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 alpha (PGF2 alpha)-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 alpha, cycloheximide and cycloheximide plus PGF2 alpha was examined. In Experiment 2, synthesis and secretion of oxytocin were evaluated in luteal slices incubated with colchicine and PGF2 alpha 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 alpha induced significant secretion of oxytocin that was not inhibited by cycloheximide. Tissue levels of oxytocin after incubation with cycloheximide and/or PGF2 alpha 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 alpha to induce significant secretion of this nonapeptide. Tissue concentrations of oxytocin after incubation with colchicine and/or PGF2 alpha 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.

Serially determined plasma alpha-tocopherol concentrations and results of the oral vitamin E absorption test in clinically normal horses and in horses with degenerative myeloencephalopathy.

Plasma alpha-tocopherol (vitamin E) values were monitored serially in 9 foals sired by a stallion with equine degenerative myeloencephalopathy (EDM) and in 5 age-matched control foals (sired by a clinically normal stallion) raised in the same environment for the first year of life. Clinical evaluation determined that 8 of the 9 foals sired by the stallion with EDM had neurologic deficits consistent with the disease on one or more occasions during the study period, whereas control foals had normal gait. From 6 weeks to 10 months of age, plasma alpha-tocopherol values in foals with signs of EDM were significantly (P less than 0.001) lower than those in control foals. An oral vitamin E absorption test was performed, and results for 8 of the affected horses and the affected stallion were compared with results for 4 of the monitored control horses and 4 additional control horses. Significant differences were not evident in any of the absorption indices. On the basis of data from this study and supported by reported prophylactic and therapeutic benefits of supplemented vitamin E, low plasma concentration of vitamin E is concluded to be a factor in the development of EDM in the first year of life of hereditarily predisposed foals. It was also concluded that the significantly lower alpha-tocopherol values seen in the foals in this study did not reflect a primary gastrointestinal tract absorption problem.

Clinical, viral, and genetic evaluation of equine degenerative myeloencephalopathy in a family of Appaloosas.

A clinical, viral, hematologic , and genetic study was conducted over a 4-year period on a family of Appaloosas with high incidence of clinical ataxia and pathologic features of equine degenerative myeloencephalopathy. Marginal to deficient serum vitamin E (alpha-tocopherol) and blood selenium values were the only other consistent antemortem abnormalities in the affected horses. Members of this family were all descendants of a clinically normal mare and were raised in 3 separate environments with variable quality of feed. All horses had access to pasture grasses. Normal chromosomal karyotypes were found in 11 affected and/or related horses examined. Equine herpesvirus type 2 was isolated from 4 of the horses, but evidence for a role of this virus in the pathogenesis of the disease was not found. The role of antioxidant deficiency in the pathogenesis of neurologic dysfunction in this equine family and in others reported to be affected with equine degenerative myeloencephalopathy remains speculative.

Stimulation of bovine luteal oxytocin secretion in vitro by a phorbol ester and calcium ionophore.

Experiments were conducted to examine the in vitro effects of a phorbol ester and a calcium ionophore on bovine luteal oxytocin (OT) secretion and synthesis and progesterone secretion. Corpora lutea removed from beef heifers on d 8 of an estrous cycle were sliced and incubated for 2 h with .81 nM 12-O-tetradecanoylphorbol-13-acetate (TPA), 1.62 nM TPA or .3 microM calcium ionophore A23187. Both concentrations of TPA increased (P less than .01) OT secretion (ng.g-1.2 h-1; control, 407.1; .81 nM TPA, 494.7; 1.62 nM TPA, 528.1; SE = 21.2). Increased secretion of OT was accompanied by a corresponding increase (P less than .02) in synthesis of the hormone (ng.g-1.2 h-1; control, 368.5; .81 nM TPA, 427.6; 1.62 nM TPA, 492.1; SE = 25.7). Phorbol ester also induced (P less than .025) progesterone secretion (ng.g-1.2 h-1; control, 1,056.2 vs .81 nM TPA, 1,333.3; SE = 86.4). Calcium ionophore increased (P less than .01) OT secretion (ng.g-1.2 h-1; control, 248.9 vs A23187, 327.4; SE = 16) and there was a trend (P = .09) toward increased synthesis of OT in response to the ionophore (control, 124.4 vs A23187, 165.6; SE = 16.4). Because TPA can activate protein kinase C and A23187 increases intracellular calcium, these intracellular constituents probably are involved in promoting secretion of OT and progesterone.

