Use of the ADVIA 120 for differentiating extracellular and intracellular hemoglobin.

BACKGROUND: The use of cell-free hemoglobin (Hgb) solutions, such as Oxyglobin (Biopure Corp, Cambridge, MA, USA), as a blood substitute for the treatment of acute anemias is increasing in veterinary medicine. These solutions interfere with colorimetric tests, which do not discriminate between cellular Hgb (Hgb-cell) from the patient and extracellular Hgb (Hgb-delta) from the Oxyglobin, and therefore make the monitoring of anemia, based on Hgb concentration, difficult. The ADVIA 120 hematology analyzer (Bayer Diagnostics, Tarrytown, NY, USA) evaluates Hgb by 2 methods, a standard cyanmethemoglobin colorimetric method and flow cytometry, and therefore might provide the means to differentiate extracellular and intracellular Hgb. OBJECTIVE: The objective of this study was to determine the accuracy and precision of the ADVIA 120 in differentiating extracellular from intracellular Hgb. METHODS: Anticoagulated whole blood samples from 10 healthy dogs were analyzed in triplicate on the ADVIA 120. Hgb-delta concentration was determined by adding Oxyglobin (13 g/dL) to the whole blood samples at dilutions of 1:1, 1:2, 1:4, 1:8, 1:16, and 1:32. Hgb-cell and Hgb-total values were calculated and compared with actual values by linear regression. Analyses were done in triplicate and repeated 9 consecutive times to evaluate intra-assay precision of Hgb-total and Hgb-cell determinations. RESULTS: Correlation between Hgb values obtained by colorimetric (Hb-total) and flow cytometric (Hgb-cell) methods on whole blood samples was high (R(2) = .99; n = 10) with a slope of 0.96 and intercept of 0. Correlation between actual and predicted Hgb-cell values also was high (R(2) = .99), with a small positive bias (0.289 +/- 0.185; n = 60). Intra-assay precisions were high, with most coefficients of variation <2%. CONCLUSION: The ADVIA 120 is capable of differentiating Hgb-cell from Hgb-delta. The flow cytometric method is accurate and precise when compared with the cyanmethemoglobin method. A small bias between the results is unlikely to be clinically significant but may affect the ability of the ADVIA to differentiate small quantities (<0.3 g/dL) of Hgb-delta.

Validation and diagnostic efficacy of a lipase assay using the substrate 1,2-o-dilauryl-rac-glycero glutaric acid-(6' methyl resorufin)-ester for the diagnosis of acute pancreatitis in dogs.

BACKGROUND: Increased serum lipase activity has been used historically to support the diagnosis of acute pancreatitis, a common disease in dogs. Most of the lipase assays that are currently in use lack optimum sensitivity and specificity. OBJECTIVE: The objectives of this study were to 1) validate the 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6'-methylresorufin) ester (DGGR) assay for determination of lipase activity in canine serum and 2) compare results, reference intervals, sensitivity, and specificity of the DGGR assay with a standard 1,2-diglyceride (1,2 DiG) assay for diagnosing acute pancreatitis in dogs. METHODS: Precision, linearity, and interference studies were performed for method validation on a Hitachi 911 analyzer. Lipase results from the DGGR and 1,2 DiG assays were compared by linear regression analysis. Sensitivity, specificity, and diagnostic efficacy were determined for both assays on a population of 30 dogs, 15 of which had acute pancreatitis based on history, clinical signs, and ultrasound findings. RESULTS: Within-run and within-day coefficients of variation (CVs) were low (<3%), with higher day-to-day CVs (< or =14 %). The assay was linear between 8 and 2792 U/L. No significant interference by hemolysis and lipemia was found. Poor correlation was found between the assays (r(s)=0.84). The lipase reference interval was 8-120 U/L for the DGGR assay and 30-699 U/L for the 1,2 DiG assay. Sensitivity and specificity for the diagnosis of pancreatitis were 93% and 53%, respectively, for the DGGR assay and 60% and 73% for the 1,2 DiG assay. Receiver operating characteristic curve analysis showed similar areas under the curve. CONCLUSIONS: On the basis of this study, the DGGR method is considered adequate for assaying serum lipase activity in dogs. The high sensitivity of the DGGR assay suggests it may be a useful screening test for canine pancreatitis.

