Search Results

You are looking at 1 - 2 of 2 items for

  • Author or Editor: Andrew D. Woolcock x
  • Refine by Access: All Content x
Clear All Modify Search

Abstract

OBJECTIVE To evaluate use of cortisol concentration prior to ACTH stimulation (baseline) to monitor efficacy of twice-daily administration of trilostane to dogs with pituitary-dependent hyperadrenocorticism (PDH).

DESIGN Retrospective case series.

ANIMALS 22 dogs with PDH.

PROCEDURES The database of a veterinary hospital was searched to identify dogs with PDH that were treated with the FDA-approved veterinary formulation of trilostane twice daily between January 1, 2008, and December 31, 2012. For each dog, signalment and details regarding each hospital visit including comorbidities, electrolyte concentrations, and clinical signs were extracted from the record. For each ACTH stimulation test performed, the respective correlations between baseline cortisol concentration and the cortisol concentration after ACTH stimulation (ACTH-stimulated cortisol concentration) and resultant decision regarding trilostane dose adjustment were determined. Excessive suppression of cortisol production was defined as an ACTH-stimulated cortisol concentration < 2.0 μg/dL. The ability of various baseline cortisol concentrations to predict whether a dog had excessive suppression of cortisol production was determined.

RESULTS 109 ACTH stimulation tests were performed for the 22 dogs. A baseline cortisol concentration > 3.2 μg/dL predicted that ACTH-stimulated cortisol concentration would be ≥ 2.0 μg/dL with 100% certainty; however, 14 of 64 tests with a baseline cortisol concentration > 3.2 μg/dL had an ACTH-stimulated cortisol concentration ≤ 3.2 μg/dL, which was suggestive of inadequate adrenocortical cortisol reserves.

CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that baseline cortisol concentration should not be used as the sole monitoring tool for management of dogs with PDH treated with trilostane twice daily. (J Am Vet Med Assoc 2016;248:814–821)

Restricted access
in Journal of the American Veterinary Medical Association

Abstract

OBJECTIVE

To validate the use of a flow cytometric assay that uses 2‘,7‘-dichlorodihydrofluorescein diacetate (DCFH-DA) to measure reactive oxygen species in the erythrocytes of healthy dogs.

ANIMALS

50 healthy adult dogs.

PROCEDURES

Erythrocytes were incubated with DCFH-DA or a vehicle control (dimethyl sulfoxide), then incubated with (stimulated) or without (unstimulated) hydrogen peroxide. The flow cytometric assay was evaluated for specificity with increasing concentrations of DCFH-DA and hydrogen peroxide, and a polynomial regression line was applied to determine optimal concentrations. For precision, samples were analyzed 5 consecutive times for determination of intra- and interassay variability. Stability of samples stored at 4°C for up to 48 hours after blood collection was determined with flow cytometric analysis. Coefficient of variation (CV) was considered acceptable at 20%. Baseline measurements were used to determine an expected range of median fluorescence intensity for unstimulated erythrocytes incubated with DCFH-DA.

RESULTS

Erythrocytes were successfully isolated, and stimulated samples demonstrated higher median fluorescence intensity, compared with unstimulated samples. The intra-assay CV was 11.9% and 8.9% and interassay CV was 11.9% and 9.1% for unstimulated and stimulated samples, respectively. Unstimulated samples were stable for up to 24 hours, whereas stimulated samples were stable for up to 48 hours.

CONCLUSIONS AND CLINICAL RELEVANCE

Flow cytometry for the measurement of reactive oxygen species in the erythrocytes of healthy dogs by use of DCFH-DA had acceptable specificity, precision, and stability. Flow cytometry is a promising technique for evaluating intraerythrocytic oxidative stress for healthy dogs.

Restricted access
in American Journal of Veterinary Research