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CMWM633 WESTERN MEDICAL LABORATORY & IMAGING Credit hour: 3 Credits, 45 Hours Prerequisite: BS -525 Pathology Concurrent: BS-524 Microbiology Contents Composition of Blood Production of Blood cells Functions of Blood cells Hemostasis Hemogram(CBC) The lab test

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CMWM633

WESTERN MEDICAL

LABORATORY & IMAGING

Credit hour: 3 Credits, 45 Hours

Prerequisite: BS-525 Pathology

Concurrent: BS-524 Microbiology


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Contents

  • Composition of Blood

  • Production of Blood cells

  • Functions of Blood cells

  • Hemostasis

  • Hemogram(CBC)



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BLOOD STUDIESHematology and Coagulation



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Composition of Blood

Average person circulates about 5L of Blood , of which 3L is plasma and 2L is cells. Plasma Fluid derives from the intestines and lymphatic systems and provides a vehicle for cell movement.


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  • Adult ♂ contains 5-6L

  • Adult ♀ contains 4-5L

  • T is about 100.4 F

    • Is this higher or lower than normal body T?

  • 5 times as viscous as water

    • Is it more or less resistant to flow than water?

    • What accounts for its viscosity?

  • pH ranges from 7.35 – 7.45 (slightly alkaline)

  • Color ranges from scarlet (oxygenated blood) to a deep red (deoxygenated blood).


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BLOOD is composed of plasma (~55%), and the formed elements which are:

  • The erythrocytes (RBCs) (~45%)

    • Contain hemoglobin

    • Function in the transport of O2 and CO2

  • The Leukocytes (WBCs) and platlets (thrombocytes) (~1%)

    • Leukocytes are involved in the body’s defense against the invasion of foreign antigens.

    • Platlets are involved in hemostasis which forms a barrier to limit blood loss at an injured site.



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Components of Blood which are:

  • 55% plasma

  • 45% cells

    • 99% RBCs

    • < 1% WBCs and platelets


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Plasma which are:

  • Liquid part of blood

    • Pale yellow made up of 91% water, 9% other

  • Colloid: Liquid containing suspended substances that don’t settle out

    • Albumin: Important in regulation of water movement between tissues and blood

    • Globulins: Immune system or transport molecules

    • Fibrinogen: Responsible for formation of blood clots


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Formed Elements which are:

  • Red blood cells (erythrocytes)

  • White blood cells (leukocytes)

    • Granulocytes

      • Neutrophils

      • Eosinophils

      • Basophils

    • Agranulocytes

      • Lymphocytes

      • Monocytes

  • Platelets (thrombocytes)


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Formed Elements of Blood which are:

  • Red blood cells ( erythrocytes )

  • White blood cells ( leukocytes )

    • granular leukocytes

      • neutrophils

      • eosinophils

      • basophils

    • agranular leukocytes

      • lymphocytes = T cells, B cells, and natural killer cells

      • monocytes

  • Platelets (special cell fragments)


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The cells are primarily produced by bone marrow and account for blood “solids”. Blood cells are classified as white cells (leukocytes),red cells(erythrocytes) and platelets(thrombocytes)


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  • Hematopoiesis is a term describing the formation and development of blood cells.

    • Cells of the blood are constantly being lost or destroyed. Thus, to maintain homeostasis, the system must have the capacity for self renewal. This system involves:

      • Proliferation of progeny stem cells

      • Differentiation and maturation of the stem cells into the functional cellular elements.

      • In normal adults, the proliferation, differentiation, and maturation of the hematopoietic cells (RBCs, WBCs, and platlets) is limited to the bone marrow and the widespread lymphatic system and only mature cells are released into the peripheral blood.


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  • Hematopoiesis begins as early as the development of blood cells.19th day after fertilization in the yolk sac of the embryo

    • Only erythrocytes are made

    • The RBCs contain unique fetal hemoglobins

  • At about 6 weeks of gestation, yolk sac production of erythrocytes decreases and production of RBCs in the human embryo itself begins.


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  • The fetal liver development of blood cells. becomes the chief site of blood cell production.

    • Erythrocytes are produced

    • The beginnings of leukocyte and thrombocyte production occurs

  • The spleen, kidney, thymus, and lymph nodes serve as minor sites of blood cell production.

  • The lymph nodes will continue as an important site of lymphopoiesis (production of lymphocytes) throughout life, but blood production in the other areas decreases and finally ceases as the bone marrow becomes the primary site of hematopoiesis at about 6 months of gestation and continues throughout life.

    • When the bone marrow becomes the chief site of hematopoiesis leukocyte and thrombocyte production become more prominent.


