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Topic:Platelets
Platelets are small, anucleated cell fragments derived from megakaryocytes in the bone marrow, playing a key role in hemostasis and thrombosis in horses. They contribute to the formation of blood clots by adhering to the site of vascular injury, aggregating with one another, and facilitating the coagulation process. In equine medicine, platelet count and function are important parameters in assessing bleeding disorders and thrombotic conditions. Variations in platelet count can indicate underlying health issues, such as inflammation, infection, or bone marrow disorders. This page compiles peer-reviewed research studies and scholarly articles that explore the physiology, pathology, and clinical implications of platelets in equine health.
Effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation. To investigate the effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation. Methods: Blood samples from 6 Thoroughbreds. Methods: 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...
Isoxsuprine hydrochloride in the horse: a review. Isoxsuprine hydrochloride has been suggested for use in horses for treatment of navicular syndrome and laminitis. The drug has been shown to be a beta-adrenoreceptor antagonist with beta-adrenoreceptor agonistic properties, with both characteristics contributing to vasodilation and uterine relaxation. In addition, the drug is capable of decreasing blood viscosity and platelet aggregation. Studies have shown i.v. isoxsuprine to have a plasma half-life of <3 h with a large apparent volume of distribution. Cardiovascular effects resolve rapidly following i.v. administration, but are absent wit...
Measurement of the activation of equine platelets by use of fluorescent-labeled annexin V, anti-human fibrinogen antibody, and anti-human thrombospondin antibody. 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. Methods: Platelets obtained from 6 Thoroughbreds. Methods: 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...
The effect of three different doses of sodium pentosan polysulphate on haematological and haemostatic variables in adult horses. To evaluate the effects of three different doses of sodium pentosan polysulphate (PPS) on haematological and haemostatic variables in adult horses. Methods: Eight adult standardbred horses were used. All horses received a single injection of 0, 3, 6, and 10 mg/kg of PPS at the beginning of each treatment week for 4 weeks so that by the end of the study all horses had received all four doses of PPS. Blood samples were collected at 0, 1, 2, 3, 4, 6, 8, 12, 24, 48, and 168 h after each weekly injection of PPS. Variables measured were packed cell volume, haemoglobin, red blood cell count, mean cor...
Effects of sodium citrate, low molecular weight heparin, and prostaglandin E1 on aggregation, fibrinogen binding, and enumeration of equine platelets. To investigate the effects of sodium citrate, low molecular weight heparin (LMWH), and prostaglandin E1 (PGE1) on aggregation, fibrinogen binding, and enumeration of equine platelets. Methods: Blood samples obtained from 4 Thoroughbreds. Methods: Blood was collected into syringes in the ratio of 9 parts blood:1 part anticoagulant. Anticoagulants used were sodium citrate, LMWH, sodium citrate and LMWH, or 300 nM PGE1/ml of anticoagulant. Platelet aggregation in response to ADP, collagen, and PGE1 was assessed, using optical aggregometry. Platelet activation was evaluated, using flow cytometry, ...
Assessment of Equine Autoimmune Thrombocytopenia (EAT) by flow cytometry. RATIONALE: Thrombocytopenia is a platelet associated process that occurs in human and animals as result of i) decreased production; ii) increased utilization; iii) increased destruction coupled to the presence of antibodies, within a process know as immune-mediated thrombocytopenia (IMT); or iv) platelet sequestration. Thus, the differentiation of the origin of IMT and the development of reliable diagnostic approaches and methodologies are important in the clarification of IMT pathogenesis. Therefore, there is a growing need in the field for easy to perform assays for assessing platelet morpho...
Lipid phase separation correlates with activation in platelets during chilling. When human platelets are chilled below 22 degrees C, they spontaneously activate, a phenomenon that severely limits their storage life. It has previously been proposed that there is a correlation between cold-induced platelet activation and passage of the membranes through a liquid-crystalline to gel phase transition. Because animal models are essential for developing methods for cold storage of platelets, it is necessary to investigate such a correlation in animal platelets. In this work, horse platelets were used as a model, and it was found that cold-induced morphological activation is rela...
Endotoxin and dietary amines may increase plasma 5-hydroxytryptamine in the horse. Uptake of 5-hydroxytryptamine (5-HT) into platelets is an important mechanism by which low plasma concentrations are maintained, and platelet activation may therefore result in significant release of this vasoconstrictor. The present study examined the kinetics of active uptake of radiolabelled [3H]5-HT by washed equine platelets in vitro, and investigated the effects on this process of 4 other naturally occurring monoamines which may be released from the caecum in conditions of carbohydrate overload. The release of [3H]5-HT by platelets was also studied, since platelet accumulation and activa...
