Characterisation of monoclonal antibodies to ovine interleukin-6 and the development of a sensitive capture ELISA

A purified recombinant ovine (rOv) interleukin-6 (IL-6) was used to generate specific murine monoclonal antibodies (mAbs) and a polyclonal rabbit antisera to this cytokine. From the 31 initial hybridoma cell lines generated, three stable clones were established which secreted mAbs to rOvIL-6, as judged by a direct enzyme-linked immunosorbent assay (ELISA) and Western blotting. Their specificity was further confirmed by demonstrating that none of the mAbs recognised any of the six other irrelevant recombinant ovine cytokines tested by direct ELISA. All three mAbs displayed cross-reactivity with human and African green monkey IL-6 as demonstrated by direct ELISA and Western blotting. In contrast, the polyclonal antibodies only cross-reacted with bovine IL-6 and not with either of the human or monkey homologues. By combining a mAb with the polyclonal antisera a sensitive, IL-6-specific, capture ELISA was developed that had a sensitivity of 150 pg/ml. This detection system was unequivocally validated by demonstrating that native OvIL-6 could be detected in efferent lymph draining from a stimulated popliteal lymph node. In addition, one of the mAbs was shown to allow the detection of OvIL-6 by intracellular cytokine staining and flow cytometry.     

Challenges for Multilevel Stakeholder Engagement in Public Trust Resource Governance

The trend in wildlife management over the last two decades has been to develop locally based approaches for responsiveness to local conditions, but some state wildlife agencies are finding the amount of staff time required to service this approach prohibitive. Although local engagement strategies have been lauded as assuring that public trust obligations of state government to citizens are met, we can expect that states with a local focus as their operational level of stakeholder engagement may opt to change their approach to reflect their resource limitations. We argue for comprehensive regional level effort to understand stakeholders augmented with local engagement processes where needed to deal with special circumstances in smaller areas within a region. Such an approach can be anticipated to have implications for stakeholder engagement and human dimensions research needs, which we discuss in the context of public trust resource administration and good governance of wildlife resources.     

Evaluation of microwave-thawed canine plasma for transfusion

Units of fresh-frozen canine plasma were thawed rapidly in a microwave oven, using a mean of 15.4 thawing periods of 10 seconds each. The activated partial thromboplastin times, the one-stage prothrombin times, concentrations of fibrinogen, factor-VIII coagulant activity, and von Willebrand factor antigen of microwave-thawed units were not significantly different from those measured in small aliquots of the same units thawed in a warm water bath (n = 5). Five dogs were given a unit of their own microwave-thawed plasma. During the 24 hours after infusion, no significant changes were measured in temperature, heart rate, or respiration rate. In addition, significant changes in PCV, total protein concentrations, estimates of platelet numbers, total RBC counts, total WBC counts, and differential WBC counts were not measured in blood specimens obtained before infusion and 24 hours after the infusion of microwave-thawed plasma.     

Pseudogout in two dogs

Extract

Pseudogout is a common condition in elderly people in which sudden onset of intense joint pain and synovitis develop. Peri-synovial calcium crystal deposition is the cause of the synovitis rather than the uric acid deposition in common gout. It is a rare condition in dogs, having only been reported twice before. Two dogs presenting with acute lameness and pyrexia showed many macrophages and occasional neutrophils containing intracellular crystals on cytological examination of synovial fluid. The crystals were single or multiple, small to large, clear, square or rhomboid-shaped, weakly birefringent and resembled those described for pseudogout. Occasional crystals were also present free in the matrix. Non-steroidal anti-inflammatory drugs that are effective in reducing pain in people with pseudogout provided no relief to either patient, but twice daily oral prednisolone therapy was successful. Dosage was gradually tapered as clinical signs improved over a period of 6 weeks.     

Plasma cortisol responses to three cortico-trophic preparations in normal dogs

Objective To compare cortisol responses to three corticotrophic preparations in normal dogs.
Animals Eight clinically normal dogs (four intact males, four intact females) of medium size.
Procedures Each dog received four treatments on four separate occasions in a duplicated Latin square pattern. Treatments were two adrenocorticotrophin (ACTH) preparations given intramuscularly at 2.2 U/kg, one of the ACTH preparations given intramuscularly at 1 U/kg and a synthetic polypeptide with ACTH-like activity (tetracosactrin, cosyntropin) given intravenously at 5 μg/kg. Plasma samples were taken for cortisol assay before and at 0.5, 1, 2 and 4 h after treatment.
Results Plasma cortisol concentrations were similar with the two ACTH preparations and at both dose rates. Tetracosactrin produced smaller mean peak cortisol concentrations, which tended to occur earlier than with ACTH, and smaller values for the area under the curve of plasma cortisol concentration from zero time to 4 h.
Conclusion The findings suggest that canine adrenal function can be tested adequately by giving ACTH intramuscularly at 1 U/kg and measuring plasma cortisol in samples taken at 0 and 2 h, or by giving tetracosactrin intravenously at 5 μg/kg and determining cortisol concentration at 0 and 1 h.     

Possible parachordoma in a dog

Abstract

CASE HISTORY: A 15-mm diameter mass developed in the S/C tissue overlying the right lateral gluteal muscle of a 6½-year-old female Flat-coated Retriever.

PATHOLOGICAL FINDINGS: Cytological preparations following aspiration of the mass were highly cellular and consisted of a population of large polygonal cells containing single to multiple nuclei, large prominent nucleoli, and intracytoplasmic vacuoles. Histologically, the neoplasm consisted of similar large cells surrounded by thick fibrous connective tissue trabeculae. The large polygonal cells reacted positively with antibodies against vimentin, low- and high-molecular-weight variants of cytokeratin (AE1/AE3), but not with antibodies to desmin or glial fibrillary acidic protein (GFAP).

DIAGNOSIS: The clinical, gross, histological and immunohistochemical findings are similar to those reported for parachordomas in humans. Neither recurrence nor metastases were noted 18 months after surgical excision of the mass.

CLINICAL RELEVANCE: This is the first reported case of a possible parachordoma in a dog, a benign tumour with cytological features of malignancy.     

Sulfhydryl groups and estradiol-receptor interaction

The characteristic ability of rat uteri to take up tritiated estradiol in vitro or to retain estradiol previously incorporated either in vivo or in vitro is destroyed by treating the tissue with various sulfhydryl-blocking reagents. The two radioactive estradiol-receptor complexes, observed in uterine homogenates in the supernatant fraction and in an extract of the nuclear fraction, respectively, are disrupted by brief exposure to organic mercurials in the cold. Sulfhydryl groups of uterine receptor substances apparently play a vital role in estradiol binding, perhaps indirectly through contribution to receptor conformation.