Hema week 1 Flashcards by Potato Patata

What determines the type of hemoglobin?

Globin chains

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Globin chains of gower 1 (EZ)

2 Zeta 2 Epsilon

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Globin chains of portland hemoglobin (GratZ)

2 Zeta 2 Gamma

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Globin chain of gower 2 hemoglobin (AE)

2 Alpha 2 Epsilon

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As early as the 19th day of gestation in the blood islands of the yolk sac of the human embryo

Blood islands remain active for 8 to 12 weeks

Mesoblastic

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3rd month, yolk sac discontinues its role, fetal liver becomes active

Spleen, thymus, and lymph nodes are active

Primitive cells disappears by the end of the 4th month, with an increase in the definitive erythroblast, granulocytes, and megakaryocytes

Hemoglobin production: Hb F, Hb A1 and Hb A2

Hepatic

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Globin chain composition of Hb F (F AG)

2 alpha 2 gamma

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Globin chain composition of Hb A1 (ewan BA)

2 alpha 2 beta

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Globin chain composition of Hb A2 (eto DA)

2 alpha 2 Delta

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Primary adult hemoglobin

Hemoglobin A1

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Primary site of hematopoiesis in adult

Sternum

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Safest, most accessible site for bone marrow aspiration and biopsy

Iliac crest

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A continuous regulated process of blood cell production that includes cell renewal, proliferation, differentiation, and maturation of the cell

Hematopoietic

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Haematopoietically active marrow consisting of the developing blood cells and their progenitors

Red marrow

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Haematopoietically inactive marrow composed primarily of ADIPOCYTES and fat cells with undifferentiated MESENCHYMAL CELLS AND MACRHOPHAGES

Yellow marrow

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Composition of stroma

Endothelial cells, Adipocytes, Osteoblasts, Osteoclasts, Reticular cells (Fibroblasts), and Macrophages

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Regulates the flow of particles entering and leaving hematopoietic spaces

Endothelial cells

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Secrete various steroids that influence erythropoiesis and maintain bone integrity and regulates the volume of marrow

Adipocytes

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Function in phagocytosis and secretion of various cytokines that regulate hematopoiesis

Macrophages

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Bone forming cells-water bug or comet appearance

Osteoblasts

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Bone resorbing cells or destroying cells

Osteoclasts

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Supports the vascular sinuses and developing hematopoietic cells

Reticular cells (Fibroblasts)

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Composition of Extracellular matrix of bone marrow

For regulation of the hematopoietic stem cells and progenitors

For survival and differentiation of cells

For adhesions

Proteoglycans/Glycosaminoglycans
Fibronectin
Collagen
Laminin
Hemonectin
Thrombospondin

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Major site of blood cell production during the second trimester of fetal development (Hepatic phase)

Capable of extra medullary hematopoiesis

Functions:

Protein synthesis and degradation, coagulation factor synthesis (except F IV), carbohydrate and lipid metabolism

Drug and toxin clearance

Iron recycling and storage

Hemoglobin degradation

Liver

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Largest lymphoid organ in the body Located directly beneath the diaphragm behind the fundus of the stomach in the upper left quadrant of the abdoment Vital but not essential for life Function: Indiscriminate filter of the circulating blood Storage for platelets 30% spleen 70% Circulation

Spleen

Disease related to autosplenectomy

Sickle Cell Anemia, Spherocytosis

Regions of spleen

1. White pulp - consists of scattered follicles with germinal center containing lymphocytes, macrophages, and dendritic cells 2. Red pulp - Composed primarily of vascular sinusoids and sinuses separated 3. Marginal zone - Surrounds the white pulp and forms a reticular meshwork containing blood vessels, macrophages, and specialized B cells

Two methods for removing senescent or abnormal RBCs from circulation

Culling and Pitting

Cells are phagocytized with subsequent degradation of cell organelles

Culling

Splenic macrophages remove inclusions or damaged surface membrane from the circulating RBCs

Pitting

Bean-shaped structures (1-5mm) Functions: Plays a role in the formation of new lymphocytes from germinal centers Involved in the processing of specific immunoglobulin Involved in the filtration of particulate matter, debris, and bacteria entering the lymph node via the lymph

Lymph nodes

Region of lymph nodes

Cortex, Paracortex, and Medulla

Outer region of lymph node Contains follicles of B cells Proliferation termed germinal centers

Cortex

Inner region Consists primarily of T lymphocytes and plasma cells

Medulla

Region between cortex and medulla Contains predominantly T cells and numerous macrophages