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Hematopoiesis development of blood cells.

6


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  • Hematopoiesis development of blood cells.in the bone marrow is called medullary hematopoiesis

  • Hematopoiesis in areas other then the bone marrow is called extramedullary hematopoiesis

    • Extramedullary hematopoiesis may occur in fetal hematopoietic tissue (liver and spleen) of an adult when the bone marrow cannot meet the physiologic needs of the tissues. This can lead to hepatomegaly and/or splenomegaly (increase in size of the liver of spleen because of increased functions in the organs).

      Hematopoietic tissue includes tissues involved in the proliferation, maturation, and destruction of blood cells


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Bone marrow – is located inside development of blood cells.spongy bone

  • In a normal adult, ½ of the bone marrow is hematopoietically active (red marrow) and ½ is inactive, fatty marrow (yellow marrow).

  • The marrow contains both Erythroid (RBC) and Myeloid (WBC) precursors as well as platlet precursors. The normal M:E ratio is 1.5:1 to 4:1.

  • Early in life most of the marrow is red marrow and it gradually decreases with age to the adult level of 50%.


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Glassy hyaline cartilage development of blood cells.

Compact+spongy

(remanent of plate-hyaline cartilage

Causes the lengthwise growth)

(red marrow)

(yellow marrow-fat)

Compact bone

Fibrous connective tissue


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  • In certain pathologic states the bone marrow can increase its activity to 5-10X its normal rate.

    • When this happens, the bone marrow is said to be hyperplastic because it replaces the yellow marrow with red marrow.

    • This occurs in conditions where there is increased or ineffective hematopoiesis.

    • The degree to which the the bone marrow becomes hyperplastic is related to the severity and duration of the pathologic state.

    • Pathologic states that cause this include:

      • Acute blood loss in which there is a temporary replacement of the yellow marrow

      • Severe chronic anemia – erythropoiesis (RBC production) may increase to the extent that the marrow starts to erode the bone itself.

      • Malignant disease – both normal red marrow and fatty marrow may be replaced by proliferating abnormal cells.


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The hematopoietic tissue may also become inactive or hypoplastic.

This may be due to:

  • Chemicals

  • Genetics

  • Myeloproliferative disease that replaces hematopoietic tissue with fiberous tissue



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Production of Formed Elements hypoplastic.

  • Hematopoiesis or hemopoiesis: Process of blood cell production

  • Stem cells: All formed elements derived from single population

    • Proerythroblasts: Develop into red blood cells

    • Myeloblasts: Develop into basophils, neutrophils, eosinophils

    • Lymphoblasts: Develop into lymphocytes

    • Monoblasts: Develop into monocytes

    • Megakaryoblasts: Develop into platelets


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Blood cell differentiation hypoplastic.

Megakaryoblast

Magakaryocyte

Platelets

Lymphoblast

Bcells Tcells

plasma Active

cell Tcell

Myeloblast

Neutrophil Eosinophil Basophil

Promyelocyte

Myelocyte

Metamyelocyte

Stab Cell

Netrophil Eosinophil Basophil

Monoblasts

Monocyte

Proerytroblast

Reticulocyte

Erythrocyte


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Hematopoiesis summary hypoplastic.


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Hematopoiesis hypoplastic.


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Hematopoiesis hypoplastic.


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Functions of Blood hypoplastic.

  • Transport of:

    • Gases, nutrients, waste products

    • Processed molecules

    • Regulatory molecules

  • Regulation of pH and osmosis

  • Maintenance of body temperature

  • Protection against foreign substances

  • Clot formation


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Red blood cells transport oxygen hypoplastic.

  • Red blood cells contain hemoglobin

    • Hemoglobin enables the transport of O2

Figure 23.14


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White blood cells help defend the body hypoplastic.

  • White blood cells function both inside and outside the circulatory system

    • They fight infections and cancer

Basophil

Eosinophil

Monocyte

Neutrophil

Lymphocyte


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Thrombocytes hypoplastic.

  • Cell fragments pinched off from megakaryocytes in red bone marrow

  • Important in preventing blood loss

    • Platelet plugs

    • Promoting formation and contraction of clots


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Hemostasis hypoplastic.

  • Arrest of bleeding

  • Events preventing excessive blood loss

    • Vascular spasm: Vasoconstriction of damaged blood vessels

    • Platelet plug formation

    • Coagulation or blood clotting


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Blood clots plug leaks when blood vessels are injured hypoplastic.

  • When a blood vessel is damaged, platelets respond

    • They help trigger the formation of an insoluble fibrin clot that plugs the leak



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1 hypoplastic.

