Hematology PPT Flashcards Unit 4 What is Hemostasis and Thrombosis? What is hemostasis? What is thrombosis? What are four components of hemostasis? What are three important component of protein in Hemostasis? What are several functionally processes are involved in hemostasis of a small blood vessel? What is the function of Arteries? What is the function of Veins? What is the function of capillaries? What are the characteristics of Arterioles? How were arterioles operated inside the capillaries? How was the microcirculation formed? What is the function of microcirculation? What is vasculature physiology role in hemostasis? The maintenance of circulation hemostasis is achieved through the process of balancing bleeding (hemorrhage) and clotting (Thrombosis) Prevent from bleeding or hemorrhage Clotting – Vascular system – Platelets (thrombocytes) – Blood coagulation factors – Fibrinolysis and ultimate tissue repair Complement system is a cascade produced in the liver. Kinin systems plays a role in inflammation, coagulation, sodium homeostasis. Protease inhibitors – Blood vessel spasm – Formation of a platelet plug – Contact between damaged blood vessel, blood platelet, and coagulation proteins – Development of a blood clot around the injury – Fibrinolytic removal of excess hemostatic material to reestablish vascular integrity Blood away from heart, arteries have the thickest walls of the vascular system. Blood come into the heart, veins are larger and have More irregular lumen than arteries Gas exchange Arterioles are the microscopic continuation of arteries that give off branches called metarterioles, which in turn join the capillaries. The walls become thinner as the arterioles approach the capillaries, with the wall of a very small arteriole consisting only of an endothelial lining and some smooth muscle surrounded by a small amount of connective tissue. The capillaries, arterioles, and venules constitute the major vessels of the microcirculation. As a unit, the microcirculation functions as the link between the arterial and the venous circulations. • The role of vasoconstriction in hemostasis 1 Hematology PPT Flashcards Unit 4 How does the vasoconstriction play a role in hemostasis? What is the endothelium? How does endothelium play a role in hemostasis? What was endothelium role in lysis of fibrin clots hemostasis? What is the prostacyclin role in hemostasis? How was the primary phase hemostasis activated after disruption of the endothelium? How was the second phase hemostasis activated after disruption of the endothelium? How was the third phase hemostasis activated after disruption of the endothelium? How was the final phase hemostasis activated after disruption of the endothelium? How many factors are required to maintain of Vascular Integrity hemostasis? What are the essential factors of vascular integrity in hemostasis? What will be happened if there is lack of • The role of the endothelium • Maintenance of vascular integrity When a smaller vessel, such as an arteriole, venule, or capillary, is injured, contraction occurs to control bleeding. This contraction of the blood vessel wall is called vasoconstriction. The endothelium contains connective tissues such as collagen and elastin. The endothelium is highly active metabolically and is involved in the clotting process by producing or storing clotting components. It is also rich with plasminogen activator, which, if appropriately stimulated, is released and activates plasminogen, which ensures rapid lysis of fibrin clots. Additionally, the endothelium elaborates prostacyclin, which is synthesized by the endothelium from prostaglandin precursors and strongly inhibits platelet aggregation and adhesion. Initially, rapid vasoconstriction for up to 30 minutes reduces blood flow and promotes contact activation of platelets and coagulation factors In the second phase, platelets adhere immediately to the exposed subendothelial connective tissue, particularly collagen. The aggregated platelets enhance sustained vasoconstriction by releasing thromboxane A2 and vasoactive amines, including serotonin and epinephrine In the third phase, coagulation is initiated through both the intrinsic and extrinsic systems Finally, fibrinolysis occurs following the release of tissue plasminogen activators (t-PAs) from the vascular wall. Fibrinolytic removal of excess hemostatic material is necessary to reestablish vascular integrity Vascular integrity or the resistance to vessel disruption requires three essential factors Circulating functional platelets Adrenocorticosteroids Ascorbic acid A lack of these factors produces fragility of the 2 Hematology PPT Flashcards Unit 4 essential factors in vascular integrity hemostasis? What will be the initial series observation of megakaryocytic cell? What will be the second series observation of megakaryocytic cell? What is the third series observation of megakaryocytic cell? What will be the last series observation of megakaryocytic cell? How were the platelets (thrombocytes) role in hemostasis of megakaryocytic development? vessels, which makes them prone to disruption. General characteristics of megakaryocytic development The developmental sequence of platelets Cellular ultrastructure of a mature platelet Platelet kinetics, life span, and normal values Mature platelets (thrombocytes) (metabolically active cell fragments) are the second critical component in the maintenance of hemostasis Where were the platelets (thrombocytes) These anuclear cells circulate in the peripheral found? blood after being produced from the cytoplasm of bone marrow megakaryocytes, the largest cells found in the bone marrow What are the stages of being formed platelets? 1) Megakaryoblast 2) Promegakaryocyte 3) Megakaryocyte a. Platelets Where were the bone marrow Bone marrow megakaryocytes are derived from megakaryocytes found? pluripotential stem cells How were the platelets formed inside the The sequence of development from cytoplasmic membrane? megakaryocytes to platelets is thought to progress from the proliferation of progenitors to polyploidization, that is, nuclear endoreduplication, and finally to cytoplasmic maturation and the formation of platelets Where was the initial process of Megakaryocytopoiesis proceeds initially megakaryocytopoiesis take place? through a phase characterized by mitotic division of a progenitor cell; Followed by a wave of nuclear endoreduplication. What is endoreduplication process of Endoreduplication is the process in which progenitors? chromosomal material (DNA) and the other events of mitosis occur without subsequent division of the cytoplasmic membrane into identical daughter cells What are two classes of progenitors that have 1) Burst-forming-unit megakaryocyte (BFU-M) been identified in the early development of 2) Colony-forming-unit megakaryocyte (CFUplatelets? M) What is the function of Burst-forming-unit The BFU-M is the most primitive progenitor megakaryocyte (BFU-M)? cell committed to megakaryocyte lineage What is the second stage of megakaryocyte The next stage of megakaryocyte development development of platelets? is a small, mononuclear marrow cell that 3 Hematology PPT Flashcards Unit 4 expresses platelet-specific phenotypic markers but is not morphologically identifiable as a megakaryocyte How well were the mononuclear marrow cells These transitional cells represent 5% of of platelets visible in cytoplasmic? marrow megakaryocyte elements. Have the transitional immature Some transitional immature megakaryocyte megakaryocyte cells been undergoing cells may be capable of cellular division, but endomitosis? most are nonproliferating while actively undergoing endomitosis. Described the final stage of megakaryocyte The final stage of megakaryocyte development development: is the morphologically identifiable megakaryocyte. These cells are readily recognizable in the marrow because of their large size and lobulated nuclei. These cells are polyploid. What is the size of a mature platelet? Platelets have an average diameter of 2 to 4 mm, with younger platelets being larger than older ones Does a platelet have a nucleus? In contrast to megakaryocytes, platelets have no nucleus. What is the characteristic of a platelet? The cytoplasm is light blue, with evenly dispersed, fine red-purple granules. What is the characteristic of an inactive An inactive or unstimulated platelet circulates as platelet? a thin, smooth-surfaced disc. What is platelet adhesion? When vascular injury exposes the endothelial surface, underlying collagen platelets adhere to the subendothelial collagen fibers, spread pseudopods along the surface, and clump together (aggregate). How quickly does platelet adhesion to 1-2 minutes after a break in the endothelium. subendothelial connective tissue, especially collagen happen? What’s the best test to use to determine Platelet aggregation. platelet function? What is a thrombus? When the bridges formed by fibrinogen in the presence of calcium produce a sticky surface on platelets, resulting in aggregation and then reinforced by fibrin is considered a thrombus. What substances can cause a blockage of a Prostaglandin E (PG E), adenosine, and pathway by an aggregation of platelets? nonsteroidal anti-inflammatory agents (e.g., aspirin). How are permanently anchored platelet plugs given additional consolidation and stabilization? With fibrinogen, under the influence of small amounts of thrombin. This process involves the precipitation of polymerized fibrin around each platelet resulting in a fibrin clot that produces an 4 Hematology PPT Flashcards Unit 4 Why is a platelet count a fundamental component in the evaluation of a patient? What are other fundamental evaluations of platelets? What are blood coagulation factors? What is blood coagulation a sequential process of? What are circulating factors that participate in the coagulation process designated by? How are the roman numerals effected if the enzymatic factor is activated? What are 3 assays that are conducted by specialized coagulation laboratories? What specimen is used for the One-stage quantitative assay for factors II, V, VII, and X? What factors does age serum contain? What factors does absorbed plasma contain? Who is the PT for One-stage quantitative assay for factors II, V, VII, and X conducted? Why is the percentage of factor activity plotted? For One-stage quantitative assay for factors VIII, IX, XI, and XII, what procedure is it based on? Who are the results expressed in One-stage quantitative assay for factors VIII, IX, XI, and XII? What is the factor VIII antibodies in irreversible platelet plug. Examination of the peripheral blood smear for platelet number and morphology is critical because many clinical clues may be obtained from an evaluation of platelet quantity and morphology. Platelet aggregation, platelet adhesion, and antiplatelet antibody assays. Blood coagulation factors are one of the most important of the assays. Basic concepts of blood coagulation, common characteristics of coagulation factors, characteristics of individual factors, the mechanism of coagulation, fibrin formation, laboratory assessment of blood coagulation factors, and assays for fibrin formation. Chemical reactions, involving plasma proteins, phospholipids, and calcium ions. Roman numerals. It’s a roman numeral followed by the suffix –a and the inactive enzymatic factor, zymogens are the roman numeral alone. 