In situ hybridization analysis of ovarian prostaglandin endoperoxide synthase mRNA throughout the periovulatory period of the ewe.

Cellular alterations in level of expression of mRNA encoding for prostaglandin endoperoxide synthase were quantified within ovarian tissues of sheep obtained before, during and after induction of the preovulatory surge of LH and ovulation with LHRH. This was accomplished by isotopic in situ hybridization using a selective cRNA probe to ovine prostaglandin endoperoxide synthase mRNA. A significant elevation in mRNA was detected within the theca interna of the preovulatory follicle at 8, 16 and 24 hr following administration of LHRH. Very close to the time of ovulation (ie., at 24 hr post-LHRH) a marked rise in mRNA was observed in association with epithelial cells covering the apical surface of the follicle. Ovarian cyclooxygenase metabolites of arachidonic acid produced during the ovulatory process in the ewe originate within the thecal layer and germinal epithelium of the follicle destined to ovulate.     

Acute infection of encephalomyocarditis (EMC) virus in Syrian hamsters.

One-month-old Syrian hamsters of the APA and Std: golden strains were inoculated intraperitoneally with 10(5) PFU/head of the D variant of encephalomyocarditis (EMC) virus and examined virologically and pathologically up to 7 days after inoculation. APA hamsters developed apparent hyperglycemia due to pancreatic islet cell damage while Std:golden hamsters did not. Hamsters of both strains showed clear histopathologic changes in the testis with prominent viral replication as well as in the brain, heart and exocrine pancreas. The susceptibility to EMC virus-infection was higher in males than in females and in APA than in Std: golden hamsters.     

Free-running rhythms of adrenocorticotropic hormone (ACTH), cortisol and melatonin in pigs.

In the domestic pig, a circadian rhythm of plasma cortisol occurs, with greatest concentrations in the morning and lowest concentrations in the afternoon. However, photic entrainment of the rhythms of ACTH and melatonin in pigs have not been defined clearly. This experiment was designed to evaluate free-running rhythms of ACTH, cortisol and melatonin in pigs housed in constant light (LL) and constant darkness (DD). Twelve crossbred barrows, maintained under ambient photoperiod, were catheterized and tethered individually in two environmentally controlled rooms, one with LL and the other with DD. For animals in LL, fluorescent lights provided 202 +/- 15 (mean +/- standard deviation) lux of light at 65 cm above the floors. Incandescent nightlights equipped with 7 watt red bulbs provided 7 +/- 2 lux and were illuminated continuously in both rooms. Pigs were given at least 14 d exposure to LL and DD, then samples of plasma and serum were obtained at hourly intervals for 48 hr. Plasma was assayed for ACTH, and serum for cortisol and melatonin. Periodograms were constructed to analyze the data. For this type of analysis, a statistic, Qp, is calculated, and circadian periodicity is suggested if maximum Qp (Qp max) occurs at or near 24 hr. The period of the free-running rhythms (tau) at Qp max for ACTH, cortisol and melatonin for pigs in LL (23.80 +/- .01, 23.78 +/- .01, and 23.21 +/- .02 hr, respectively) did not differ significantly from those for pigs in DD (23.39 +/- .01, 23.20 +/- .01, and 22.55 +/- .02 hr, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Humoral immune responses of black-footed penguins (Spheniscus demersus) after DNA-mediated immunization with a beta-galactosidase reporter gene.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a beta-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-beta, containing the beta-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-beta-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to beta-galactosidase were detected in the sera of pCMV-beta-inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding beta-galactosidase and will mount a humoral response against the in vivo-expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.     

Characterization of and radioimmunoassay for canine thyroglobulin

Canine thyroglobulin (cTg) has been isolated and purified. It has similar electrophoretic patterns as Tg from other mammalian species. The main fraction had a MW of 660,000, whereas also fractions of a MW of approximately 1,300,000 (dimer) and 330,000 (subunit) were present. The iodine content was 0.8 to 1.0 % (w/w). cTg did not cross-react with antibodies against human Tg to a degree that would allow the use of a radioimmunoassay for human Tg for the determination of cTg in serum or plasma. Therefore a polyclonal antiserum was raised against cTg and a homologous radioimmunoassay was developed, which was sensitive (0.4 μg/l) and specific (cross-reactivity in cTg assay of human Tg, goat Tg, T₄, T₃, and DIT < 0.01 %).

Plasma Tg levels in normal dogs of both sexes and aged 3–15 years amounted to 192 ± 73 μg/l (mean ± SD, n=30). There was no relation between plasma Tg and T₄ levels.