantly during the following seven weeks. Cats given MA (n = 7) developed suppression of basal and ACTH-stimulated cortisol concentrations and fasting hyperglycaemia during treatment. Effects on cortisol persisted for two weeks after MA dosage ceased. In cats given PRG (n = 7), basal cortisol concentrations were reduced overall, but only three cats had persistently suppressed post-ACTH cortisol concentrations. Adrenal suppression continued for 14 weeks in one of these and for at least 22 weeks in two cats. Fasting blood glucose concentrations were unchanged in PRG-treated cats.     

A new method for recording and analysing evoked motor potentials from dogs

Normal values for motor conduction in the tibial, ulnar and fibular nerves of dogs have been determined using a new method for recording and analysing evoked motor potentials. The use of an alligator clip as a roving surface electrode for recording, and a personal computer to analyse the evoked potentials has facilitated faster and more reproducible motor conduction studies. Compound muscle action potential (CMAP) and motor nerve conduction velocity data are in good agreement with previous studies. Normal values for CMAP area, residual latency and proximal to distal ratios for CMAP area and amplitude are presented for the first time.     

Iopamidol myelography in the horse

The use of the non-ionic, water-soluble contrast agent iopamidol for myelography in seven horses is described. Contrast columns of diagnostic quality were produced in all seven cases and the procedure did not invoke any adverse reactions in the five cases which were recovered from general anaesthesia. It is concluded that iopamidol is a safe and effective contrast agent for myelography in the horse.     

Plasma cortisol concentrations in dogs given cortisone or placebo by mouth

Fasted normal dogs (n=8) were given fixed doses of cortisone acetate orally as 5 mg and 25 mg tablets; plasma cortisol concentrations were determined, and C_max, t_max and area under the curve of plasma cortisol concentration plotted against time from zero to 12 hours were compared for the two preparations. In addition, these variables were compared when 25 mg tablets were administered with and without food. No significant difference in cortisol availability was noted for the two preparations and feeding did not apparently affect cortisone absorption. The findings in two hypoadrenocorticoid dogs were similar. Plasma cortisol concentrations in placebo-treated dogs similarly sampled showed minor fluctuations and were generally within accepted reference limits for normal dogs.     

A Retrospective Survey of Diseases of Feedlot Cattle in Alberta

A survey of feedlot managers was used to obtain information on disease occurrence, management practices and preventive techniques employed in feedlots in Alberta. Respiratory diseases were reported to be the most frequent causes of sickness and death. Costs associated with disease occurrence were estimated to be C $15.6 million in feedlots in Alberta annually during the period under study.