1) One-stage quantitative assay for factors II, V, VII, and X. 2) One-stage quantitative assay for factors VIII, IX, XI, and XII. 3) Factor VIII antibodies in hemophiliacs. Aged serum and absorbed plasma in the PT. Factors VII, IX, X, IX, and XII. Factors V, VIII, XI, and XIII Using specific factor-deficient plasma with specific dilutions of patient plasma. To construct an activity curve. The APTT assay. And it is based on the results of patient plasma to correct specific factor-deficient plasma. In percent activity on an activity curve. An ELISA technique that uses the binding of 5 Hematology PPT Flashcards Unit 4 hemophiliacs assay? What is Warfarin? Why do Warfarin drugs cause incomplete coagulation? Biological activity is significantly1)______ (decreased or increased) as revealed by the PT. The onset of action of most warfarin derivatives is between how many hours? The maximum affect occurs in approximately how many hours? What is the duration of action of Warfain? What is the mainstay of immediate therapy for acute PE. How does Heparin acts as an anticoagulant? Why is Heparin termed an antithrombin? What can Heparin cause? Before initiating heparin, what should patients be screened for? What should the baseline laboratory evaluation include? When is Heparin anticoagulation used? What are 2 other Antithrombin-Dependent inhibiters? What 2 new types of drugs are direct thrombin inhibitors? What are the new types of thromboplastins for measuring the PT? What are Fibrinogen assays useful for? antibodies in the plasma to solid phase antigen, which is subsequently detected by a human polyclonal IgG labeled with the alkaline phosphatase-p-nitrophenyl phosphate substrate system. The traditional oral anticoagulant (Coumadin). Warfarin drugs are vitamin K antagonists that interfere with the normal synthesis of factors II, VII, IX, and X as well as proteins C and S. Because they lack calcium-binding sites and cannot form enzyme substrate complexes. Thus, these factors are unable to function as procoagulants or anticoagulants. Decreased 8 and 12 hours 36 hours Approximately 72 hours. Heparin anticoagulation Heparin has no anticoagulant activity of its own but acts as an anticoagulant by accelerating the binding of antithrombin to target enzymes (e.g., thrombin and factor Xa). Because it helps prevent new thrombus formation and buys time for endogenous fibrinolytic mechanisms to lyse the clot. Bleeding, thrombocytopenia, and osteopenia. Clinical evidence of active bleeding CBC, platelets, PTT, PT, stool analysis for occult blood, and urine dipstick for hematuria. During percutaneous transluminal coronary angioplasty (PTCA) and cardiopulmonary bypass (CPB) to prevent clot formation. 1) Danaparoid (Orgaran) – 2) Fondaparinux (Arixtra) 1) Hirudin, lepirudin (Refludan) 2) Argatroban Mixtures of phospholipids and recombinantly derived human tissue factor. Detecting deficiencies of fibrinogen and alterations in the conversion of fibrinogen to 6 Hematology PPT Flashcards Unit 4 The normal value of 200 to 400 mg/dL may be decreased in? What is the normal titer of fibrinogen? What does the thrombin time test determines? What is the normal value of thrombin? What is the Reptilase time assay similar to? What is the D-Dimer? What are the 3 major hemostatic stages that must be functioning in order for D-dimers to form? All 3 of these stages require adequate formation of? What are 4 normal protective mechanisms against Thrombosis? In the blood circulation, the predisposition to thrombosis depends on? Several important biological activities normally protect the body against thrombosis. What are these activities? What does the normal flow of blood prevent? This mechanism reduces the chance of? What is another normal mechanism against inappropriate thrombosis? What does this process, along with the naturally occurring inhibitors do? Why is the removal of particulate material by the cells of the mononuclear phagocytic system also important? fibrin. Liver disease or the consumption of fibrinogen owing to accelerated intravascular clotting 1:128 to 1:256. The rate of thrombin-induced cleavage of fibrinogen to fibrin monomers and the subsequent polymerization of hydrogen-bonded fibrin polymers to form an insoluble fibrin clot. The normal value is less than 20 seconds. The thrombin time assay. Is a specific fragment generated from two crosslinked fibrin molecules after a clot has formed. 1) Coagulation to form the clot 2) Covalent cross-linking of fibrin by activated factor XIII 3) Fibrinolysis to dissolve the fibrin clot into smaller fragment Thrombin 1) Normal blood flow 2) Removal of activated clotting factors and particulate material 3) The natural anticoagulant systems 4) Cellular regulators The balance between procoagulant and anticoagulant factors. -The normal flow of blood -The removal of activated clotting factors and particulate material -Natural anticoagulant systems known to be operative in vivo: * Antithrombin III (AT-III) * Heparin cofactor II (HC-II) * Protein C and its cofactor, protein S -Cellular regulators The accumulation of procoagulant material. Local fibrin formation. The removal of activated clotting factors by hepatocytes from the blood. Limits intravascular clotting and fibrinolysis by inactivation of such factors as XIa, IXa, Xa, and IIa. It prevents the initiation of coagulation. 7 Hematology PPT Flashcards Unit 4 Why is the in vivo existence of natural anticoagulant systems essential? What are the natural anticoagulant systems? What are AT-III and HC-II? When activated, protein C is capable of degrading which activated factors? What is considered the major inhibitor of coagulation? What are the serpin superfamily of serine proteinase inhibitors? To prevent thrombosis. Antithrombin III, heparin cofactor II, and protein C and its cofactor, protein S Serine-protease inhibitors. V (Va) and VIII (VIIIa) in the presence of the cofactor protein S. Antithrombin III (AT-III) III (AT-III), alpha-1 antitrypsin, C1 inhibitor, alpha-2 antiplasmin, heparin cofactor II, and plasminogen activator inhibitor. Heparin is produced endogenously by? Mass cells. Heparin-like molecules are found in the? Endothelium. What are the two heparin-dependent thrombin AT-III heparin cofactor and heparin cofactor II, inhibitors present in human plasma? previously referred to as heparin cofactor A. The inhibitory activity of heparin cofactor II Heparin. is accelerated by? What does the inhibition of thrombin by The activity of thrombin or fibrinogen; also heparin cofactor II not limited to? inhibited are thrombin-induced platelet aggregation and release. In addition to thrombin, heparin cofactor II Chymotrypsin also inhibits? Heparin cofactor II does not significantly Blood coagulation factors IXa, Xa, and XIa or inhibit? plasmin. What 2 proteins are involved in one of the Protein C and S major natural anticoagulation systems in the body? Deficiency and/or alteration in either protein Predisposition to thrombosis. (C and/or S) have been clearly associated with? What does Protein C (A vitamin K–dependent A natural anticoagulant formed in response to plasma protein synthesized in the liver) thrombin generation. represents What is APC resistance (A hypercoagulable A point mutation in the factor V gene [factor V condition discovered in 1993) is associated (Leiden)]. with What does the mutation results in? The replacement of Arg506 with Gln(Q) in the factor V protein. This mutation slows the inactivation of what Factor Va by APC factor causing a hypercoagulable state? What is the most common cause of The Inherited thrombophilia. presence of factor V (Leiden)? It accounts for ____? % to _____? % of the 20% to 50% cases. Protein S is another vitamin K–dependent APC to express an anticoagulant effect. 8 Hematology PPT Flashcards Unit 4 plasma protein that is an essential cofactor for? Protein S does not require 1) ____? modification to function, but it can be regulated by 2) ____? What is becoming recognized as essential to the maintenance of hemostasis and thrombosis? What are 2 powerful mechanisms to limit the formation or spread of clotting and the reliquefication of clots? What have been demonstrated to be the major pathway for activation of coagulation? What has also been discovered about thrombi? What is the key enzyme in the conversion of soluble fibrinogen into fibrin What is required for the activation of the protein C system, which downregulates hemostasis? What is required for the activation of thrombin-activatable fibrinolysis inhibitor (TAFI), which is involved in the downregulation of the fibrinolytic process? What has led to the development of alternative schematic representations of the pathways that better represent the normal in vivo process of coagulation? What may cause abnormal bleeding? What is purpura? Purpura is characterized by? Purpura may be produced by a variety of vascular abnormalities such as? 1) Proteolytic 2) proteolysis. Cellular activities related to thrombosis Plasma possesses, and plasmin Tissue factor and factor VIIa in the extrinsic pathway That it has a far more important role as an activator and an inhibitor. Thrombin Thrombin Thrombin The cumulative result of new discoveries in coagulation has led. Disorders of the microcirculation, platelets, or plasma proteins. Abnormal bleeding involving the loss of red blood cells from the microcirculation. Hemorrhages into the skin, mucous membranes, and internal organs. 