Paracortex

Originates from endodermal and mesenchymal tissues Populated initially by lymphocytes from the yolk sac and the liver An efficient, well-developed organ at birth that consists of two lobules each measure 0.5 to 2cm in diameter Organ responsible in the conditioning of T lymphoctes

Thymus

Cells that have extensive proliferative capacity - ability to give rise to new stem cell - Ability to differentiate into any blood cell lines Hematopoietic stem cells are Bone marrow cells that are capable of producing all types of blood cells They differentiate into one or another type of committed stem cells( Progenitor cells)

Stem cells

Stimulates proliferation, growth, and differentiation of erythroid precursors and may have minor effects on megakaryocytes Target cells are pronormoblast and CFU-Erythroid cells Source: KIDNEY

Erythropoietin (EPO)

The entry of mature blood cells into the intravascular space relies upon:

1. Multiplication of developing cells 2. Gradual maturation 3. Orderly release of cell from bone marrow

3 Possible activities of hematopoietic stem cells

1. Self- renewal 2. Differentiation 3. Apoptosis

Normal blood cells maturation

1. Cytoplasmic changes 2. Nuclear changes 3. Reduction in cell size

Abnormal cell maturation

Characterized by persistent cytoplasmic basophilia and late hemoglobinization Inclusion bodies may be found

Cells are present in the first few hours after an ovum is fertilized Most versatile type of stem cell, can develop into any human cell type, including development from embryo into fetus

Totipotential stem cells

Cells are present several days after fertilization Can develop into any cell type except into a fetus

Pluripotential stem cells

Derived from pluripotent stem cells Found in adults but are limited to specific types of cells to form tissues

Multipotential stem cells

Size: Large cell with high Nucleus:Cytoplasm ratio Cytoplasm: Very dark blue (Increase in RNA) and small in amount in comparison to the size of nucleus. No granular is present Nucleus: Large in size as compared to the size of cytoplasm. Chromatic which is reddish purple and indicated predominance of DNA

Blast

Present only in the bone marrow Nucleus Cytoplasm ratio - 8:1 Contains one or two nucleoli Cytoplasm is dark blue because of the concentration of ribosomes Cellular activity: Accumulates the components necessary for hemoglobin production Enzymes and proteins necessary for iron uptake and protoporphyrin synthesis are produced Globin production begins Stage last more than 24 hours Mitosis is present = 2 prorubricyte

Pronormoblast/Rubriblast

Nucleus: Chromatin begins to condense N:C ratio decrease to about 6:1 Chromatin stains deep purple red Nucleoli may be present early in the stage but disappear later Mitosis is present = produces 4 rubricyte Bone marrow is the location Detectable hemoglobin synthesis occurs. (1st stage of hemoglobin synthesis) Mistaken as Rubriblast

Prorubricyte / Basophilic normoblast

Differentiation of prorubricyte to rubricyte

1. Coarse Chromatin 2. Nucleoli are absent. (Present in the early stage but disappear later)

Nucleus: Chromatin pattern varies during this stage of development, showing some openness early in the stage but becoming condense The condensation of chromatin reduces the diameter of the nucleus considerably so the N:C ratio decreases from 4:1 to about 1:1 by the end of the stage NO nucleoli are present Mitosis is present = 2 metarubricyte LAST STAGE OF MITOSIS First stage in which cytoplasm is turning pink because Hemoglobin synthesis increases, and the accumulation begins to be visible in the color of the cytoplasm Last approximately 30 hours Confused with lymphocyte

Polychromatic normoblast/ Rubricyte

Rubricyte and Lymphocyte differentiation

Lymphocyte - Crushed velvet nucleus Cytoplasm - Sky blue Rubricyte - Nucleus - Checkerboard Cytoplasm - Muddy/Gray

Nucleus: Completely condensed N:C ratio is low or approximately 1:2 Cytoplasm: Increase in the salmon-pink color of the cytoplasm reflects the nearly complete hemoglobin production Division: Not capable of division due to the condensation of the chromatin Location: Present only in the bone marrow in healthy states

Orthochromic normoblast / Metarubricyte

No nucleus but has mitochondria and ribosomes Last stage to synthesize hemoglobin Last stage in bone marrow before release to the blood Location: Polychromatic erythrocyte resides in the bone marrow for 1 day or longer and then moves into the peripheral blood for about 1 day before reaching maturity 0.5% - 1.5% in adult 2% - 6% in newborn Indicators of bone marrow functions Also known as polychromatophilic erythrocytes Diffusely basophilic erythrocytes Polychromatophilic macrocytes