Effects of intravenous administration of formaldehyde on platelet and coagulation variables in healthy horses. To assess safety and determine effects of IV administration of formaldehyde on hemostatic variables in healthy horses. Methods: 7 healthy adult horses. Methods: Clinical signs and results of CBC, serum biochemical analyses, and coagulation testing including template bleeding time (TBT) and activated clotting time (ACT) were compared in horses given a dose of 0.37% formaldehyde or lactated Ringer's solution (LRS), IV, in a 2-way crossover design. In a subsequent experiment, horses received an infusion of 0.74% formaldehyde or LRS. In another experiment, horses were treated with aspirin to impai...
Equid herpesvirus 1: platelets and alveolar macrophages are potential sources of activated TGF-B1 in the horse. Cell mediated responses to Equid herpesvirus 1 (EHV-1) are of short duration in vivo and require considerable expansion to be detected in vitro. Raised serum levels of active transforming growth factor B (TGF-B1) have been shown to depress proliferative T cell responses in experimental infections with EHV-1 in ponies. The present work indicates that latent transforming growth factor B (TGF-B1) is present in circulating platelets, lymph node, bronchial epithelium and alveolar macrophages. Activation of platelets in vitro by thrombin resulted in the release of latent TGF-B1 from platelets, with ...
Differential activation of platelets from normal and allergic ponies by PAF and ADP. Altered platelet responsiveness has been demonstrated in human atopic dermatitis. This study has compared the in vitro function of platelets from normal ponies and those with the allergic skin disease, sweet itch. Methods: Ponies with a clinical history of sweet itch and normal ponies were used as blood donors. Methods: PAF and ADP-induced platelet aggregation was measured and TxB2 production quantitated at the time of maximal aggregation; 12-HETE was additionally measured in some samples. Agonist-induced release of 3[H]5-HT was also studied. Results: Although both PAF and ADP caused equine pl...
Actions and interactions of ADP, 5- HT, histamine and PAF on equine platelets. Platelets are thought to play a role in equine diseases such as acute laminitis and verminous arteritis and may be involved in allergic disease. Mediators implicated in the pathophysiology of these conditions activate platelets and responses may be enhanced by interactions between mediators. The present study compared platelet aggregation, thromboxane production and release of radiolabelled [(3)H]5- HT in response to 5- HT, histamine, ADP and PAF alone and in combination in vitro.PAF caused concentration-related aggregation, [(3)H]5- HT release and thromboxane production. In contrast, ADP caus...
The effect of supramaximal exercise on equine platelet function. When blood is collected into sodium citrate in the proportion of 9 parts blood:1 part sodium citrate, the concentration of plasma sodium citrate in the sample will depend on the packed cell volume (PCV) of the blood sample. This difference in plasma sodium citrate concentration secondary to alterations in PCV significantly affects human platelet aggregation responses. Since horses attain a high PCV in response to high-intensity exercise we investigated the effect of differences in sample plasma sodium citrate concentration on equine platelet aggregability. In addition, low molecular weight hep...
Detection and effects on platelet function of anti-platelet antibody in mule foals with experimentally induced neonatal alloimmune thrombocytopenia. Horse mares carrying mule foals were immunized during the last trimester of pregnancy with whole acid-citrate-dextrose-anticoagulated donkey blood to experimentally induce neonatal alloimmune thrombocytopenia. Thrombocytopenia occurred in the neonatal mule foals born to immunized horse mares within 24 hours after ingestion of their dams' colostrum. Mule foals born to mares not immunized with donkey blood did not develop thrombocytopenia. These findings suggest that antibodies may have been directed against a donkey platelet antigen present in the mule foals but not present in their dams. The o...
Platelets from thrombocytopenic ponies acutely infected with equine infectious anemia virus are activated in vivo and hypofunctional. Thrombocytopenia is a consistent finding and one of the earliest hematological abnormalities in horses acutely infected with equine infectious anemia virus (EIAV), a lentivirus closely related to human immunodeficiency virus. Multifactorial mechanisms, including immune-mediated platelet destruction and impaired platelet production, are implicated in the pathogenesis of EIAV-associated thrombocytopenia. This study was undertaken to investigate whether regenerative thrombopoiesis and platelet destruction occurred in ponies acutely infected with EIAV. Circulating large, immature platelets were in...
