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Platelet-poor plasma PRP Platelet-rich plasma a. SysLoc safety A.V. fistula needle set, JMS Singapore PTE Ltd, Singapore, Singapore. b. ACD-A, Baxter-Fenwal, Deerfield, Ill. c. Cell-Dyn 3700, Abbott Diagnostics, Abbott Park, Ill. d. Scil Vet

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in American Journal of Veterinary Research

venipuncture and immediately transferred into a commercial specimen tube e containing acid-citrate-dextrose to achieve a 1:7 acid-citrate-dextrose–to–blood ratio. Platelet-rich plasma as well as platelet-rich plasma with 6% DMSO was prepared for subsequent

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in American Journal of Veterinary Research

SUMMARY

Ten adult dogs (5 Beagles and 5 mixed-breed dogs) were inoculated IV with canine platelets containing Ehrlichia platys. Inclusions and morulae of E platys developed in platelets of infected dogs at 10 to 14 days after inoculation, followed by marked thrombocytopenia at 14 to 21 days. Parasitemia and marked thrombocytopenia recurred at 24 to 28 days after inoculation. Increased numbers of megakaryocytes were observed in marrow aspirate smears from infected dogs, indicative of regenerative thrombocytopenia. Prior to infection, platelet-rich plasma from these dogs was determined to have similar aggregatory response to arachidonate. After infection with E platys, the aggregatory response of platelet-rich plasma to collagen or 3 dilutions of adenosine diphosphate was evaluated. A statistically significant (P < 0.05) inhibition of platelet aggregatory response to the lowest dilution of adenosine diphosphate was detected for mixed-breed dogs, whereas aggregation responses were unchanged in Beagles. Results indicate that platelet activation may occur in dogs with acute ehrlichial infection.

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in American Journal of Veterinary Research

Abstract

Objective

To evaluate a method for detecting thiazole orange-positive (TO+, reticulated) platelets in equine blood, using flow cytometry.

Animals

16 healthy, equine infectious anemia virus (EIAV)-negative horses and ponies; 9 thrombocytopenic, ElAV-positive horses and ponies; and 2 thrombocytopenic, ElAV-negative horses.

Procedure

Blood from healthy and thrombocytopenic horses was collected by jugular venipuncture. Appropriate sample requirement and incubation time for the assay were evaluated, using blood anticoagulated with EDTA or sodium citrate, or platelet-rich plasma in sodium citrate. The sample of blood or platelet-rich plasma was incubated with thiazole orange, and flow cytometric analysis was performed. Percentage of circulating TO+ platelets was determined from fluorescence (FL-1) logarithmic histograms.

Results

Healthy ponies (n = 9) had 1.28 to 2.83% (mean ± SD, 2.03 ± 0.50%) and horses (n = 7) had 0.9 to 3.44% (2.12 ± 1.14%) TO+ platelets in circulation. Thrombocytopenic ponies (n = 7) had 11.14 to 48.41 % (26.51 ± 11.99%) and thrombocytopenic horses (n = 4) had 2.33 to 8.52% (6.19 ± 2.68%) TO+ platelets in circulation. Mean platelet counts for the thrombocytopenic ponies and horses were 24,400 ± 20,500 and 39,300 ± 13,500 platelets/μl, respectively (reference range, 94,000 to 232,000 platelets/μl).

Conclusion

Thiazole orange-positive platelets can be detected in equine blood and percentages of TO+ platelets are increased in thrombocytopenic horses.

Clinical Relevance

Enumeration of TO+ platelets may prove to be a helpful noninvasive clinical measurement of bone marrow platelet production and aid in the assessment of platelet kinetics in thrombocytopenic horses. (Am J Vet Res 1997;58:1092–1096)

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in American Journal of Veterinary Research

Abstract

Objective

To establish the existence of platelet-derived proteins in equine plasma, with the future goal of developing an assay for the detection of in vivo platelet activation.

Animals

5 mature healthy horses.

Procedure

Platelet-rich plasma and platelet-poor plasma were prepared from anticoagulated blood. Platelets were separated from plasma proteins by gel filtration, then activated with 0.5 μM platelet-activating factor. Protease inhibitors were added, and the released platelet proteins were harvested. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed on the released platelet proteins and platelet-poor plasma, and the resultant silver-stained bands were compared. Immunoblot analysis was performed on released platelet proteins, using an antibody to human thrombospondin; human platelet-derived proteins served as the positive control for the antibody.

Results

Released platelet proteins in the presence of β-mercaptoethanol (reduced samples) contained several proteins that were not observed in plasma including (mean ± SEM) 194 ± 2, 159 ± 2, 151 ±2, 104 ± 2, and 95 ± 1 kd. Immunoblots of released platelet proteins had a prominent 180 ± 2-kd protein in reduced samples that was recognized by an antibody to human thrombospondin, and with prolonged color development, 2 additional less prominent proteins (166 ± 1 and 155 ± 1 kd) were observed.

Conclusions

Several proteins are released from activated equine platelets that are not detectable in normal equine plasma. Thrombospondin is one of the high molecular mass proteins released by activated equine platelets.

Clinical Relevance

An assay can be developed for detection of thrombospondin in equine plasma and may be useful for detection of in vivo platelet activation in horses. (Am J Vet Res 1997;58:954–960)

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in American Journal of Veterinary Research

PMPs in agonist-stimulated equine leukocyte- and platelet-rich plasma before and after clopidogrel administration to healthy horses. 17 The aims of the study reported here were to use a bead-calibrated flow cytometric assay to standardize enumeration

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in American Journal of Veterinary Research

Born method, which is the most commonly used method for platelet function testing) has several disadvantages. Disadvantages of the Born method include the need for preparation of platelet-rich plasma, which results in separation of other blood cells

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in American Journal of Veterinary Research

is used to determine interactions of platelet agonists with either whole blood or platelet-rich plasma. The strong (collagen, arachidonate, and thrombin) or weak (ADP, epinephrine, and ristocetin) platelet agonists may stimulate platelet activation

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in American Journal of Veterinary Research

with PRP and platelet concentrate. Platelet-rich plasma was prepared by centrifugation of citrated blood at 850 × g for 10 minutes at room temperature. The PRP was then aspirated, and the platelet count was determined. d The PRP was allowed to rest

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in American Journal of Veterinary Research

thromboxane A2 formation in cat platelet rich plasma . Pharmacol Res Commun 1978 ; 10 : 1 – 12 . 10.1016/S0031-6989(78)80057-5 3. Ting HJ , Murad JP , Espinosa EV , et al. Thromboxane A2 receptor: biology and function of a peculiar receptor

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in American Journal of Veterinary Research