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).     

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.     

Serum Progesterone Concentrations and Blood Neutrophil Alkaline Phosphatase Activity in Cows Following Normal and Abnormal Parturition

Serum progesterone concentrations and alkaline phosphatase (ALP) activity of blood neutrophils were determined in 3 groups of cows (n = 5 each) on days 1 and 2 and then at 3-day intervals up to 32 days post-partum. Group I cows had a normal delivery, Group II cows had dexamethasone-induced parturition and Group III cows were subjected to a caesarian section. All cows in Group III and 2 cows in Group II retained their fetal membranes. Mean serum progesterone concentrations declined the second day after calving (to < 0.67 ng/ml) and remained at low levels (< 0.54 ng/ml) throughout the observation period, except for the values in Group III, which were elevated on day 16 (0.94 ng/ml), declined again on day 26 (0.46 ng/ml) and peaked (1.05 ng/ml) on day 32 portpartum. Significant (P < 0.01) differences were found between serum progesterone concentrations on day 1 and on each of the other sampling days in Groups I and III. Day X parturition group interaction was significant (P < 0.05) for the progesterone concentrations. No significant differences were found between the overall means of ALP activity of blood neutrophils in the 3 parturition groups nor between days of the experiment. No significant correlation was found between serum progesterone concentrations and ALP activity values of blood PMN during the first 32 days post-partum. Inhalt: Serum Progesteron Konzentrat und Aktivität der alkalischen Phosphatase in neutrophilen Blutzellen bei Küken mit normalem und abnormalen Geburtsverlauf Serum-Progesteronkonzentrationen und alkalische Phosphatase- (ALP) Aktivität von neutrophilen Granulozysten, nach der Abkalbung am Tag 1 und 2 und dann im Abstand von 3 Tagen bis zum 32. Tag p.p. bei Kühen untersucht, die in drei Untersuchungsgruppen (n = 5) eingeteilt waren. Gruppe I kalbte normal ab, in Gruppe II wurde die Geburt durch Dexamethason eingeleitet, und Kälber der Gruppe III wurden via Kaiserschnitt gewonnen. Alle Tiere der Gruppe III und 2 Kühe der Gruppe II wiesen Nachgeburtsverhaltungen auf Der mittlere Serum-Progesterongehalt sank am 2. Tag p.p. auf < 0,67 ng/ml ab und blieb auf niederigem Niveau (< 0,54 ng/ml) während der gesamten Untersuchungs-periode. Lediglich in Gruppe III, mit erhöhten Werten (0,94 ng/ml) an Tag 16 p.p., sanken die Werte wieder an Tag 26 p.p. (0,46 ng/ml) und erreichten einen Maximalwert an Tag 32 p.p. mit 1,05 ng/ml. Signifikante (P 0,01) Unterschiede in der Serum-Progesteronkonrentration wurden zwischen Tag 1 p.p. und allen anderen Untersuchungstagen in den Gruppen I und III gefunden. Interaktionen zwischen den Gruppen für Tag X waren signifikant (P 0,05). Keine signifikanten Unterschiede wurden für den Gesamtmittelwert der ALP-Aktivität in den neutrophilen Granulorysten zwischen den Gruppen und im Vergleich der Untersuchungstage gefunden. Es wurden auch keine signifikanten Korrelationen zwischen den Serum-Progesteronkonzentrationen und der ALP-Aktivität der PMN während der ersten 32 Tage p.p. nachgewiesen.     

Critical and controlled tests of activity of a macrocyclic lactone (compound F28249-alpha) against natural infections in internal parasites of equids

Thirteen critical tests (n = 11 horses and 2 ponies) and 4 controlled tests (n = 4 donkeys and 6 horses) were performed to evaluate the activity of the experimental macrocyclic lactone compound F28249-alpha against internal parasites of equids. In the critical tests, activity was determined mainly against the large parasites, but 1 critical test also included benzimidazole-resistant small strongyles. In the controlled tests, evaluation of drug activity included large parasites and stomach worms in all 4 tests, and lungworms in 2 tests. The period between treatment and euthanasia was 6 to 9 days for the critical tests and 14, 17, or 52 days for the controlled tests. The compound was administered by stomach tube at dose rates of 1, 2, 3, 3.5, or 4 mg/kg of body weight. In the critical tests, removal at all 5 dose rates was 100% for Gasterophilus nasalis (2nd and 3rd instars), Parascaris equorum (mature), Strongylus vulgaris, and Strongulus edentatus from the gastrointestinal tract. For Gasterophilus intestinalis in the stomach, mean removals of 2nd instars were 88% at the rate of 2 mg/kg and 93% to 100% at rates greater than or equal to 3 mg/kg. For 3rd instars, mean removals were 7% at 1 mg/kg, 77% at 2 mg/kg, 90% at 3 mg/kg, and 98% at 3.5 mg/kg. Discharge of G intestinalis in feces was typically a slow, prolonged process and probably higher removal values, especially at lower dose rates, would have attended a longer interval after treatment before necropsy examination.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum amyloid A protein concentration measured by radial immunodiffusion in Abyssinian and non-Abyssinian cats

Serum amyloid A (SAA) protein concentration was determined by use of radial immunodiffusion (RID) in 4 groups of cats: Abyssinian cats with amyloidosis, healthy Abyssinian cats without clinical evidence of amyloidosis, hospitalized non-Abyssinian cats, and clinically normal non-Abyssinian cats. Mean SAA concentration in Abyssinian cats with amyloidosis was significantly (P = 0.05) higher than mean SAA concentration in healthy Abyssinian cats without clinical evidence of amyloidosis and in hospitalized non-Abyssinian cats. Mean SAA concentration in clinically normal non-Abyssinian cats was significantly (P = 0.05) lower than mean SAA concentration in healthy Abyssinian cats without clinical evidence of amyloidosis and in hospitalized non-Abyssinian cats. Affected and healthy Abyssinian cats, however, could not reliably be distinguished on the basis of SAA concentration, because of the wide range of SAA values in these 2 groups of cats.