Shear-induced platelet activation and platelet-neutrophil aggregate formation by equine platelets. To determine whether platelets become activated and form platelet-platelet or platelet-neutrophil aggregates, or both, when subjected to shear. Methods: Blood obtained from 3 Thoroughbreds. Methods: Blood, with PCV adjusted to 32 (low hematocrit) or 60 (high hematocrit)%, was subjected to shear rates of 11.25, 22.5, 45, 90, 225, and 750/s for 3 minutes by use of a cone-plate viscometer. Flow cytometric techniques were used to identify activated platelets, platelet-platelet aggregates, and platelet-neutrophil aggregates. Results: Shear resulted in decreased platelet count, increased mean platel...
Dextran-70 inhibits equine platelet aggregation induced by PAF but not by other agonists. Dextrans of mean molecular weight 70 kDa (dextran-70) have had clinical use as anti-thrombotics in man. A major part of the anti-thrombotic action is mediated via inhibition of platelet function. Greatorex (1975, 1977) treated thromboembolic colic in horses with infusions of dextran-70 and reported a 90% recovery rate, but this treatment is nonetheless rarely used. We have used an in vitro method to examine the effect of dextran-70 on equine platelet suspensions, in the hope that understanding the mechanism of action of dextran-70 might lead to the development of alternative therapeutic agents...
Effect of a competitive inhibitor of platelet aggregation on experimentally induced laminitis in ponies. To determining whether inhibition of platelet aggregation prevents development of carbohydrate overload-induced alimentary laminitis. Methods: 22 healthy adult ponies. Methods: Acute laminitis was induced by oral administration of corn starch/wood flour to 16 ponies, 8 of which were treated with a synthetic analogue of the platelet fibrinogen receptor antagonist peptide (RPR) RGDS (arginine-glycine-aspartic acid-serine) 110885; 6 ponies served as negative controls. Blood was collected before and at 4, 8, 12, 24, 28, and 32 hours after administration of carbohydrate overload, and PCV, total pla...
Evaluation of a haematological analyser (Sysmex F-800) with equine blood. A semiautomatic electronic blood cell counter (Sysmex F-800) was evaluated with equine blood, according to the protocol of the International Committee for Standardization in Haematology (ICSH, 1984). The precision and overall reproducibility were acceptable for all the parameters studied except for the platelet count, in which a coefficient of variation of 18.8% and 21.7% was obtained for within and between batch precision and 26.76% for overall reproducibility. Carry-over for the haematocrit value and platelet count was unsatisfactory, thus the use of a blank diluent sample between different ...
Evaluation of platelet activation and platelet-neutrophil aggregates in Thoroughbreds undergoing near-maximal treadmill exercise. To determine whether platelets become activated and form platelet-neutrophil aggregates during near-maximal treadmill exercise in horses. Methods: 4 Thoroughbreds. Methods: Horses were subjected to 4 standardized exercise tests on a treadmill, and blood samples were collected before exercise, at treadmill speed of 12 m/s, and 5 minutes after exercise. Flow cytometric techniques were used to identify activated platelets, and flow cytometric and microscopic techniques were used to identify platelet-neutrophil aggregates. Results: Platelet-neutrophil aggregates increased from 2.8 +/- 0.4% at rest...
Plasma 5-hydroxytryptamine constricts equine digital blood vessels in vitro: implications for pathogenesis of acute laminitis. Cumulative concentration response curves to 5-hydroxytryptamine (5-HT; 10(-10)-10(-4) mol/l) were constructed using isolated rings of equine digital, facial, tail and coronary arteries (endothelium intact). 5-HT was 17.7 and 41 times more potent as a vasoconstrictor of digital arteries than facial and tail arteries respectively. Removal of the endothelium increased the vasoconstrictor potency of 5-HT in the facial artery by 3.7-fold (P<0.05) but did not alter the sensitivity of digital arteries to 5-HT. Coronary arteries failed to contract to 5-HT. Coronary arteries pre-contracted with U440...
Neonatal alloimmune thrombocytopenia in a quarter horse foal. Neonatal alloimmune thrombocytopenia is recognized as a spontaneous disease of human infants, piglets, and possibly mules, but it has not been previously reported in horses. A 1-day-old Quarter Horse foal presented to Michigan State University Large Animal Clinic with severe thrombocytopenia of unknown origin. Immunoglobulins that bound to the foal's platelets were identified in the mare's plasma, serum, and milk by indirect assays. The immunoglobulins were further shown to recognize platelets from the foal's full brother, born 1 year earlier. These findings, coupled with the clinical course o...