Injury to lining of bloodvessel exposes connectivetissue; platelets adhere

2

Platelet plug forms

3

Fibrin clot trapsblood cells

Connectivetissue

Plateletplug

Platelet releases chemicalsthat make nearby platelets sticky

Clotting factors from:

Platelets

Calcium andother factorsin blood plasma

Damaged cells

Prothrombin

Thrombin

Fibrinogen

Fibrin

Figure 23.16A


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Platelet plug formation hypoplastic.

  • Platelets store a lot of chemicals in granules needed for platelet plug formation

    • ADP, Ca+2, serotonin, fibrin-stabilizing factor, & enzymes that produce thromboxane A2

  • Steps in the process

    • (1) platelet adhesion (2) platelet release reaction (3) platelet aggregation


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    1. Platelet Adhesion hypoplastic.

    • Platelets stick to exposed collagen underlying damaged endothelial cells in vessel wall


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    2. Platelet Release Reaction hypoplastic.

    • Platelets activated by adhesion

    • Extend projections to make contact with each other

    • Release thromboxane A2, serotonin & ADP activating other platelets

    • Serotonin & thromboxane A2 are vasoconstrictors decreasing blood flow through the injured vessel. ADP causes stickiness


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    3. Platelet Aggregation hypoplastic.

    • Activated platelets stick together and activate new platelets to form a mass called a platelet plug

    • Plug reinforced by fibrin threads formed during clotting process


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    Coagulation hypoplastic.

    • Stages

      • Activation of prothrombinase

      • Conversion of prothrombin to thrombin

      • Conversion of fibrinogen to fibrin

    • Pathways

      • Extrinsic

      • Intrinsic


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    Blood Clotting hypoplastic.

    • Blood drawn from the body thickens into a gel

      • gel separates into liquid (serum) and a clot of insoluble fibers (fibrin) in which the cells are trapped

    • If clotting occurs in an unbroken vessel is called a thrombosis

    • Substances required for clotting are Ca+2, enzymes synthesized by liver cells(clotting factors) and substances released by platelets or damaged tissues

    • Clotting is a cascade of reactions in which each clotting factor activates the next in a fixed sequence resulting in the formation of fibrin threads


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    Clot Formation hypoplastic.


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    Plasminogen (plasmin) hypoplastic.

    • Plasminogen is a glycoprotein, synthesized in the liver, present in plasma.Under normal conditions,plasminogen is part of any clot because of the tendency of fibrin to absorb plasminogen from plasma.When plasminogen activators perform their function plasmin is formed within the clot;this gradually dissolves the clot while leaving time for tissues repair,free plasmin is also released to the plasma;however,antiplasmins there immediately destroy any plasmin released from the clot.


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    • FUNCTION: hypoplastic. Determines plasminogen activity in persons with thrombosis or DIC, in these conditions excessive free plasmin is released to the plasma, In these situations antiplasmin is depleted and plasmin begins destroying components other than fibrin,including fibrinogen, factors V and VIII, and other factors


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    D-Dimer hypoplastic. is an assay used to assess possible intravascular clotting. D-Dimer is a cleavage product of fibrin and is elevated in disseminated intravascular coagulation, in thromboembolic events and after surgery.


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    DIC hypoplastic.

    • Activation of coagulation cascade leading to microthrombi and global consumption of platelets,fibrin,and coagulation factors.Causes: obstetric complications, gram-negative sepsis,transfusion,trauma,malignancy.Lab findings: ^PT^PTT, fibrin split products <platelet count.


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    Bleeding time hypoplastic.

    • Measures the primary phase of hemostasis:

    • the interaction of platelets with the blood vessels wall and the formation of a hemostatic plug.

    • It is the best single screening test for platelets function disorders and is one of the primary screening tests for a coagulation disorder.

    • principal use of this test is in the diagnosis of Von Willebrands and inherited defective molecule of factor VIII

    • Normal 3-10minutes


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    HEMOGRAM hypoplastic.

    Consists of a WBC count , RBC count,hemoglobin (Hb) ,Hemotocrit (Hct), red blood cells indices and a platelet count.

    A complete blood cell count (CBC) consists of a hemogram plus a differential WBC.


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    Diagnostic Blood Tests hypoplastic.

