The laboratory diagnosis of platelet disorders: an algorithmic approach. (CAP Laboratory Improvement Programs).
Abstract: * Objective.--To provide both a detailed description of the laboratory tests available in the diagnosis of platelet disorders and a testing algorithm, based on platelet count, that can be used to direct the evaluation of platelet disorders.

Data Sources.--A literature search was conducted using the National Library of Medicine database.

Study Selection.--The literature on laboratory testing of platelet function was reviewed.

Data Extraction and Data Synthesis.--Based on the literature review, an algorithm for platelet testing was developed.

Conclusions.--A history of mucocutaneous bleeding often indicates abnormal platelet function that can be associated with a normal, increased, or decreased platelet count. Multiple laboratory procedures can now be used to determine the underlying pathologic condition of platelet dysfunction when other deficiencies or defects of the coagulation cascade or fibrinolysis are ruled out. Simple procedures, such as platelet count, peripheral blood smear, and a platelet function screening test, will often lead the investigator to more specific analyses. Although platelet function testing is often limited to larger medical centers with highly trained technologists, newer technologies are being developed to simplify current procedures and make platelet function testing more accessible. This review provides an algorithm for platelet testing that may be of benefit to pathologists and physicians who deal with hemostatic disorders.
Subject: Blood platelet disorders (Diagnosis)
Diagnosis, Laboratory (Methods)
Authors: Kottke-Marchant, Kandice
Corcoran, George
Pub Date: 02/01/2002
Publication: Name: Archives of Pathology & Laboratory Medicine Publisher: College of American Pathologists Audience: Academic; Professional Format: Magazine/Journal Subject: Health Copyright: COPYRIGHT 2002 College of American Pathologists ISSN: 1543-2165
Issue: Date: Feb, 2002 Source Volume: 126 Source Issue: 2
Geographic: Geographic Scope: United States Geographic Code: 1USA United States
Accession Number: 83551778
Full Text: Platelets are small (2-[micro]m-diameter), nonnucleated blood cells produced in the bone marrow from megakaryocytes. Platelets are activated rapidly after blood vessel injury or blood exposure to the artificial surfaces of implanted devices, and they are a crucial component of the primary hemostatic response. In their inactivated state, platelets are roughly discoid in shape and contain cytoplasmic organelles, cytoskeletal elements, invaginating open-canalicular membrane systems, and platelet-specific granules, called alpha and dense granules. Platelets have numerous intrinsic glycoproteins embedded in the outer surface of their plasma membrane that are receptors for ligands ranging from fibrinogen, collagen, thrombin, and thrombospondin to von Willebrand factor (VWF) and fibronectin. (1-3) Platelets promote hemostasis by 4 interconnected mechanisms: (1) adhering to sites of vascular injury or artificial surfaces, (2) releasing compounds from their granules, (3) aggregating together to form a hemostatic platelet plug, and (4) providing a procoagulant surface for activated coagulation protein complexes on their phospholipid membranes (Figure 1).

[FIGURE 1 OMITTED]

Platelet adhesion to the subendothelium is the initial step in platelet activation. The subendothelium is composed of extracellular matrix proteins, such as collagen, fibronectin, VWF, thrombospondin, and laminin, (4) many of which are ligands for receptors on the platelet surface. These adhesive proteins are exposed when the endothelial layer is disrupted. Because of the large number of extracellular matrix proteins and a high density of platelet surface receptors, platelet adhesion to areas of vascular injury is extremely rapid. VWF, a large, multimetric protein secreted into the extracellular matrix from endothelial cells, facilitates platelet adhesion by binding to platelet surface glycoprotein Ib/IX/V, especially at high shear rates. (5-7) Platelets can also adhere to vascular wall-associated fibrin or fibrinogen through interaction with platelet surface glycoprotein IIb/IIIa. (8,9)

After adhering to the subendothelium, platelets undergo a cytoskeletal activation that leads to a shape change with development of pseudopods. Intracellular signaling processes lead to increased cytoplasmic calcium and then initiate a secretory release reaction, whereby products from the alpha granules (platelet factor 4, [beta]-thromboglobulin, thrombospondin, platelet-derived growth factor, fibrinogen, VWF) and dense granules (adenosine diphosphate [ADP], serotonin) are released into the surrounding milieu. (10) The granule membranes contain many integral glycoproteins on their inner leaflet, such as P-selectin (CD62p) in the alpha granule and gp53 (CD63) in the lysosome, which become expressed on the outer platelet membrane after the release reaction. (11) The release of ADP from the dense granules, together with calcium mobilization, leads to a conformational change of the fibrinogen receptor, the glycoprotein IIb/IIIa receptor complex (integrin [[alpha].sub.IIb][[beta].sub.3]). (12) This conformational change of the fibrinogen receptor initiates the process of aggregation, whereby a glycoprotein IIb/IIIa receptor on one platelet is bound in a homotypic fashion to the same receptor on adjacent platelets via a central fibrinogen molecular bridge. Beside ADP, other agonists, such as epinephrine, thrombin, collagen, and platelet-activating factor, can initiate platelet aggregation by interaction with membrane receptors. This platelet release reaction and aggregation lead to the recruitment of many other platelets to the vessel wall with the formation of a hemostatic platelet plug.

Activated platelets also play a vital procoagulant role that serves as a link between platelet function and coagulation activation. Platelet membrane phospholipids undergo a rearrangement during activation with a transfer of phosphatidyl serine from the inner table to the outer table of the platelet membrane, providing a binding site for phospholipid-dependent coagulation complexes that activate both factor X and prothrombin. (13)

LABORATORY TESTS USED IN THE EVALUATION OF PLATELET FUNCTION

Clinical History

A careful clinical and family bleeding history should be taken before beginning a laboratory evaluation of platelet function. The history should include an assessment of the duration, pattern, and severity of bleeding problems, including whether the bleeding is spontaneous or associated with trauma or surgery. A lifelong bleeding diathesis may suggest a congenital platelet dysfunction, but an onset in adulthood does not necessarily exclude a congenital problem. In obtaining a history of bleeding pattern, it is necessary to determine whether a true hemorrhagic disorder exists. In this regard, it is often helpful to assess if the bleeding is out of proportion to the degree of trauma, or whether blood transfusions were required for relatively minor surgical procedures, such as tooth extractions.

Platelet-mediated bleeding disorders usually result in a mucocutaneous bleeding pattern, with ecchymosis, petechiae, purpura, epistaxis, and gingival bleeding commonly observed. (4) This pattern is in contrast to that observed with coagulation protein disorders, in which deep tissue bleeding and hemarthroses are more common. Von Willebrand disease, an abnormality of VWF, has bleeding symptoms very similar to platelet dysfunction, and evaluation for von Willebrand disease should be included in the initial evaluation of a possible platelet disorder. (14) Bleeding diatheses due to vascular malformations may give a bleeding pattern similar to platelet disorders, but is often more focal than diffuse. Acquired purpuras, such as those seen with disseminated intravascular coagulation, vasculitis, or infections, can usually be distinguished from platelet dysfunction, because platelet disorders usually cause bleeding from mucous membranes ("wet" purpura), whereas vascular purpura is usually confined to the skin ("dry" purpura). (15)

Many drugs and foods can affect platelet function (Table 1), so a complete drug history should be obtained. (16) It is important to remember that aspirin, an irreversible inhibitor of platelet function, is an ingredient of many over-the-counter and prescription medications, such as cold or flu remedies. Platelet dysfunction is associated with many systemic disorders, such as renal disease, hepatic failure, connective tissue disorders, myeloproliferative disorders, myelodysplastic disorders, malignancy, and cardiovascular disease. Additionally, some clinical features, such as albinism, deafness, nephritis, and susceptibility to infections, may help in the differential diagnosis of the inherited platelet disorders. (17)

Platelet Count and Peripheral Blood Smear

The accepted normal range of the platelet count is generally between 150 to 400 X [10.sup.3]/[micro]L of blood, although values much lower than this can be quite adequate for hemostasis. Many clinicians will refrain from platelet repletion in a stable patient until counts drop below 10 to 20 X [10.sup.3]/[micro]L. (18) Initial evaluation of the platelet count must take into consideration any pseudothrombocytopenia. Pseudothrombocytopenia is often due to cold-reacting platelet agglutinins or platelet binding to neutrophils (platelet satellitism). The agglutinins are often seen in patients with high immunoglobulin levels or infections and usually only bind platelets when calcium is chelated, such as in an EDTA blood collection tube. (19) A pseudothrombocytopenia associated with the glycoprotein IIb/IIIa antagonist drug abciximab has also been reported. (20) Pseudothrombocytopenia can be diagnosed by examining a peripheral smear, where large aggregates of platelets are observed, often around the feathered edge. A more accurate platelet count can be established by collecting the blood sample in either citrate or heparin anticoagulants. Giant platelets observed with macrothrombocytopenia syndromes can give false low platelet counts, because the large platelets may be counted as leukocytes by automated cell counters. (21)

The mean platelet volume (MPV) is an indication of platelet size. Normal MPV ranges are approximately 7 to 11 fL. The MPV can be an indication of platelet turnover, because platelets newly released from the bone marrow are larger and tend to decrease in size with age in the circulation. (22) In patients with rapid turnover, the platelets will, in general, be larger because of the larger size of newly produced platelets. True congenital macrothrombocytopenias usually have uniformly large platelets; often the platelets are at least twice the normal size and may be as large as erythrocytes. Newer techniques based on messenger RNA detection in platelets (reticulated platelets) may also be helpful to indicate the rate of thrombopoiesis. (23,24)

