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.     

Evaluation of sodium thiosulfate as an extracellular water marker in cattle.

Two studies were conducted to determine whether sodium thiosulfate (THS) can estimate extracellular water (ECW) in beef cattle in conjunction with empty body water (EBW) estimation by urea space. Experiment 1 used 24 steers (366 kg) to determine the clearance parameters for THS and urea. Blood samples were taken over 1 h. A two-component curve, Y = A1ek1(t) + A2ek2(t), (t = hours after infusion) fit the clearance of both markers; intercepts (A1, A2) and clearance coefficients (k1, k2) were 44.8, 44.4, -25.8, and -2.24 mg/dL, respectively, for THS (r2 = .98, Sy.x = 2.72, animal effects removed and 24.4, 10.5, -21.7, and -.71 mg/dL, respectively, for urea (r2 = .98, Sy.x = 1.49). Sodium thiosulfate equilibrated with ECW 5 to 10 min after infusion. Experiment 2 consisted of 22 steers (483 kg) infused with a combination solution of 20% urea, 10% THS, and 4% sodium thiocyanate (SCN; equilibration time = 28 min); half the steers were implanted with estradiol. Empty body water increased with implantation (P less than .01). Extracellular water tended to increase in implanted steers as measured by THS (12 min, P = .14) and SCN (P = .10). The estimation of ECW at 12 min was not different (P greater than .2) from the SCN estimate at 28 min (SCN = 3.7 + .873 THS; r2 = .70; P less than .001). Sodium thiosulfate gave reasonable estimates of ECW (22 to 26% of BW) and required only 0- and 12-min blood samples.(ABSTRACT TRUNCATED AT 250 WORDS)     

Selective measurement of lipoprotein lipase and hepatic triglyceride lipase in heparinized plasma from horses.

Affinity chromatography on heparin sepharose was used to identify 2 lipolytic enzymes in heparinized plasma from horses. One enzyme was typical of hepatic triglyceride lipase (HTGL), because it was resistant to inactivation by high concentrations of NaCl, and it did not require the addition of serum for activity. The other enzyme was identified as lipoprotein lipase (LPL), because of its inactivation at NaCl concentrations in excess of 0.2M, and its dependency on addition of serum as a source of apolipoprotein C-II activator. The enzymes were purified by 347-(HTGL) and 442- (LPL) fold, with yields of 54 and 58%, respectively. The partially purified enzymes were used to design incubation conditions that gave optimal activities for each enzyme in vitro. A selective assay was then developed for direct measurement of LPL and HTGL activities in heparinized plasma from horses. Analysis of HTGL took advantage of the almost complete inactivation of LPL when serum cofactor was excluded from the assay at the NaCl concentration that gave optimal HTGL activity. Prior incubation of heparinized plasma with sodium dodecyl sulfate to inhibit HTGL was necessary for measurement of LPL, because HTGL retained 67% of its activity at the NaCl concentration required for optimal LPL activity. Activity of each enzyme was measured in heparinized plasma from 12 Shetland ponies. The mean activity +/- SD for LPL was 3.22 +/- 1.04 mumol of fatty acids/ml of heparinized plasma/h (mumol of FA/ml/h. The mean activity for HTGL was 4.9 +/- 1.56 mumol of FA/ml/h. The performance of the assay was assessed by replicate analysis of pools of each enzyme with high and low activities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevalence of antibody to caprine arthritis-encephalitis virus in goats in the United States.

Goats from 28 states were tested for antibodies to caprine arthritis-encephalitis virus. Of 3,790 goats, 1,175 (31%) tested positive, and of 196 herds tested, 143 (73%) had 1 or more seropositive members. This prevalence, based on serum samples from all goats in the participating herds, was lower than most rates reported in other studies. Such studies were based on fewer samples, incomplete sampling of herds, or smaller geographic base. Prevalence was highest in western Pacific and northern plains regions, increased with age to 3 years, was highest among goats on family-owned farms, and was lowest in the Angora breed. Differences in prevalence was not related to gender or size of herd.     

Infectious load of Aeromonas salmonicida subsp. salmonicida during the initial phase of a cohabitant infection experiment with Atlantic salmon, Salmo salar L.

Abstract The abundances of Aeromonas salmonicida subsp. salmonicida in the water and in the surface microlayer was studied during the initial phase of a cohabitant infection experiment with Atlantic salmon, Salmo salar L., smolt. Aeromonas salmonicida was detected in the water samples only until the intraperitoneally infected smolt were dead and had been removed. In the lipid rich surface microlayer, A. salmonicida was detected in high concentrations from the day of the first fish mortality and throughout the rest of the experiment. The significance of the high cell surface hydrophobicity is discussed as a possible reason for enrichment of A. salmonicida at the air-water interface.     

A review of the usefulness of myelography in 50 dogs.

Fifty dogs showing clinical signs of spinal disease were investigated by myelography, using iopamidol. In 27 cases the technique was considered worthwhile. Of the 19 dogs not subjected to surgery or euthanasia as a result of the findings, three suffered seizures during recovery from anaesthesia, eight deteriorated in neurological condition and one suffered permanent respiratory arrest as a result of extensive subarachnoid haemorrhage.