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  • Author or Editor: Joseph L. Dorner x
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SUMMARY

Corticosteroid-induced alkaline phosphatase (calp) and intestinal alkaline phosphatase (ialp) from dogs were purified to homogeneity, as determined by polyacrylamide gel electrophoresis. Purification involved an uninterrupted system using deae-cellulose, concanavalin Aagarose, and monoclonal antibody affinity columns. The monoclonal antibody was prepared by use of ialp as the antigen. The 2 isoenzymes were compared, using molecular weight determinations, amino acid analyses, peptide mapping, N-terminal sequencing of the first 10 amino acids, carbohydrate analyses, and recognition by anti-ialp)monoclonal antibody. The data indicated that canine ialp and (calp are identical with regard to recognition by monoclonal antibody and N-terminal amino acid sequence, nearly identical in amino acid content and peptide maps, but different in carbohydrate content. It was concluded that (calp is a product of the same gene as ialp and that differences in glycosyl transferase activities between liver and intestines or the presence of glycosidase activities in or around the intestinal mucosae result in the marked difference in carbohydrate content.

Free access
in American Journal of Veterinary Research

Abstract

Quantitative determination of the corticosteroid-induced isoenzyme of alkaline phosphatase (cap) was evaluated as a screening test for hyperadrenocorticism (hac) in dogs. A series of 40 dogs with hac (cap range, 96 to 14,872 U/L), 30 clinically normal dogs (cap range, 0 to 38 U/L), and 80 dogs with various diseases (non-hac) and without history of exogenous glucocorticoid exposure for a minimum of 60 days (cap range, 0 to 1163 U/L) were used to evaluate the test. Sensitivity and specificity of cap was calculated at various cutoff points for absolute cap activity and for cap activity expressed as a percentage of total alkaline phosphatase activity. A cutoff point of 90 U/L was selected as optimal for use of this assay as a screening test for hac. A prevalence survey then was done of all canine serum samples submitted to our diagnostic laboratory over a 3-month period, to calculate the predictive values of a positive and a negative test result in a clinical population and to determine the relative frequency and magnitude of cap activity in dogs that had received glucocorticoids. The predictive values of a positive and a negative test result at the 90 U/L cutoff value were 21.43% (95% confidence limits, 8.3 to 40.95%) and 100% (95% confidence limit > 96%), respectively. It was concluded that cap isoenzyme activity, determined by routine biochemical analysis by an automated levamisole-inhibition assay, could function as a screening test for hac, however, (the predictive value of a positive test result was too low to recommend the assay as a diagnostic test. The cap activity was > 90 U/L in 52% of dogs receiving glucocorticoids, emphasizing the importance of treatment history in the use of this test. To help eliminate possible workup bias, an additional set of serum samples from 30 dogs diagnosed as having hac at a second institution also were analyzed. Sensitivity of the assay in this sample group, using the 90 U/L cutoff value, was 83.3% (95% confidence limits, 65.3 to 94.4%), suggesting that test sensitivity may vary, depending on the institution in which the assay is used and the selection of cases for diagnostic workup.

Free access
in Journal of the American Veterinary Medical Association

SUMMARY

Assay procedures for determining serum haptoglobin concentration and ceruloplasmin oxidase activity in dogs were validated, and reference values were established. Serum haptoglobin concentration is reported as milligrams per deciliter of cyanmethemoglobin binding capacity, whereas serum ceruloplasmin oxidase activity was determined by use of p-phenylenediamine as substrate. Both assays were used to analyze serum samples from 288 dogs. In each dog’s case record, clinical history and final diagnosis were evaluated to determine whether the dog had an inflammatory condition. Complete blood cell counts were performed in 265 dogs, using simultaneously collected blood samples. Plasma fibrinogen concentration was determined for 161 dogs. A positive correlation (P < 0.01) was found for serum haptoglobin concentration and for ceruloplasmin oxidase activity, compared with wbc counts, segmented neutrophil and band neutrophil counts, and plasma fibrinogen concentration. Ceruloplasmin oxidase activity and haptoglobin concentration were up to 6 times more sensitive than fibrinogen concentration or leukocyte counts in detecting inflammation. Specificity of ceruloplasmin oxidase activity was comparable to fibrinogen concentration and leukocyte counts, whereas haptoglobin concentration was found to be slightly less specific. Specificity of haptoglobin concentration improved slightly (from 0.82 to 0.88) when dogs with a history of glucocorticoid administration were excluded from analysis. Predictive value of a negative test result (haptoglobin concentration < 125 mg/dl; ceruloplasmin oxidase activity < 20 IU/L) and predictive value of a positive test result for haptoglobin concentration and ceruloplasmin activity were comparable to or better than fibrinogen concentration or various oxidase leukocyte counts in detection of inflammation in a variety of disease conditions. We concluded that serum haptoglobin and ceruloplasmin oxidase assays could be used as adjuncts for diagnosis of the inflammation in dogs.

Free access
in American Journal of Veterinary Research