Platelet disorders can be associated with varying platelet appearances. In von Willebrand disease, Glanzmann thrombasthenia, and myeloproliferative disorders, the platelets have typical morphologic features, whereas giant platelets are seen in Bernard-Soulier disease and other macrothrombocytopenia syndromes. (17,25) In patients with Wiskott-Aldrich syndrome, the platelets may be small. (26) Platelets in the gray platelet syndrome, an alpha granule deficit, are characteristic for being pale, gray, and hypogranular on a Wright-stained blood smear. (27) Some platelet storage pool disorders (SPDs) may have morphologically normal platelet counts by light microscopy, but may have decreased alpha and/or dense granules by electron microscopy. (28)

Platelet Function Screening Tests or Bleeding Time

In the initial evaluation of platelets, it is desirable to perform a screening test to evaluate platelet function. For nearly a century, the bleeding time was the only platelet function screening test available. (29) The bleeding time is a test that is fraught with variability and involves the creation of a standardized cut in the skin and measurement of the time it takes for bleeding to stop. The initial Duke bleeding time used a small incision in the earlobe, and the similar Ratnoff method uses an incision in the ball of the finger. (29,30) The most commonly used bleeding time is the Ivy bleeding time, where a standardized incision is made on the volar surface of the forearm with a spring-loaded device, using venostatic pressure applied on the upper arm by a sphygmomanometer. (31) The bleeding time result depends not only on platelet number and function, but also on fibrinogen concentration, adequate vascular function, orientation and size of the incision, site of the incision, skin quality, skin temperature, operator technique, and patient cooperation.

Although procedural variability affects the bleeding times, bleeding time has been included traditionally as a screening test for suspected bleeding disorders. (32) The bleeding time has little use as a presurgical screen for hemostatic competence in individuals without a history of bleeding and is not useful in discerning platelet dysfunction in thrombocytopenic patients. (33) Many laboratories have stopped performing the bleeding time test completely because of its variability, poor reproducibility, and lack of correlation with intraoperative bleeding. Newer automated whole blood platelet function screening assays, such as the Platelet Function Analyzer-100 (PFA-100, Dade Behring, Marburg, Germany), are gaining popularity as initial screens for platelet function even though they do not measure the vascular component of the bleeding time. (34) These are described later in the section entitled "Newer Methods of Platelet Evaluation."

Bone Marrow Examination

Examination of the bone marrow can be performed to evaluate both thrombocytopenia or thrombocytosis, but it has little role in the evaluation of platelet dysfunction with a normal platelet count. The bone marrow examination may be helpful to ascertain whether thrombocytosis is due to reactive or myeloproliferative disorders. Thrombocytosis can often accompany iron deficiency; evaluation of serum iron studies together with the erythrocyte indices on the complete blood cell count can usually diagnose the condition and obviate the need for a bone marrow evaluation. In a thrombocytopenic patient, when no other reason for low platelet counts can be determined, the bone marrow examination is useful for determining the presence or absence of megakaryocytes; absence indicates dysfunctional marrow, whereas increased numbers suggest peripheral destruction with attempted bone marrow compensation. Bone marrow examination can also detect myelophthisic disorders, such as acute leukemia, lymphoma, or metastatic malignancy, which could explain a patient's thrombocytopenia.

Platelet Aggregation

Platelet aggregation studies measure the ability of agonists to cause in vitro platelet activation and platelet-platelet binding. Platelet aggregation studies can be performed in whole blood by an impedance technique or in platelet-rich plasma by a turbidimetric technique. (35,36) Platelet aggregation techniques using a microtiter plate or flow cytometer have also been described, but are not widely performed. (37,38) Whole blood platelet aggregation can be combined with studies of dense granule adenosine triphosphate release with a lumiaggregometer. (39) Turbidimetric platelet aggregation studies require platelet-rich plasma prepared from a whole blood specimen. Many factors can affect the platelet aggregation results, such as the platelet count, processing temperature, stirring rate, and processing time (testing should be completed within 4 hours of phlebotomy). (40) In addition, clinicians who order the tests should advise patients to discontinue using, if possible, any medication that may interfere with the results of the test (Table 1).

In the turbidimetric platelet aggregation assay, platelet aggregation is measured spectrophotometrically by the increase in light transmission after addition of an aggregation agonist. (35) The agonists typically used in the assay include ADP, collagen, arachidonic acid, and epinephrine. Optimal platelet aggregation shows a biphasic pattern for the agonists ADP and epinephrine; the initial increase in aggregation is due to primary aggregation in response to activation of the glycoprotein IIb/IIIa platelet membrane receptor, whereas the second wave of aggregation is the result of platelet degranulation with recruitment of additional platelet aggregates. Other agonists, such as arachidonic acid, thrombin receptor agonists, and collagen, usually show only a single wave of aggregation.

Another important reagent used in the evaluation of platelet function by aggregation is the antibiotic ristocetin, which facilitates the binding of VWF to the glycoprotein Ib/IX/V complex. (41) Ristocetin-induced platelet aggregation evaluates aggregation after the addition of various concentrations of ristocetin. This dose response allows testing for both increased and decreased sensitivity to ristocetin. For a normal result, the patient requires the presence of both functional VWF and normal glycoprotein Ib/ IX/V, so ristocetin-induced platelet aggregation is an assay that can detect both von Willebrand disease and some platelet dysfunctions, such as Bernard-Soulier syndrome.

Coagulation Testing and von Willebrand Assays

Platelet dysfunction does not directly affect the coagulation proteins; however, the laboratory evaluation of platelet dysfunction should also include some basic coagulation assays, such as the prothrombin time (PT) and activated partial thromboplastin time (APTT), to exclude a coagulopathy as the reason for bleeding.

Von Willebrand disease is not strictly a platelet dysfunction, but it is often considered in the differential diagnosis of bleeding disorders with long bleeding times or abnormal platelet function screening test results. (14) Patients with von Willebrand disease will usually have a family history of a bleeding diathesis and present with mucocutaneous bleeding. The underlying pathologic features of von Willebrand disease are due to decreased levels or defective function of VWF. Von Willebrand disease is protean in manifestation, but the various types have been grouped into 3 general categories (types 1, 2, and 3). Patients with von Willebrand disease may have decreased VWF antigen, decreased ristocetin cofactor activity, decreased ristocetin aggregation, decreased factor VIII levels, normal or increased APTT, and increased bleeding times with normal platelet counts. (14)

Newer Methods of Platelet Evaluation

New assay systems to assess platelet function have recently become clinically available. (42) These include the PFA-100, the Ultegra (Accumetrics, San Diego, Calif), and the Plateletworks (Helena, Beaumont, Tex). Most of these devices are small, stand-alone devices that can be used at the patient's bedside or in laboratories that otherwise could not perform platelet function studies.

The PFA-100 is a device that measures platelet-related primary hemostasis in a citrated whole blood specimen. (34) It uses 2 disposable cartridges that contain a membrane with a central aperture (147 [micro]m) coated with aggregation agonists (collagen and epinephrine and collagen and ADP), through which platelets are passed at high shear rates (5000-6000 [s.sup.-1]). The instrument measures the "closure time" required for platelets to adhere to the membrane, aggregate, and occlude the aperture. The collagen-epinephrine cartridge is the primary screening cartridge; it detects platelet dysfunction induced by intrinsic platelet defects, von Willebrand disease, or platelet-inhibiting agents. The collagen-ADP cartridge usually produces abnormal results with platelet disorders and von Willebrand disease, but produces a normal closure time with aspirin-like drugs because of the high ADP concentration. Von Willebrand disease, intrinsic platelet dysfunction, and nonaspirin drugs may produce an abnormal closure time with both cartridges. (34) Samples collected for analysis on the PFA-100 are stable for up to 5 hours. The PFA-100 results can be affected by low platelet counts and low hematocrits, but are not affected by heparin. (43)

The Ultegra, a rapid platelet function assay, is an automated turbidimetric whole blood assay designed to assess platelet aggregation based on the ability of activated platelets to bind fibrinogen. (44) Fibrinogen-coated polystyrene microparticles agglutinate in whole blood in proportion to the number of available platelet glycoprotein IIb/IIIa receptors. (44) This test uses a whole blood specimen and can be used at the patient's bedside without highly trained laboratory personnel. Because of the use of agonist-activated platelets and fibrinogen-coated microparticles, the Ultegra is designed to measure specifically the effect of glycoprotein IIb/IIIa antagonist drugs, such as abciximab, tirofiban, or eptifibatide, and may be of use to monitor the effect of these potent antiplatelet drugs in the cardiac catheterization laboratory or intensive care unit. (42,45,46) It is not sensitive to drugs such as aspirin or the thienopyridines (clopidogrel and ticlopidine), and it is not designed to detect platelet functional disorders or von Willebrand disease.

Recently, a rapid platelet aggregometer has become available (Plateletworks), which is designed to determine the percentage of platelet aggregation in fresh whole blood samples taken during interventional cardiac procedures. It measures the change in the platelet count due to the aggregation of functional platelets in the blood sample. This is the first bedside test to simultaneously measure both platelet count and platelet aggregation.

An experimental device, a dynamic clot retractometer (Hemodyne), measures platelet force development by exerting a tensile force on a platelet clot. (47) The device is based on the principle of clot retraction and measures both the tensile properties of the platelet cytoskeleton and the integrity of the glycoprotein IIb/IIa receptor--fibrinogen linkage. Decreased platelet force development has been detected with this device in uremia and with glycoprotein IIb/IIIa antagonists, (48) and increased force development has been detected in patients with coronary artery disease.

