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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.
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...
Oncotic, hemodilutional, and hemostatic effects of isotonic saline and hydroxyethyl starch solutions in clinically normal ponies. To evaluate the oncotic, hemodilutional, and hemostatic effects of IV infusions of a large volume of isotonic saline solution and 2 doses of 6% hydroxyethyl starch (HES) in clinically normal ponies. Methods: 12 adult ponies. Methods: Ponies were assigned to 3 treatment groups and received the following IV infusions: 80 ml of 0.9% sodium chloride/kg; 10 ml of 6% HES (in 0.9% sodium chloride)/kg; or 20 ml of 6% HES (in 0.9% sodium chloride)/kg. Blood samples were collected for determination of colloid oncotic pressure (COP), PCV, plasma total protein concentration, platelet count, von Willebrand...
Effect of antigen challenge on the activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease. The effect of antigen challenge on the state of activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease (COPD) has been determined by measuring neutrophil superoxide anion formation. Prior to a seven-hour antigen challenge superoxide anion production by neutrophils from asymptomatic horses with COPD and normal horses in response to platelet activating factor (PAF) (with and without cytochalasin B), serum treated zymosan (STZ) and phorbol myristate acetate (PMA) was similar. Agonist-induced superoxide production by neutrophils from symptomatic COPD and ...
Thrombocytosis in 24 horses (1989-1994). The records of horses presented to the Veterinary Teaching Hospital of North Carolina State University College of Veterinary Medicine between January 1, 1989 and April 30, 1994 were evaluated to determine risk factors associated with thrombocytosis. Of the 2,346 horses for which a CBC was performed, 24 (1.0%) had a platelet count > 400,000/microL. Demographic, diagnostic, physical examination, and clinicopathologic variables from these cases were compared with a reference population consisting of 189 horses with a normal platelet count presenting during the same period. Infectious/ inflamma...
Clinical and laboratory alterations in horses during immunization with snake venoms for the production of polyvalent (Crotalinae) antivenom. Six horses were immunized with the venoms of Bothrops asper, Crotalus durissus durissus and Lachesis muta stenophrys for the production of polyvalent (Crotalinae) antivenom. During the immunization, clinical and laboratory alterations were evaluated in these animals, and the development of humoral immune response was followed. Only moderate local tissue changes (edema, abscesses, fistules and fibrosis) were observed in these animals, whereas no systemic alterations occurred. Regarding laboratory tests, there was a drop in hemoglobin concentration and hematocrit, together with an increment in t...
The role of ADP in endotoxin-induced equine platelet activation. We have shown previously that endotoxin induces platelet aggregation in equine heparinised whole blood in a platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) dependent manner. ADP is an agonist of platelets and is present in platelet dense granules with ATP in high concentrations. An investigation was carried out to establish whether endotoxin-induced platelet activation was associated with release of platelet ATP and ADP. ADP-scavenging enzyme systems significantly inhibited endotoxin-induced aggregation. Plasma levels of adenine nucleotides were measured using ...
A primary production deficit in the thrombocytopenia of equine infectious anemia. The purpose of this study was to identify the mechanisms responsible for the thrombocytopenia that develops following infection of horses by the lentivirus equine infectious anemia virus (EIAV). Immunocompetent Arabian foals and Arabian foals with severe combined immunodeficiency (SCID), which lack functional B and T lymphocytes, were experimentally infected with EIAV. Levels of viremia and a number of clinical and hematologic parameters were examined prior to and following infection. Thrombocytopenia was not dependent on the immune response: SCID foals were affected as severely as immunocompe...
Evaluation of thrombin-antithrombin complexes and fibrin fragment D in carbohydrate-induced acute laminitis. Carbohydrate-induced laminitis has been associated with decreased platelet survival, decreased blood flow to the hoof wall and with the deposition of platelets and microthrombi within venules in the dermal laminae. To evaluate further the systemic prothrombotic events occurring in the prodromal stages of laminitis, plasma samples from control and laminitis-affected ponies and horses were tested for the presence of thrombin-antithrombin (TAT) complexes and fibrin fragment D (D-dimer). No statistically significant differences between the control and laminitis-affected animals were observed for e...
Effects of platelet activating factor on the distribution of radiolabelled leucocytes and platelets in normal horses and asymptomatic horses with chronic obstructive pulmonary disease. Antigen challenge is known to cause the recruitment of neutrophils to the lungs of horses with chronic obstructive pulmonary disease (COPD). To evaluate a possible role of platelet activating factor (PAF) in this process, the effects of PAF on the distribution of radiolabelled neutrophils were compared in normal horses and asymptomatic horses with COPD. Changes in lung function, heart rate and the distribution of platelets and eosinophils were also measured. PAF (5 ng kg-1 intravenously) caused immediate but transient increases in the number of radiolabelled neutrophils in the lungs and a conc...