    • Type and crossmatch

    • Complete blood count

      • Red blood count

      • Hemoglobin measurement

      • Hematocrit measurement

    • White blood count

    • Differential white blood count

    • Clotting


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    Withdraw hypoplastic. blood

    Centrifuge

    Place in tube

    PLASMA 55%

    CONSTITUENT

    MAJOR FUNCTIONS

    CELLULAR ELEMENTS 45%

    Solvent forcarrying othersubstances

    CELL TYPE

    NUMBER(per mm3 of blood)

    FUNCTIONS

    Water

    Erythrocytes(red blood cells)

    Salts

    5–6 million

    Transport ofoxygen (and carbon dioxide)

    Sodium

    Potassium

    Calcium

    Magnesium

    Chloride

    Bicarbonate

    Osmotic balance,pH buffering, andregulation ofmembranepermeability

    Leukocytes(white blood cells)

    Defense andimmunity

    5,000–10,000

    Plasma proteins

    Albumin

    Fibrinogen

    Immunoglobins(antibodies)

    Osmotic balance,pH buffering

    Clotting

    Immunity

    Lymphocyte

    Basophil

    Eosinophil

    Substances transported by blood

    Monocyte

    Nutrients (e.g., glucose, fatty acids, vitamins)

    Waste products of metabolism

    Respiratory gases (O2 and CO2)

    Hormones

    Neutrophil

    Platelets

    250,000–400,000

    Blood clotting

    Figure 23.13


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    CBC hypoplastic.

    • CONSISTS OF A SERIES OF TESTS THAT DETERMINE NUMBER ,VARIETY, PERCENTAGE, CONCENTRATIONS,AND QUALITY OF BLOOD CELLS;WBC, RBC, Hct, Hb, MCV, MCH, MCHC, MPV


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    White Blood Cells: hypoplastic.


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    • Tests of White Blood Cells hypoplastic. White Blood Cell Count (CBC)LeukocytesAre divided into two main groups :Granulocytes ; neutrophils,basophils,eosinophils also called PMNsAgranulocytes; lymphocytes and monocytes also called mononuclears

    • Normal values:Adult 3.200 – 10,000 cells/mm3


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    Function of Lekocytes hypoplastic.


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    Leukocytes hypoplastic.

    • Protect body against microorganisms and remove dead cells and debris

    • Movements

      • Ameboid

      • Diapedesis

      • Chemotaxis

      • Passive Immunity

      • Active Immunity

      • Antigen – Antibody

    • Types

      • Neutrophils: Most common; phagocytic cells destroy bacteria (60%)

      • Eosinophils: Detoxify chemicals; reduce inflammation (4%)

      • Basophils: Alergic reactions; Release histamine, heparin increase inflam. response (1%)

      • Lymphocytes: Immunity 2 types; b & t Cell types. IgG-infection, IgM-microbes, IgA-Resp & GI, IgE- Alergy, IgD-immune response

      • Monocytes: Become macrophages


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    Leukocytes tissue injury or infection during the innate immune response


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    fight infections and defend the body by a process called phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)


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    Leukocytosis phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)occurs in acute infections, in which the degree of increase of leukocytes depends on the severity of the infection. In certain disease (measles,pertussis,sepsis) the increase of leukocytes is so great that the blood pictures suggests leukemia ( in leukemia the leukocytosis is permanent and progressive)Other causes:Trauma or tissue injuryMalignant neoplasm's (bronchogenic carcinoma)Drugs (ether,epinephrine)Acute hemolysis


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    Leukopenia: WBC < 4000 mm3 phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)CAUSESViral infections, some bacterial infections, overwhelming bacterial infectionsHypersplenismBone Marrow depression caused by intoxication'sPrimary bone disorders: leukemia, pernicious anemia


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    Clinical Implications phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Leukocytosis: WBC > 11,000mm3 It is usually caused by the increase of only one type of leukocyte, and it is given the name of the type of cell that shows the main increase.Neutrophilic leukocytosisLymphocytic leukocytosisMonocytic leukocytosisBasophilic leukocytosisEosinophilic leukocytosis


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    ALERT/ Panic values phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody) WBC < 500/mm3 WBC > 30,000/mm3


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    Functions of circulating WBC According to leukocyte type. phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)


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    NEUTROPHILS phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Neutrophils, or neutrophil polymorphonuclear leucocytes, Acute Inflamatory response cell .40-70% WBC’c Phagocyitic . Multilobed.Large,sperical, azurophilic 1 granules(called lysosomes) contain hydrolytic enzymes,lysozyme,meyloperoxidase)


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    Neutrophils phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody) Pyogenic infections


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    LYMPHOCYTES phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Lymphocytes are produced within bone marrow (a primary lymphoid organ). If they achieve immune-competence within the bone marrow, they are known as B cells, or if in the thymus (also a primary lymphoid organ), they are known as T cells. Organized lymphoid tissue elsewhere is known as secondary lymphoid tissue, and includes lymph nodes, adenoids, tonsils and mucosa associated tissue (MALT). Small amount of pale cytoplasm,small ,round , densely staining nucleus. B lymphocytes produce antibodies. T lymphocytes manifest in the cellular immune response as well as regulate B lymphocytes and macrophages.