Flow cytometry has been used to study platelet structure and function, but this technique is used only in specialized centers. Flow cytometric analysis is based on the detection of cell surface proteins with fluorescently labeled antibodies. It has been used in the detection of platelet activation by using antibodies to proteins newly expressed on the platelet surface during activation, such as P-selectin or thrombospondin, or by detecting new epitopes on glycoprotein IIb/IIIa induced by binding fibrinogen (ligand-induced binding sites). (49) Flow cytometric evaluation of P-selectin (CD62P) expression has been used to distinguish heparin-induced thrombocytopenia (HIT) from HIT with thrombosis, which may allow for early intervention for prevention-of thrombotic complications. (50) Flow cytometry, using fluorescently labeled abciximab, can be used to determine the number of inhibited glycoprotein IIb/IIIa receptors after the infusion of glycoprotein Ib/IIIa inhibitors. (51) It can also measure platelet activation, which may correlate with thrombotic risk in certain clinical situations. (52) Platelet flow cytometry can be used to diagnose deficiencies of platelet surface glycoproteins. It has been used to detect the absence of glycoprotein IIb/IIIa receptors in patients with Glanzmann thrombasthenia and has been used to study deficiencies of glycoprotein Ia, Ib, IIb, IV, and IX. (53) Flow cytometric methods have also been used to measure dense granules (mepacrine uptake or release), aggregation, microparticle formation, and platelet procoagulant activity. (54)

Another use of flow cytometry is the detection of platelet autoantibodies in patients with idiopathic thrombocytopenic purpura and drug-induced thrombocytopenias, which is sensitive but not specific. (55) This test can be made more specific for drug-induced antibodies by incubating the platelets in the presence of the drugs in question or by using activation-dependent tests, such as [sup.51]Cr release (56) or [sup.14]C serotonin release. (57) Antigen capture assays, such as monoclonal antibody immobilization of platelet antigens (MAIPA), have improved specificity further by being able to detect antibody binding to specific platelet surface glycoproteins. (58)

Platelets with increased RNA content (reticulated platelets) can be measured by flow cytometry using the dye thiazole orange, which binds to RNA and DNA. (23,59) This technique is gaining acceptance as a diagnostic tool to evaluate whether thrombocytopenia is due to increased platelet destruction or decreased platelet production, since platelets newly released from bone marrow have increased RNA content. This assay has recently been automated on the Cell Dyn instruments (Abbott, Abbott Park, III). It is anticipated that implementation of reticulated platelet counts may help to avoid bone marrow examination in some individuals with thrombocytopenia.

Electron microscopy may be used for the ultrastructural evaluation of platelets, particularly in patients with suspected SPDs, showing a decrease or absence of the organelles (cytoplasmic dense granules) that store adenine nucleotides, serotonin, and calcium. (28) Giant platelet disorders also have characteristic electron microscopic findings. (28,60) Other specialized methods of platelet evaluation, such as crossed immunoelectrophoresis, will be discussed in the following section with the individual disorders for which they are useful.

DIAGNOSTIC CATEGORIES OF PLATELET-DERIVED BLEEDING DIATHESIS

Platelet disorders may be divided into 3 major categories: platelet dysfunction associated with normal, decreased, or increased platelet counts, as shown in the algorithms in Figures 2, 3, and 5. In all of the disorders discussed herein, the results of the coagulation screening tests PT and APTT should be considered normal.

[FIGURES 2-3 and 5 OMITTED]

Platelet Dysfunction With Normal Platelet Count

Platelet dysfunction with a normal platelet count usually indicates a qualitative platelet disorder. In following the algorithm in Figure 2, these disorders would be evaluated in a patient with a normal PT, APTT, and platelet count. The results of a platelet function screening test would be abnormal and test results for von Willebrand disease would be normal. Platelet aggregation studies would then be used to distinguish the following disorders, followed by more specific tests, if required. Most drug-induced platelet dysfunction will also demonstrate platelet dysfunction with a normal platelet count, so it is extremely important to take a careful drug history. (16) A list of drugs that cause platelet dysfunction can be found in Table 1. Platelet aggregation abnormalities typically found with drugs such as aspirin, glycoprotein IIb/IIIa antagonists, or the thienopyridines can be found in Table 2.

Glanzmann thrombasthenia is a congenital deficiency or dysfunction of glycoprotein IIb/IIIa, the receptor for fibrinogen responsible for mediating platelet aggregation. (61,62) It is an autosomal recessive disorder that manifests in lifelong mucocutaneous bleeding. Glanzmann thrombasthenia can be classified according to the amount of glycoprotein IIb/IIIa: type I, 0% to 5% of normal; type II, 6% to 20% of normal; and variant disease, 50% to 100% of normal with abnormal fibrinogen binding. (63) Mutations of both glycoprotein IIb and glycoprotein IIIa have been implicated. (63) In patients with Glanzmann thrombasthenia, the bleeding time or platelet function screening test results will be abnormal. No aggregation response will be seen on addition of ADP, collagen, epinephrine, and arachidonic acid-aggregating agents, whereas the ristocetin-induced aggregation is normal. (40) This finding is virtually diagnostic of Glanzmann thrombasthenia, but the disorder can be confirmed by platelet flow cytometry or crossed immunoelectrophoresis of platelet membrane proteins (Table 2). (64) Afibrinogenemia, a rare deficiency of fibrinogen, can present with similar initial platelet aggregation results, but the aggregation defect in afibrinogenemia is restored with addition of fibrinogen to the specimen. Additional laboratory studies in patients with Glanzmann thrombasthenia will show decreased platelet-associated fibrinogen, defective fibrinogen binding to platelets, and decreased clot retraction. (65)

Bernard-Soulier disease is a congenital deficiency of the platelet glycoprotein Ib[alpha]/Ib[beta]/IX/V receptor, the surface receptor for VWF-mediated platelet aggregation. (66) The disorder is inherited as an incompletely recessive autosomal trait with severe bleeding. Many patients with Bernard-Soulier disease have moderately severe thrombocytopenia with large platelets, and this disorder is included with the macrothrombocytopenia syndromes discussed herein. Most of the Bernard-Soulier genetic defects are due to mutations of the GPIb[alpha] gene, but may also be due to defects of the GPIb[beta] or GPIX genes. (67,68) Glycoprotein Ib is expressed on the demarcation membrane system in the megakaryocytes that is responsible for platelet fragmentation, so it is postulated that glycoprotein Ib plays a role during megakaryopoiesis and maintenance of platelet size. (17) Normal platelet aggregation is noted with exposure to ADP, collagen, epinephrine, and arachidonic acid, but aggregation is absent with the addition of ristocetin. (65) Adhesion of platelets to subendothelium or immobilized VWF is markedly reduced at all shear rates in patients with Bernard-Soulier syndrome; this finding may have direct clinical consequences. (69) The glycoprotein abnormality can be confirmed with flow cytometry or crossed immunoelectrophoresis. (70) Additional laboratory studies show normal VWF antigen and ristocetin cofactor activity to distinguish Bernard-Soulier syndrome from von Willebrand disease.

Abnormalities of platelet secretion can be due to either deficiency of platelet granules or defects in the signal transduction events that regulate secretion or aggregation. (71) Platelet SPDs can be congenital or acquired and are the result of either a deficiency of granules (alpha and/or dense granules) or defective granule release on platelet activation. (72) Dense granule SPDs ([delta]-SPDs) can be seen as a singular clinical entity or as part of other hereditary disorders, such as Chediak-Higashi, Hermansky-Pudlak syndrome, thrombocytopenia-absent radius syndrome, or Wiskott-Aldrich syndrome. (72-74) Often [delta]-SPD shows decreased aggregation response to ADP, epinephrine, and collagen with normal aggregation to arachidonic acid and ristocetin. Decreased adenosine triphosphate release by lumiaggregometry and decreased mepacrine uptake or release by flow cytometry are observed. Ultrastructural abnormalities in these disorders usually show decreased dense granules. In addition, [alpha]-SPD (Gray platelet syndrome) has decreased alpha granules and is usually considered a macrothrombocytopenia. (75) A rare [alpha]/[delta]-SPD has been described that has features of both disorders. (72) Acquired platelet storage pool disorders can be seen with underlying myeloproliferative disorders in which the platelet degranulation is defective as a result of the disease. Circulating "exhausted" platelets simulating SPDs can be observed in clinical scenarios where there is ongoing in vivo platelet activation, such as cardiopulmonary bypass, disseminated intravascular coagulation, and thrombotic thrombocytopenic purpura or hemolytic uremic syndrome.

In addition to the SPDs, platelet release defects can be seen with defects of platelet signal transduction. In general, these disorders are poorly defined, but may constitute a significant percentage of patients with abnormal secondary wave of aggregation and decreased granule release, in whom alpha and dense granules are not deficient. (71) Defects of the platelet receptors for thromboxane A2, collagen, ADP, and epinephrine are included in this category. (76-78) Defects of the collagen and epinephrine receptors can be distinguished because usually they demonstrate a selective defect in aggregation to a single agonist. (76,77) These disorders can be confirmed by flow cytometry, where a deficiency of a surface glycoprotein is identified. Defects of the signaling pathways, including G protein activation, phospholipase C activation, calcium mobilization, pleckstrin phosphorylation, and tyrosine phosphorylation, have also been described. (71) In general, these patients show decreased primary aggregation and decreased granule release without granule deficiency. Identification of the exact defect requires detailed biochemical studies, which are not available in most laboratories. Defects of thromboxane A2 synthesis have been described, including defective liberation of arachidonic acid from the platelet membrane, cyclooxygenase enzyme deficiency, or thromboxane synthase deficiency. (79) These individuals will display an aspirin-like defect in aggregation despite having never used aspirin therapy.