Reticulocytes

No nucleus is present Cytoplasm: Biconcave disc measure 7 to 8 mm in diameter with a thickness of about 1.5 to 2.5 mm On a stained blood film it appears as a salmon pink-staining cell with a central pale are. The central pallor is about one third the diameter of the cell Division: No division (Stops at rubricyte) Remains active in the circulation for 120 days Aging leads to their removal at the spleen and by the spleen Delivers oxygen to tissues, releases it, and returns to the lungs to be reoxygenated

Erythrocytes

- important in terminal erythroid differentiation in terms of - cell division, - & cell motility Actin - Contraction and relaxation of membrane or RBC

Tubulin and actin in reticulocytes membrane

Main function of RBC membrane

Facilitates Iron transport Maintains the membrane integrity Maintains the membrane deformability

Constantly changes as it moves through the circulation Soft and pliable Biconcave shape (Maximum Surface area) Consists of a membrane skeleton protein lattice and lipid bilayer More than 50 transmembrane proteins have been identified and more than half carries blood group antigens

Mature Red Blood cell membrane

Rbc membrane composed of three comoponents

40% Lipids mostly phospholipids, cholesterol 8% carbohydrate linked to lipid or protein 52% glycoproteins

Structure of RBC membrane Membrane lipids

Outer layer: Phosphatidyl choline Sphingomyelin Inner layer: Phosphatidyl ethanolamine Phosphatidyl serine

Structure of RBC membrane Membrane proteins

Integral protein: Band 3 (Anion exchanger protein) Glycophorin Aquaphorin Peripheral protein: Spectrin Actin Protein 4.1 Pallidin (Band 4.2) Ankyrin Adducin Tropomycin Tropomodulin

Peripheral protein is responsible for

Cell shape and structural deformability

Band 3 protein is responsible for

Prevention of Surface Area loss Binding site of enzyme and cytoplasmic membrane Anion transport - Exchanges bicarbonate for chloride Linkage of lipid bilayer to underlying membrane skeleton - Interaction w/ ankyrin and protein 4.2, secondarily through binding to protein 4.1

Vertical interaction

Stabilizes the lipid bilayer membrane

Horizontal interaction

Support the structural integrity of RBC

Glycophorin

Imparts a negative charge to the cell Glycophorin A carries MN, Gerbich blood group antigen Glycophorin C, Glycophorin A important for P. Falciparum invasion and development in RBC

Aquaporin 1

Selective pores for water transport Allows RBC to remain in osmotic equilibrium with ECF

Red cell membrane skeleton

Hexagonal lattice with 6 spectrin molecules Each linked to multiple spectrin tetramers Composed of spectrin, actin, 4.1 Ankyrin links the lipid bilayer to membrane via interaction with band 3

Spectrin is responsible for

Flexible, rod-like molecule Biconcave shape for red blood cell Important factor in RBC integrity (Binds to other peripheral proteins e.g actin, ankyrin, adducin) then forms a skeletal network of microfilaments Two sub-units (Alpha and Beta) Beta spectrin: Attachment for ankyrin near C terminus (which binds cytoplasmic tail of band 3) thus attachment of skeleton to lipid bilayer At N terminus: Attachment for 4.1 protein (associated with glycophorin C) - second anchor point with lipid membrane Binding sites for actin filaments and protein 4.1 - forming a junctional complex

Skeletal Network of microfilaments

Strengthen the membrane Controls the biconcave shape Controls the deformability of the cell Provide stability of the RBC

Most abundant peripheral protein

Spectrin

Actin and its function

Contraction and retraction of membrane Short, uniform filaments Length modulated by tropomyosin/Tropomodulin Approximately 6 spectrin ends interface with one actin filament stabilized by protein 4.1

Skeletal RBC protein Regulates actin proliferation

Tropomyosin

Controls the actin filaments (paghaba)

Tropomodulin

Stabilizes actin-spectrin interaction

Protein 4.1

Stabilizes interaction of spectrin with actin Influenced by calmodulin (Calcium binding protein) Promotes spectrin actin interactions

Adducin

Interacts with band 3 and spectrin to achieve linkage between bilayer and skeleton Augmented by protein 4.2 Anchors the lipid bilayer via spectrin and band 3

Ankyrin

Red cell mechanics

RBC survival Deformability is an important property of red cell function Influenced by: Cell shape - Ratio of cell surface area to cell volume Cytoplasmic viscosity - Regulated by MCHC and thus cell volume Membrane deformability and stability

Red Blood Cell shape

Biconcave disc shape creates and advantageous surface area/ volume relationship Facilitates deformation while maintaining constant surface area Progressive loss of intracellular and membrane components results in biconcave shape and improved deformability SA/V ration alteration will result in a spherical shape with less redundant surface area, thus less capacity for deformability and diminished survival. Membrane loss = reduced SA Increase in cell water content = increased volume