    Lymphocytes are only slightly larger than red blood cells (small lymphocytes) and they have a relatively large nucleus:cytoplasm ratio. 


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    Lymphocytes phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Viral infections


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    B Lymphocyte: Part of the humoral immune response. phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Arises from stem cells in bone marrow.Matures in marrow.Migrates to the peripheral tissue (follicles of lymph nodes,white pulp of spleen,unencapsulated lymphoid tissue.When an Antigen is encountered, B cells differentiate into plasma cells and produce antibodies.Has a memory, Functions as antigen presenting cell via MHCmajor histocompatibility complex


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    Plasma cells phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody) (fully differentiated B cells) are similar in appearance to lymphocytes, even though they are not normally present in this blood test measures the number of lymphocytes in the peripheral blood.Off-center nucleus, clock face chromatin distribution, abundant RER and well developed Golgi apparatus.Multiple Myeloma is a plasma cell neoplasm


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    The T cells the master immune cells include phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)CD4+ helpers T cells CD8+ suppressor T cellsMediates cellular immune response,Originates from stem cells in the bone marrow but matures in the thymus.T cells differentiate in:1- cytotoxic T cells (MHC I,CD8) 2-helper T cells,delayed hypersensitivity Tcell.MHC=Mayor histocompability complex


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    Clinical Implications: phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)1-Inmmunosuppressive drugs therapy usually decreases lymphocyte total2-Patients with an absolute helper T lymphocyte count <200mm3 are at greatest risk for developing clinical AIDS3-Decrease T cells occur in congenital immunodeficincy(DiGeorge syndrome,thymic hypoplasia)4-A marked increase in B cells occurs in lymphoproliferative disorders (chronic lymphocyitc leukemia)


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    Lymphocytosis > 4000 phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)/mm3 in adultsLymphatic leukemiaInfectious mononucleosisCytomegalovirusMeasles,mumps,chickenpoxHepatitis


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    Lymphopenia < 1000cells phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)/mm3ChemotherapyAfter administration of ATCHIncrease loss of G-I tract owing to obstruction of lymphatic drainageAplastic anemia


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    BASOPHILS phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Basophils are non-phagocytic cells which, when activated, release numerous compounds from the basophilic granules within their cytoplasm. They play a major role in allergic responses, particularly type I hypersensitive reactions.


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    Basophils phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Parasitic infections,some allergic disorders


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    MONOCYTES phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Monocytes circulate in the peripheral blood prior to emigration into the tissues. Within certain organs they have special names, e.g. in liver they are known as Kupfer cells, in brain as microglia, in kidney as mesangial cells, and in bone as osteoclasts. Elsewhere they are referred to as tissue macrophages. Phagocytosis is mediated by macrophages and polymorphonuclear leucocytes


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    Monocytes phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)severe infections by phagocytosis


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    EOSINOPHILS phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)These cells have the purpose of giving large parasites such as helminths, a hard time. The eosinophils release various substances from their eosinophilic granules. The granule contents are capable of damaging the parasite membrane.


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    EOSINOPHIL phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    Allergic Processes, asthma

    Parasites

    Tumors, Collagen vascular disease


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    Lymphocyte Neutrophils phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    70%

    30%

    Bacterial disorders

    Viral disorders


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    Breath……relax phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)


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    WBCs in order of abundance: phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    Never(neutrophils

    Let(lymphocytes)

    Monkeys(monocytes)

    Eat(eosinophils)

    Bananas(basophils)

    How do I remember the

    relative percentages?

    60 + 30 + 6 + 3 +1

    (i.e., 60% neutrophils, 30% lymphocytes, 6% monocytes, 3% eosinophils & 1% basophils)


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    STAINS FOR LEUKEMIAS phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)


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    Leukemias phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)^leukocytesbone marrow

    Acute Leukemias Chronic Leukemias

    Blast predominante

    children-elderly

    short and drastic courses

    More mature cells

    midlife age range

    longer,less devastating course

    CLL CML

    ALL AML

    Children more

    responsive to therapy

    Auer rods

    adults

    Older adults

    PAS

    SBB

    TDT


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    SBB STAINS (Sudan Black B) phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)The SBB stain aids in differentiation of the immature cells of acute leukemias, specially acute myeloblastic leukemias (AML). The SBB stains a variety of fats and lipids that are present in myeloid leukemias but not present in the lymphoid leukemias.


