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Biomedical subjects

K Jane Wardrop

Publications and source records attributed to K Jane Wardrop.

10 recordsLinked to original sources

Detection of activated platelets in canine blood by use of flow cytometry.

OBJECTIVE: To evaluate whether markers of platelet activation, including P-selectin expression, phosphatidylserine exposure, platelet-leukocyte aggregates, and microparticle formation, could be measured in nonstimulated and stimulated canine blood samples and develop a standardized protocol for detection of activated platelet markers in canine blood. SAMPLE POPULATION: Blood samples from 10 dogs. PROCEDURE: Platelet activation was determined by flow cytometric measurement of platelets with P-selectin expression, platelet-leukocyte aggregates, platelet microparticles, and platelets with phosphatidylserine exposure. Changes in specific markers of platelet activation in nonstimulated versus stimulated samples were assessed by use of varying concentrations of 2 platelet agonists, platelet-activating factor (PAF) and adenosine diphosphate. Flow cytometry was used to detect platelet CD61 (glycoprotein IIIa), CD62P (P-selectin), and the leukocyte marker CD45. Annexin V was used to identify exposed phosphatidylserine. RESULTS: A significant difference was detected in the percentages of platelets with P-selectin, plateletleukocyte aggregates, microparticles, and platelets with annexin V exposure (phosphatidylserine) in samples stimulated with 10nM PAF versus the nonstimulated samples, with platelet-leukocyte aggregates having the greatest difference. CONCLUSIONS AND CLINICAL RELEVANCE: Platelet activation is essential for thrombus formation and hemostasis and may be potentially useful for evaluation of dogs with suspected thromboembolic disease. Prior to development of a thrombotic state, a prothrombotic state may exist in which only a small number of platelets is activated. Identification of a prothrombotic state by use of activated platelets may help direct medical intervention to prevent a thromboembolic episode.

Analysis of Variance↗

The Coombs' test in veterinary medicine: past, present, future.

The Coombs' test, also known as the antiglobulin test, is used most frequently in veterinary medicine as an aid in the diagnosis of immune-mediated hemolytic anemia. The test also is used widely in human medicine to screen for red blood cell alloantibodies. Polyspecific reagents for veterinary use typically contain anti-IgG, anti-IgM, and anti-C3. Monospecific reagents also are available. False-positive and false-negative test results can be obtained. Inadequate sensitivity of the standard test in human and veterinary medicine has necessitated development of alternate, more sensitive technologies.

Anemia, Hemolytic↗

Measurement of the activation of equine platelets by use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody.

OBJECTIVE: To investigate the potential use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody for detection of the activation of equine platelets by use of flow cytometry. SAMPLE POPULATION: Platelets obtained from 6 Thoroughbreds. PROCEDURE: Flow cytometry was used to assess platelet activation as indicated by detection of binding of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-thrombospondin antibody to unactivated and ADP-, collagen-, platelet activating factor (PAF)-, and A23187-activated equine platelets. Human platelets were used as control samples. Determination of 14C-serotonin uptake and release was used to assess the extent of platelet secretion. RESULTS: Anti-human thrombospondin antibody failed to bind to equine platelets. Annexin V bound to platelets activated with PAF or A23187 when platelets had undergone secretion. Anti-human fibrinogen antibody bound to ADP-, PAF-, and A23817-activated platelets, but binding was not dependent on platelet secretion. The extent of binding of anti-fibrinogen antibody was less in equine platelets, compared with that for human platelets, despite maximal stimulation. CONCLUSIONS AND CLINICAL RELEVANCE: Activation of equine platelets can be detected by use of fluorescent-labeled annexin V and anti-human fibrinogen antibody but not by use of anti-human thrombospondin antibody. These flow cytometric techniques have the potential for detection of in vivo platelet activation in horses at risk of developing thrombotic disorders.

Animals↗

Effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation.