1) Purpura associated with direct endothelial cell damage. Endothelial damage may result from physical or chemical injury to the tissue caused by microbial agents such as in rickettsial disease or immunological antibody-mediated injury. Bacterial toxins produce deendothelialization induced by an endotoxin. Antibody vascular injury, vasculitis, may be induced by drug reactions, insect bites, or the activation of complement. 2) Purpura associated with an inherited disease of the connective tissue. Alterations of the vascular supportive framework can occur in disorders such as diabetes. 3) Purpura associated with decreased mechanical strength of the microcirculation. 9 Hematology PPT Flashcards Unit 4 Decreased strength can be seen in conditions such as scurvy and amyloidosis. 4) Purpura associated with mechanical disruption of small venules. The principal cause of this type of purpura is increased intraluminal pressure. This condition can be observed around the ankles with prolonged standing and may be caused by the presence of abnormal proteins in macroglobulinemias or hyperviscosity disorders. 5) Purpura associated with microthrombi (small clots). This type of disorder is associated with abnormal intravascular coagulation conditions. 6) Purpura associated with vascular malignancy. Purpura of this origin is observed in Kaposi sarcoma and vascular tumors. What are the disorders of platelet size? What is Wiskott-Aldrich syndrome? What is May-Hegglin anomaly? What is Alport syndrome? What is Bernard-Soulier syndrome? 1) Wiskott-Aldrich syndrome 2) May-Hegglin anomaly 3) Alport syndrome 4) Bernard-Soulier syndrome Is the smallest platelets seen. It is characterized by the presence of large platelets and the presence of Döhle-like bodies in the granulocytic leukocytes. Exhibits giant platelets and thrombocytopenia Is the largest platelets seen and is also referred to as giant platelet syndrome. 150 x 109/L to 450 x 109/L What is the normal range of circulating platelets? When the quantity of platelets decreases to Thrombocytopenia levels below 150 x 109/L to 450 x 109/L a condition known as _____? exists. Is the quantity of platelets increases then what Thrombocytosis is the result? Disorders of platelets can be classified as? Quantitative (thrombocytopenia or thrombocytosis) or qualitative (thrombocytopathy). If platelets are absent or severely decreased The presence of petechiae or purpura. below 100 x 109/L, what clinical symptoms are usually seen? What is Petechiae? Small purplish hemorrhagic spots on the skin or mucous membranes. What is Purpura? It is characterized by extensive areas of red or dark-purple discoloration. Most thrombocytopenic conditions can be 1) Disorders of production of platelets may be classified into major categories such as? caused by hypoproliferation of the megakaryocytic cell line or ineffective thrombopoiesis caused by acquired conditions 10 Hematology PPT Flashcards Unit 4 What is the most drug-induced thrombocytopenia? What are the predominant clinical symptoms of HI? What are the two types of HIT that exist? What replace the term idiopathic thrombocytopenic purpura? What is immune thrombocytopenia (ITP)? What can contribute to the increased utilization of platelets? What rapidly consumes platelets? What are some examples of disorders that can trigger the accelerated consumption of platelets? What can contribute to the thrombocytopenia? Why do vascular injuries (vasculitis) causes a decrease in platelets? What is thrombotic thrombocytopenic purpura (TTP)? What are the diagnostic of the diseases TTP? In TTP, coagulation testing will demonstrate 1) ____ prothrombin and 2) _____ partial thromboplastin time but 3) ______D-dimer and fibrinogen levels. TTP is in contrast to DIC, which demonstrates 1) _____ prothrombin time and 2) _____ partial thromboplastin time. What 3 types of TTP have been identified? or hereditary factors. 2) Disorders of destruction, including decreased megakaryocytopoiesis and ineffective platelet production, and disorders of utilization. It may result from a number of mechanisms—drugs, bacterial sepsis, etc. 3) Disorders of platelet distribution and dilution Heparin-Induced Thrombocytopenia (HIT) Thrombocytopenia and thrombosis 1) Nonimmune HIT: Type I 2) Immune HIT: Type II New standard nomenclature, immune thrombocytopenia (ITP) ITP is an acquired immune-mediated disorder characterized by isolated thrombocytopenia (platelet count <100 x 109/L) and the absence of any obvious initiating and/or underlying cause of the thrombocytopenia. Intravascular coagulation, vascular injury or occlusion, and tissue injury. DIC (disseminated intravascular coagulation) Trauma, obstetrical complications, and microbial sepsis. In the case of bacterial sepsis, thrombin-induced platelet aggregation in vivo contributes to thrombocytopenia. Because of the direct consumption of platelets at the sites of endothelial loss without appreciable depletion of clotting factors such as fibrinogen. Is a clinical syndrome with a high mortality rate that is characterized by formation of microthrombi in the microvasculature. The hematological findings of thrombocytopenia and red blood cell schistocytes. 1) Normal 2) Activated 3) Elevated 1) Abnormal 2) Activated 1) Idiopathic 11 Hematology PPT Flashcards Unit 4 A platelet distribution disorder can result from? This type of thrombocytopenia develops when? Disorders that may produce splenomegaly with resultant splenic pooling or delayed intrasplenic transit include? 2) Secondary 3) Inherited (Upshaw-Shculman) A pooling of platelets in the spleen, which is frequent if splenomegaly is present. More than a double or triple increase in platelet production is required to maintain the normal quantity of circulating platelets. Alcoholic or posthepatic cirrhosis with portal hypertension, lymphomas, and leukemias and lipid disorders, for example, Gaucher disease. Generally defined as a substantial increase in circulating platelets over the normal upper limit of 450 x109/L. Thrombocytosis can be classified into three major categories which are? 1) Hereditary or familial thrombocytosis associated with germline mutations of the thrombopoietin (THPO) gene in the thrombopoietin receptor (MPL) gene 2) Thrombocytosis associated with myeloproliferative neoplasms and/or myelodysplastic disorders (clonal thrombocytosis associated with somatic mutations of JAK2 [V617F], MPL, and additional currently unknown genes) 3) Reactive (secondary thrombocytosis) If platelets are normal in number but fail to perform effectively then….? In addition to both individual and family medical histories, laboratory tests are critical for? What do Laboratory tests of platelet function include? A platelet dysfunction exists. Three separate categories of platelet dysfunction can be identified based on? Hyperactive platelets associated with hypercoagulability and thrombosis make up an additional category of what? Examples of inherited disorders include? Determining a platelet dysfunctional diagnosis. bleeding time, clot retraction, platelet aggregation, platelet adhesiveness, and antiplatelet antibody assay. Etiology- the more common acquired causes and the less frequent hereditary causes. Abnormal platelet function. Bernard-Soulier syndrome and Glanzmann thrombasthenia. Vascular response and platelet plug formation. What is responsible for the initial phases of hemostasis. Subsequent to these activities what is initiated The clotting factors to form the fibrin clot? Fibrin formation can occur if ? The activity of various factors is at least 30% to 12 Hematology PPT Flashcards Unit 4 40% of normal. A coagulation factor Bleeding and defective fibrin clot formation are frequently related to what? Disorders of the blood coagulation factors can 1) Defective production may be related to a be grouped into three categories, which are? deficiency of vitamin K. Classic hemophilia (hemophilia A) and von Willebrand disease are examples of hereditary disorders that represent functionally inactive factor VIII. 2) Excessive destruction 3) Pathological inhibition What is hemophilia ? When a clotting factor is either missing or is not working to its full ability causing for a greater chance of prolonged bleeding as well as the blood taking longer to clot Is there a cure for hemophilia No cure is a life long disease but can be treated and managed What causes hemophilia ? It is an X-linked autosomal recessive disease this means that the women are the carries and the men are affected What is hemophilia A? Clotting factor 8 is either deficient or not present What is hemophilia B? Clotting factor 9 is deficient or not present What is Von willebrand Disease ? This is when vonon willebrand clotting factor is not present this can affect both men and women What is mild severity of hemophilia? What are manifestation of hemophilia ? What types of tests are done to diagnose hemophilia? True or false ? von willebrand disease may be an acquired or inherited disorder ? What distinguish von willebrand disease from classic factor V111: C ? Von wildebrand’s disease occurs in ____of population _________is a major relatory mechanism of hemostasis Pts who have herediatary protein Cor S 6%-50% of the clotting factor is active Hemarthrosis- bledding in the joint Hematuria- blood in the urine Ecchymosis-bruising Partial thromboplastin time (ptt) Typically prolonged Prothrombin time (pt) Bleeding time Fibrinogen level Platelet count Factor 8,9 and van willebrand assay Genetic testing True A deficiency of factor V111 procoagulant activity 1-2% Protein C (PC) Skin necrosis 13 Hematology PPT Flashcards Unit 4 deficiency have of what if take Coumadin? Is homozygous antithrombin deficiency compatible with life? What is DIC ? What include triggering events that may predispose patients to DIC ? What are some examples of clinical situations in which tissue thromboplastin can activate coagulation ? Through what 4 ways does the body stop the clotting process ? How does Heparin help inhibit the clotting process ? How does TFPI (Tissue Factor Pathway Inhibitor) work ? How does fibrinolysis happen ? What is disseminated intravascular coagulation What are some characteristics of DIC What is DIC caused by? Is DIC a primary or secondary? No DIC is a complication or intermediary phase of many diseases and does not constitute a disorder in itself Alterations in the endothelium direct activation of fibrinogen release of thromboplastin-like substances and erythrocyte or platelet destruction Extravascular trauma abruption placentae advanced malignancy leukemia and retained fetal synf=rome TFPI antithrombin III Protein C / Protein S / Thrombomodulin