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.     

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.     

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)     

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.     

Metabolic response of horses to a high soluble carbohydrate diet: effects of low-intensity submaximal exercise and sodium bicarbonate supplementation.

Four mares fed a low fiber, high soluble carbohydrate diet were used in a crossover design to evaluate the effects of dietary sodium bicarbonate (NaHCO3) supplementation during daily low-intensity submaximal working conditions. Mares were fed the diet at 1.7 times the maintenance energy requirement for mature horses at work. The horses tolerated the diet well and had no clinical abnormalities. Resting venous blood bicarbonate (HCO3), standard HCO3, and base excess (BE) concentrations significantly (P less than 0.05) increased with NaHCO3 supplementation, but no significant changes in resting venous blood pH or carbon dioxide tension (PCO2) were recorded. Venous blood HCO3, standard HCO3, BE, hemoglobin, and heart rate were significantly (P less than 0.05) increased and plasma lactate concentration was significantly (P less than 0.05) decreased in the control horses and in the horses given the NaHCO3 supplement during low-intensity submaximal exercise. There were no significant changes in venous blood pH, PCO2, or plasma protein concentration with exercise. Venous blood HCO3, standard HCO3, and BE concentrations were significantly (P less than 0.05) greater during submaximal exercise in horses given the NaHCO3 supplement. There were no significant differences in plasma lactate or total protein concentrations, blood pH, PCO2, or hemoglobin concentration between the 2 groups during exercise.     

Use of 64Copper Measurements to Diagnose Canine Copper Toxicosis

Inherited canine copper toxicosis is a serious problem in Bedlington terriers and West Highland White terriers, and may also be a problem in other less-studied breeds. Affected dogs become ill at midlife with progressive and ultimately fatal liver disease. Treatments for removal of copper and prevention of copper accumulation are available, but are most effective if begun before the dog becomes ill. Until recently diagnosis has not been available until the dog is 1 year of age, and then only by an invasive liver biopsy with determination of liver copper concentration. The authors studied the use of 64copper for early diagnosis of canine copper toxicosis. Two procedures were evaluated. The first involved measuring the concentration of 64copper in blood 24 hours after oral administration of the radioisotope. At this time, 64copper was associated primarily with ceruloplasmin secreted into the blood by the liver. This procedure is useful in the diagnosis of the human counterpart, Wilson's disease. However, the authors found it to be nondiscriminatory between affected and unaffected dogs. In contrast, the second procedure, which involved measuring 64copper excreted in stool during 48 hours after an intravenous dose of radioisotope, yielded results that differentiated most affected and unaffected dogs.     

Effect of hot-fat trimming on factors associated with the subprimal yield of beef carcasses.

Thirty-two crossbred cattle (steers = 17; heifers = 15) exhibiting an ultrasound fat thickness at the 12 to 13th rib region of at least 10 mm were selected from a slaughter shift at a commercial packing plant. After splitting, alternating sides of each carcass were trimmed of 1) subcutaneous fat in excess of 6.4 mm; 2) all kidney, pelvic, and heart fat; and 3) all cod or udder fat and fat in the flank region. Both sides of each carcass were fabricated into subprimals (final trim level of 6.4 mm) according to normal industry procedures. Effect of hot-fat trimming, yield grade (3, 4, and 5), and gender on hot-fat trim, fabrication fat trim, major subprimal, and total subprimal yield of untrimmed and trimmed carcasses were determined. Higher numerical yield grade (YG) corresponded with higher (P less than .05) percentages of hot-fat trim. Hot-fat trimming increased (P less than .05) the difference in fabrication fat trim between steers and heifers and between YG 3 and YG 5. Steers and heifers differed (P less than .05) in percentage of major subprimals and total subprimals when processed conventionally, whereas hot-fat trimming eliminated this difference (P less than .05). Untrimmed YG 3 carcasses had 3.1 and 5.0% higher major subprimal yield (P less than .05) than untrimmed YG 4 and YG 5 carcasses, respectively, whereas hot-fat trimming reduced this difference to 2.5% for YG 4 and to 3.7% for YG 5.(ABSTRACT TRUNCATED AT 250 WORDS)