Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate

Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein-associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I.     

Correlation of DNA ploidy to tumor histologic grade, clinical variables, and survival in dogs with mast cell tumors.

By using flow cytometry, a retrospective analysis of the DNA content of 40 primary canine mast cell tumors and seven lymph nodes that contained metastatic mast cell tumor from 44 dogs of various breed, sex, and age was performed on formalin-fixed, paraffin-embedded samples of the tumors and nodes. These samples were chosen according to the following criteria: samples contained sufficient well-preserved tumor tissue in the paraffin block for processing, sufficient patient history data were available, clean and homogeneous cell suspensions were obtained after processing, and interpretable DNA histograms were produced on analysis. The ploidy data obtained were compared with the histopathologic grade, the anatomical site of occurrence, the clinical stage of the tumors, and the survival of the dogs. Over 70% (29/40) of the mast cell tumors were diploid. Three metastatic mast cell tumors in lymph nodes had the same ploidy status as their corresponding primary tumors. In five dogs, mast cell tumors from multiple sites in each dog displayed similar ploidy status. Of 26 dogs evaluated for survival times, 69% (18/26) had diploid tumors and 31% (8/26) had aneuploid tumors. When numbers of diploid versus aneuploid tumors were compared, no significant difference was found between any two grades, clinical stages, or anatomic sites. A significant difference (P = 0.02) was found, however, between aneuploid and diploid tumors when comparing Stage I and non-Stage I disease. The Kaplan-Meier survival plot indicated a tendency towards an increased survival within the first year in dogs with diploid versus aneuploid tumors (P = 0.06).     

Dense Cultivation and Fertilization for Higher Yield of Thyme (Thymus vulgaris L.)

Thyme ( Thymus vulgaris L.) plants were spaced at 15, 30 or 45 cm distances in a clay-loamy soil. They received different levels of nitrogen and phosphorus, besides a constant level of potassium. The results showed, that the wider spacing promoted the growth and production of herb and oil per plant, however dense cultivation significantly increased the yields of herb and oil per unit area. In all cases, the applied fertilization treatments significantly increased the productivity per unit area. Dense cultivation accompanied with higher levels of fertilization proved to be very usefull. The essential oil content was not influenced by either the plant spacing or fertilization treatments applied in this study.     

Effects of atherosclerosis on mean and daily variation of arterial pressure in conscious WHHL rabbits.

Effects of atherosclerosis on the mean value and daily variation of arterial pressure were studied in 12 Watanabe-heritable hyperlipidemic (WHHL) rabbits aged 12 to 35 months and 25 normal Japanese white rabbits aged 6 to 30 months. A pressure catheter was inserted through the left subclavian artery under pentobarbital anesthesia. A few days after the catheterization, the mean arterial pressure (MAP) of the rabbits, which were active and in a good state of appetite, was recorded by an analogue-to-digital converter every second for about 6 hrs and stored in a computer. The mean (M) and standard deviation (SD) in the WHHL rabbit, calculated from each successive MAP record, ranged widely from 85.8 to 131.4 mmHg and 5.6 to 12.6 mmHg, respectively. There was no significant correlation between M and SD in the WHHL rabbit. M and variance (V) of MAP in the WHHL rabbit were significantly higher than those in the normal rabbit. M did not show any significant change with increasing ages, whereas SD increased significantly with aging in the WHHL rabbit. Concentrations of serum total cholesterol and triglyceride in the WHHL rabbit were 475 and 328 mg/dl, which were about nine and seven times as high as those in the normal rabbit, respectively. Macroscopic and histopathological examinations of the aorta revealed development and spread of sclerotic lesions with aging in the WHHL rabbit. We can conclude that development of atherosclerosis with aging in the WHHL rabbit causes malfunction of the baroreceptors, which contributes to hypertension and lability of arterial pressure.     

Growth activity of bovid herpesvirus 1 in bovine follicular oocytes with cumulus cells.

Bovine follicular oocytes collected from bovine ovaries were exposed to bovid herpesvirus 1 (BHV-1). After washings, these oocytes were cultured to mature. As a result BHV-1 could not be removed from the oocytes and could replicate in the oocytes with cumulus cells, but not in the oocytes without the cells. Moreover, the specific fluorescence for BHV-1 was detected in the cumulus cells by a indirect immunofluorescent technique. Therefore these findings suggested BHV-1 could be absorbed in the oocytes but the replication of BHV-1 was done in the cumulus cells.     

Anti-erythrocyte membrane antibodies detected in sera of dogs naturally infected with Babesia gibsoni.

Due to the potential for anti-erythrocyte membrane antibodies as possible enhancers of erythrocyte destruction, the presence of serum anti-erythrocyte membrane antibodies in 31 dogs with Babesia gibsoni infection admitted to a veterinary hospital was investigated by an enzyme linked immunosorbent assay (ELISA) and immunoblotting analyses. This infection resulted in an increase of anti-erythrocyte membrane antibodies in 84% (IgG) and 74% (IgM) of 31 infected dogs, respectively. This was confirmed by the similarity in the protein profiles of the erythrocyte membrane antigens immunoblotted with rabbit antiserum to dog erythrocyte membrane antigens and infected dog serum. These results suggest the production of anti-erythrocyte membrane antibodies was induced by B. gibsoni infection.     

Laryngeal paralysis in immature and mature dogs as one sign of a more diffuse polyneuropathy

Six dogs with laryngeal paralysis had clinical, electrophysiologic, and pathologic evidence of a more generalized polyneuropathy. Three of the dogs were young Dalmatians, one was a young Bouvier des Flandres, and two were older, large-breed dogs. The results of this study suggest that laryngeal paralysis in dogs may frequently be one clinical sign of an underlying, more generalized polyneuropathy. Two forms of this generalized polyneuropathy may exist: an early form, as seen in young dogs with congenital or hereditary disease, and a delayed-onset form that is usually found in older dogs with so-called idiopathic laryngeal paralysis, some of which may have hypothyroidism.