Response of the bovine corpus luteum to increased secretion of luteinizing hormone induced by exogenous gonadotropin releasing hormone.

Two experiments were conducted to investigate the response of the bovine corpus luteum to surges of luteinizing hormone (LH) induced by natural gonadotropin-releasing hormone (GnRH) administered twice during the same estrous cycle. In experiment 1, eight mature beef cows, each cow serving as her own control, were injected intravenously (iv) with saline on days 2 and 8 of the cycle (day of estrus = day 0 of the cycle), then with 100 micrograms GnRH on days 2 and 8 of the subsequent cycle. Jugular blood samples were taken immediately prior to an injection and at 15, 30, 45, 60, 120 and 240 min postinjection, to quantitate changes in serum luteinizing hormone. Blood was also collected on alternate days after an injection until day 16 of the cycle, to characterize changes in serum progesterone concentrations. Although exogenous GnRH caused release of LH on days 2 and 8 of the cycle, the quantity of LH released was greater on day 8 (P less than .025). Serum levels of progesterone after treatment with GnRH on day 8 of the cycle did not differ significantly from those observed during the control cycles of the heifers. Because exposure of the bovine corpus luteum to excess LH, induced by GnRH early during the estrous cycle, causes attenuated progesterone secretion during the same cycle, these data suggest that a second surge of endogenous LH may ameliorate the suppressive effect of the initial release of LH on luteal function. Duration of the estrous cycle was not altered by treatment (control, 20.4 +/- .5 vs. treated, 20.4 +/- .4 days).(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of the potential for interference by dimethyl sulfoxide (DMSO) in drug detection in racing animals.

Dimethyl sulfoxide (DMSO) had been postulated to be a 'masking agent' when used concurrently with therapeutic or prohibited drugs in racing animals. Eight drugs (flunixin, furosemide, caffeine, apomorphine, phenylbutazone, lidocaine, cocaine, and acepromazine maleate) were administered to six horses singly and with concurrent intravenous DMSO. Urine samples were analyzed for the presence of the drugs and/or their metabolites by thin layer chromatography. Direct comparison of thin layer chromatograms of extracts of positive urine samples with and without DMSO verified that DMSO did not interfere with the detection of these drugs.

Pharmacokinetic disposition of dimethyl sulfoxide administered intravenously to horses.

Dimethyl sulfoxide (DMSO) was administered IV to 6 Thoroughbred horses at 2 dosages: 1.0 g/kg and 0.1 g/kg. The pharmacokinetics seemed linear, with biological half-lives of 8.6 +/- 0.3 hours and 9.8 +/- 2.2 hours for the 1.0 g/kg and 0.1 g/kg dosages, respectively. This was further substantiated by mean residence times of 9.8 +/- 0.44 hours and 13.8 +/- 4.25 hours, areas under the curve of 12.55 +/- 1.42 mg/ml/hr and 1.63 +/- 0.49 mg/ml/hr, and the clearances of 0.081 +/- 0.009 L/kg/hr and 0.066 +/- 0.022 L/kg/hr for the large and small dosages, respectively. At 12 hours after 1.0 g/kg was administered, 26.6% of the DMSO dose was excreted unchanged into the urine; at 12 hours after 0.1 g/kg was administered, 25.3% of the DMSO dose was excreted unchanged into the urine. It was predicted that 29.4% and 40.6% of the total DMSO dose would be excreted into the urine for the 1.0 g/kg and 0.1 g/kg dosages, respectively. A 10% DMSO concentration in normal saline solution was safe to give as rapid IV infusion. Slow administration is recommended for more concentrated solutions. Based on the half-life, DMSO should be administered 2 times a day IV for the treatment of increased intracranial pressure and/or cerebral edema in horses.