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    Periodic Acid-Schiff (PAS) Stain. phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)The PASS stian aid in the diagnosis of acute lymphoblastic leukemias (ALL) Early myeloid precursors and erythrocyte precursors are negative. As granulocytes mature,they increase in PAS positivity, whereas mature RBC’s stay negative. The PAS stain cannot be used to distinguish between ALL and AML or between benign and malignant lymphocytic disorders.


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    Terminal Deoxynucleotidyl Transferase phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)(TDT)The thyme is the primary site of TDT-positive cells, and TDT is found in the nucleus of the more primitive T cells. TDT is increase in >90% of cases of ALL of childhoodTDT positive blasts are predominant in some cases of chronic myelogenous leukemia (CML) , relating to the development of an acute blast phase.


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    Red Blood Cells phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)


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    Erythrocytes phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    • Structure

      • Biconcave, anucleate

    • Components

      • Hemoglobin

      • Lipids, ATP, carbonic anhydrase

    • Function

      • Transport oxygen from lungs to tissues and carbon dioxide from tissues to lungs


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    Red Blood Cells or Erythrocytes phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    • Contain oxygen-carrying protein hemoglobin that gives blood its red color

      • 1/3 of cell’s weight is hemoglobin

    • Biconcave disk

      • increased surface area/volume ratio

      • flexible shape for narrow passages

      • no nucleus or other organelles

      • no mitochondrial ATP formation

    • Normal RBC count

      • male 5.4 million/drop ---- female 4.8 million/drop

      • new RBCs enter circulation at 2-3 million/second


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    RBC Life Cycle phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    • RBCs live only 120 days

      • wear out from bending to fit through capillaries

      • no repair possible due to lack of organelles

    • Worn out cells removed by fixed macrophages in spleen & liver

    • Breakdown products are recycled


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    RED BLOOD CELL COUNT phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)(RBC,Erythrocyte count)Many test look at the red blood cells:their number and size,amount of Hb , rate of production and percent composition of the blood.The red blood cell count(RBC), hemotocrit(Hct) and hemoglobin(Hb) are closely related ,the same conditions cause an increase (or decrease) in each of these indicators


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    The main function phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody) of the RBC is to carry oxygen from the lungs to the body tissues and to transfer dioxide from the tissues to the lungs , by the Hb in the RBC which combines easily with oxygen and carbon dioxide. This test is an important measurement in the evaluation of anemia or polycytemia,determines the total number of erythrocytes in a microliter of blood.

    O2

    CO2


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    Erythropoiesis phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    • Production of red blood cells

      • Stem cells proerythroblasts early erythroblasts intermediate late reticulocytes

    • Erythropoietin: Hormone to stimulate RBC production


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    1-Decrease values occur in: phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)AnemiaAcute and chronic disorders


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    Erythrocytosis phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)a-primary (polycythemia vera, Erythemic Erythrocytosis)b-secundary(renal disease, high altitud etc..)


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    Hematocrit (Hct) phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)The word means separate blood , the mechanism of the test separates by centrifugation the plasma and the blood cells.Indirectly measures the RBC mass, important for determination of anemia or polycythemia.NormalWomen; 36%-48%Men 42%-52%


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    Hematocrit (Hct) phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    • Packed cell volume diagram


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    Hb x 3 = Hct phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)12 x 3 = 36


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    Hemoglobin phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)

    • Consists of:

      • 4 globin molecules: Transport carbon dioxide (carbonic anhydrase involved), nitric oxide

      • 4 heme molecules: Transport oxygen

        • Iron is required for oxygen transport


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    Hemoglobin (Hb) phagocytosis, also produce, transport and distribute antibodies as part of the immune response to a foreign susbstance (antibody)Hb the main component of erythrocytes , serves as the vehicle for the transportation of O2 and C02. It is composed of amino acids that form a single protein called globulin and a compound called heme, which contains iron atoms and the red pigment porphyrin. It is the Iron pigment that combines with oxygen and gives blood its characteristic red color


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    The Hb determination is part of the CBC. It is used to screen for disease associated with anemia, to determine the severity of anemia, to monitor the response to treatment for anemia and to evaluate polycytemia.