Plasminogen catalyzes antithrombin III's inactivation of IX, X, XI, and thrombin complexes with factors VIIa, TF, and Xa, and inactivates Xa TPA (Tissue Plasminogen Activator), which is on the surface of endothelial cells, converts plasminogen--> plasmin. Plasmin is a general protease that loves blasting proteins w/lysine residues. Fibrin has a lot of lysine residues, so it looks particularly sexy to plasmin (high affinity). It turns what was once a cold-hearted insoluble clot all soluble and easygoing, and Ddimers float around. A life-threatening uncontrolled activation of the coagulation system 1. Widespread small vessel thrombosis 2. Consumption of platelets and factors 3. Uncontrolled bleeding due to depletion of platelets and coagulation factors 4. organ failure Various clinical conditions that activate clotting mechanisms: Infections Hemorrhage Shoc k Septic shock DIC is never a primary condition. It is always secondary to another problem (ex: a person in septic shock would have DIC as a consequence 14 Hematology PPT Flashcards Unit 4 Describe the onset and end result of DIC Why is DIC considered a consumptive colaguopathy ? What does consumption of coagulation factors and platelets result in? Why does hemorrhage occur? What occurs in all preestablished clots? How type of cycle is DIC? Stimulation of clottiing --> Thrombosis emboli formation --> Vascular occlusion --> Organ/tissue ischemia --> Infarction --> Necrosis Because there's a primary stimulus that stimulates the body to think that it has a major issue where it needs to form clots (may be true in some cases for a trauma patient that is actively bleeding) As the small clots consume coagulation proteins and platelets, normal coagulation is disrupted and abnormal bleeding occurs from the skin Bleeding Because platelets and coagulation factors are no longer present Clot is programmed to break down and Fibrinolysis starts Vicious cycle of clotting and bleeding What are two abnormal coagulation profiles that decrease? Thrombocytopenia (decrease in platelets) and Decreased clotting factors (any Roman numeral, fibrinogen, or thrombin) What are two abnormal coagulation profiles that increase? What are some treatments for DIC? Increased clotting measurements/ amount of time (Prothrombin time, Partial prothrombin time, bleeding times) and Increased Fibrin Degradation Products (FDPs) or Fibrin Split Products (FSPs) -->D-Dimer is specifically measurable with FDP to indicate Fibrinolysis Monitor CBC for evolving and worsening anemia *The anemia will normally be Normocyctic normochromic from acute hemorrhage situation What are some treatments for DIC? What is responsibility of hematology Remove percipitating primary) problem (because DIC is secondary consequence Restore balance between coagulation and fibrinolysis Maintain organ viability Administer anti- coagulants (heparin) Gross physical examination, total cell count, How should DIC be evaluated? 15 Hematology PPT Flashcards Unit 4 technicians and technologists? microscopic examination, and other special tests Where are chemical analyses and microbial and cytological examinations? Examinations are generally performed in the chemistry, microbiology, and cytology departments, respectively Where sterile body fluid can be found? It found in various body cavities under normal conditions. In diverse disorders and disease processes, the quantity of these fluids can increase significantly. Fluid specimens aspirated from different anatomical sites and examined for the total number of cells, differentiation of cell types, chemical composition, and microbial contents. How all body fluids are should be handled with caution? Standard precautions must be practiced. What are the type of examination performed on the body fluid depends on? the body fluid depends on the source of the specimen. However, a portion of the examination of cerebrospinal fluid (CSF); serous fluids from the pleural, pericardial, and peritoneal cavities; synovial fluid; and seminal fluid is frequently performed in the hematology laboratory. What the most is commonly analyzed body fluid? Cerebrospinal Fluid How cerebrospinal fluids (CSF) are acts? CSF acts as a shock absorber for the brain and spinal cord, circulates nutrients, lubricates the central nervous system (CNS), and may also contribute to the nourishment of brain tissue. Where cerebrospinal fluids (CSF) are circulates through? The CSF circulates through the ventricles and subarachnoid space that surrounds both the brain and the spinal cord. The ventricles consist of four hollow, fluid-filled spaces inside the brain. 16 Hematology PPT Flashcards Unit 4 What CSF is production? What the plexus is composed of two layers? What make constitute the anatomical structure of the blood-brain barrier? Where CSF is found? How many the total maximum volume of CSF in adults is? How to measure CSF? How a specimen of CSF is examined? A lateral ventricle lies inside each hemisphere of the cerebrum Primarily a function of the choroid plexus, with a smaller proportion being derived from the ependymal lining and perivascular spaces The ependyma (the lining epithelium of the ventricle) and the pia mater. The plexuses in the lateral ventricles are the largest and produce most of the CSF. The choroid plexus epithelium and the endothelium of capillaries in contact with CSF. Inside all the ventricles, in the central canal of the spinal cord, and in the subarachnoid space around both the brain and the spinal cord. About 150 mL. The maximum volume in neonates is approximately 60 mL. Introducing a needle into the subarachnoid space makes it possible to measure CSF pressure and to obtain fluid for analysis. A specimen of CSF is examined visually and microscopically. The total number of cells can be enumerated, and the types of cells can be morphologically distinguished. .Gross physical examination • • • • What normal CSF is contains? How many WBC count are low or high? Turbidity Color Viscosity Microscopic enumeration examination: cellular A few mononuclear cells (lymphocytes and monocytes) and rare ependymal cells. Very few leukocytes should be seen in normal CSF. The generally accepted total number of WBCs in the CSF ranges from 0 to 5 cells/mL or 0 to 5 × 106/L. A value of up to 20 x106/L can be considered within the range of normal for children and adults. Neonates have a higher normal range, 0 to 30 cells × 106/L. Occasionally, the WBC count may be very low (0 to 20 × 106/L), but the CSF is turbid because of a high concentration of bacteria. In acute, untreated bacterial meningitis, the CSF leukocyte count usually ranges from 100 to 10,000 cells × 106/L. Very high WBC counts 17 Hematology PPT Flashcards Unit 4 Cells Unique to the Cerebrospinal Fluid • Body fluids that are analyzed in the hematology laboratory. What is an effusion? What transudates generally? What exudates are? What is pleural fluid? How difference color between transudates and exudates? Erythrocyte and leukocyte counts How WBC count are effect transudates and exudates Cell types that can be encountered in the examination of a Wright-Giemsa–stained specimen include the following: (greater than 50,000 × 106/L) are unusual and suggest intraventricular rupture of a brain abscess. Ependymal cells Choroidal cells Pleural, Peritoneal, and Pericardial Fluids An effusion is an abnormal accumulation of fluid in a particular space of the body. Effusions in the pleural, pericardial, and peritoneal cavities are divided into transudates and exudates. Fluid has accumulated because of the presence of a systemic disease. Associated with disorders such as inflammation, infection, and malignant conditions involving the cells that line the surfaces of organs (e.g., lung or abdominal organs). normally produced by the parietal pleura and absorbed by the visceral pleura as a continuous process. Although healthy individuals form 600 to 800 mL of fluid daily, the normal volume of fluid in each pleural space is estimated at less than 10 mL. Aspiration of pleural fluid is referred to as thoracentesis. Transudates are usually clear, are pale yellow, and do not clot. In comparison, exudates can display a range of colors depending on the associated disorder. They are of limited value in the differential diagnosis of pleural effusions. A massively bloody (hemorrhagic) effusion in the absence of trauma almost always suggests malignancy, or occasionally pulmonary infarct. Extremely elevated total leukocyte (WBC) counts of 50.0 × 109/L or higher are consistent with a diagnosis of empyema. In general, WBC counts less than 1.0 × 109/L are associated with transudates, and WBC counts greater than 1,000 × 109/L are associated with exudates. Neutrophils (PMNs) Eosinophils Basophils Lymphocytes Plasma cells Mononuclear phagocytes (monocytes, 18 Hematology PPT Flashcards Unit 4 What is peritoneal Fluid? How about an abnormal amount of fluid (an effusion)? What are result of the collection of fluid in the peritoneal cavity, ascites? What are causes of Peritoneal Effusions? How can effusions that may conform with the definition of transudates? What the laboratory are criteria for distinguishing transudates from exudates? How difference baboratory Analysis of transudates and exudates? What includes of an evaluation of ascitic fluid in? What is total WBC count higher than 0.3 × 109/L? histiocytes, and macrophages) Mesothelial cells (normal, reactive, atypical, and malignant) Metastatic tumor (malignant) cells A body fluid that is analyzed in the hematology laboratory. It can accumulate in the peritoneal cavity if the balance between fluid formation and reabsorption is altered by a disease process. Results from increased hydrostatic pressure in the systemic circulation, increased peritoneal capillary permeability, decreased plasma oncotic pressure, or decreased fluid reabsorption by the lymphatic system. The causes of peritoneal effusions range from disorders and diseases that directly represent involvement of the peritoneum, such as bacterial peritonitis, to abdominal conditions that do not directly involve the peritoneum, such as hepatic cirrhosis, cirrhosis, congestive heart failure, Budd-Chiari syndrome, hypoalbuminemia (caused by nephrotic syndrome or protein-losing enteropathy malnutrition), and miscellaneous disorders such as myxedema, ovarian diseases, pancreatic disease, and chylous ascites. Can be associated with congestive heart failure, hepatic cirrhosis, and hypoproteinemia. less clearly defined for peritoneal (ascitic) fluid than