Membrane deformability/Stability

During pressure upon RBC, spectrin molecules undergo reversible change in conformation: some uncoiled and extended, others compressed and folded During extreme or sustained pressure, membrane exhibits permanent plastic deformation Deformability can be reduced by increases in associations between skeletal proteins or between skeletal and integral proteins (esp band 3)

Cytoplasmic characteristics

Cytoplasmic contents of RBCs include: Potassium ions, Sodium ions, glucose, intermediate products of glycolysis and enzymes Embded-meyerhof pathway utilizes 90% of RBC total glucose Efficient cellular metabolism depends on long-lived enzymes

Major source of the essential cellular energy Glucose undergoes glycolysis (Glucose to lactate) to form ATPs Maintains pyridine nucleotides in a reduced state to permit their function in oxidation-reduction reactions within the cell Deficiencies to production of ATP can be exhibited by: Premature cell death due to inherited defects in glycolysis Loss of viability during the storage of blood for transfusion

Embden-Meyerhof Pathway

Oxidative catabolism of glucose with reduction of NADP (nicotinamide-adenine dinucleotide phosphate) to NADPH (reduced form of NADP) which is required to reduce glutathione Pathway's activity is increased w/ increased oxidation of glutathione If pathway is defective, amount of reduced glutathione becomes insufficient to neutralize oxidants - causes denaturation of globin (Heinz bodies)

Oxidative pathway or Hexose monophosphate shunt

Depends on embden-meyerhof pathway for the reduced pyridine nucleotides that keeps hemoglobin in a reduced state Prevents the oxidation of heme iron Requires the reducing action of NADH and the enzyme methemoglobin reductase Maintains hemoglobin in FERROUS STATE to bind to oxygen Methemoglobin (ferric state) can't bind to oxygen

Methemoglobin reductase pathway

Important in the oxygen carrying capacity of RBCs Mechanism is low in energy consumption Capable of regulating oxygen transport even with hypoxia and acid-base disorders Permits accumulation of 2,3 DPG Increased in deoxyhemoglobin results to binding of 2,3 DPG which stimulates glycolysis

Leubering-Rapoport pathway

Summary of metabolic pathways in the erythrocyte

Embden meyerhof - Maintains cellular energy by generating ATP Oxidative or hexose-monophosphate shunt - Prevents denaturation of globin of the hemoglobin molecule by oxidation Methemoglobin reductase - Prevents oxidation of heme iron Leubering-Rapoport - Regulates oxygen affinity of hemoglobin

Term that describes the dynamics of RBC production and destruction

Erythrokinetics

Name given to the collection of all stages of erythrocytes throughout the body, developing precursor in the bm and the circulating rbc in peripheral blood

Erythron

Hormone produced in the kidney in response to tissue hypoxia Actions: 1. Induces committed progenitor cells in the bone marrow to differentiate and proliferation into pronormoblast 2. Shortens the generation time of pronormoblast 3. Promotes the early release of reticulocytes to the peripheral blood

Erythropoietin (EPO)

Elevated EPO levels are observed in

Erythroid hyperplasia Polycythemia Hemorrhages Inclusion RBC destruction

Decreased EPO levels are observed in

Anemia End stage renal disease (Kidney)

Mechanism of red cell destruction Loss of a portion of the erythrocytes membrane, accompanied by loss of cellular contents, including hemoglobin

Fragmentation

Mechanism of red cell destruction Passing of water into the red cell as to ultimately burst it

Osmotic lysis

Mechanism of red cell destruction Ingestion of whole red cells by circulating monocytes or neutrophil or by fixed macrophages of the mononuclear phagocyte system

Erythrophagocytosis

Mechanism of red cell destruction Complement has the ability to attach itself to the cells and induce lysis

Complement induced cytolysis

Mechanism of red cell destruction When hemoglobin is exposed to oxidant stress and the mechanism to protect the cell from such damage fails to work, denatured hb precipitates forming inclusion bodies known as heinz bodies

Hemoglobin denaturation

Lysis of erythrocytes which occurs within the CIRCULATION throughout the classic pathway. It is the usual outcome of sensitization of erythrocytes with complement. 10% of aged red cell undergo the destruction

Intravascular hemolysis

Lysis of erythrocytes OUTSIDE of CIRCULATION, in the RES (Reticuloendothelial System) of the cell liver, spleen. Usually happen through phagocytosis About 90% of aged red cells are destroyed