Platelets play an important procoagulant role with assembly of coagulation complexes on activated platelet membranes that are rich in phosphatidyl serine. A rare congenital platelet functional disorder is Scott syndrome, due to defective "flip" of phosphatidyl serine to the outer table of the platelet membrane. (80) These patients will have normal platelet aggregation studies, but have abnormal platelet procoagulant activity (platelet factor 3) and microparticle formation.

Other significant disorders of platelet function with platelet counts in the normal range are usually acquired with the presence of another disease or drug therapy. These are, by far, more common than the disorders described herein. Platelet dysfunction is often observed with chronic renal failure or liver disease, in patients experiencing a variety of myeloproliferative and lymphoproliferative disorders (ie, polycythemia vera, myelofibrosis, paroxysmal nocturnal hemoglobinuria, acute myelogenous leukemia, and hairy cell leukemia). Platelet dysfunction also may be associated with a variety of clinical scenarios, such as previous cardiopulmonary bypass, implantation of prosthetic materials such as vascular grafts and prosthetic heart valves, and ventricular assistance devices. (81, 82) Platelet dysfunction in these disorders is usually difficult to characterize because nonspecific defects of platelet aggregation are usually observed.

Platelet Disorders With Thrombocytosis

Patients with elevated platelet counts may have clinical bleeding, but may also be asymptomatic or have thrombosis. In these patients, laboratory evaluation should be primarily aimed at elucidating the cause of the thrombocytosis and should include a complete blood cell count, peripheral blood smear, bone marrow evaluation, cytogenetic study, and platelet aggregation study. In general, platelet function screening tests have little usefulness in evaluating these disorders and do not necessarily correlate with further platelet function tests. In patients with thrombocytosis, the differential diagnosis is primarily between a reactive thrombocytosis and a myeloproliferative process (essential thrombocytosis, chronic myelogenous leukemia, polycythemia vera, and myelofibrosis). The algorithmic approach to the diagnosis of thrombocytosis is shown in Figure 2. In general, patients with a myeloproliferative disorder often have platelet counts greater than 1 X [10.sup.6]/[micro]L, and patients with reactive thrombocytoses have counts less than this, but there is a great deal of overlap. For myeloproliferative disorders, characteristic features of a specific disease can be discerned by examination of the peripheral blood smear, bone marrow, and cytogenetic studies.

Platelet aggregation studies alone can suggest an underlying myeloproliferative disorder, particularly when epinephrine-induced aggregation alone is reduced or absent. (83,84) The decreased epinephrine-induced aggregation is thought to be due to down-regulation of [[alpha].sub.2]-adrenergic receptors. (83) This pattern of platelet aggregation is also observed in patients with a congenital defect of the [[alpha].sub.2]-adrenergic receptors, (76) but these patients usually have a normal platelet count. Other patterns of platelet dysfunction with myeloproliferative disorders include decreased platelet aggregation to ADP or collagen, dense-granule storage pool pattern, abnormal platelet morphologic structure, abnormalities of the arachidonic acid pathway, and decreased receptors for prostaglandin [D.sub.2]. (83,85) Additionally, some patients may show increased aggregation with various agonists or may have spontaneous aggregation without added agonists. (86) In the clinical evaluation of patients with myeloproliferative disorders, both bleeding and thrombosis can be observed in these patients, and the results of the platelet functional tests will not necessarily distinguish whether the patient is at risk for bleeding or thrombosis. (87)

In contrast to patients with myeloproliferative disorders, patients with reactive thrombocytosis usually have normal platelet function. A reactive, or secondary, thrombocytosis can be associated with many clinical entities, such as iron deficiency, inflammatory and infectious disorders after splenectomy in malignancies such as carcinomas or lymphomas, myelodysplastic disorders, smoking, or exercise. It can also be observed as a rebound thrombocytosis following splenectomy, treatment for idiopathic thrombocytopenic purpura, pernicious anemia, or cessation of myelosuppressive drugs.

Platelet Disorders With Thrombocytopenia

Disorders in which the platelet count is decreased can be divided, for evaluation purposes, by the size of the platelets. Thrombocytopenias can be congenital or acquired, but they have been grouped by platelet size in- this discussion. See Figure 3 for an algorithmic approach to small and large platelets and Figure 5 for an approach to the diagnosis of normal-sized platelets.

Thrombocytopenia with small platelets can be seen in patients with Wiskott-Aldrich syndrome. This is an X-linked recessive disorder characterized by recurrent infections, eczema, and thrombocytopenia. These individuals will have absent immunologic responses to polysaccharide antigens and progressive decline in T-lymphocyte function. The MPV is often low (approximately half the normal size) and lymphocytes are deficient in CD43 (sialophorin). (88) Platelet dysfunction is severe; the platelets are unable to aggregate and a storage pool-like pattern is often seen. (89) Patients with thrombocytopenia due to marrow aplasia may also have small platelets, but the MPV is usually low to normal not decreased.

The rare macrothrombocytopenia disorders are all congenital in nature and most are inherited in an autosomal dominant fashion. They are usually due to congenital defects in platelet production by megakaryocyte or demarcation membrane systems, although the structural or genetic abnormalities are known in only a few disorders (Figure 3). (17) Some patients with acquired platelet destruction and turnover, such as idiopathic thrombocytopenic purpura, may have high MPVs due to the rapid release of new platelets, but, in general, the macrothrombocytopenia syndrome platelets are much larger and more uniform in size.

Several macrothrombocytopenia disorders are characterized by the presence of neutrophilic inclusions. May-Hegglin anomaly is the most common macrothrombocytopenia and is an autosomal dominant disorder characterized by Dohle body inclusions in neutrophils with a mild bleeding disorder. (17,90) The Dohle bodies are blue, spindle-shaped inclusions in the periphery of the neutrophil cytoplasm (Figure 4). The thrombocytopenia is usually moderate, with platelet counts of 60 to 100 X [10.sup.3]/[micro]L, and the mean MPV is approximately 12.5 fL, but is often much larger. Laboratory studies will usually show normal platelet aggregation and a normal bleeding time, (91) attesting to the increased functionality of the larger platelets. Electron microscopic analysis of platelets in the May-Hegglin anomaly will often show a disorganization of the microtubules. (17,91) Electron microscopic analysis of the neutrophilic inclusions shows them to lack a limiting membrane, be free of specific granules, and contain parallel bundles of ribosomes, microfilaments, and segments of endoplasmic reticulum. (17) Platelet surface glycoproteins are usually normal. (92)

[FIGURE 4 OMITTED]

The 2 other macrothrombocytopenia disorders with neutrophilic inclusion are Fechtner syndrome and Sebastian syndrome. Fechtner syndrome can be distinguished by hereditary nephritis, deafness, cataracts (Alport syndrome), and macrothrombocytopenia with a mild-to-moderate bleeding disorder. (60) The MPV may be as large as 20 fL, and the peripheral smear shows uniformly giant platelets with pale blue, irregularly shaped inclusions in the neutrophil cytoplasm. (17) Platelet aggregation studies and platelet surface glycoprotein studies are normal. Sebastian syndrome (93) has no clinical associations like Fechtner syndrome.

There are several rare macrothrombocytopenia syndromes without neutrophil inclusions, which are generally characterized by either surface glycoprotein abnormalities or platelet functional defects. Bernard-Soulier syndrome lacks glycoprotein Ib/IX/V on platelet surfaces, as has been discussed herein. Patients who are heterozygous for the disease will show only giant platelets on the blood smear without hypoplatelet function, thrombocytopenia, or bleeding. These heterozygous patients may have associated velopharyngeal insufficiency, conotruncal heart disease, and learning disabilities together with an abnormality of glycoprotein Ib[beta], and are classified as having the velocardiofacial syndrome. (94) The Gray platelet syndrome is an autosomal dominant [alpha]-SPD characterized by mild bleeding symptoms, reticulin fibrosis of the bone marrow, variable thrombocytopenia, and large (mean, 13 fL), gray-appearing platelets on the peripheral blood smear due to decreased alpha granules. (75) Pale platelets can also be seen with ongoing platelet activation and circulating "exhausted" platelets, but in these patients there will be a mixture of normal and pale platelets. It is unclear whether the pathophysiologic origin of the Gray platelet syndrome is due to premature release of alpha granules from the cell or abnormal signal transduction or calcium flux. (17) Platelet aggregation study results may be abnormal for thrombin and collagen, and flow cytometry studies have shown increased surface P-selectin, but decreased alpha granule P-selectin. (75) Other rare macrothrombocytopenias are listed in Figure 3. Those that have known surface glycoprotein abnormalities include the glycoprotein IV abnormality and mitral valve insufficiency with abnormalities of glycoproteins Ia, Ic, and IIa. (17)

Bone marrow examination may be helpful in differentiating the underlying causes in thrombocytopenic platelet disorders with normal platelet morphologic structure and size. This group of disorders includes both congenital and acquired thrombocytopenias that are usually due to either decreased platelet production or increased platelet destruction (Figure 5). The number of megakaryocytes on the bone marrow can help to distinguish between these origins, but analysis of platelet turnover by messenger RNA analysis may also be helpful.