Biochemical analysis of neoplastic versus nonneoplastic abdominal effusions in dogs.

This prospective study of 15 dogs evaluated biochemical parameters in abdominal effusions of neoplastic and nonneoplastic origin in an attempt to identify markers for malignant effusions. Dogs in the neoplastic group had statistically significant lower glucose concentrations (mean, 72.6 mg/dL versus 110.0 mg/dL; P=0.0431) and higher lactate levels (mean, 3.81 mmol/L versus 1.68 mmol/L; P=0.0377) in their abdominal fluid than did dogs in the nonneoplastic group, indicating that low glucose and high lactate in abdominal effusions may be markers for neoplasia.

Immune-mediated hemolytic anemia: understanding the nemesis.

IMHA is one of the most common causes of anemia in small animals. Although treatment may be rewarding, many patients do not respond adequately to glucocorticoids alone and require additional immunosuppressive therapy. Some patients may succumb to acute severe anemia and die within the first few weeks of treatment; even if they survive, relapses may occur. IMHA is the nemesis; as our understanding of this disease increases and treatment options expand, it is hoped that survival rates will finally improve.

Calculation of the total plasma concentration of nonvolatile weak acids and the effective dissociation constant of nonvolatile buffers in plasma for use in the strong ion approach to acid-base balance in cats.

OBJECTIVE: To determine values for the total concentration of nonvolatile weak acids (Atot) and effective dissociation constant of nonvolatile weak acids (Ka) in plasma of cats. SAMPLE POPULATION: Convenience plasma samples of 5 male and 5 female healthy adult cats. PROCEDURE: Cats were sedated, and 20 mL of blood was obtained from the jugular vein. Plasma was tonometered at 37 degrees C to systematically vary PCO2 from 8 to 156 mm Hg, thereby altering plasma pH from 6.90 to 7.97. Plasma pH, PCO2, and concentrations of quantitatively important strong cations (Na+, K+, and Ca2+), strong anions (Cl-, lactate), and buffer ions (total protein, albumin, and phosphate) were determined. Strong ion difference was estimated from the measured strong ion concentrations and nonlinear regression used to calculate Atot and Ka from the measured pH and PCO2 and estimated strong ion difference. RESULTS: Mean (+/- SD) values were as follows: Atot = 24.3 +/- 4.6 mmol/L (equivalent to 0.35 mmol/g of protein or 0.76 mmol/g of albumin); Ka = 0.67 +/- 0.40 x 10(-7); and the negative logarithm (base 10) of Ka (pKa) = 7.17. At 37 degrees C, pH of 7.35, and a partial pressure of CO2 (PCO2) of 30 mm Hg, the calculated venous strong ion difference was 30 mEq/L. CONCLUSIONS AND CLINICAL RELEVANCE: These results indicate that at a plasma pH of 7.35, a 1 mEq/L decrease in strong ion difference will decrease pH by 0.020, a 1 mm Hg decrease in PCO2 will increase plasma pH by 0.011, and a 1 g/dL decrease in albumin concentration will increase plasma pH by 0.093.

Racing Alaskan sled dogs as a model of "ski asthma".

Athletes who play sports in cold weather, particularly skaters and cross-country skiers, have an increased prevalence of lower airway disease that is hypothesized to result from repeated penetration of incompletely conditioned air into the lung periphery. In this study, we investigated the hypothesis that canine winter athletes also suffer from increased prevalence of lung disease secondary to hyperpnea with cold air. Bronchoscopy and bronchoalveolar lavage was conducted in elite racing sled dogs 24 to 48 hours after completion of a 1,100-mile endurance race. Bronchoscopic abnormalities were classified as none, mild, moderate, or severe, based on the quantity and distribution of intralumenal debris. Eighty-one percent of the dogs (48 of 59) examined had abnormal accumulations of intralumenal debris, with 46% (27 of 59) classified as moderate or severe, indicating significant accumulation of exudate. Bronchoalveolar lavage obtained from dogs after the race had significantly higher nucleated macrophage and eosinophil counts compared with sedentary control dogs. Our findings support the hypothesis that strenuous exercise in cold environments can lead to lower airway disease and suggest that racing sled dogs may be a useful naturally occurring animal model of the analogous human disease.