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    Normal screen for disease associated with anemia, to determine the severity of anemia, to monitor the response to treatment for anemia and to evaluate polycytemia.Women:12.0-16.0g/dl

    Men:14.0-17.4g/dl


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    Decreased Hb levels are found in anemic states, the Hb must be evaluated with the Htc and the RBCIron deficiencyliver diseasehemorrhagehemolytic anemias


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    Increased Hb levels be evaluated with the Htc and the RBCPolycitemia veraCongestive heart failureCOPDVariations in Hb occurs after transfusions….


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    RED BLOOD CELLS INDICES be evaluated with the Htc and the RBC

    • The red blood cell indices define the size and Hb content of the RBC and consist of:

      MCV : the mean corpuscular volume

      MCHC: the mean corpuscular concentration

      MCH: the mean corpuscular hemoglobin


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    The RBC indices are used in differentiating anemias. When they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.In deficient states anemias can be classified by cell size as macrocytic,normocytic,microcytic, or by cells size and color , as microcytic hypochromic.


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    MCV they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Indicates the red blood cell sizenormal: small<82um3 or larger >100um3MCV= Hct% x 10 / RBCMacrocytic: (MCV>100) Megaloblastic (B12-Folic acid) Microcytic: (MCV<80) Iron Deficiency Thalasemias, lead poisoningNormocytic,normochromic: enzyme defects(G6PD def-) , RBC membrane defects, bone marrow disorders, Autoinmmune hemolytic anemias, anemias of chronic disease.


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    MCHC they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained. (Mean Corpuscular Hemoglobin Concentration)Measures the average concentration of Hb in RBC’s , used for monitoring therapy for anemias.Normal: 32-36g/dLMCHC=Hb x 100/Hct%


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    MCH ( they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Mean Corpuscular Hemoglobin)Is a measure of the average weight of Hb per RBC , this index is of value in Dx severly anemic patients.Normal:26-34pg/cell


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    RBC disorders (Anemias) : they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.

    “Anemia is decreased red cell mass affecting tissue oxygenation”

    * Low Hb <13.5 (males), <11.5 (females)

    • Onset, chronicity, presentation & pattern.

    • Congenital disorders:

      • Membrane, Hb & enzyme disorders.

    • Acquired disorders:

      • Decreased production

      • Increased loss


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    Congenital RBC Disorders: they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.

    1

    2

    3

    (Hemolysis)

    • Membrane Disorders:

      • Spherocytosis, Elliptocytosis

    • Hemoglobin Disorders:

      • Hemoglobinopathies - Sickle cell, HbC etc.

      • Thalassemia Syndromes - , , 

    • Enzyme disorders:

      • G6PD, PK deficiency


    Acquired rbc disorders l.jpg
    Acquired RBC disorders : they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.

    (Deficiency, Marrow disorder, Blood loss & Hemolytic)

    • Decreased Production:

      • Deficiency anemias Iron, B12, Folate etc.

      • Aplastic, Hypoplastic anemias

      • Marrow disease, malignancy, radiation

    • Increased loss/destruction:

      • Blood loss anemias - parasites, bleeding

      • Hemolytic anemias - Autoimmune (cold & warm antibody) mechanical, drugs & toxins.


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    Anemias they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Microcytic MCV<80Iron deficiencyThalassemiasLead poisoningMacrocytic MCV>100Megaloblastic:B12/folate def.Drugs that block DNA: Methotrexate-MTXNormocytic-normochromic:Enzyme defects,RBC membrane defects,bone marrow disorders, hemolytic,chronic disease.


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    OH NO !!!! they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.How to make diagnosis?


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    Microcytic Anemia (IDA) they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.


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    Macrocytic Anemia (Meg.): they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.


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    = they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.B12

    MEGALO B LASTIC

    = IRON

    M I CROCYTIC


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    Reticulocyte Count. they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Young immature nonnucleated RBC , normally a small number of these cells are found in the circulating blood , it is used to differentiate anemias caused by bone marrow failure from those caused by hemorrhage or hemolysis , to check the effectiveness of treatment in pernicious anemia and folate and iron deficiency, to asses the recovery of bone marrow function in aplastic anemia, and to determine the effects of radioactive substances on exposed workers.


    Slide129 l.jpg

    Normal: 0.5%-1.5% of total erythrocytes. they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Increased : means that increase production RBC is occurring as the bone marrow replaces cells lost or prematurely destroyed. Identification of reticulocytosis may lead to the recognition of an otherwise occult disease , such as hidden chronic hemorrhage or unrecognized hemolysis(eg sickle cell anemia, thalassemia).


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    Test for Hemoglobin disorders they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.