The finding of adequate or increased megakaryocytes on the bone marrow or increased reticulated platelets suggests peripheral platelet destruction. Platelet functional tests are usually not helpful in differentiating between the entities in this class of disorders, since most functional studies will give abnormal results simply because of the low platelet number. The overall MPV is usually normal with destructive thrombocytopenia, but there is usually a range of platelet size and many large platelets are seen, indicating the rapid platelet turnover. These disorders are invariably acquired and an underlying abnormality should be sought. In general, the clinical scenario is the most helpful in classifying these disorders.

Idiopathic thrombocytopenic purpura is known to be due to platelet sensitization, with autoantibodies leading to platelet destruction in the reticuloendothelial system; peripheral smears may show variable macrothrombocytopenia and autoantibodies to specific surface glycoproteins can be detected by flow cytometry or immunoassay, (95) although diagnosis is largely from clinical findings. Another immune thrombocytopenia is posttransfusion purpura, where immune destruction of both transfused and recipient platelets is seen, usually beginning 5 to 12 days after transfusion. Patients often have abnormal platelet antigens and are frequently [PL.sup.A1] negative ([PL.sup.A2] homozygous or HPA-1b). (96) Anti-[PL.sup.A1] antibodies can be detected both in plasma and attached to the platelet surface. Neonatal alloimmune thrombocytopenia occurs when the mother lacks a platelet antigen present on the neonates platelets. Maternal antibodies cross the placenta and cause severe thrombocytopenia shortly after birth. Several platelet antigen systems may be responsible for this scenario, but often the mother is [PA.sup.A1] negative ([PL.sup.A2] homozygote). (97) Interestingly, this same [PL.sup.A2] genetic polymorphism may impart a risk for coronary artery disease. (98)

The thrombocytopenia of thrombotic thrombocytopenic purpura is thought to be secondary to deficiency of a VWF-cleaving metalloproteinase in many patients, leading to diffuse thrombus formation in small vessels and a decline of circulating platelets. (99) These patients will show characteristic clinical symptoms with renal failure, mental status changes, fever, and hemolysis with prominent schistocytes on the peripheral blood smear, but normal screening coagulation test results. (100) Thrombotic thrombocytopenic purpura has also been associated with Shiga toxin-producing strains of Escherichia coli and with drugs such as cyclosporine, quinine, ticlopidine, and clopidogrel. (100-103) An assay for the VWF-cleaving metalloproteinase is now available, but because it is based on the VWF-multimer assay, is not readily available in most laboratories. (99)

Drug-induced thrombocytopenias due to immunologic platelet destruction can be seen with many drugs, but the most common offenders are quinidine, quinine, heparin, sulfonamide drugs, and gold salts. (104) This syndrome has also been described with the glycoprotein IIb/IIIa antagonist abciximab, but because this drug is based on a monoclonal antibody to the fibrinogen receptor (Fab' fragment), the thrombocytopenia may not be alloimmune in origin. (105) Drug-induced thrombocytopenias can be diagnosed by detecting the presence of platelet-associated antibody by flow cytometry, although this is a nonspecific finding that can also be seen with infections and autoimmune disorders. The drug dependence of the antibody binding can be demonstrated by incubating platelets with patient plasma in the presence of the drug. (55)

Heparin-induced thrombocytopenia is a distinctive drug-induced thrombocytopenia associated with heparin therapy, where antibodies are formed to heparin-platelet factor 4 complexes, leading to platelet aggregation, platelet microparticle formation, endothelial injury, and paradoxical thrombosis. (106) The thrombotic complications can be either venous or arterial and may cause death, limb loss, and pulmonary thrombosis. Bleeding secondary to thrombocytopenia is rare in HIT. The thrombocytopenia is often delayed 5 to 12 days after starting heparin therapy and usually resolves after heparin therapy is stopped. Specific laboratory tests, such as heparin-induced platelet aggregation, serotonin release, and heparin-platelet factor 4 enzyme-linked immunosorbent assay, are available to positively diagnose this disorder. (106,107)

Thrombocytopenias due to decreased platelet production include both rare congenital and more common acquired causes (Figure 3). Congenital thrombocytopenias due to decreased megakaryocytes with normal platelet size include thrombocytopenia-absent radius syndrome, X-linked megakaryocytic thrombocytopenia, and Fanconi anemia. This last disorder is distinguished by erythroid hypoplasia and DNA instability. Acquired megakaryocytic hypoplasia is seen with myelophthisic disorders, viral illnesses, and drug-induced hypoplasia. (104)

ACTIVATED PLATELET SYNDROMES

Some patients with myeloproliferative disorders will show evidence of activated platelets or hyperplatelet function. Although most patients with essential thrombocythemia and polycythemia vera will display hypoaggregation, some will actually show an increased response to various agonists, excluding epinephrine. (85) Some of these patients will also exhibit spontaneous platelet aggregation in vitro. In vitro hypoplatelet or hyperplatelet function in patients with myeloproliferative disorders does not necessarily correlate with clinical symptoms of bleeding or thrombosis.

The sticky platelet syndrome is a poorly defined disorder associated with arterial and venous thromboembolic events characterized by hyperaggregability of platelets on exposure to ADP and epinephrine. The molecular mechanism of this disorder is not known, but it has been suggested that emotional stress may be a precipitating factor for systemic thrombosis. (108)

Platelet activation is known to occur in patients with concomitant cardiovascular disease, (109) hypertension, (110) and diabetes, (111) most likely due to platelet activation secondary to vascular injury. Additionally, abnormal glycation is thought to contribute to abnormalities of platelet function in diabetic patients. (112) Older patients with hypertension are found to have a higher prevalence of thromboembolic tendencies-associated with platelet hyperactivity. (113) Genetic polymorphisms of the glycoprotein IIIa platelet membrane receptor gene (the [Pl.sup.A2] genotype) may predispose patients with this allele to a higher risk of acute coronary and cerebrovascular events, although the mechanism for the thrombotic tendency is uncertain. (98) Additional genetic polymorphisms of other platelet surface proteins are being investigated as risk factors for cardiovascular disease.

Various techniques for the diagnosis of in vivo platelet activation or increased platelet function are available. Spontaneous in vitro platelet aggregation or the finding of increased aggregation response to low concentrations of platelet agonists suggests increased platelet function. The detection of circulating platelet aggregates may be made either by flow cytometry or by collecting a whole blood sample into a fixative. (114) Platelet flow cytometry can also be used to detect circulating activated platelets due to neoexpression of new surfaces markers (ie, P-selectin or CD63) or by detecting proteins bound to platelet surface glycoproteins (ie, fibrinogen, thrombospondin). (49,52) Some caution must be exercised in the diagnosis of increased platelet activation, because persistent in vivo activation can lead to degranulation of platelets with the detection of hypoaggregation in the laboratory.

CONCLUSION

Multiple causes exist for platelet-derived bleeding diatheses. The laboratory evaluation of these disorders can range from simple to complex as outlined herein, but should initially include a thorough evaluation of the patient's medical history, concentrating on personal and familial bleeding disorders and all current medications. With this as a starting point, the clinician and/or pathologist may find algorithms such as those presented herein helpful to guide investigation toward elucidating the underlying origin for platelet-derived bleeding.

References

(1.) Brass LF, Vassallo RR Jr, Belmonte E, et al. Structure and function of the human platelet thrombin receptor: studies using monoclonal antibodies directed against a defined domain within the receptor N terminus. J Biol Chem. 1992; 267:13795-13798.

(2.) Kieffer N, Phillips DR. Platelet membrane glycoproteins: functions in cellular interactions. Annu Rev Cell Biol. 1990;16:329-357.

(3.) Peerschke EIB. Platelet membrane glycoproteins: functional characterization and clinical applications. Am J Clin Pathol. 1992;98:455-463.

(4.) Marcus AJ. Platelets and their disorders. In: Ratnoff OC, Forbes CD, eds. Disorders of Hemostasis. 3rd ed. Philadelphia, Pa: WB Saunders; 1996:79-137.

(5.) Meyer D, Baumgartner HR. Role of von Willebrand factor in platelet adhesion to the subendothelium. Br J Haematol. 1983;54:1-9.

(6.) Tracy PR. Role of platelets and leukocytes in coagulation. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice. 4th ed. Philadelphia, Pa: JB Lippincott Co; 2001:575-596.

(7.) Siedlecki CA, Lestini BJ, Kottke-Marchant K, Eppell SJ, Wilson DL, Marchant RE. Shear dependent changes in the three dimensional structure of human von Willebrand factor. Blood. 1996;88:2939-2950.

(8.) Savage B, Saldivar E, Rugged ZM. Initiation of platelet adhesion by arrest onto fibrinogen or translocation on von Willebrand factor. Cell. 1996;84:289-297.

(9.) Ruggeri ZM, Savage B. Biological functions of von Willebrand factor. In: Ruggeri ZM, ed. Von Willebrand Factor and the Mechanisms of Platelet Function. Berlin, Germany: Springer-Verlag; 1998:79-109.

(10.) Fukami MH, Holmsen H, Kowalski MA, Niewiarowski S. Platelet secretion. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice. 4th ed. Phladelphia, Pa: JB Lippincott Co; 2001:561-574.

(11.) McEver RP, Beckstead JH, Moore KL, Marshall-Carlson L, Bainton KF. GMP-140, a platelet ct-granule membrane protein, is also synthesized by vascular endothelial cells and is localized in Weibel-Palade bodies. J Clin Invest. 1989; 84:92-99.