for pleural fluid. Transudates are usually clear and pale yellow. Exudates are cloudy or turbid because of an increased concentration of leukocytes, elevated protein levels, and occasionally microorganisms. Exudates may be seen in peritonitis, cases of perforated or infarcted intestine, and pancreatitis. gross inspection, total cell count, microscopic examination of sediment for cell differentiation, cytological studies, chemical analysis for constituents such as total protein and lactic dehydrogenase, and microbial culture. Total WBC counts are of limited value in differential diagnosis, but a total WBC count higher than 0.3 × 109/L is considered to be 19 Hematology PPT Flashcards Unit 4 What is A wide variation in the peritoneal WBC count? An abnormal accumulation of fluid in the cavity, a ?, is most frequently caused by damage to the lining of the cavity and increased capillary permeability. In addition, in acute pericarditis, interference with ? venous and lymphatic drainage predisposes the patient to effusion development. A wide variety of diseases and disorders can produce ? effusion. Normal fluid is 1 and 2. Hemorrhagic (bloody) effusions may result from a variety of abnormal conditions or from aspiration of intracardiac blood into the specimen. On visible examination, a hemorrhagic effusion should not form clots in a plain (nonanticoagulant) tube, but aspirated blood usually exhibits clotting. A milky-appearing effusion may be a true or ? fluid. Erythrocyte and leukocyte cell counts are of limited value in the differential diagnosis of a ?. A ? is a body fluid that is analyzed in the hematology laboratory. Seminal fluid (semen) is examined 1 and 2. Semen analysis is the primary test for the evaluation of male infertility. Sperm concentration, motility, and morphology can be used to classify men as what other 3 categories ? also can be analyzed for a variety of reasons, including artificial insemination protocols, postvasectomy assessment, and evaluation of probable sexual assault. A fresh specimen is needed. It is mandatory that the specimen be kept at ?°C and examined within 1 hour of collection. After 60 minutes of storage in a plastic container, sperm motility is significantly reduced. Is the following macroscopic or microscopic? A fresh specimen should be examined for abnormal. It seen in patients with chronic liver disease because of extracellular shifts in fluid associated with ascites formation or resolution. pericardial effusion pericardial pericardial 1. Transparent 2. pale yellow pseudochylous pericardial effusion Seminal fluid 1. macroscopically 2. microscopically subfertile, of indeterminate fertility, or fertile. Semen 37°C Macroscopic 20 Hematology PPT Flashcards Unit 4 coagulum formation, color, pH, volume, and viscosity. Is the following macroscopic or microscopic? microscopic Enumeration of the number of sperms and examination of the morphological characteristics of the cells are routinely performed procedures. Other microscopic procedures include motility, viability, and agglutination studies. What type fluid is a transparent, viscous fluid Synovial (joint) secreted by the synovial membrane. This fluid is found in joint cavities, bursae, and tendon sheaths Arthrocentesis constitutes a liquid biopsy joint of the ?. Disorders such as gout, calcium pyrophosphate dihydrate (CPPD) deposition disease, and septic arthritis can be diagnosed definitively by synovial fluid analysis and may allow for consideration or exclusion of rheumatoid arthritis and ? Synovial fluid is readily obtained by ? from most joints. Frequent sites of ? include the knee, shoulder, elbow, wrist, interphalangeal joints, hip, and ankle. Routine analysis of synovial fluid should include microscopic examination of a wet preparation, crystals, Gram stain, and ? culture. If the fluid is very turbid, or if septic arthritis is considered for other reasons, the specimen should be sent for 1. And 2.? True or False? A Gram stain is needed if a high likelihood of infection exists. Other observations or procedures can include volume and appearance, viscosity, mucin test, chemical analysis for protein, and glucose. This is what type of examination? Normal, noninflammed joints have small quantities of clear or transparent fluid. This fluid is viscous and slightly alkaline and, if normal, does not clot. This describes what? The best use of the ? test is to distinguish the anatomical origin of bloody or other fluids. The presence of a mucin clot implies that it is systemic lupus erythematosus (SLE). aspiration microbiological 1. Gram stain 2. culture – True – Gross – appearance – mucin clot 21 Hematology PPT Flashcards Unit 4 synovial fluid. A differential cell count on a body fluid – hemocytometer should be performed on stained smears prepared from a concentrated preparation— not in a ?. Some of the techniques of sediment preparation and staining are different for body fluids than for blood. Morphological descriptions of cells 1. Wright encountered in body fluids reflect their 2. Wright-Giemsa stain. microscopic appearance with 1. or 2? The coloration of cells with Papanicolaou • Cytological stain is somewhat different. Papanicolaou stain is a commonly used what type of stain? The following describe what type of CLSI Pricedural Format format? • Procedure title and specific method • Test principle including type of reaction and the clinical reasons for the test • Specimen collection and preparation • Reagents, supplies, and equipment • Calibration of a standard curve • Quality control • Procedure • Calculations • Reporting results (normal values) • Procedure notes including sources of error, clinical applications, and limitations of the procedure • References T or F? All specimens should be treated with caution. All blood, tissues, and blood derivatives should be considered potentially infectious. A stained smear is examined to determine the percentage of each type of leukocyte present and assess the erythrocyte and platelet ?. Increases in any of the normal leukocyte types and the presence of immature leukocytes or erythrocytes in peripheral blood are important diagnostically in a wide variety of inflammatory disorders and ? Erythrocyte abnormalities are clinically True morphology leukemia. anemias. 22 Hematology PPT Flashcards Unit 4 important in various ?. Platelet size irregularities are suggestive of particular what type of disorders? Disorders associated with increases or decrease in normal leukocyte types? • • Neutrophils Bacterial infections, inflammation, stress, and chronic leukemia • Lymphocytes Viral infections, whooping cough, and chronic leukemia • Monocytes Tuberculosis, rheumatoid arthritis, and fever of unknown origin • Eosinophils Active allergies and invasive parasites • Basophils Ulcerative colitis and hyperlipidemia thrombocyte Increase The packed cell volume (PCV) is a measurement of the ratio of the volume occupied by the RBCs to the volume of whole blood in a sample of ?. Following centrifugation, this ? is measured and expressed as a percentage or decimal fraction. The ? is used to calculate the mean corpuscular volume (MCV) and the mean corpuscular hemoglobin concentration (MCHC). True or False? The PCV is used to detect anemia, polycythemia, hemodilution, or hemoconcentration. capillary or venous blood Name the 11 elements that the CLSI Procedural Format must have. • • • • • • • • • • • ratio PCV True Procedure title and specific method Test principle Specimen collection and preparation Reagents, supplies, and equipment Calibration of a standard curve Quality control Procedure Calculations Reporting results (normal values) Procedure notes References 23 Hematology PPT Flashcards Unit 4 What must be included in Test Principle? • What must be included in the Procedure Notes? How must all specimens be treated? • • Type of reaction and Clinical reasons for the test Sources of error, clinical applications, and limitations of the procedure. With Caution What specimens should be considered potentially infectious? What are the 5 basic procedures in Hematology? • All blood, tissues, and blood derivatives. What is the principle of Leukocyte Differential Count? Increases in any of the normal leukocyte types and the presence of immature leukocytes or erythrocytes in peripheral blood are important diagnostically in what? What are erythrocyte abnormalities are clinically important in? Platelet size irregularities are suggestive of what? Disorders associated with Neutrophils are? Disorders associated with Lymphocytes are? Disorders associated with Monocytes are? Disorders associated with Eosinophils are? Disorders associated with Basophils are? How to you read a slide when doing a differential count? What is packed cell volume (PCV)? How is the ratio is measured and expressed? What does the PCV calculate? What are the PCV Reference values? What is the PCV is used to detect? What are the 3 calculations of RBC Indices? • Leukocyte differential count • Packed cell volume of whole blood • Red blood cell indices • Reticulocyte count • Sedimentation rate of erythrocytes To determine the percentage of each type of leukocyte present and assess the erythrocyte and platelet morphology A wide variety of inflammatory disorders and leukemia. Various anemias Thrombocyte disorders Bacterial infections, inflammation, stress, and chronic leukemia Viral infections, whooping cough, and chronic leukemia Tuberculosis, rheumatoid arthritis, and fever of unknown origin Active allergies and invasive parasites Ulcerative colitis and hyperlipidemia Up, Across, Down, Across A measurement of the ratio of the volume occupied by the RBCs to the volume of whole blood in a sample of capillary or venous blood As a percentage or decimal fraction. The mean corpuscular volume (MCV) and the mean corpuscular hemoglobin concentration (MCHC). Males, 0.47 L/L ± 0.07 Females, 0.42 L/L ± 0.05 Anemia, polycythemia, hemodilution, or hemoconcentration Mean corpuscular volume 24 Hematology PPT Flashcards Unit 4 What does Mean Corpuscular Volume (MCV) do? What is the MCV Reference value? What does Mean Corpuscular Hemoglobin (MCH) do? What is MCH directly proportional to? What is the MCH Reference value? What does Mean Corpuscular Hemoglobin Concentration (MCHC) do? What does the MCHC defined as the ratio of? What is the MCHC Reference value? How do you calculate MCHC? What is a reticulocyte? What is Renumeration of reticulocytes important in? What type of stain is new methylene blue N? What does new methylene blue N cause? What is Reticulum? What is the Reference value of Reticulocytes in methyl blue? Name 2 disorders associated with abnormal reticulocyte results. What does Sedimentation rate measures? What does Sedimentation rate depend on? What is the clinical value of this procedure? What are 2 other names for Sedimentation Rate of Erythrocytes? What is ESR directly proportional to? Which sediments more slowly, microctytes or Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration It expresses the average volume (size) of an erythrocyte 80 to 96 fL It expresses the average weight (content) of hemoglobin in an average erythrocyte. The amount of hemoglobin and the size of the erythrocyte. 