Nontraditional applications in clinical pathology.

Most published reviews of preclinical toxicological clinical pathology focus on the fundamental aspects of hematology, clinical chemistry, coagulation, and urinalysis in routine toxicology animal species, for example, rats, mice, dogs, and nonhuman primates. The objective of this continuing education course was to present and discuss contemporary examples of nonroutine applications of clinical pathology endpoints used in the drug development setting. Area experts discussed bone turnover markers of laboratory animal species, clinical pathology of pregnant and growing laboratory animals, clinical pathology of nonroutine laboratory animal species, and unique applications of the Siemens Advia(®) hematology analyzer. This article is a summary based on a presentation given at the 31st Annual Symposium of the Society of Toxicologic Pathology, during the Continuing Education Course titled "Nontraditional Applications of Clinical Pathology in Drug Discovery and Preclinical Toxicology."

Enumeration of nucleated red blood cells with the ADVIA 2120 Hematology System: an International Multicenter Clinical Trial.

The accurate, reproducible, and timely reporting of nucleated red blood cells (NRBC) is an important function of the clinical hematology laboratory. We used 960 samples from 5 worldwide sites to evaluate a new NRBC enumeration method for the ADVIA 2120 Hematology System. The method showed excellent correlation with microscopy (r = 0.93). Sensitivity and specificity for the presence of NRBC for all samples analyzed was 77.3% and 74.6%, respectively. Almost all false negative samples were at NRBC counts 10/100 WBC. The NRBC method automatically corrects WBC counts and differential results for the presence of NRBC; the uncorrected counts are available to the user on the run screen. This method allows the enumeration of NRBC with the ADVIA 2120 Hematology System in CBC/DIFF sample mode without need for additional hardware, sample preparation, or reagents.

Prognostic relevance of large-platelet counts in patients with immune thrombocytopenic purpura.

In this preliminary study, the value of different platelet parameters, measured by the ADVIA120 Analyzer, in predicting the immediate response to intravenous immunoglobulin or intravenous anti-RhoD was assessed in 31 patients with immune thrombocytopenic purpura. The number of large platelets pre-treatment was the only independent predictor of the 24 hour-platelet increase.

A rapid, automated flow cytometric method to measure activated degranulated platelets by density determination.

Platelet activation is reported to correlate with acute coronary syndromes. A platelet analysis method on the ADVIA 120 Hematology System provides rapid analysis of platelet density, reported as mean platelet component (MPC) concentration, utilizes routine hematology specimens, requires no pre-treatment, and thirty seconds to generate results. Sub-populations of platelets separated by density gradients showed excellent correlation with the ADVIA 120 MPC parameter (r = 0.997). Platelet activation induced by thrombin treatment resulted in a shift of platelets into the lowest density fraction (d

Evaluation of the anticoagulants EDTA and citrate, theophylline, adenosine, and dipyridamole (CTAD) for assessing platelet activation on the ADVIA 120 hematology system.

BACKGROUND: Monitoring of platelet activation by the ADVIA 120 Hematology System requires an anticoagulant and protocol that ensures that platelets are sphered and their activation status is not altered artifactually in vitro. METHODS: Blood from healthy controls was collected into tripotassium EDTA; citrate, theophylline, adenosine, and dipyridamole (CTAD); or a combination of both (E/C) and stored at ambient temperature or at 4 degrees C (E/C only) and then analyzed between 0 and 180 min later on the ADVIA 120. In addition, immunofluorescent flow cytometry was used to identify activated platelets and platelet-leukocyte aggregates. RESULTS: In blood stored with all three anticoagulants, the platelet count changed little, but the mean platelet volume (MPV) at first decreased and then increased, whereas the mean platelet component (MPC; an indicator of activation) changed in a reciprocal manner. The changes in MPV and MPC, which reflect platelet sphering and swelling, were greatest between 30 and 60 min in blood stored at ambient temperature, irrespective of which anticoagulant was used, and between 60 and 180 min when blood anticoagulated with E/C was stored at 4 degrees C. In all anticoagulants, the percentages of platelets expressing CD62P and of leukocytes in platelet-leukocyte aggregates increased significantly (P <0.01) over 180 min at ambient temperature. Only minimal (<2%) increases occurred when blood with E/C was stored at 4 degrees C. CONCLUSIONS: When determining platelet activation ex vivo on the ADVIA 120, blood should be collected into E/C, stored at 4 degrees C, and analyzed between 60 and 180 min later; these conditions ensure maximum platelet sphering without concurrent artifactual platelet activation.

Automated analysis of feline platelets in whole blood, including platelet count, mean platelet volume, and activation state.

A new whole-blood flow cytometric method has been developed for counting and sizing platelets in samples from cats, a species in which platelet and red blood cell sizes overlap significantly. The method is a modified version of the two-angle laser light scattering technology used by Bayer H*System hematology analyzers. The new method provided accurate platelet counts and mean platelet volumes (MPV, fl) for cats. The method also measured mean platelet component concentration (MPC, g/dl), a parameter which was shown to be sensitive to platelet activation state, and which decreased in value as activation progressed.