(12.) Plow EF, Ginsberg MH. The molecular basis of platelet function. In: Hoffman R, Benz EJ Jr, Shattil SJ, et al, eds. Hematology: Basic Principles and Practice. New York, NY: Churchill Livingstone; 1991:1165-1176.

(13.) Zwaal RFA, Comfurius P, Bevers EM. Mechanisms and function of changes in membrane-phospholipid asymmetry in platelets and erythrocytes. Biochem Soc Trans. 1993;21:248-252.

(14.) Sadler JE, Gralnick HR. Commentary: a new classification for von Willebrand Disease. Blood. 1994;84:676-679.

(15.) Lowe GDO. Vascular disease and vasculitis. In: Ratnoff OC, Forbes CD, eds. Disorders of Hemostasis. 3rd ed. Philadelphia, Pa: WB Saunders; 1996:489-504.

(16.) George JN, Shattil SJ. The clinical importance of acquired abnormalities of platelet function. N Engl J Med. 1991;324:27-39.

(17.) Mhawech P, Saleem A. Inherited giant platelet disorders: classification and literature review. Am J Clin Pathol. 2000;113:176-190.

(18.) Norfolk DR, Ancliffe PJ, Contreras M, et al. Consensus Conference on Platelet Transfusion, Royal College of Physicians of Edinburgh, 27-28 November 1997--synopsis of background papers. Br J Haematol. 1998;101609-617.

(19.) Cunningham VL, Brandt JT. Spurious thrombocytopenia due to EDTA-independent cold-reactive agglutinins. Am J Clin Pathol. 1992;97:359-362.

(20.) Christopoulos CG, Machin SJ. A new type of pseudothrombocytopenia: EDTA-mediated agglutination of platelets bearing Fab fragments of a chimaeric antibody. Br J Haematol. 1994;87:650-652.

(21.) White JG. Structural defects in inherited and giant platelet disorders. In: Harris H, Hirschhorn K, eds. Adv Hum Genet. 1990;19:133-234.

(22.) Corash L, Chen HY, Levin J, et al. Regulation of thrombopoiesis: effects of the degree of thrombocytopenia on megakaryocyte ploidy and platelet volume. Blood. 1987;70:177-185.

(23.) Ault KA, Rinder HM, Mitchell J, Carmody MB, Vary CPH, Hillman RS. The significance of platelets with increased RNA content (reticulated platelets): a measure of the rate of thrombopoiesis. Am J Clin Pathol. 1992;98:637-646.

(24.) Romp KG, Peters WP, Hoffman M. Reticulated platelet counts in patients undergoing autologous bone marrow transplantation: an aid in assessing marrow recovery. Am J Hematol. 1994;46:319-324.

(25.) Murphy S. Hereditary thrombocytopenia. Clin Haematol. 1972;1:359-368.

(26.) Baldini MG. Nature of the platelet defect in the Wiskott-Aldrich syndrome. Ann N Y Acad Sci. 1972;201:437-444.

(27.) Toledano SL, Cane JP, Breton-Gorius J, et al. Gray platelet syndrome: [alpha]-granule deficiency. J Lab Clin Med. 1981;98:831-848.

(28.) White JG. Use of the electron microscope for diagnosis of platelet disorders. Semin Thromb Hemost. 1998;24:163-168.

(29.) Duke WW. The relation of blood platelets to hemorrhagic disease: description of a method for determining the bleeding time and coagulation time and report of three cases of hemorrhagic disease relieved by transfusion. JAMA. 1910; 55:1185-1192.

(30.) Ratnoff OD. Bleeding Syndromes: A Clinical Manual. Springfield, Ill: Charles C Thomas; 1960.

(31.) McGlasson DL, Strickland DM, Hare RJ, Reilly PA, Patterson WR. Evaluation of three modified Ivy bleeding time devices. Lab Med. 1988;19:645-648

(32.) Burns ER, Lawrence C. Bleeding time: a guide to its diagnostic and clinical utility. Arch Pathol Lab Med. 1989;113:1219-1224.

(33.) Lind SE. The bleeding time does not predict surgical bleeding. Blood. 1991;77:2547-2552.

(34.) Mammen EF, Comp PC, Gosselin R, et al. PFA-100 system: a new method for assessment of platelet dysfunction. Semin Thromb Hemost. 1998;24:195-202.

(35.) Born GVR. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature. 1962;194:927-929.

(36.) Riess H, Braun G, Brehm G, Hiller E. Critical evaluation of platelet aggregation in whole human blood. Am J Clin Pathol. 1986;85:50-56.

(37.) Fratantoni JC, Poindexter BJ. Measuring platelet aggregation with micro-plate reader: a new technical approach to platelet aggregation studies. Am J Clin Pathol. 1990;94:613-617.

(38.) Ault KA, Rinder HM, Mitchell JG, Rinder CS, Lambrew CT, Hillman RS. Correlated measurement of platelet release and aggregation in whole blood. Cytometry. 1989;10:448-455.

(39.) White MM, Foust JT, Mauer AM, Robertson JT, Jennings LK. Assessment of lumiaggregometry for research and clinical laboratories. Thromb Haemost. 1992; 67:572-577.

(40.) Triplett DA, Harms CS, Newhouse P, Clark C. Platelet Function: Laboratory Evaluation and Clinical Application. Chicago, Ill: ASCP Press; 1978.

(41.) Howard MA, Firkin BG. Ristocetin--a new tool in the investigation of platelet. Thromb Diath Haemorrh. 1971:26:362-369.

(42.) Nicholson NS, Panzer-Knodle SG, Haas NF, et al. Assessment of platelet function assays. Am Heart J. 1998;135(5 pt 2 suppl):S170-S178.

(43.) Kottke-Marchant K, Powers JB, Brooks L, Kundu S, Christie DJ. The effect of antiplatelet drugs, heparin and preanalytical variables on platelet function detected by the platelet function analyzer (PFA-100TM). Clin Appl Thromb Hemost. 1999;5:122-130.

(44.) Coller BS, Lang D, Scudder LE. Rapid and simple platelet function assay to assess GPIIb/IIIa receptor blockade. Circulation. 1997;95:860-867.

(45.) Coller BS. Monitoring platelet GP IIb/IIIa antagonist therapy. Circulation. 1998;97:5-9.

(46.) Steinhubl SR, Kottke-Marchant K, Moliterno DJ, et al. Attainment and maintenance of platelet inhibition through standard dosing of abciximab in diabetic and nondiabetic patients undergoing percutaneous coronary intervention. Circulation. 1999;100:1977-1982.

(47.) Carr ME, Zekert SL. Measurement of platelet-mediated force development during plasma clot formation. Am J Med Sci. 1991;302:13-18.

(48.) Carr ME, Carr SL, Hantgan RR, Braaten J. Glycoprotein IIb/IIIa blockade inhibits platelet-mediated force development and reduces gel elastic modulus. Thromb Haemost. 1995;73:499-505.

(49.) Abrams C, Shattil SJ. Immunological detection of activated platelets in clinical disorders. Thromb Haemost. 1991;65:467-175.

(50.) Jy W, Mao WW, Horstman LL, Valant PA, Ahn YS. A flow cytometric assay of platelet activation marker P-selectin distinguishes heparin induced thrombocytopenia(HIT) from HIT with thrombosis (HITT). Thromb Haemost. 1999;82: 1255-1259.

(51.) Liu C-Z, Hur B-T, Huang T-F. Measurement of glycoprotein IIb/IIIa blockade by flow cytometry with fluorescein isothiocyanate-conjugated crotavirin, a member of disintegrins. Thromb Haemost. 1996;76:585-591.

(52.) Tschoepe D, Schultheis HP, Kolarov P, et al. Platelet membrane activation markers are predictive for increased risk of acute ischemic events after PTCA. Circulation. 1993;88:37-42.

(53.) Seidl C, Siehl J, Kirchmaier CM, Seifried E. Analysis of glycoprotein Ia, IIa, IIb, and IV RNA in platelets: quantitative determination using fluorescence-based polymerase chain reaction. Haemostasis. 1997;27:131-139.

(54.) Wall JE, Buijs-Wilts M, Arnold JT, Wang W, White MM. A flow cytometric assay using mepacrine for study of uptake and release of platelet dense granule contents. Br J Haematol. 1995:89:380-38

(55.) McFarland JG. Laboratory investigation of drug-induced immune thrombocytopenias. Transf Med Rev. 1993;7:275-287.

(56.) Cimo PL, Pisciotta AV, Desai RG, et al. Detection of drug-dependent antibodies by the 51Cr platelet lysis test: documentation of immune thrombocytopenia induced by diphenyl-hydantoin, diazepam, and sulfisoxazole. Am J Hematol. 1977;2:65-72.

(57.) Hirshman RJ, Shulman NR. The use of platelet serotonin release as a sensitive method for detecting anti-platelet antibodies and a plasma anti-platelet factor in patients with idiopathic thrombocytopenic purpura. Br J Haematol. 1973; 24:793-802.

(58.) Kiefel V, Santoso S, Weisheit M, Mueller-Eckhardt C. monoclonal antibody-specific immobilization of platelet antigens (MAIPA): a new tool for the identification of platelet-reactive antibodies. Blood. 1987;70:1722-1726.

(59.) Kienast J, Schmitz G. Flow cytometric analysis and Thiazole Orange uptake by platelets: a diagnostic aid in the evaluation of thrombocytopenic disorders. Blood. 1990;75:116-121.

