Serum 5'nucleotidase activity in rats: a method for automated analysis and criteria for interpretation

A manual kit for determining serum 5'nucleotidase (5'NT, EC 3.1.3.5) activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within-run C.V. = 2.14%; between-run C.V. = 5.5%). A comparison of the new automated method with a manual and semi-automated method gave regression statistics of y = 1.18X -3.66 (Sy. x = 4.54), and y = 0.733X + 1.97 (Sy. x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9-week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.     

Using the correct regression analysis technique in method comparison studies

Regression analysis frequently is used to evaluate a new clinical laboratory method. Results from the new method are compared to results from an existing more established method. If measurement error exists in the established method, then least squares may not be an appropriate statistical method to use for the regression analysis. An errors variable regression analysis model was used to evaluate data from five method comparison studies. Results were compared to least squares analyses performed on the same data. When significant measurement error existed in the "reference" method, the errors variable analysis provided a less biased estimate of the regression statistics, which differed markedly from the least squares results. Inappropriate use of least squares in method comparison studies can lead to erroneous conclusions about the relationship of a new analytical method to an existing method.     

Contribution of whole blood L-lactate, pyruvate, D-lactate, acetoacetate, and 3-hydroxybutyrate concentrations to the plasma anion gap in horses with intestinal disorders

Increased anion gap (AG) was due, in part, to L-lactic acidosis in 14 of 14 horses with intestinal disorders. In a few horses, increased whole blood concentrations of D-lactate made a minor contribution to the AG. However, the increase in AG was often greater than the sum of the increases in these 2 acid anions. This unexplained increase was not a result of increases in whole blood pyruvate, 3-hydroxybutyrate, or acetoacetate concentrations or serum albumin or phosphate concentrations. Identification of other anions causing increased AG could lead to better understanding, diagnosis, and treatment of metabolic imbalances in critically ill horses.     

EFFECTS OF METRIZAMIDE MYELOGRAPHY ON CEREBROSPINAL FLUID ANALYSIS IN THE DOG

The effect of metrizamide myelography on the composition of cerebrospinal fluid (CSF) was studied. Seven dogs received an intracisternal injection of metrizamide. An intracisternal puncture was performed in three additional dogs that did not receive metrizamide. CSF was collected before myelography and 24, 72, and 144 hours after myelography from all dogs. A significant increase in the percentage of neutrophils (p<0.05) and a pleocytosis noted 24 hours after myelography (p<0.02) were attributed to the effect of metrizamide. Significant increases in total protein concentration (p<0.001) and erythrocyte count (p<0.05), and a decrease in the percentage of small mononuclear cells (p<0.01) were attributed to repeated intracisternal puncture. No significant changes were observed for CSF creatine phosphokinase activity or the percentage of large mononuclear cells.     

Effect of EDTA on morphology of neutrophils of healthy dogs and dogs with inflammation

Increased basophilia and foamy vacuolation in the cytoplasm of neutrophils in smears made within 1 hour of blood collection suggest inflammatory disease. Due to the mild increase in foamy vacuolation which occurred with time in EDTA, the significance of mildly vacuolated neutrophils in smears prepared a few hours after blood collection is questionable. However, moderate or severe foamy vacuolation should be considered clinically significant. In vitro morphologic changes in neutrophils from healthy dogs in EDTA include clear, discrete vacuoles in the cytoplasm, uneven distribution of cytoplasmic granules, irregular cell membrane and pyknosis. These neutrophils generally lack basophilia, have minimal foamy vacuolation of the cytoplasm and are morphologically different from neutrophils associated with severe inflammation.