Effect of supplemental beta-carotene on incidence and responsiveness of ovarian cysts to hormone treatment.

Sixty-two multiparous and 35 primiparous Holstein cows were assigned randomly at 10 days postpartum to receive a ration with or without 300 mg beta-carotene/cow per day. Multiparous and primiparous cows were grouped separately and group-fed complete rations once daily. Incidence of ovarian cysts (26% by rectal palpation) was not affected by beta-carotene fed. Multiparous cows had greater incidence (39%) of ovarian cysts than primiparous cows (11%). Fifty-seven percent of cysts were classified follicular by rectal palpation. Progesterone concentration of milk also was used for diagnosis of type of cyst. Cows with ovarian cysts and with progesterone concentrations in milk less than 1 ng/ml were classified follicular, and those having concentration greater than 1 ng/ml were classified luteal. As determined by milk progesterone, rectal palpation was more accurate for diagnosis of luteal cysts than for diagnosis of follicular cysts. Progesterone concentrations of milk for animals with luteal and follicular cysts were 10.66 +/- 1.29 and .37 +/- .07 ng/ml. beta-Carotene did not affect response or days to respond to treatment with human chorionic gonadotropin or gonadotropin-releasing hormone. Supplemental beta-carotene was not beneficial for reducing incidence of ovarian cysts in cows receiving an adequate supply of beta-carotene in their diet.

Association of increased estradiol and progesterone blood values with altered bovine polymorphonuclear leukocyte function.

Polymorphonuclear leukocyte (PMN) function and serum concentrations of estradiol, progesterone, and cortisol (hydrocortisone) were monitored concurrently in clinically normal cows during the estrous cycle. Five parameters were used to evaluate PMN function: (i) random migration under agarose, (ii) ingestion of 125I-labeled Staphylococcus aureus, (iii) nitroblue tetrazolium (NBT) reduction, (iv) iodination, and (v) antibody-dependent cell-mediated cytotoxicity. Increased serum estradiol concentrations were associated with enhanced random migration, but had no apparent effect on NBT reduction, iodination, or ingestion of S aureus by bovine PMN. Increased serum estradiol was also associated with increased serum cortisol. Increased serum progesterone values were associated with a depression of NBT reduction and iodination by PMN, but with enhanced random migration and antibody-dependent cell-mediated cytotoxicity. These results indicate that physiologic changes in steroid hormone values during the normal estrous cycle of the cow are associated with alterations in PMN function.

Mycobacterium paratuberculosis in the semen and genital organs of a semen-donor bull.

Mycobacterium paratuberculosis, the cause of Johne's disease, was isolated from the feces of a donor bull in an artificial insemination stud. During isolation and observation for 21 months, the organism was recovered from all of 26 fecal samples and from 8 of 31 semen samples. At necropsy, it was isolated from the intestine and adjacent lymph nodes, lung, spleen, seminal vesicles, and prostate gland but not from the testicles. We concluded that routine fecal cultures at bull studs will reveal infected bulls before they become genitally infected and shed M paratuberculosis in semen.

Inhibition of respiratory virus infections of mice with aerosols of synthetic double-stranded ribonucleic Acid.

Aerosols of double-stranded complexes of polyinosinic and polycytidylic acids (poly I:C) were useful in protecting mice infected with aerosols of influenza (A(2)/Taiwan/64) and parainfluenza type 1 (Sendai) viruses. Administration of poly I:C as an aerosol offers an advantage, particularly in therapy, by eliminating the risk of pulmonary dissemination of viral infections due to intranasally instilled fluids. Treatment of mice with aerosols of poly I:C reduced the infection rate with influenza virus but did not inhibit virus multiplication in the lungs of most of those animals where infection became established. Sendai virus infection rates were undiminished in mice treated with poly I:C, but lung-virus titers were significantly suppressed as compared with those of untreated animals. The maximum poly I:C doses (40 mug) administered by aerosol produced no evidence of toxicity in the mice.