Extravascular hemolysis

Causes of intravascular hemolysis

ABO mismatched blood transfusion Cold agglutinin disease Paroxysmal cold hemoglobinuria Burns Snake Bites Bacterial - C. perfringens sepsis Parasitic infections - P. malaria Mechanical heart valves Paroxysmal nocturnal hemoglobinuria

Causes of extravascular hemolysis

Bacterial/ Viral infections Drug induced Autoimmune Microangiopathy - Malignancy DIC, TTP, Eclampsia Hemoglobinopathies Membrane defects - spherocytosis, elliptocytosis, acanthocytosis Metabolic defects - G6PD deficiency/ Oxidant drugs

An iron bearing protein contained within the erythrocytes It is synthesized by young erythroblast from the polychromatophilic normoblast stage up to reticulocytes stage One gram of this can carry 1.34 of oxygen

Hemoglobin

Functions of hemoglobin

Transport oxygen from the lungs to the tissue and carbon dioxide from the tissue Acid-base balance regulation - binding and releasing / transport of nitric oxide (regulator vascular tone)

Composition of heme

1. Protoporphyrin IX consists of a ring of : A. Carbon B. Hydrogen C. Nitrogen 2. Ferrous iron

The hemoglobin molecule can be described by its Amino acid sequence

Primary

The hemoglobin molecule can be described by its Helices and non helices

Secondary

The hemoglobin molecule can be described by its Pretzel like configuration

Tertiary

The hemoglobin molecule can be described by its Complete molecule

Quaternary

Ability of hemoglobin to bind or release oxygen. Expressed in terms of the oxygen tension at which hgb is 50% saturated

Oxygen affinity

Relationship of oxygen affinity with hemoglobin to pH which states that Increase pH (alkalosis) = Increase hemoglobin affinity for oxygen Decrease pH (acidosis) = decrease hemoglobin affinity for oxygen

Bohr Effect

Increased temp Increased 2,3 DPG Increased Hydrogen concentration Reduced oxygen affinity More oxygen released to the tissues Acidosis (dec pH)

Right shift

Decreased temp Decreased 2,3 DPG Decreased Hydrogen concentration Increase oxygen affinity Less oxygen released to the tissues Alkalosis (Inc pH)

Left shift

Synthesis of globin chains

Occurs in the cytoplasm of normoblast and reticulocytes Polypeptide chains are manufactured in the ribosomes Globin protein are made via transcription of M genetic code to mRNA and translation of mRNA

Hemoglobin in combination with oxygen Gives pinkness to the skin and mucous membrane. Seen in arterial circulation

Oxyhemoglobin

Hemoglobin with iron but no oxygen seen in venous circulation Unassociated with oxygen

Deoxyhemoglobin

Found in normal human embryos and fetuses with a gestational age of less than 3 months Absent at birth

Embryonic hemoglobin

The major hemoglobin of the fetus and the newborn Composed of 2 alpha and 2 gamma Produced FOUR MONTHS after conception

Fetal hemoglobin HbF

Normal adult hemoglobin 95% - 97% of hemoglobin in normal adults produced after one year onwards Composed of 2 alpha and 2 beta chains

Hemoglobin A or Hemoglobin A1

Constitutes less than 3% of the total hemoglobin Composed of 2 alpha and 2 delta

Hemoglobin A2

Degradation product of hemoglobin A2 Composed of 2 alpha and 2 delta

Hemoglobin A3

Primary hemoglobin in people with sickle cell disease Those with Hb S disease have two ABNORMAL BETA chains and two normal alpha chains causes the red blood cell to deform and assume a sickle shape when exposed to decrease amounts of oxygen Glutamic acid is REPLACED by VALINE in the 6TH position of beta chain

Hemoglobin S (SaGaVAL)

About 2-3% of people of west african descent are heterozygotes for hemoglobin C Instead of glutamic acid, lysine is in B6 It usually causes a minor amount of hemolytic anemia and a mild to moderate enlargement of the spleen

Hemoglobin C (CGaL)

One of the most common beta chain hemoglobin variants in the world A single copy of the hemoglobin E gene does not cause symptoms unless it is combined with another mutation, such as the one for beta thalassemia trait Mild hemolytic anemia, Microcytic red blood cells, and a mild enlargement of the spleen Substitution of glutamic acid to lysine on 26th position

Hemoglobin E (26th EGaL)

An abnormal hemoglobin that occurs in some cases of alpha thalassemia Composed of four beta globin chains and is produced in response to a SEVERE SHORTAGE OF ALPHA CHAINS

Hemoglobin H

Are acquired hemoglobin variants whose structure has been modified by drugs or environmental chemicals. Do not transport oxygen to the tissue well resulting in cyanosis

