Development of Immunologic Assays to Measure Response in Horses Vaccinated with Xenogeneic Plasmid DNA Encoding Human Tyrosinase

Xenogeneic plasmid DNA constructs have been developed and optimized for immunotherapies targeting cancer in both humans and dogs. Specifically, plasmid vectors containing the tumor antigen tyrosinase have demonstrated immunoreactivity and clinical benefit in the treatment of melanocytic tumors in these species. Overexpression of tyrosinase has also been noted in equine melanocytic tumors, supporting its role as a valid tumor antigen in the horse. Vaccination with plasmid constructs containing tyrosinase may thus have translational immunoreactivity in the treatment of equine melanomas. Here, we describe a methodology that is highly sensitive and specific for the detection of both humoral and cell-mediated immunoreactivity against tyrosinase in equine patients. These antigen-specific immunoassays are used to measure the humoral and cell-mediated responses in a cohort of horses vaccinated with xenogeneic plasmid DNA encoding human tyrosinase. Serum humoral responses were measured using standard enzyme-linked immunosorbent assay technique against the full-length recombinant human tyrosinase protein. Peripheral blood mononuclear cells were collected from vaccinated horses and stimulated with tyrosinase-specific peptides. Cell-mediated responses were then measured using a novel quantitative real-time-polymerase chain reaction technique to determine resultant interferon-γ expression. All horses developed significantly positive humoral and cell-mediated immune responses compared with their individual prevaccination values. No adverse reactions or signs of autoimmunity were detected. Vaccination with xenogeneic plasmid DNA expressing tyrosinase appears to elicit tumor antigen-specific reactivity and should be evaluated in a larger cohort of horses with melanocytic tumors.     

mRNA expression of canine ATP10C, a P4-type ATPase, is positively associated with body condition score

Mouse and human Atp10c genes are strong candidates for changes in bodyweight and glucose homeostasis. Using comparative genomic analysis, a novel canine P4-type ATPase, ATP10C, was identified. Expression of ATP10C was compared between sex-matched lean (body condition score, BCS<8; n=7) and obese (BCS?8, n=8) client-owned dogs of comparable ages. Canine ATP10C is highly expressed in visceral and subcutaneous fat at approximately 3-fold levels compared to the omental adipose depot. There was a 5-fold significant increase (P<0.0001) in mRNA expression of ATP10C in dogs with a BCS?8.     

From monospecific to mixed forest after fire?: An early forecast for the montane belt of Majella,Italy

Alternative hypotheses on post-fire successions of monospecific European beech (Fagus sylvatica L.) and black pine (Pinus nigra Arnold) forests are investigated. The research area is located in the centre of the Apennine montane belt, Italy and covers portions of the Roccamorice and Lettomanoppello municipalities within the Majella National Park. Temporal moderate-resolution (MODIS) and spatial mid-resolution (ASTER) imagery are combined for the assessment of the 2007-fire features. Tree seedlings were surveyed on the ground in post-fire forests and their unburnt equivalents during September 2008. The satellite imagery analysis provides records on spread, intensity, location and extent of the forest fires. The evidence suggests an abandoned farmland fire spreading secondarily uphill into the forests.     

Survey of heartworm prevention practices among members of a national hunting dog club

Surveillance data indicate that failures have been reported for virtually all heartworm prevention product categories. Resistance of third and fourth stage larvae of Dirofilaria immitis to macrocyclic lactones, lack of compliance, other unknown factors, or a combination of these reasons may be the cause of failure. A survey of members of a national hunting dog club was conducted to identify practices used to prevent canine heartworm infections. Questionnaires were completed by 708 dog owners. Year-round administration of heartworm preventive medication was reported by 208 (88%) respondents residing north of the 37th parallel. Dosing was based on the estimated weight of the dog by 54 (7%) respondents, 389 (55%) did not record the date prophylaxis was administered, and 89 (13%) observed the dogs spit out pills. Heartworm testing at least once per year was done by 556 (79%) respondents and test dates were spread throughout the year. Only 448 (64%) respondents tested newly acquired dogs for heartworm. These findings suggest that veterinarians should place a greater emphasis on the frequency and timing of heartworm diagnostic tests, the importance of weighing dogs, duration of administration, recording the date monthly heartworm prophylaxis is given, and observing dogs to ensure that oral medication is retained.