27 to 32 pg It expresses the average concentration of hemoglobin per unit volume of erythrocytes. The weight of hemoglobin to the volume of erythrocytes. 32% to 36% Hemoglobin (g / dL) MCHC g / dL PCV or hematocrit (L / L) An erythrocyte still possessing RNA Assessing the status of erythrocyte production in the bone marrow (erythropoiesis). Supravital stains Ribosomal and residual RNA to co-precipitate with the few remaining mitochondria and ferritin masses in living young erythrocytes to form microscopically visible dark-blue clusters and filaments. Clusters and filaments. 0.5% to 1.5%; neonates, 2.5% to 6.5% Decreased reticulocytes - Aplastic anemia Increased reticulocytes - Acute blood loss The rate of settling of erythrocytes in diluted human plasma An interrelationship of variables, such as the plasma protein composition, the concentration of erythrocytes, and the shape of the erythrocytes. It is in the diagnosis and monitoring of inflammatory or infectious states. ESR and Sedrate The weight of the cell aggregate and inversely proportional to the surface area. Microcytes 25 Hematology PPT Flashcards Unit 4 macrocytes? Name 3 factors that contribute to decrease in ESR. Name 3 irregular erythrocyte shapes. When is an increased ESR value seen What are some clinical conditions associated with increased ESR values? Give an example of tissue necrosis. Name 10 Specialized hematology procedures. What test is Acid serum lysis called? How do you test for hemolysis? Weak acid is used in specific serum cell mixtures to maximize what? The presence of hemolysis may be observed in cases such as? How is a marrow examined histologically? Who preforms a bone marrow aspiration? What does the physician examine in the bone marrow? What Stain(s) is used on bone marrow examinations? What is bone marrow examination? Give an example of a bone marrow The removal of fibrinogen by defibrination Erythrocytes with abnormalities o Erythrocytes with irregular shapes Sickle cells Spherocytes Hinder rouleaux various abnormal blood conditions: rouleaux, increased fibrinogen levels, a relative increase of plasma globulins caused by the loss of plasma albumin, and an absolute increase of plasma globulins. a) Anemia, infections, inflammation, tissue necrosis, pregnancy, and some types of hemolytic anemia b) Myocardial infarction. • Acidified serum lysis test • Bone marrow examination • Donath-Landsteiner screening test • Glucose-6-phosphate dehydrogenase activity in erythrocytes • Hemoglobin electrophoresis • Hemoglobin S screening test • Malarial smears • Monospot test • Osmotic fragility of erythrocytes • Sucrose hemolysis test HAM test Erythrocytes are incubated with fresh and heated serum Hemolytic activity. Antibody-sensitized coated erythrocytes, spherocytes, or paroxysmal nocturnal hemoglobinuria (PNH). By using a hematoxylin-eosin (H&E) stain. A physician Cellular activities of the marrow Wright-Giemsa stain Prussian blue and cytochemical stains Is valuable in the diagnosis of disorders and in the study of leukemia’s and some types of anemia specifically involving the marrow Multiple myeloma. 26 Hematology PPT Flashcards Unit 4 examination. What's the normal distribution of bone marrow cells in an adult? What is the myeloid:erythroid ratio? The Donath-Landsteiner screening antibody test requires? A positive result shows? True or False: The observed rate of appearance of bright fluorescence is proportional to the blood G6PD activity. What is one of the most prevalent hereditary erythrocyte enzyme deficiencies? A deficiency of G6PD can produce? The movement of charged particles on various media under the influence of an electric current is called? True or False: Particles move at the same speeds because of their weight and electric charge. How is hemoglobin electrophoresis useful? A hemolysate prepared from intact erythrocytes is placed on which medium? What describes a biphasic system with an upper organic phase of toulene and a lower aqueous phase of phosphate buffer, saponin, and reducing agents? How are erythrocytes lysed? The released hemoglobin is reduced by? The resulting colors of the aqueous phase and the interface phase allow for the differentiation of which hemoglobin types? True or False: Detection of the abnormal Hb S is diagnostic of sickle cell disease. The detection of this heterozygous state is important to which individuals? Thick and thin blood smears are prepared, stained and examined microscopically for which malaria types? What is the diagnosis of malaria based on? What is based on agglutination of horse erythrocytes by heterophile antibody in Rubricytes, neutrophils, lymphocytes, monocytes, megakaryocytes, reticulum cells, plasma cells, myeloid, erythroid M/E ratio 2.5:1 Cold incubation to exhibit hemolysis in the patient's serum. Paroxysmal cold hemoglobinuria(PCH), the rarest form of autoimmune hemolytic anemia TRUE Glucose-6-Phosphate Dehydrogenase (G6PD) deficiency Drug or stress induced hemolytic anemia Electrophoresis FALSE To identify and quantify hemoglobin variants and abnormal quantities of hemoglobin fractions Cellulose acetate Hemoglobin S Screening Test By toluene and saponin Sodium hydrosulfite AA, AS, and SS TRUE Long distance runners, test pilots Plasmodium malariae, P. vivax, P. falciparum, P. ovale The demonstration of the Plasmodium species in the blood. Monospot Test 27 Hematology PPT Flashcards Unit 4 infectious mononucleosis? How is the specific heterophile antibody distinguished from the Forssman type? Serum or plasma is absorbed with? What does Guinea pig kidney absorb? What does beef erythrocytes absorb? A positive reaction for heterophile antibody occurs when? Agglutination of horse RBCS by the absorbed patient specimen indicates____ for heterophile antibody Guinea pig kidney contains only the ___ and beef erythrocytes contain only the __ Whole blood is added to varying concentrations of sodium chloride solution and allowed to incubate at room temperature in the ____ test? How is the amount of hemolysis determined? The main factor in this procedure is? The shape of the erythrocyte depends on? True or False: The fragility of erythrocytes is increased when the rate of hemolysis is increased True or False: If the rate of hemolysis is increased, the erythrocytic fragility is decreased Which test demonstrates the sensitivity of erythrocytes to the protein complement? When do erythrocytes in PNH lyse? Which cells are tartrate resistant? Most of the acid phosphatase isoenzyme is inhibited by? Where are peripheral blood or bone marrow smears fixed and incubated? True or False: Lymphocytes display more activity than other leukocytes. Leukocyte alkaline phosphatase(LAP) activity is often used to distinguish? In a variety of conditions LAP activity can be By differential absorption of the patient's serum Both guinea pig kidney and beef erythrocyte stroma Only heterophile antibodies of the Forssman type Only the heterophile antibody of infectious mononucleosis Agglutination of horse RBCs by the absorbed patient specimen happens A positive reaction Forssman antigen, antigen associated with infectious mononucleosis Osmotic fragility test By examining the supernatant fluid either visually or with a spectrophotometer The shape of the erythrocyte The volume, surface area, functional state of the erythrocytic membrane TRUE FALSE Sucrose Hemolysis Test When exposed to serum solutions of low ionic strength containing complement Hairy cell leukemia, Sezary syndrome, some Tcell acute lymphoblastic leukemias L-tartaric acid In a solution of naphthol As-BI phosphoric acid and fast garnet GBC salt FALSE Leukemoid reactions and chronic granulocytic (myelogenous) leukemia Increased, normal, or decreased 28 Hematology PPT Flashcards Unit 4 ___? What is the leukocyte alkaline phosphatase (LAP) procedure often used to distinguish? Which best describes esterases? This method distinguishes what? How are positive reacting cells differentiated from monocytes? In the alpha- naphthyl acetate esterase procedure, which is incubated with naphthol AS-D chloroacetate? Naphthol compounds are coupled with what? Which represent unstable types of hemoglobin, denatured by dyes and appear as intraerythrocytic stained bodies? How are Heinz bodies formed? What is the significance of increased numbers of Heinz bodies? What happens shortly after birth to the amount of fetal hemoglobin (Hb F)? Increased amounts of Hb F are found in which conditions? What happens when leukocytes are treated with Periodic Acid–Schiff (PAS)? How is Myeloperoxidase (MP) detected? Leukemoid reactions and chronic granulocytic (myelogenous) leukemia Ubiquitous in nature and encompass a variety of different enzymes acting on selective substrates Cells of the granulocytic series from cells of the monocytic series, useful in differentiation of leukemias Sodium fluoride is incorporated with the incubation system. Blood or bone marrow smears Diazonium salt, forming highly colored deposits at the sites of enzyme activity Heinz bodies When the glycolytic enzymes in the erythrocytes are unable to prevent the oxidation of hemoglobin Unstable forms of hemoglobin in hemolytic disorders such as G6PD. It decreases to low levels Sickle cell anemia, thalassemias, and hereditary persistence of fetal hemoglobin When treated with periodic acid, glycols are oxidized to aldehydes. After reaction with Schiff’s reagent (a mixture of pararosaniline and sodium metabisulfite) a pararosaniline adduct is released that stains the glyco-containing cellular elements. Clinically, the PAS stain is helpful in recognizing some cases of erythroleukemia and acute lymphoblastic leukemia. Myeloperoxidase (MP) is detected by means of the enzyme’s interaction with diaminobenzidine (DAB), a benzidine substitute. The brown reaction product is first intensified with copper salts followed by Gill’s modified Papanicolaou stain, which results in intense gray-black granules at sites of neutrophil and monocyte myeloperoxidase activity. This procedure differentiates cells of lymphoid origin from granulocytes and their precursors and monocytes. 