Chemically modified Hemoglobins

A form of hemoglobin in its FERRIC state Brownish to bluish color and does not revert to red on exposure to oxygen Peak in the range of 620-640 nm at pH 7.1 under spectral absorption test Causes: Presence of oxidants Genetic deficiency - decrease activity of MethHB 30% - Cyanosis 50% - Coma to death Resolution: Intravenous methylene blue

Methemoglobin (HI)

Formed by the irreversible oxidation of Hemoglobin of certain drugs and chemicals Examples: Sulfonamides Phenacetin Acetanilide Formed by the addition of HYDROGEN SULFIDE to hemoglobin Has a greenish pigment If it reaches the critical level in the blood, it imparts MAUVE LAVENDER

Sulfhemoglobin

Sulfhemoglobin can be usually reported in the following situation:

Patient under prolonged treatment with sulfonamides or aromatic compounds (Acetanilide, Phenacetin) Patient with severe constipation In cases of bacteremia caused by C.perfringens In condition known as enterogenous cyanosis

Results from the binding of carbon monoxide to heme iron Hemoglobin can combine with carbon monoxide with affinity 200 times greater than that of oxygen Carbon monoxide is termed as silent killer for its colorless gas, odor, and patient becomes easily hypoxic

Carboxyhemoglobin

Hemoglobin determination

Visual methods Gasometric Method Spectronic Method Automated Other methods such as Alkaline, SG, Comparator

Hemoglobin determination Visual methods

Sahli method Dares Method Hadens method Wintrobe Haldene -Lyses RBC w/ Hypotonic solution Tallquists - Quickest method

Hemoglobin determination Spectronic Method

Oxyhemoglobin Cyanmethemoglobin

Qualitative screening test based on Specific gravity. The density of the drop of blood is directly proportional to the amount of hemoglobin it contains The principles of the test is that when the drop of donor's blood dropped into copper sulfate solution becomes encased in a sac of copper proteinate, which prevents any change in the specific gravity for 15 seconds

Copper sulfate specific gravity

Hemoglobin will combine and liberate a fixed quantity of oxygen. The blood is hemolyzed with saponin and the gas is collected and measured in a Van Slyke apparatus Research purposes Most accurate method

Gasometric Method (Oxygen capacity method)

Measures plasma hemoglobin

Oxyhemoglobin method

Standard and reference method Measure the difference type of hemoglobin except sulfhemoglobin Blood is diluted in a solution of potassium ferricyanide and potassium cyanide. The hemoglobin is oxidized to methemoglobin by the potassium ferricyanide The potassium cyanide then converts the methemoglobin to cyanmethemoglobin. The absorbance is measured spectrophotometrically at 540nm Drabkin's reagent is used

Cyanmethemoglobin

Measuring hemoglobin using cyanmethemoglobin

5mL Drabkin's reagent 20 uL whole blood Mix then stand for about 5 minutes transfer to cuvette and read at 540nm

Increased hemoglobin level Found in: Polycythemia Dehydration Changing from high to low altitudes

Hyperchromia

Decreased hemoglobin levels

Oligochromia

Hemoglobin determination test is used to

Screen for disease associated with anemia Determine the severity of anemia Follow the response to treatment for anemia Evaluate polycythemia

Decreased hemoglobin levels can be seen in the ff conditions

Anemia Iron deficiency, Thalassemia, Pernicious anemia Liver disease, hypothyroidism Hemorrhage Hemolytic anemia caused by transfusion of incompatible blood Rxn to chemical or drugs Rxn to infectious agents Various systemic disease such as Hodgkins disease Leukemia Lymphoma SLE

Increased hemoglobin levels are found in

Polycythemia vera Congestive heart failure COPD

Variations in hemoglobin levels

Occurs after transfusion, hemorrhages, burns The H and H provide valuable information in an emergency situations

Interfering factors

People living at high altitudes have increased Hb values as well as inc. Hct and RBC Excessive fluid intake cause a decrease Hemoglobin Hemoglobin is higher in infants Drugs Hemoglobin is normally decreased in pregnancy

Clinical alert

Panic hemoglobin is less than 5.0 g/dL a condition that leads to heart failure and death A value more than 20g/dl leads to clogging of the capillaries as a result of hemoconcentration

Refers to erythrocytes with normal amount of hemoglobin Possesses a central pallor which is about 1/3 of its diameter

Normochromic cell

Refers to erythrocytes wherein the central light area of the cell is larger and paler than the normal MCH and MCHC are decreased often associated with microcytosis

Hypochromic cell

Red cells which have an increase Hb content and wherein the central light area is smaller than the normal