    Slide131 l.jpg

    Fetal Hemoglobin (hbF) they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Is a normal Hb manufactured in the RBCs of the fetus and infant; it makes up to 50% to 90% of the Hb in newborn.The portion of the Hb in the newborn is made up to of HbA1 and HbA2. In adult types under normal conditions the manufacture of fetal Hb is replaced by the adults types during the first year of life, but if the FHb persists and constitutes more than 5% of the Hb after 6 moths an abnormality should be expected.


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    Hemoglobin A2 they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.HbA2 levels have special application in the diagnosis of beta thalassemias trait,which may be present even though the peripheral blood smear is normal. This measurements is used in the investigation of hemolytic anemias for hemoglobinopathies especially thalassemia (beta thalassemia)


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    Hemoglobin S (sickle cell test) they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Sickle cell disease is a term for a group of hereditary blood disorders,caused by an abnormality of the Hb,the red protein in the red blood cell that carries oxygen from the lungs to the tissues. People with sickle cell disease make abnormal Hb, hemoglobin S , this cells do not last as long as normal red blood cells, they become rigid and take a sickle or crescent shape, they are not flexible enough to squeeze through the vessels, resulting in blood vessels being blocked damaging tissues and organs.Normal:not present.


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    Sickle Cells they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.


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    Myoglobin they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Is the oxygen binding protein of striated muscles.It resembles Hb but is unable to release oxygen except in extremely low tension, injury of the muscle results in the release of the oxygen, Myoglobin is not tightly bound to protein and is rapidly excreted in urine, It is used as an early marker of muscle damage in myocardial infarction and to detect injury of the skeletal muscle


    Slide137 l.jpg

    Platelet Count they are used together with an examination of the erythrocytes on the stained smear, a clear picture of the RBC morphology may be ascertained.Mean Platelet Volume(MPV)Platelets are the smallest of the formed elements in the blood ,they are nonnucleated round or oval ,dish-shaped structures, Platelet activity is necessary for the blood clotting,vascular integrity and vasoconstriction,and the adhesion and aggregation activity .Thorombocyte development takes place in the bone marrow life is about 7.5 days


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    PANIC VALUES , a decrease in platelets to <20X10 3/mm3/L is associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.Platelets count >50x10 3/mm3 are not generally associated with spontaneous bleeding.


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    MPV associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.Is valuable in assessing bleeding disorders that occur in thrombocytopenia,uremia,liver disease or malignancies and for monitoring the course of disease associated by the bone marrow failure.The mean platelet volume is sometimes ordered in conjunction with a platelet count it indicates the uniformity of size of platelets population.It is used for differential diagnosis of thrombocytopenia.ValuesNormal: adults:140-400x 10 3/mm3


    Coagulation cascade l.jpg
    Coagulation Cascade associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.

    Intrinsic pathway Extrinsic Pathway

    endothelial disruption

    XI XI a tissue factor

    IX IX a VII VII a

    VIII(pro-ag)

    X X a

    II II a

    FIBRINOGEN FIBRIN fibrin products

    V

    Procoagulant-antigenic portions VIII


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    Platelet Aggregation associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.It is used to evaluate congenital qualitative functional disorders of adhesion,release ,or aggregation. It is rarely used to evaluate acquired bleeding disorders.


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    Prothrombin time(PT) associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis. is used to assess the extrinsic pathway Prolongation the PT is seen with warfarin use,vitamin K deficiency and liver disease.Monitors Warfarin Tx


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    Partial Thromboplastin time (PTT) associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis. is used to assess the intrinsic system.It is prolonged during heparin use, in hemophilia A,B and von Willebrand’s disease.Monitors Heparin Tx


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    TESTS OF HEMOSTASIS associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.


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    Hemorragic Disorders associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.

    Platelet count bleeding time PT PTT

    -

    -

    -

    -

    >

    >

    -

    >

    >

    >

    >

    >

    Thrombocytopenia

    hemophilia A (factor VIII)

    hemophilia B (factor IX)

    Von Willebrands (VIII)

    DIC,vitamin

    Vitamin K def(1972)

    <

    -

    -

    -

    <

    -

    >

    -

    -

    >

    -

    -


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    Blood Grouping associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.

    • Determined by antigens (agglutinogens) on surface of RBCs

    • Antibodies (agglutinins) can bind to RBC antigens, resulting in agglutination (clumping) or hemolysis (rupture) of RBCs

    • Groups

      • ABO and Rh


    Slide147 l.jpg

    ? associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.


    End of the chapter l.jpg

    End of the Chapter associated with a tendency for spontaneous bleeding prolonged bleeding time, pethechie and ecchymosis.


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