Oxytocin synthesis and secretion from bovine corpora lutea exposed in vitro to cycloheximide and colchicine

Two experiments were conducted to study the effects of cycloheximide and colchicine on prostaglandin F₂ (PGF₂)-induced secretion and synthesis of oxytocin in bovine luteal tissue in vitro. Corpora lutea were collected from beef heifers on Day 8 of the estrous cycle. In Experiment 1, incorporation of [¹⁴C]-leucine into oxytocin synthesized and secreted by luteal slices after exposure to PGF₂, cycloheximide and cycloheximide plus PGF₂ was examined. In Experiment 2, synthesis and secretion of oxytocin were evaluated in luteal slices incubated with colchicine and PGF₂ alone and in combination. Cycloheximide inhibited incorporation of labeled leucine into luteal proteins by more than 90% and no labeled oxytocin was detected in the media or tissue. Prostaglandin F₂ induced significant secretion of oxytocin that was not inhibited by cycloheximide. Tissue levels of oxytocin after incubation with cycloheximide and/or PGF₂ did not differ and were similar to those of the incubated control. Colchicine alone did not suppress oxytocin secretion and did not alter the ability of PGF₂ to induce significant secretion of this nonapeptide. Tissue concentrations of oxytocin after incubation with colchicine and/or PGF₂ did not differ. These studies indicate that secretion and replenishment of luteal oxytocin in vitro is not contingent upon de novo protein synthesis. Inability of colchicine to suppress oxytocin secretion and synthesis may have been due to the short duration of exposure of luteal tissue to the drug.     

Transforming growth factor type beta (TGF-beta) and adipogenesis in pigs

The present study was performed on s.c. adipose tissue of fetal pigs at 35 to 110 d of gestation to examine the distribution of TGF-beta-positive cells, to localize TGF-beta immunoreactivity at the cellular level using electron microscopy (EM), and to determine the effect of TGF-beta on primary cultures of pig adipose tissue cells. Tissues for EM were fixed and embedded in LR white resin. Sections then were incubated with a polyclonal antibody specific for TGF-beta and TGF-beta was located using 20 nm colloidal gold conjugated second antibody. Tissues were fixed and embedded in paraffin for localization of TGF-beta at the light microscope (LM) level. Tissues were incubated with anti-TGF-beta followed by localization using biotinylated second antibody. Using LM, only a few cells stained positively for TGF-beta within developing blood vessels at 35 d. By 50 d, more TGF-beta-positive cells were associated with forming capillary networks. Between 70 d and 110 d, positively stained adipocytes usually were clustered around blood vessels. Cells surrounding hair follicles stained positive for TGF-beta between 90 to 110 d. Electron microscopy revealed TGF-beta labeling within fat cells. Fibroblasts and endothelial cells did not exhibit TGF-beta immunoreactivity. The addition of TGF-beta to primary cultures of s.c. adipose tissue cells from newborn pigs prevented lipid filling in fat cells. This effect was dose-dependent, with half-maximal inhibition occurring at 3 pM maximum inhibition occurred at 40 pM. These results indicate that TGF-beta may regulate angiogenic activity and lipid filling in s.c. adipose tissue of fetal pigs. Although TGF-beta was present in adipocytes and in cells associated with developing capillary networks, the physiological role of TGF-beta during early adipose tissue development is not known.     

Pathology of infectious diarrhea of infant rats (IDIR) induced by an antigenically distinct rotavirus

Suckling rats were inoculated with a group B rotavirus to determine the progression of the morphologic changes induced in the intestine by this virus. Several changes were observed by light microscopy 1 day after viral inoculation: shortening of small intestinal villi, villous epithelial necrosis, and villous epithelial syncytia. The lesions were most often present in the distal small intestine, although other small intestinal segments were affected to a lesser degree. By day 3 post-inoculation, epithelial necrosis, and syncytia were no longer present; however, the villous epithelium was disorganized and irregularly vacuolated, and intestinal crypt epithelium was hyperplastic. Alterations in villous height to crypt depth ratios were present in portions of the small intestine for the remainder of the 12-day study period. Epithelial syncytia appeared to form by the breakdown of the lateral interdigitating membranes of the absorptive villous epithelium. Viral particles, abundant in the syncytia, appeared to form from amorphous or reticular arrays of viral precursor material. Group B rotaviral antigens, as detected by indirect immunofluorescence, were present in large amounts in the small intestinal villous epithelium only on the first day after viral inoculation. These studies show that two important diagnostic features of group B rotaviral infections of rats, epithelial syncytia and viral antigen as determined by immunofluorescence, are present only on the first day of disease. These findings should be taken into consideration when attempting to diagnose disease induced by this agent.     

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).     

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.     

Relationship between colloid osmotic pressure and plasma protein concentration in cattle, horses, dogs, and cats.

The relationship between colloid osmotic pressure (COP) and protein concentration was investigated for purified proteins and plasma samples obtained from cattle, horses, dogs, and cats. At equivalent concentrations, bovine albumin exerted a COP that exceeded that of gamma-globulins by a mean factor of 4.4. Similar relationships between COP and protein were observed in the other species. Consequently, for a given total protein concentration, COP was dependent on the albumin/gamma-globulins ratio. A commonly used nomogram for estimating COP from protein concentration, the Landis-Pappenheimer equation, did not provide reliable results for plasma samples from these species.     

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.     

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)