Evaluation of platelet activation and platelet-neutrophil aggregates in ponies with alimentary laminitis. To determine whether platelets are hyperaggregable or form platelet-neutrophil aggregates during the prodromal stages of acute laminitis of ponies. Methods: Healthy adult ponies: 8 experimental and 6 control. Methods: Acute laminitis was induced by oral administration of corn starch and wood flour to 8 ponies, and indices of platelet activation were evaluated. Blood samples were collected before and at 4, 8, 12, 24, 28, and 32 hours after carbohydrate administration, and PCV, total plasma protein concentration, platelet count, activated clotting time, whole blood recalcification time, spontane...
Flow cytometric method for detecting thiazole orange-positive (reticulated) platelets in thrombocytopenic horses. To evaluate a method for detecting thiazole orange-positive (TO+, reticulated) platelets in equine blood, using flow cytometry. Methods: 16 healthy, equine infectious anemia virus (EIAV)-negative horses and ponies; 9 thrombocytopenic, EIAV-positive horses and ponies; and 2 thrombocytopenic, EIAV-negative horses. Methods: 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 ...
Influence of antigen challenge on platelet responsiveness in horses with chronic obstructive pulmonary disease. A role for platelets in allergic airways disease has been postulated and changes in the responsiveness of circulating platelets have been demonstrated following antigen challenge of asthmatic human subjects. In this study agonist-induced aggregation of equine platelets in vitro has been compared before and after exposure of horses to a controlled hay and stray challenge. Prior to challenge the response of platelets, from horses with chronic obstructive pulmonary disease (COPD) and normal animals, to adenosine diphosphate (ADP) and platelet activating factor (PAF) did not differ. Five hours aft...
Suppression of megakaryocyte colony growth by plasma from foals infected with equine infectious anemia virus. Foals infected with equine infectious anemia virus become thrombocytopenic 7 to 20 days after virus inoculation, and within a few days following the onset of detectable viremia. The thrombocytopenia is associated with suppression of platelet production. Possible mediators of suppression of thrombopoiesis include tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta), cytokines that are released during inflammation. To assess effects of plasma or serum from infected foals on megakaryocyte (MK) growth and maturation in vitro, equine low-density bone marrow cells w...
Identification of thrombospondin as a high molecular mass protein released from activated equine platelets. 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. Methods: 5 mature healthy horses. Methods: 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 microM 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 platele...
Detection of activated platelets and platelet-leukocyte aggregates in horses. To determine the potential usefulness of tests for detection of platelet activation and platelet-leukocyte aggregates in horses. Methods: Blood from 3 healthy Thoroughbreds. Methods: Microscopic and flow cytometric assays were used to evaluate spontaneous platelet aggregation, platelet activation, and platelet-leukocyte aggregates. Platelet activation was detected by evaluation of binding of anti-human fibrinogen to unactivated and ADP-, thrombin-, thrombin agonist receptor peptide-, and platelet activating factor-activated platelets. Platelet-leukocyte aggregates were evaluated microscopicall...
Protonophoric activity of NADH coenzyme Q reductase and ATP synthase in coupled submitochondrial particles from horse platelets. A method to prepare coupled submitochondrial particles from horse platelets is described. The method allowed us to study the protonophoric activities of both complex I and complex V following the fluorescence quenching of the monoamine 9-amino-6-chloro-2 methoxyacridine (ACMA), a probe highly sensitive to the generation of a transmembrane delta pH. We carried out a kinetic analysis of each enzyme complex studying the proton translocation and the electron transfer activities of complex I as well as the proton translocation and the ATP hydrolytic activities of complex V. A micromethod to prepare...
Evaluation of arginine-glycine-aspartate-containing peptides as inhibitors of equine platelet function. To determine whether synthetic peptides containing the arginine-glycine-aspartate (RGD) sequence inhibit equine platelet function. Methods: For in vitro studies of blood, 3 healthy Thoroughbreds; for in vivo and ex vivo studies of administration of RGD-containing peptides, 4 young adult pony mares. Methods: Blood was incubated with and without addition of aspirin or RGD-containing peptides (RGDS, RPR 110885) and platelet aggregation responses and platelet adhesion to subendothelial collagen were determined. RPR 110885 was administered IV, and platelet function was evaluated. Platelet aggregati...
