The Influence of Opioid Peptides on Steroidogenesis in Porcine Granulosa Cells

The present studies were undertaken to examine the influence of mu (beta-endorphin, DAMGO, FK 33-824), delta (met-enkephalin, leu-enkephalin, DPLPE) and kappa opioid receptor agonists (dynorphin A, dynorphin B, U 50488) used at different doses (1-1000 nM) alone and in combination with LH (100 ng/ml) on steroidogenesis in porcine granulosa cells derived from large follicles. The effects of mu, delta and kappa receptor agonists on both basal and LH-induced progesterone (P4) secretion were negligible. Agonists of mu opioid receptors reduced basal androstenedione (A4), testosterone (T) and oestradiol (E2) release. Co-treatment with LH entirely abolished the inhibitory effect of these agonists on A4 and E2 secretion and resulted in an increase in T release. The addition of delta receptor agonists was followed by a decrease in basal A4, T and E2 secretion. The cells incubated in the presence of LH increased the androgen production and abrogated the inhibitory effect of delta agonists on E2 output. Basal A4, T and E2 release was also suppressed by kappa receptor agonists. The presence of LH in culture media extended the inhibitory effect of these opioids on E2 output and caused either abolition of the inhibitory influence of kappa agonists or even augmentation of both androgen release in response to the opioids. In conclusion, these data support the involvement of three major types of opioid receptors in the regulation of porcine granulosa cell steroidogenesis.     

Traumatic elbow luxation repair in a common squirrel monkey (Saimiri Sciureus) and a bonnet macaque (Macaca radiata).

Traumatic elbow luxation was diagnosed in a common squirrel monkey (Saimiri sciureus) and a bonnet macaque (Macaca radiata). In the common squirrel monkey, the luxation was complicated by an ulnar fracture and would not stay reduced when attempts at closed reduction were made. The luxation was surgically reduced and a transarticular type I external skeletal fixator was placed. In the bonnet macaque, closed reduction was performed and the elbow was stabilized using a cast. Both monkeys regained good function of the injured elbows.     

Intramedullary Intervertebral Disk Extrusion in a Cat

Intervertebral disk extrusion is rarely reported in the cat. In this case, the clinical, radiologic, and magnetic resonance imaging findings of an acute intramedullary intervertebral disk extrusion in a 5-year-old domestic shorthaired cat are described. Radiographically, there was mineralized disk material within the vertebral canal. On magnetic resonance images, the disk material was found to be within the spinal cord. A linear disk trail extending from the disk space into the spinal cord may be specific for intramedullary disk extrusion.     

Molecular epidemiology of two fowl cholera outbreaks on a free-range chicken layer farm.

Two outbreaks of fowl cholera on a multiage free-range egg farm were investigated. The outbreaks occurred in 1994 and 2002. A total of 22 strains of Pasteurella multocida were available for study, 11 from the 1994 outbreak and 11 from the 2002 outbreak. Lesions typical of acute fowl cholera were seen in the 1994 outbreak, whereas both acute and chronic fowl cholera occurred in the 2002 outbreak. The isolates were examined in an extended phenotypic typing methodology, by a P. multocida-specific polymerase chain reaction (PCR), by the Heddleston somatic serotyping scheme, and by restriction endonuclease analysis (REA) typing using the enzyme HpaII. All 22 strains had the same phenotypic properties, all were confirmed as P. multocida by PCR, all were Heddleston serovar 4, and all had the same REA pattern. The results indicate that these 2 outbreaks were caused by the same clone of P. multocida--despite the 8-year time period between the outbreaks.     

Cloning and characterization of a bovine genomic fragment homologous to epidermal growth factor genes

Epidermal growth factor (EGF) is a potent mitogen for a variety of cell types. The 53-amino acid mature EGF protein is encoded by sequences in exons 20 and 21 of a gene spanning over 110 kb. In this study, we report the cloning and characterization of 7.5 kb of bovine genomic sequence homologous to exon 19 through 21 from EGF genes from other mammalian species. The cloned gene fragment had an unusual sequence composition in the form of an in-frame TGA codon in the coding sequence. The sequence was expressed at low levels in kidney tissue and the corresponding cDNA contained the TGA codon. The level of similarity between the bovine exonic sequence and the human, porcine, murine, feline, and canine corresponding sequences varied from 64% to 73%; however, when only sequences encoding the mature EGF protein were compared, the level of similarity between the bovine sequence and the sequence from these species was 59% to 66%. The sequence similarity of the deduced mature protein was lower (34% to 39%) than the sequence similarity of the deduced propeptide. Although the cloned sequences could originate from a bovine EGF pseudogene, the possibility exists that they originate from the functional EGF gene. An as yet unidentified mechanism to by-pass the stop codon would allow the synthesis of a functional EGF protein. Alternatively, the cloned sequence could originate from an EGF-like gene.     

Bayesian comparison of test-day models under different assumptions of heterogeneity for the residual variance: the change point technique versus arbitrary intervals

Test‐day milk yields from Spanish Holstein cows were analysed with two random regression models based on Legendre polynomials under two different assumptions of heterogeneity of residual variance which aim to describe the variability of temporary measurement errors along days in milk with a reduced number of parameters, such as (i) the change point identification technique with two unknown change points and (ii) using 10 arbitrary intervals of residual variance. Both implementations were based on a previous study where the trajectory of the residual variance was estimated using 30 intervals. The change point technique has been previously implemented in the analysis of the heterogeneity of the residual variance in the Spanish population, yet no comparisons with other methods have been reported so far. This study aims to compare the change point technique identification versus the use of arbitrary intervals as two possible techniques to deal with the characterization of the residual variance in random regression test‐day models. The Bayes factor and the cross‐validation predictive densities were employed for the model assessment. The two model‐selecting tools revealed a strong consistency between them. Both specifications for the residual variance were close to each other. The 10 intervals modelling showed a slightly better performance probably because the change point function overestimates the residual variance values at the very early lactation.     

FC-17 Cutaneous papillomatosis and carcinomatosis in the highly endangered Western Barred Bandicoot

Newcastle disease virus (NDV) has antineoplastic and immunostimulatory properties, and it is currently being clinically tested in anticancer therapy. In order to analyse the immunostimulatory effects of NDV on bovine papillomatosis, we inoculated 14 cows subcutaneously with an attenuated vaccine containing the LaSota strain of NDV (LS-NDV). Four cows with papillomatosis served as controls. Serum samples were collected from each animal 1 h preinoculation and 7 and 21 days postinoculation. In inoculated cows on days 7 and 21, the mean antibody titres were log₂ 2.43 ± 0.92 and 5.57 ± 0.72, respectively, by haemagglutination inhibition, and the mean levels of TNFα were 5.80 ± 4.19 and 5.39 ± 2.66 ng/mL, respectively, by WEHI-164 cytotoxicity assay. Significant differences between inoculated and control animals were evident for antibody titres on day 21 and clinical scores on day 60. A correlation was evident between the TNFα activities and clinical scores on day 21. The clinical observations at day 60 showed that the papillomas in five cows had completely resolved (36%), one animal had no alterations on clinical appearance of the tumour (7%), and papillomas in eight cows had regressed (57%). In conclusion, these results demonstrated that inoculation of LS-NDV vaccine stimulates an antibody response and a limited increase in TNFα activity and may enhance clinical recovery in bovine papillomatosis.
Funding: Scientific Research Council of Akdeniz University.