OBJECTIVE: To investigate the effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation. SAMPLE POPULATION: Blood samples from 6 Thoroughbreds. PROCEDURE: The degree of fluorescence associated with binding of fluorescein isothiocyanate (FITC)-conjugated anti-human fibrinogen antibody and FITC-annexin V in unactivated and adenosine diphosphate (ADP)-, platelet activating factor (PAF)-, and A23187-activated platelet samples in unfixed and 0.5, 1.0, and 2.0% formaldehyde-fixed samples was assessed by use of flow cytometry. RESULTS: In samples incubated with FITC-anti-human fibrinogen antibody prior to fixation, addition of 2.0% formaldehyde resulted in a 30% increase in total fluorescence in ADP- and PAF-activated samples and a 60% increase in A23187-activated samples. Fixation for 24 hours prior to addition of antibody resulted in reduced fluorescence of samples containing antihuman fibrinogen antibody for all 3 concentrations of formaldehyde in PAF-activated samples. The addition of all 3 concentrations of formaldehyde after incubation with FITC-annexin V resulted in significant increases in fluorescence in unactivated and activated platelet samples. As length of fixation time increased, there was a gradual increase in fluorescence that was significant at 24 hours. CONCLUSIONS: Because fixation with 2.0% formaldehyde results in significant changes in fluorescence in activated platelet samples containing anti-fibrinogen antibody, lower concentrations of formaldehyde should be used to fix equine platelet samples. Formaldehyde-fixed platelet samples should be analyzed within 12 hours of fixation to avoid artifactual increases in fluorescence. Fixation of samples containing FITC-annexin V should be avoided because of significant increases in fluorescence that may interfere with interpretation of results.

Adenosine Diphosphate↗

Effects of a polymerized ultrapurified bovine hemoglobin blood substitute administered to ponies with normovolemic anemia.

The development of ultrapurified hemoglobin-based oxygen carriers has eliminated many problems associated with whole-blood transfusions in other species. We hypothesized that the administration of polymerized ultrapurified bovine hemoglobin (PUBH) would result in improved hemodynamic parameters in ponies with normovolemic anemia without adverse effects on renal function or coagulation times. Normovolemic anemia was induced in 6 healthy adult ponies. Over a 3-day period, at least 45 mL/kg of whole blood was withdrawn from each pony until a target PCV of <12% was attained. Plasma was separated from the red blood cells via centrifugation and readministered to the ponies on each day. After the final plasma transfusion, 15 mL/kg of hetastarch (control, n = 6) or 15 mL/kg of PUBH (treatment, n = 6) was administered at 10 mL/kg/h IV. Administration of PUBH at a rate of 10 mL/kg/h was not associated with any adverse effects in 5 of the 6 ponies. One pony experienced an anaphylactoid reaction during infusion of PUBH. The reaction, characterized by intense pruritus, tachycardia, and tachypnea resolved shortly after stopping the infusion. Ponies receiving PUBH had significantly lower cardiac indices (P = .03) and heart rates (P = .002) than control animals. A significantly greater increase in central venous pressure was observed in the PUBH group compared to the hetastarch group (P = .02). No adverse renal or coagulation effects were observed with PUBH infusion. These results suggest that PUBH improves hemodynamics and oxygen transport parameters in horses experiencing normovolemic anemia. Patients should be monitored closely during infusion for any adverse reactions.

Anemia↗

Systemic Mycobacterium avium infection in a dog diagnosed by polymerase chain reaction analysis of buffy coat.

Dogs may be infected by Mycobacterium (M.) tuberculosis, M. bovis, and M. avium complex, and the clinical signs associated with each of these infections may be indistinguishable. Rapid speciation of the infecting organism is desirable because of the public health concerns associated with M. bovis and M. tuberculosis infections. A mycobacterial infection was suspected in the dog of this report based on acid-fast staining of organisms in macrophages obtained from liver aspirates and buffy-coat preparations. Polymerase chain reaction (PCR) analysis of a buffy-coat preparation identified M. avium.

Animals↗

Canine and feline blood donor screening for infectious disease.

Thousands of blood transfusions are performed each year on dogs and cats, and the demand for blood products continues to grow. Risks associated with transfusions include the risk of disease transmission. Appropriate screening of blood donors for bloodborne infectious disease agents should be performed to lessen this risk. Geographic restrictions of disease, breed predilection, and documentation of actual disease transmission by transfusion all are factors that might need to be considered when making a decision on what screening program to use. In addition, factors involving general health care and management of blood donors should be employed to further ensure blood safety.

Animals↗