Temporal dynamic changes in synthesis of chondroitin sulfate isomers in canine articular chondrocyte culture.

To investigate temporal dynamic changes in the synthesis of chondroitin 6-sulfate (CS6) and chondroitin 4-sulfate (CS4) in vitro, normal articular cartilage of femoral heads was harvested from three dogs. Chondrocytes were isolated and cultured in alginate microspheres for 21 days. On days 7, 14 and 21, DNA content was quantified by fluorometric assay using Hoechst 33258. On days 14 and 21, proteoglycans were extracted, and the amounts of CS6 and CS4 were quantified after chondroitinase ABC digestion using capillary electrophoresis. The DNA content and amounts of CS6 and CS4 increased during the culture period. The amounts of CS6 and CS4 divided by DNA content revealed that the synthesis of CS6 was more up-regulated than CS4.

Influence of dietary calcium and phosphorus content in a fixed ratio on growth and development in Great Danes.

OBJECTIVE: To study the musculoskeletal development of Great Dane puppies fed various dietary concentrations of calcium (Ca) and phosphorus (P) in fixed ratio by use of dual energy x-ray absorptiometry (DEXA), determination of serum insulin-like growth factor 1 and parathyroid hormone concentrations, radiography, and blood chemistry analysis results. ANIMALS: 32 purebred Great Dane puppies from 4 litters. PROCEDURE: At weaning, puppies were assigned randomly to 1 of 3 diets. Blood was collected for biochemical analyses and hormone assays, and radiography and DEXA were performed through 18 months of age. Changes in body weight, bone mineral content, fat tissue weight, lean mass, result of serum biochemical analyses, hormonal concentrations, and radius lengths were analyzed through 18 months of age. RESULTS: Bone mineral content of puppies correlated positively with Ca and P content of the diets fed. Significant differences between groups in bone mineral content, lean mass, and body fat were apparent early. The disparity among groups increased until 6 months of age and then declined until body composition was no longer different at 12 months of age. Accretion rates for skeletal mineral content, fat, and lean tissue differed from each other and by diet group. CONCLUSIONS AND CLINICAL RELEVANCE: Ca and P concentrations in the diet of young Great Dane puppies are rapidly reflected in the bone mineral content of the puppies until 5 to 6 months of age, after which hormonal regulation adjusts absorption and excretion of these minerals. Appropriate Ca and P concentrations in diets are important in young puppies < 6 months of age.

Effects of exercise on canine skeletal muscle proteolysis: an investigation of the ubiquitin-proteasome pathway and other metabolic markers.

The effects of long-term athletic training are associated with excessive skeletal muscle turnover attributable to increased rates of myofibrillar protein synthesis and proteolysis, which are mechanisms poorly understood in the athletic dog. A physiologic field study using 44 English pointers and Labrador retrievers that had been purposely bred for bird hunting and retrieving was conducted to examine changes in the ubiquitin-proteasome (UP) pathway, which has been implicated in exercise-induced proteolysis. Muscle biopsy samples were collected from all dogs in September (preseason, pretraining) and February (peak season, peak activity). Western blot analysis was used to assess changes in expression of various components of the UP pathway in the biopsy samples. Citrate synthase and glycogen levels were also measured in a subset of these samples. Results across the population indicated pronounced up-regulation of ubiquitinated conjugates and the p31 regulatory capping subunit during the peak hunting period compared with the preseason period. In contrast, the catalytic core of the proteasome (beta-subunits) showed no apparent up-regulation in response to increased physical activity. Increased physical activity during the hunting season was associated with increased muscle glycogen levels and citrate synthase activity in these dogs. Overall, up-regulation of specific components of the UP pathway was an indication that it plays a role in the proteolytic process associated with skeletal muscle turnover during long-term athletic training, as previously believed.