Hyperchromic cell

Condition wherein the red cell are stained with various shades of blue with tinges of pink This is due combination of the affinity of hemoglobin to acid stain and the affinity of RNA to the basic dye Slightly microcytic (RBC are smaller) Indicates reticulocytotic

Polychromasia

Polychromasia grading

Slight - 1% 1+ - 3% 2+ - 5% 3+ - 10% 4+ - >11% percentage = percent of rbcs that are polychromatophilic

Condition where in the red cells appear pale 2 possible causes: Decrease Hemoglobin concentration Abnormal thinness of the cells: Iron deficiency anemia Sideroblastic anemia Thalassemia

Hypochromasia

Hypochromasia grading

1+ - area of central pallor is 1/2 of cell diameter 2+ - Area of central pallor is 2/3 of cell diameter 3+ - Area of central pallor is 3/4 of the cell diameter 4+ - Thin rim of hemoglobin

Condition wherein the red cell are deeply stained to abnormal thickness of cells Macrocytosis Spherocytosis Megaloblastic anemia

Hyperchromasia

Condition where in the red cell vary in size both macrocytes and microcytes coexist on the same smear Associated with acute post hemorrhagic anemia, hemolytic anemia and aplastic anemia

Anisocytosis

6-8 um in diameter (Normal)

Normocyte

Larger than normal, greater than 8um in size round in shape MCV > 100 FL Defect: Abnormal nuclear maturation but normal cytoplasmic maturation Associated disease: Non-megaloblastic anemia myelodysplastic syndrome Chronic Liver Disease BM failure Reticulocytosis

Macrocyte

Cell which is less than 6 um in size MCV less than 80FL Defect: Abnormal cytoplasmic maturation but normal nuclear maturation Found in: IDA Thalassemia Hemolytic anemia Hb E disease Inflammation Chronic post hemorrhagic anemia Sideroblastic anemia

Microcyte

Large oval-shaped red cell which is 9-12 um Defect: Abnormal nuclear maturation but normal cytoplasmic maturation Megalocytosis is found in: Megaloblastic anemias like pernicious anemia Vit B12 deficiency anemia or vit b12 def D. latum infection

Megalocyte

Red cells exhibit variation in shape

Poikilocytosis

Normal cell with a biconcave disc shape with increased surface volume Associated disease Normal condition Acute post hemorrhagic anemia Aplastic anemia

Discocyte

Small dense RBC with few irregularly spaced projections of varying length Defect: Abnormal membrane defect caused by an increase sphingomyelin and decrease in cholesterol and phospholipid Associated diseases: Neuroacanthocytosis (Abetalipoproteinemia, Mcleod syndrome) Sever liver disease (Spur cell anemia)

Acanthocyte (Spur cell)

Cell assumes a pocket book roll appearance or biscuit shape Defect: Cell membrane is folded Associated disease: HbSC disease (hemoglobin sickle C disease) HbCC disease

Biscuit cell (Folded RBC)

Cell with irregularly spaced blunt processes. resembles crenated RBC Defect: Abnormal LIPID content of the membrane Associated disease: Uremia, MAHA, Liver disease, DIC, TTP, Pyruvate Kinase deficiency

Burr cell (Echinocyte)

Cell w/ eccentric vacuoles due to the plucked out heinz body Defect: G6PD deficiency resulting to accumulation of heinz body Associated disease: G6PD deficiency Hemolytic urine Syndrome Microangiopathic hemolytic anemia

Blister cell (Bite Cell)

Sea urchin cells If pathological: Due to abnormal lipid content of the membrane If artifactual: ATP deficiency due to prolonged storage of anticoagulated blood Associated disease: Uremia Bleeding ulcers Gastric carcinoma Hepatitis Cirrhosis

Echinocytes or crenated cells

Target cell / Mexican Hat / Cells with bull's eye appearance Cell w/ central area of hemoglobin surrounded by colorless area and a peripheral ring Defect: Deficiency in cholesterol, phospholipid membrane. Deficiency in Lecithin cholesterol acyl transferase (LACT) Associated with: Thalassemia Liver disease Hemolytic anemia HbSS HbCC LCAT deficiency

Codocyte or Leptocyte

Cells appear in the shape of a teardrop or pear with a single short or long protrusion Defect: Abnormal maturation squeezing and fragmentation during splenic passage Associated disease: Hemolytic anemia Megaloblastic anemia Myelofibrosis w/ Myeloid metaplasia Tennis Racquet

Tear drop cell (Dacryocyte/Dacrocyte)