A study of the effect of a platelet activating factor (PAF) receptor antagonist on antigen challenge of horses with chronic obstructive pulmonary disease. Antigen challenge involving exposure to straw and mouldy hay for 7 h produced lung function changes and neutrophil recruitment to the lungs in horses with chronic obstructive pulmonary disease (COPD). During the challenge, an increase in radiolabelled neutrophils in the lungs occurred, together with increased respiratory rate and pleural pressure. The role of platelet activating factor (PAF) in antigen-induced neutrophil accumulation, and increased pleural pressure and respiratory rate was investigated by administering the PAF receptor antagonist WEB 2086 to asymptomatic COPD horses prior to a...
Thrombocytopenia in horses: 35 cases (1989-1994). The records of 3,952 equine patients presenting to the Veterinary Teaching Hospital at North Carolina State University College of Veterinary Medicine were evaluated to determine risk factors associated with thrombocytopenia. Of 2,346 horses from which a CBC was obtained, 35 (1.49%) were thrombocytopenic (platelet count < 75,000/microL). A reference population of 189 horses with normal platelet counts (75,000 to 300,000/microL) was also studied. Standardbred horses were at increased risk for thrombocytopenia, but age and gender were not identified as significant risk factors. Horses with inf...
A morphometric study of bone marrow megakaryocytes in foals infected with equine infectious anemia virus. Morphometric evaluation of bone marrow core biopsies was used to determine megakaryocyte (MK) numbers and MK size in nine foals with equine infectious anemia virus (EIAV)-induced thrombocytopenia. Both immunocompetent normal foals and foals with severe combined immunodeficiency (SCID) were used. Platelet counts were made three times weekly following viral infection. Bone marrow core biopsies were taken from the ilium of each foal prior to experimental infection, immediately after the onset of thrombocytopenia, and at necropsy. All foals developed thrombocytopenia by 23 days postinfection. The ...
Platelet-activating factor and not thromboxane A2 is an important mediator of endotoxin-induced platelet aggregation in equine heparinised whole blood in vitro. Endotoxin has previously been shown to induce platelet aggregation in equine heparinised whole blood. This study aimed to determine whether platelet-activating factor or products of cyclo-oxygenase metabolism (thromboxane A2 or prostaglandins) were important in mediating the response of platelets to endotoxin. The effects of the following drugs on endotoxin-induced aggregation were investigated: aspirin, flunixin meglumine and carprofen (non-steroidal anti-inflammatory drugs); CV-3988 and WEB2086 (platelet-activating factor receptor antagonists); quinacrine (phospholipase A2 inhibitor). The ef...
The inhibition of adenylate cyclase in equine platelets by collagen and by platelet-activating factor. Equine platelet aggregation was stimulated by collagen fibres or platelet-activating factor. The action of both ligands was blocked by forskolin or prostaglandin E(1) agents which are known to activate adenylate cyclase. Equine platelet membranes were found to contain adenylate cyclase activity which was inhibited in dose-dependent fashion by both collagen and platelet-activating factor. Platelet-activating factor-induced inhibition was antagonised by WEB2086.
Protein tyrosine phosphorylation in equine platelets: the effect of stimulation by thrombin and platelet-activating factor (PAF). Protein tyrosine phosphorylation (PTP) in thrombin- and platelet-activating factor (PAF)-stimulated equine platelet activation was investigated in the absence and presence of 2 protein tyrosine kinase inhibitors (PTKIs), methyl 2,5-dihydroxycinnamate (MDHC) and genistein. Washed equine platelets aggregated irreversibly in response to thrombin or PAF in an agonist concentration dependent fashion. MDHC produced an MDHC concentration and time dependent inhibitory effect on rate and extent of thrombin- and PAF-induced aggregations, whereas the effect of genistein on the same parameters was only ge...
Actions of PAF receptor antagonists in horses with the allergic skin disease sweet itch. Platelet activating factor (PAF) mimics the effects of Culicoides antigen by inducing oedema and inflammatory cell accumulation in the dermis of horses with the allergic skin disease, sweet itch. PAF could therefore contribute to antigen-induced inflammatory changes in these horses. We now report that intravenous administration of the PAF receptor antagonist WEB 2086 (3 mg kg-1), at a dose that inhibited the vascular and cellular responses to PAF in sweet itch horses, reduced Culicoides antigen-induced oedema at 1 h by 73% and at 8 h by 71% (p < 0.05). Neutrophil accumulation and eosinophil...