29 Hematology PPT Flashcards Unit 4 What is the Prussian blue reaction? What is a sideroblast ? When does a ring sideroblast occurs? When is a Sudan Black B Stain performed? What is a Terminal Deoxynucleotidyl Transferase Test and where is it found? What is a Specimen quality procedure? What is a Special collection techniques? The Prussian blue reaction precipitates free iron into small blue or blue-green granules in erythrocytes. Free iron is not identifiable on Wright-stained or Wright-Giemsa–stained blood smears. A sideroblast or siderocyte is an immature or mature erythrocyte containing free iron. Increased numbers of siderocytes are seen in disorders such as thalassemia major or in patients after a splenectomy. If the iron granules encircle the nucleus of the erythrocyte, it is referred to as a ring sideroblast. Although alcoholism is the most common cause of ring sideroblasts, they may also be seen in cases of lead poisoning or anemia. Following fixation, blood or bone marrow films are immersed in a buffered Sudan black B solution. After rinsing, slides are counterstained with Mayer hematoxylin. Cells are examined microscopically for the presence of blue-black discrete granulation. Cells committed to the lymphoid pathway display negative staining reactions, whereas myeloid and monocytoid forms display characteristic positive reactions. The Sudan black B staining pattern usually parallels the myeloperoxidase stain and is useful in the identification of myelogenous and myelomonocytic leukemias. Terminal deoxynucleotidyl transferase (TdT) is a non–template-directed DNA polymerase that catalyzes the irreversible addition of deoxynucleotides to the 3-hydroxy groups on the end of DNA. The primary methods of detection are immunofluorescence and immunoperoxidase using a monoclonal antibody. TdT is a cell marker found on immature and neoplastic cells frequently seen in leukemic states. All coagulation testing critically depends on the quality of the specimen. Minimum tissue trauma and the avoidance of hemolysis are essential. Proper phlebotomy techniques must be strictly followed. To reduce the possibility of introducing tissue 30 Hematology PPT Flashcards Unit 4 What are some general sources of error? How is quality control maintained? List some of the Coagulation Procedures Describe the principle of the Activated Partial Thromboplastin Time procedure How is the aPTT used? How are the aPTT results reported? What are the Clinical applications of the aPTT? thromboplastin into a whole blood sample and the subsequent utilization of certain factors with clot formation, certain techniques should be followed. All procedural directions must be strictly followed because variations in pH, reagent concentration, and temperature are major sources of error. Water baths and heat blocks must be monitored continually for accurate temperatures. Refrigerators and freezers should also be monitored to ensure the stability of specimens and test reagents. Reagents must not be beyond their stated expiration date. Procedural technique must be consistent and appropriate. Activated partial thromboplastin time, Antithrombin III, Bleeding time, Circulating anticoagulants, D-dimer assay, Euglobulin lysis time, Substitution studies and factor assays,Fibrin split products, Fibrinogen assay The aPTT procedure measures the time required to generate thrombin and fibrin polymers via the intrinsic pathway. Although a partial thromboplastin time test can be performed, contact factors can be activated more thoroughly by the addition of substances such as kaolin in the activated form of this assay. In the aPTT, calcium ions and phospholipids that substitute for platelet phospholipids are added to blood plasma. The generation of fibrin is the end point. –Clinically, the aPTT is used to identify and quantitate deficiencies in the intrinsic clotting system and to control anticoagulant therapy. Reference values are dependent on the activator and phospholipid reagents used; however, 20 to 35 seconds is typically normal. In some laboratories, ranges may be from 28 to 42 seconds, with 42 to 46 seconds being marginal. The aPTT is widely advocated as the test of choice for the control of heparin therapy. It is also important in the screening profile of prekallikrein; high-molecular-weight kininogen; factors XII, XI, IX, VIII, X, V, II, and I; and 31 Hematology PPT Flashcards Unit 4 Describe the principle of the Antithrombin (AT III) What is the advantage of the Bleeding Time test? What is a circulating anticoagulant? How can a circulating anticoagulant be detected? How can a circulating anticoagulant be reported? What are some of the anticoagulants that may interfere with coagulation? inhibitors against these factors. In the presence of heparin, thrombin is neutralized at a rate that is proportional to the antithrombin (AT III) concentration. Following defibrination, plasma is assayed in a two-stage procedure that utilizes standardized amounts of heparin, fibrinogen, and thrombin. The resulting clotting time is interpreted using a calibration curve. Clinically, the AT III assay is useful prior to and subsequent to treatment with heparin in cases of disseminated intravascular coagulation (DIC). The bleeding time test is an in vivo measurement of platelet adhesion and aggregation on locally injured vascular subendothelium. This test provides an estimate of the integrity of the platelet plug and thereby measures the interaction between the capillaries and the platelets. Clinically, the bleeding time is prolonged in thrombocytopenia, qualitative platelet disorders such as von Willebrand disease, aspirin ingestion, or the presence of vascular problems When coagulation deficiencies are caused by inhibitors to specific factors rather than the lack of a factor, these inhibitors are sometimes referred to as circulating anticoagulants . To detect a circulating anticoagulant, the aPTT and the PT that were originally abnormal are repeated using various dilutions of patient plasma and normal plasma. The detection of an inhibitor may show up immediately or may require incubation of the normal plasma in the presence of the inhibitor. Differentiation between a coagulation factor deficiency and a circulating anticoagulant is important. If the abnormality is that of a deficiency, a normal plasma sample will correct the assay results to a reference value. If the abnormality is caused by a circulating anticoagulant (inhibitor), a greater correction is demonstrated as the ratio of normal plasma increases in the mixture. Various types of anticoagulants may interfere with coagulation at different stages, especially 32 Hematology PPT Flashcards Unit 4 What is principle of the D-dimer assay? Explain the principle of the Euglobulin Lysis Time Describe Substitution studies Assay and how they can be performed What is the principle of Specific factor assays? When conducting a Fibrin Split Products (FSP) test, what is the purpose of adding thrombin as well as soy bean enzyme inhibitors to the whole blood sample? What is demonstrated from FSP elevated results? factor VIII, heparin-like activity, and antithromboplastins. Most acquired anticoagulants are autoantibodies (usually IgG, sometimes IgM) directed against specific coagulation factors. The fundamental principle of the D-dimer assay has remained largely unchanged: recognition of the unique neoepitope in D-dimer by specific antisera. D-dimer contains a neoepitope that is formed following the cross-linking of adjacent D domains by factor XIIIa. It is this epitope that is recognized by specific antisera used in clinical assays. The euglobulin fraction of plasma contains plasminogen, fibrinogen, and activators with the potential for transforming plasminogen to plasmin. This fraction is precipitated with 1% acetic acid and resuspended in a borate solution. The euglobulins are then clotted by the addition of thrombin. The clot is incubated and the time of lysis is reported. Clinically, the euglobulin lysis test is a screening procedure for fibrinolytic activity. Substitution studies and factor assays can be used to identify specific coagulation factor deficiencies. Substitution studies may be performed using adsorbed plasma and aged serum with the aPTT to identify deficiencies of blood coagulation. Substitution studies may also be performed using adsorbed plasma with the PT to identify a factor VII deficiency. Factor assays are based on the ability of the plasma in question to correct a factor-deficient substrate, such as factor VIII. The actual assay is the same as the aPTT with the exception of the factor-deficient substrate. Identification of specific factor deficiencies is valuable in both the diagnosis and the treatment of patients. Thrombin is added to ensure complete clotting of the sample, and the soy bean enzyme inhibitors are added to prevent any breakdown of fibrin. Elevated results demonstrate that the activation of plasmin has occurred or is occurring, such as in primary fibrinolysis and DIC with secondary 33 Hematology PPT Flashcards Unit 4 What is the principle behind a fibrinogen assay? What is the use of the simple protein, protamine? What is the indication of a positive result from a Protamine Sulfate Assay? What is being measured when conducting a prothrombin time? What are some situations in which prothrombin time test is used? What is indicated from prolonged results in a prothrombin time? What is determined from a thrombin time test? What might cause the abnormal result of a thrombin time test? How is cell size and quantity determined based on the use of an electrical impedance procedure? Which method of cell counting uses a laser light? How does the diluted blood specimen interact with the laser light during optical detection? fibrinolysis. When plasma is diluted and clotted with excess thrombin, the fibrinogen concentration is inversely proportional to the clotting time Fibrinogen assays are useful in detecting deficiencies of fibrinogen and in detecting an alteration in the conversion of fibrinogen to fibrin. – Protamine is used to neutralize the effects of heparin. – Inappropriate presence of intravascular fibrin monomers – Length of time required to form a fibrin clot (measured in