Also known as ovalocyte Appear as oval or elliptical Egg shape, Cigar, Rod, Pencil form, sausage form Hemoglobin appears to be concentrated at the two ends of the cell leaving a normal central area of pallor Life span is shortened Can be found in healthy person Defect: Abnormal membrane due to defective SPECTRIN, defective in band protein 4.1 Associated with Megaloblastic anemia Hypochromic anemia Hereditary ovalocytosis

Elliptocytes

Small round dense cell which LACKS the central pallor area usually microcytic and sphere shaped Defects: Primary: Spectrin deficiency Secondary: Defective interaction of spectrin with other skeletal protein Associated with: Hereditary spherocytosis Chronic Lymphocytic leukemia Immune hemolytic anemia due to ABO incompatibility

Spherocyte

Cells are smaller and denser with increase hemoglobin content and become less deformable with age Shortened survival time because they can be sequestered in the spleen and destroyed Associated with Hereditary spherocytosis Immune hemolytic anemia Extensive burns (along w/ schistocytes)

Spherocyte

Crescent shape cell due to abnormal aggregation of HbS which gives a tendency for the cell to assume a sickle shape Sickle cells are thin and elongated with pointed ends and are well filled with hemoglobin They may be curved or straight or have S, V, or L shaped Found in sickle anemia and sickle cell trait Also known as Menisocyte

Sickle cells (Drepanocytes)

Irregularly contracted cell; fragmented cell Defects: Cell fragmentation due to trauma caused by physical and mechanical agents Associated with Microangiopathic hemolytic anemia Thrombotic Thrombocytopenic Purpura Hemolytic urine syndrome Uremia

Schistocytes

Mouth cells / Hydrocyte Characterized by an elongated or slit-like area of central pallor Caused by osmotic changes due to cation imbalance (Na, K) Associated with Alcoholic cirrhosis Hereditary stomatocytosis Hepatobiliary disease Rh null syndrome

Stomatocyte

Supravital stain: Dark blue granules and filaments in cytoplasm Wright stain: Bluish tinge throughout cytoplasm Composition of inclusion: RNA Associated disease: Hemolytic anemia After treatment for iron, vitamin B12, or Folate deficiency

Diffuse basophilia

Supravital stain: Dark blue-purple, fine or coarse punctate granules distributed throughout cytoplasm Wright stain: Same with supravital stain Composition of inclusion: Precipitated RNA Associated disease: LEAD POISONING Thalassemia Hemoglobinopathies Megaloblastic anemia Myelodysplastic syndrome

Basophilic stippling

Supravital stain: Dark blue-purple dense, round granule, usually one per cell; occasionally multiple Wright stain: Same with SS Composition of inclusion: DNA (nuclear fragment) Associated disease: Hyposplenism Postspelenctomy Megaloblastic anemia Hemolytic anemia thalassemia Myelodysplastic syndrome

Howell-Jolly bodies

Supravital stain: Round, Dark blue-purple granule attached to inner RBC membrane Wright stain: Not visible Composition of inclusion: Denatured hemoglobin Associated disease: G6PD deficiency Unstable hemoglobins Oxidant drugs/chemicals

Heinz body

Supravital stain: Irregular cluster of small, light to dark blue granules often near periphery of the cell Wright stain: Same with SS Composition of inclusion: Iron Iron stain: Prussian blue (Perl's prussian blue) Associated disease: Sideroblastic anemia Hemoglobinopathies Thalassemia megaloblastic anemia Myelodysplastic syndrome Hyposplenism Post-splenectomy

Pappenheimer bodies

Sideroblastic anemia

Blockage in protoporphyrin = many pappenheimer's bodies

Supravital stain: Rings or figure-eights Wright stain: BLUE rings of figure-eights Composition of inclusion: Remnant of mitotic spindle Associated disease: Megaloblastic anemia Myelodysplastic syndromes Tall hat

Cabot ring

Supravital stain: Fine, evenly dispersed, dark blue granules; imparts "golf ball" appearance to RBCs Wright stain: NOT VISIBLE Composition of inclusion: Precipitate of b-globin chains of hemoglobin Associated disease: Hb H disease

Hemoglobin H inclusions

Stacks of coins appearance Maybe pathologic Pangit smear High protein (Multiple myeloma, waldenstrom macroglobulinemia)

Rouleaux formation

Red cell is colored red

Acid stain of erythrocytes

Red cells are dirty gray

Alkaline stain of erythrocyte

Caused by fat or oil on the slide ahead of the spreader during the smear prep

Design formation of RBC

Extraction mistake

Partially hemolyzed RBC

Hema week 1 Flashcards

(185 cards)