(60.) Peterson LC, Rao KV, Crosson JT, White JG. Fechtner syndrome--a variant of Alport's syndrome with leukocyte inclusions and macrothrombocytopenia. Blood. 1985;65:397-406.

(61.) Glanzmann E. Hereditar hamorrhagische thrombasthenie: Ein Beitrag zur Pathologie der Blutplattchen. J Kinderkranken. 1918;88:113-117.

(62.) Vinciguerra C, Trzeciak MC, Philippe N, et al. Molecular study of Glanzmann thrombasthenia in 3 patients issue from 2 different families. Thromb Haemost. 1994;74:822-827.

(63.) Peretz H, Rosenberg N, Usher S, et al. Glanzmann's thrombasthenia associated with deletion-insertion and alternative splicing in the glycoprotein IIb gene. Blood. 1995;85:414-420.

(64.) Johnston GI, Heptinstall S, Robins RA, Price MR. The expression of glycoproteins on single blood platelets from healthy individuals and form patients with congenital bleeding disorders. Biochem Biophys Res Commun. 1984;123: 1091-1098.

(65.) Nurden AT, George JN. Inherited disorders of the platelet membrane: Glanzmann thrombasthenia, Bernard-Soulier syndrome, and other disorders. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice, 4th ed. Phladelphia, Pa: JB Lippincott Co; 2001:921-944.

(66.) Bernard J, Soulier JP. Sur une nouvelle variete de dystrophie thrombocytair hemorragipar congenitale. Semin Hop Paris. 1948;24:3217-3223.

(67.) Li C, Pasquale DN, Roth GJ. Bernard-Soulier syndrome with severe bleeding: absent platelet glycoprotein Ib alpha due to a homozygous one-base deletion. Thromb Haemost. 1996;76:670-674.

(68.) Noris P, Arbustini E, Spedini P, et al. A new variant of Bernard-Soulier syndrome characterized by dysfunctional glycoprotein (gp) Ib and severely reduced amounts of gpIX and gpV. Br J Haematol. 1998;103:1004-1013.

(69.) Olson JD, Moake JL, Collins MF, Michael BS. Adhesion of human platelets to purified solid-phase yon Willebrand factor: studies of normal and Bernard-Soulier platelets. Thromb Res. 1983;32:115-XX.

(70.) Bunescu A, Lindahl T, Solum NO, et al. Partial expression of GP Ib measured by flow cytometry in two patients with Bernard-Soulier syndrome. Thromb Res. 1994;76:441-450.

(71.) Rao AK, Gabbeta J. Congenital disorders of platelet signal transduction. Arterioscler Thromb Vasc Biol. 2000;20:285-289.

(72.) White JG. Inherited abnormalities of the platelet membrane and secretory granules. Hum Pathol. 1987;18:123-139.

(73.) Gahl WA, Brandy M, Kaiser-Kupfer MI, et al. Genetic defects and clinical characteristics of patients with a form of oculocutaneous albinism (Hermansky-Pudlak syndrome). N Engl J Med. 1998;338:1258-1264.

(74.) Apitz-Castro R, Cruz MR, Ledezma E, et al. The storage pool deficiency in platelets from humans with the Chediak-Higashi syndrome: study of six patients. Br J Haematol. 1985;59:471-483.

(75.) Lages B, Sussman II, Levine SP, Coletti D, Weiss H. Platelet alpha granule deficiency associated with decreased P-selectin and selective impairment of thrombin-induced activation in a new patient with the gray platelet syndrome ([alpha]-storage pool deficiency). J Lab Clin Med. 1997;129:364-375.

(76.) Rao AK, Willis J, Kowalska MA, Wachtfogel Y, Colman RW. Differential requirements for epinephrine induced platelet aggregation and inhibition of adenylate cyclase: studies in familial [alpha] 2-adrenergic receptor defect. Blood. 1988; 71:494-501.

(77.) Nieuwenhuis HK, Akkerman JWN, Houdijk WPM, Sixma JJ. Human blood platelets showing no response to collagen fail to express surface glycoprotein la. Nature. 1985;318:470-472.

(78.) Cattaneo M, Lecchi A, Randi AM, McGregor JL, Mannucci PM. Identification of a new congenital defect of platelet function characterized by severe impairment of platelet responses to adenosine diphosphate. Blood. 1992;80: 2787-2796.

(79.) Rao AK. Congenital disorders of platelet function. Hematol Oncol Clin North Am. 1990;4:65-87.

(80.) Sims PJ, Wiedmer T, Esmon CT, et al. Assembly of the platelet prothrombinase complex is linked to vesiculation of the platelet plasma membrane: studies in Scott syndrome: an isolated defect in platelet procoagulant activity. J Biol Chem. 1989;264:17-49.

(81.) Rodgers GM. Overview of platelet physiology and laboratory evaluation of platelet function. Clin Obstet Gynecol. 1999;42:349-359.

(82.) Weerasinghe A, Taylor KM. The platelet in cardiopulmonary bypass. Ann Thorac Surg. 1998;66:2145-2152.

(83.) Schafer Al. Bleeding and thrombosis in the myeloproliferative disorders. Blood. 1984;64:1-12.

(84.) Raman BKS, van Slyck EJ, Riddle J, Sawdyk MA, Abraham JP, Saeed SM. Platelet function, and structure in myeloproliferative disease, myelodysplastic syndrome, and secondary thrombocytosis. Am J Clin Pathol. 1989;91:647-655.

(85.) Rao AK. Acquired qualitative platelet defects. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice. 4th ed. Philadelphia, Pa: JB Lippincott Co; 2001: 905-920.

(86.) Baldiuni CL, Bertolino G, Noris P, Piletta GC. Platelet aggregation in platelet-rich plasma and whole blood in 120 patients with myeloproliferative disorders. Am J Clin Pathol. 1991;95:82-86.

(87.) Ravandi-Kashani F, Schafer Al. Microvascular disturbances, thrombosis and bleeding in thrombocythemia: current concepts and perspectives. Semin Thromb Hemost. 1997;23:479-488.

(88.) Shelley CS, Remold-O'Donnell E, Davis AE Ill, et al. Molecular characterization of sialophorin (CD43), the lymphocyte surface sialoglycoprotein defective in Wiskott-Aldrich syndrome. Proc Natl Acad Sci U S A. 1989;86:2819-2823.

(89.) Marone G, Albini F, di Martino L, et al. The Wiskott-Aldrich syndrome: studies of platelets, basophils and polymorphonuclear leukocytes. Br J Haematol. 1986;62:737-745.

(90.) Noris P, Spedini P, Belletti S, et al. Thrombocytopenia, giant platelets and leukocyte inclusion bodies (MHA): clinical and laboratory findings. Am J Med. 1998;194:355-360.

(91.) Hamilton RW, Shaikh BS, Ottie JN, Storch AE, Saleem A, White JG. Platelet function, ultrastructure and survival in the May-Hegglin anomaly. Am J Clin Pathol. 1980;74:663-668.

(92.) Coller BS, Zarrabi MH. Platelet membrane studies in the May-Hegglin anomaly. Blood. 1981;58:279-284.

(93.) Greinacher A, Nieuwenhuis HK, White JG. Sebastian platelet syndrome: a new variant of hereditary macrothrombocytopenia with leukocyte inclusions. Blut. 1990;61:282-288.

(94.) Ryan AK, Goodship JA, Wilson DI, et al. Spectrum of clinical features associated with interstitial chromosome 22q11 deletions: a European collaborative study. J Med Genet. 1997;34:798-804.

(95.) Winiarski J, Ekelund E. Antibody binding to platelet antigens in acute and chronic idiopathic thrombocytopenia purpura: a platelet membrane ELISA for the detection of antiplatelet antibodies in serum. Clin Exp Immunol. 1986;63:459-465.

(96.) Mueller-Eckhardt C, Lechner K, Heinrich D, et al. Post-transfusion thrombocytopenic purpura: immunological and clinical studies in two cases and review of the literature. Blut. 1980;40:249-257.

(97.) Murphy MF, Manley R, Roberts D. Neonatal alloimmune thrombocytopenia. Haematologica. 1999;84:110-114.

(98.) Weiss EJ, Bray PF, Tayback M, et al. A polymorphism of a platelet glycoprotein receptor as an inherited risk factor for coronary thrombosis. N Engl J Med. 1996;334:1090-1094.

(99.) Furlan M, Robles R, Galbusera M, et al. von Willebrand factor--cleaving protease in thrombotic thrombocytopenic purpura and the hemolytic-uremic syndrome. N Engl J Med. 1998;339:1578-1584.

(100.) George JN, Vesely SK, Rizvi MA. Thrombotic thrombocytopenic purpurahemolytic uremic syndrome. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and Thrombosis: Basic Principles and Clinical Practice. 4th ed. Phladelphia, Pa: JB Lippincott Co; 2001:1235-1242.

(101.) Boyce TG, Swerdlow DL, Griffin PM. Escherichia coil O157:h7 and the hemolytic-uremic syndrome. N Engl J Med. 1994;333:364-368.

(102.) Bennett CL, Connors JM, Carwile JM. Thrombotic thrombocytopenic purpura associated with clopidogrel. N Engl J Med. 2000;342:1773-1777

(103.) Atkinson K, Biggs JC, Hayes J, et al. Cyclosporin A associated nephrotoxicity in the first 100 days after allogeneic bone marrow transplantation: three distinct syndromes. Br J Haematol. 1983;54:59-67.