seconds) to monitor oral anticoagulant therapy as a screening test in the diagnosis of coagulation deficiencies – as a component of a liver profile assessment Deficiency of one or more factors in the extrinsic pathway: factors VII, X, and V and factor II or I. The presence of an inhibitor will also produce prolonged values. – Determines the rate of thrombin-induced cleavage of fibrinogen to fibrin monomers and the subsequent polymerization of hydrogenbonded fibrin polymers Extremely low fibrinogen levels Abnormal fibrinogen thrombin inhibitors High concentrations of immunoglobulin (e.g., myeloma proteins) Presence of heparin High concentrations of fibrin-fibrinogen degradation products Cell counting and sizing are based on the detection and measurement of changes in electrical impedance (resistance) produced by a particle as it passes through a small aperture. Optical Detection method A diluted blood specimen passes in a steady stream through which a beam of laser light is focused. As each cell passes through the sensing zone of 34 Hematology PPT Flashcards Unit 4 the flow cell, it scatters the focused light. Scattered light is detected by a photodetector and converted to an electrical pulse. What is represented by the number of The number of pulses generated is directly electrical pulses detected in an optical proportional to the number of cells passing detection method? through the sensing zone in a specific period. What are the three operational processes of Diffraction and the bending of light light amplification generated by emission of around corners with the use of small radiation? angles Refraction and the bending of light because of a change in speed with the use of intermediate angles Reflection and light rays turned back by the surface or an obstruction with the use of large angles How is Radio Frequency (RF) used to detect – High-voltage electromagnetic current is cell size? used to detect cell size, based on the cellular density. Why is laser light the most common light – Because of the properties of intensity, source used in flow cytometers? stability, and monochromatism. What is flow cytometry? – The simultaneous measurement of multiple physical characteristics of a single cell, as the cell flows in suspension through a measuring device. How can flow cytometry rapidly classify – Flow cytometry combines the groups of cells? technologies of fluid dynamics, optics, lasers, computers, and fluorochromeconjugated monoclonal antibodies that rapidly classify groups of cells with heterogeneous mixtures. In flow cytometry, what intrinsic properties of – Forward- and right-angle light scatter, a cell are detected? which correlate with size and granularity of a cell, respectively. Regarding scattered light, what is a forward – Light scattered along the axis of the laser scatter? beam Regarding scattered light, what is a side/ – Light scattered perpendicular to the axis orthogonal scatter? Forward scatter is proportional to? – Cell size Side scatter is proportional to? – Cytoplasmic granularity Define parameter: – Statistical term that refers to any numerical value that describes an entire population. Major types of automation are representations Representative of the ways that blood cells can of what? be counted, leukocytes differentiated, and other components (e.g., MCH and MCHC) calculated How does hemoglobin being measured? Measured by the traditional cyanmethemoglobin 35 Hematology PPT Flashcards Unit 4 flow-cell method at 525 and 546 nm, depending on the instrument manufacturer What are histograms? Histograms are graphic representations of cell frequencies versus sizes. In a homogeneous cell population, the curve A symmetrical bell-shaped or gaussian assumes what shape or distribution? distribution. In a histogram, a wide or more flattened curve A wide or more flattened curve is seen when the is seen in what? standard deviation from the mean is increased. True or false. True Histograms not only provide information about erythrocyte, leukocyte, and platelet frequency and their distribution about the mean but also depict the presence of subpopulations. True or False True Histograms provide a means of comparing the sizes of a patient’s cells with normal populations. True or False. True In a histogram, shifts in one direction or the other can be of diagnostic importance. Tue or False False (x-axis represents the size) In the Coulter system, the size (volume in femtoliters) is represented on the y-axis. In analysis of instrumental data output The erythrocyte histogram includes what? Quantitative descriptors of erythrocytes The leukocyte histograms Platelet histograms Derived platelet parameters What does an erythrocyte histogram reflects? The erythrocyte histogram reflects the native size of erythrocytes or any other particles in the erythrocyte size range. The erythrocyte histogram in the Coulter 1) 24 fL system displays cells as small as __(1)___fL., 2) 36 fL but only those greater than ___(2)___fL are counted as erythrocytes. In a erythrocyte histogram, what is the It allows for the detection of erythrocyte advantage of the extension of the lower end fragments, leukocyte fragments, and large of the scale from 36 to 24 fL? platelets An expression of erythrocyte size is the RDW _______ in the Coulter series. This term refers to variation in erythrocyte size. Correlations between the _______ and the RDW and the MCV _________exist for various types of anemia. What does size-referenced leukocyte The classification of leukocytes according to histograms displays? size following lysis. It does not display the 36 Hematology PPT Flashcards Unit 4 What does a platelet histogram reflects? What is the measure of the average volume of platelets in a sample? True or False The MPV is analogous to the erythrocytic MCV. True or False Mean platelet volume calculation is derived from the same data as the platelet count. True or False In healthy patients, there is an inverse relationship between platelet count and size. What are the three steps in a laser technology? RBC/platelet channel uses a laser-based optical assembly that is shared with what channel? Where is the platelet histogram derived? What is the mode of the measured platelet volumes? Most flow-cell instruments can simultaneously analyze multiple parameters at what rate? In flow cytometry, the cellular analysis yields what properties of individual cells? Major advances in this technology are owing to several factors, what are these factors? native cell size Platelet counting and sizing in both the electrical impedance and the optical systems reflect the native cell size. MPV True True True Cytochemical reactions prepare the blood cells for analysis. A cytometer measures specific cell properties. Algorithms convert these measurements into familiar results for cell classification, cell count, cell size, and hemoglobinization. The basophil/lobularity channel. The platelet histogram is derived from measurements made with the high-angle detector. MPV at the rate of 5,000 to 10,000 cells/sec. The cellular analysis yields quantitative data about the chemical and physical properties of individual cells. The ability to produce monoclonal antibodies resulted in the subsequent development of specific surface markers for various subpopulations of cells. The development of new fluorescent probes for DNA, RNA, and other cellular components increased the variety of possible applications at the molecular and cellular levels. The expansion of computer applications has improved the instrumentation technology, 37 Hematology PPT Flashcards Unit 4 In hematology, automated differentials can be based on a variety of principles which includes what? Because single-cell suspensions of peripheral blood and bone marrow are easy to obtain, most clinical applications of flow cytometry are in the specialties of __________ and ______________. Reticulocytes Enumeration by flow cytometry is more ____________. Platelet counts can provide an estimate of young, reticulated platelets by counting platelets that stain with what kind of dye? What kind of typing identified by a cluster designation (CD), are used in most flow cytometry immunophenotyping. A cellular application that has become an important tool in the diagnosis and classification of hematologic neoplasia by immunophenotyping. This techniques with bone marrow cells are applicable to DNA cell cycle analysis.What is this technique? What kind of new technology that referred to as automated digital cell morphology, provides an unprecedented level of efficiency and consistency. In its simplest form, automated digital cell morphology is a process where blood cells are automatically located and preclassified into categories of blood cell? This is the earliest instruments to detect blood clotting. This uses a methodology to measure the conduction or impedance of an electrical current by the formation of fibrin. What kind of instrumental methodology is this? This principle/methodology measures is the change in light transmission as optical density (absorbance) versus time can be used to quantitatively determine the activity of various coagulation stages or factors. _______ is defined as the resistance that a material has to a change in its form. making it easier to operate and more practical for use in clinical as well as research laboratories. – Determination of cell volume by electrical impedance or forward light scatter, cytochemistry or peroxidase staining, and VCS technology. – hematology and immunology – accurate, precise, and cost-effective. – with an RNA dye. – Immunophenotyping monoclonal antibodies – Flow Cytometry – Digital Technology – Electromechanical methods – Photo-optical Method – Viscosity 38 Hematology PPT Flashcards Unit 4 This principle usesthe natural thickening (viscosity) and monitored by the motion (amplitude of an oscillating steel ball in a specially designed cuvette), as a change in form takes place. What type of platelet agglutination method measures the ability of a patient’s plasma to agglutinate formalin-fixed platelets in the presence of ristocetin? The principle of the test is that platelet-rich plasma is treated with a known aggregating agent. What is this test? A new type of automated system that incorporates a high shear flow system to simulate the in vivo hemodynamic conditions of platelet adhesion and aggregation as encountered at a vascular lesion. – Viscosity-Based Detection System (VDS) – ristocetin cofactor assay – Platelet Aggregation – Siemens PFA-100 39