(104.) Jandl JH. Disorders of platelets. In: Blood: Textbook of Hematology. 2nd ed. Boston, Mass: Little Brown; 1996:1301-1360.

(105.) Berkowitz SD, Harrington RA, Rund MM, Tcheng JE. Acute profound thrombocytopenia after c7E3 Fab (Abciximab) therapy. Circulation. 1997;95:809-813.

(106.) Warkentin TE, Chong BH, Greinacher A. Heparin-induced thrombocytopenia: towards consensus. Thromb Haemost. 1998;79:1-7.

(107.) Greinacher A, Amiral J, Dummel V, Vissac A, Kiefel V, Mueller-Eckhardt C. Laboratory diagnosis of heparin-associated thrombocytopenia and comparison of platelet aggregation test, heparin-induced platelet activation test, and platelet factor 4/heparin enzyme-linked immunosorbent assay. Transfusion. 1994;34:381-385.

(108.) Mammen EF. Sticky platelet syndrome. Semin Thromb Hemost. 1999;25: 361-365.

(109.) Knight CJ, Panesar M, Wright C, et al. Altered platelet function detected by flow cytometry: effects of coronary artery disease and age. Arterioscler Thromb Vasc Biol. 1997;17:2044-2053.

(110.) Blann AD, Lip GY. Platelets and hypertension; the pressure is on the clot. J Hum Hypertens. 1997;11:763-764

(111.) Winocour PD. Platelet abnormalities in diabetes mellitus. Diabetes. 1992; 41:26-31.

(112.) Mustard JF, Packham MA. Platelets and diabetes mellitus. N Engl J Med. 1984;311:665-667.

(113.) Riondino S, Pignatelli P, Pulcinelli FM, et al. Platelet hyperactivity in hypertensive older patients is controlled by lowering BP. J Am Geriatr Soc. 1999; 47:943-947.

(114.) Wu KK, Hoak JC. A new method for the quantitative detection of platelet aggregates in patients with arterial insufficiency. Lancet. 1974;2:924-926.

Accepted for publication September 7, 2001.

From the Department of Clinical Pathology, The Cleveland Clinic Foundation, Cleveland, Ohio.

Reprints: Kandice Kottke-Marchant, MD, PhD, Department of Clinical Pathology, The Cleveland Clinic Foundation, L30, 9500 Euclid Ave, Cleveland, OH 44195 (e-mail: marchak@ccf.org).
Table 1. Drugs That Affect Platelet Function *

Nonsteroidal anti-inflammatory      Psychotropics and anesthetics
  drugs (NSAIDs)                     Tricyclic antidepressants
 Aspirin                              (ie, imipramine)
 Ibuprofen                           Phenothiazines (ie,
 Mefenamic acid                       chlorpromazine)
 Indomethacin                        Local and general anasthesia
 Cox-2 inhibitors                     (ie, halothane)

Antimicrobials                      Chemotherapeutic agents
 Penicillins                         Mithramycin
 Cephalosporin                       Daunorubicin
 Nitrofurantoin                      Carmustine
 Hydroxychloroquine
 Amphotericin                       Miscellaneous agents
                                     Dextrans
Cardiovascular agents                Radiographic contrast
 [beta]-Adrenergic blockers          Quinidine
  (ie, propanol)                     Ethanol
 Vasodilators (ie, nitropursside,
  nitroglycerin)                    Foods
 Diuretics (ie, furosemide)          Caffeine
 Calcium channel blockers            Garlic
                                     Cumin
Anticoagulants                       Turmeric
 Heparin
 Coumadin                           Antiplatelet drugs
 Lepirudin                           Phosphodiesterase inhibitors
 Argatroban                           Dipyridamole
 Bivalirudin                          Cilostazole

Thrombolytic agents                 Adenosine diphosphate receptor
 Streptokinase                       antagonists
 Urokinase                            Ticlopidine
 Tissue plasminogen activator         Clopidogrel

                                    Glycoprotein IIb/IIIA antagonists
                                      Abciximab
                                      Eptifibatide
                                      Tirofiban

* Adapted from George and Shattil. (16)
Table 2. Aggregation Characteristics *

                                      ADP

        Disorder            Primary       Secondary          AA

VWD                       N              N              N

Glanzmann throm-          [down arrow]   [down arrow]   [down arrow]
 basthenia                 or Abs         or Abs         or Abs
Bernard-soulier syn-      N              N              N
 drome
Dense granule plate-      N              [down arrow]   N
 let storage pool
 disorder ([delta]-SPD)
[alpha]-SPD               Var            Var            N
Signal transduction       N              [down arrow]   Var [down
 disorders                                               arrow]
Aspirin-like drug or      N              [down arrow]   [down arrow]
 defects of throm-                       [down arrow]    or Abs
 boxane synthesis
Myeloproliferative        N              N              N
 disorder
Scott syndrome            N              N              N
Uremia                    N              [down arrow]   [down arrow]
Thienopyridines (ti-      [down arrow]   Abs            N
 clopidine and clo-
 pidogrel)
GP IIb/IIIa antago-       [down arrow]   [down arrow]   [down arrow]
 nists                     or Abs         or Abs         or Abs

        Disorder              EPI          Collagen      Ristocetin

VWD                       N              N              [down arrow],
                                                         N or  [up
                                                         arrow]
Glanzmann throm-          [down arrow]   [down arrow]   N
 basthenia                 or Abs         or Abs
Bernard-soulier syn-      N              N              [down arrow]
 drome                                                   or Abs
Dense granule plate-      N or [down     N or [down     N
 let storage pool          arrow]         arrow]
 disorder ([delta]-SPD)
 [alpha]-SPD               N              Var [down     N
                                          arrow]
Signal transduction       Var [down      Var [down      N
 disorders                 arrow]         arrow]
Aspirin-like drug or      [down arrow]   [down arrow]   N
 defects of throm-                        or Abs
 boxane synthesis
Myeloproliferative        [down arrow]   N              N
 disorder                  or Abs
                           delayed lag
Scott syndrome            N              N              N
Uremia                    N/[down        N/[down        N
                           arrow]         arrow]
Thienopyridines (ti-      N              N              N
 clopidine and clo-
 pidogrel)
GP IIb/IIIa antago-       [down arrow]   [down arrow]   N
 nists                     or Abs         or Abs

        Disorder                  Other Studies

VWD                      Factor VIII:C, VWF antigen, VWF
                          risocetin cofactor, VWF multimers

Glanzmann throm-         Deficiency of GP IIb and/or GP
 basthenia                IIIa by flow cytometry
Bernard-soulier syn-     Macrothrombocytopenia
 drome                   Deficiency of GP Ib/IX/V (one or
                          more) by flow cytometry
Dense granule plate-     Decreased ATP release by
 let storage pool         lumiaggregometry
 disorder ([delta]-SPD)  [down arrow] Dense granule
                          mepacrine up-take and release
                          by flow cytometry
                         Decreased dense granules by
                          TEM
                         Abnormally high ATP/ADP ratio
                         Acquired SPD "exhausted platelets"
                          (CPB, DIC, TTP, HUS,
                          MPD)
                         Albinism in Hermansky-Pudlak
                          and Chediak-Higashi
                         Infections, small platelets seen
                          with Wiskott-Aldrich
[alpha]-SPD              Pale platelets on smear, [down
                          arrow] alpha granules by TEM,
                          [down arrow] P-selectin
Signal transduction      Decreased granule release with
 disorders                normal number of granules
                         Receptor defects may show [down
                          arrow] aggregation to EPI,
                          collagen only
                         [down arrow] G-protein activation,
                          tion, phospholipase C activation,
                          calcium mobilization,
                          pleckstrin, or tyrosine
                          phosphorylation
Aspirin-like drug or     Normal aggregation with prostaglandin
 defects of throm-        [G.sub.2] seen with
 boxane synthesis         aspirin or cyclooxygenase
                          deficiency
                         Decreased or absent prostaglandin
                          [G.sub.2] aggregation with
                          thromboxane synthetase deficiency

Myeloproliferative       Other abnormalities: [alpha] or
 disorder                 [delta]-SPD, cyclooxygenase
                          abnormality, other surface GP
                          derangements, [down arrow] or
                          [up arrow] aggregation to ADP,
                          collagen, spontaneous
                          aggregation
Scott syndrome           Defective platelet procoagulant
                          activity (ie, PF3)
                         Defective microparticle formation

Uremia                   Abnormal creatinine, BUN
                         Decreased PF3
Thienopyridines (ti-     History of clopidogrel or
 clopidine and clo-       ticlopidine therapy
 pidogrel)
GP IIb/IIIa antago-      History of treatment; abciximab,
 nists                    tirofiban, or eptifibatide
                         Increased receptor occupancy
                          by flow cytometry

* ADP indicates adenosine diphosphate; AA, arachidonic acid;
EPI, epinephrine; VWD, von Willebrand disease; Abs, absent;
GP, glycoprotein; ATP, adenosine triphosphate; TEM,
transmission electron microscopy; CPB, cardiopulmonary bypass;
DIC, disseminated intravascular coagulation; TTP, thrombotic
thrombocytopenic purpura; HUS, hemolytic uremic syndrome;
MPD, myeloproliferative disease; Var, variable response; PF3,
platelet factor 3; and BUN, blood urea nitrogen. Up and down
arrows indicate increased and decreased, respectively.
Gale Copyright: Copyright 2002 Gale, Cengage Learning. All rights reserved.