Exercise 25: 25: COMPLETE BLOOD COUNT
BLOOD - Flows and circulates within Cardiovascular system - Connective tissue - Has a liquid matrix * serum- clotted blood; w/o fibrinogen *plasma- unclotted blood
BLOOD TESTS - For clinical evaluation of patients - Blood Samples are collected
HEMOCYTOMETER - Counting the cellular elements of the blood (RBC, WBC & Platelets) - What to look for (besides count): RBC: color, size, borders, parasites WBC : early forms , contour granulation, parasites/ inclusions •
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Hematocrit Hematocrit (Hct) / packed cell volume - %RBCs in plasma of peripheral blood - Can also be calculated: Hct = (RBC x MCV)/10 - Increased in: Polycthemia (reactive or neoplastic), shock, severe dehydration - Decreased in: Leukemia, Anemia, Hyperthyroidism, Cirrhosis Male: 47 +/- 7% v/v Female: 42 +/- 5% v/v
At Birth: 56 +/- % v/v8 Adam’s Micromethod
Hemoglobin o o o o o
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capillary tube – ¾ filled w/ blood cover with sealing clay microhematocrit centrifuge 10,000 rpm for 4-5 mins microhematocrit reader
Red pigmented protein Transports oxygen and Carbon dioxide Measured as Oxyhemoglobin pigment in RBC that has affinity to O 2 blood: the darker color, the more hemoglobin
Hgb
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Hgb concentration on whole blood (g/dL) Increased in: High Altitudes, COPD, CHF, Polycthemia Decreased in: Severe Hemorrhage , Anemia, Kidney Disease, Hyperthyroidism, Liver cirrhosis Sahli’s apparatus Male: 14-16.5 g/100mL Female: 12-15g/100ml
Acid-Hematin Method
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a. 0.01N HCl : 2 mark : Sahli’s tube b. 0.02 mL blood : Sahli’s pipette combine a + b adding distilled H 2O until the mixture matches the color of the comparator block
WHITE AND RED BLOOD CELL COUNT RBCs or WBCs/ µL of blood • •
RBCs: -Increased: Polycthemia, Globin synthesis Problem -Decreased: Anemia
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WBCs - Increased: Leukocytosis (non/neoplastic) - Decreased: Leukopenia
Red Blood Cell / Erythrocyte O2 transport o CO2 disposal o
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Blood Flow, BP Buffer
White Blood Cell/ Leukocyte Granulocytes Neutrophils 60-70%
Eosinophils
2-4%
Basophils
0.5-1 %
Acute infections ▪ Phagocytosis ▪ Bacterial Destruction ▪
Allergic Responses ▪ Phagocytize antigen-antibody complexes ▪ Destroys some parasitic worms ▪
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Aggranulocytes Lymphocytes
20-25 %
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Monocytes
3-8 %
Chronic infections Heparin, H1, 5-HT release (allergy Antibody reactions B cells and plasma cells T cells (cytotoxic & helper) Natural killer cells
Chronic infections ▪ Becomes Macrophage ▪
Differential WBC Count • Neutrophil (60-70%) - Inc: acute infections, Cancer - Dec: certain cancers, chemo, other medications •
Eosinophil (2-4%) - Inc: Allergic reaction, Parasites
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Basophil (0.5-1%) - Heparin, H1, 5-HT release (allergy) - Inc: allergies, Myeloproliferative, Hypothyroidism, Chronic Infections
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Lymphocyte (20-25%) - Inc: antibody reactions - Dec: HIV
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Monocyte (3-8%) - Becomes Macrophage - Inc: Chronic Infections
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Absolute Neutrophil Count (ANC) - WBC Count x % (PMN + Bands) * PMN = poly (polymorphonuclear), aka neutrophils * bands= immature poly o
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Neutropenia - Mild- ANC ≥ 1000 and < 1500 cells/microL - Moderate- ANC ≥ 500 and < 1000 cells/microL - Severe- ANc < 500 cells/microL *Benign ethnic neutropenia (BEN) ANC < 1000 - 1200 cells/microL
EXERCISE 26: BLOOD GROUPS RBCs have 300 + blood group antigens, each with own blood g roup system • Antibodies (aka immunoglobulins ) – Y-shaped proteins used by immune system to o neutralize pathogen; in the plasma ; agglutinins Antigen: sugars/ proteins on cell membrane surface (linked by membrane o proteins/lipids); on surface of RBC ; agglutinogens ; inherited
*ABO *Rh *Glob *I *MNSs *Kell *Duffy *Kidd *Lutheran
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ABO groupings Anti-A antiserum (blue) o Anti-B antiserum (yellow) o
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Rh Grouping o
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Anti-D antiserum (colorless)
Importance: - Blood transfusion and hemolytic transfusion reaction (ABO, Rh) ➢ RBC (ABO, Rh, etc antigens ) ➢ Platelets (ABO antigens) ➢ Plasma (ABO antibodies) - Tissue/ Organ Transplant - Risk for Infections (E. coli , malaria, etc) - Hemolytic disease of the fetus and newborn (HDFN/HDN) •
ABO
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Rh – over 45 antigens - Most common: D, C, c, E and e - Rh-negative: no Rh (D) antigen - Rh –negative mother carrying the Rh – positive baby can develop antibodies for Rh (D) → Rh (D) alloimmunization
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Consequences of Mismatch - Acute hemolytic transfusion reactions (clerical errors) → life-threatening - Hemolytic disease in fetus and newborn → usually mild - Organ rejection
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Antigen (Ag) –agglutinogen (blue)
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Antibody (Ab) – agglutinin (yellow)
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Reaction : Agglutination (colorless)
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There has to be antigen and antibody to have reaction → agglutination
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Agglutination = incompatibility
EXERCISE 27&28: COAGULATION TIME & BLEEDING TIME PLATELETS PLATELETS: 40,000-440,000 cells/mm3 w/ circulating life span of 5 – 10 days • •
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THROMBOCYTOPENIA: < 100,000 cells/mm3 Bleeding Risk substantially elevated < 20,000 cells/mm 3
HEMOSTASIS Platelet plug formation • - Platelet adhesion- deposition of platelets on subendothelial matrix
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Platelet aggregation-platelet-platelet cohesion (via GPIIb/IIIA br idged by fibrinogen)
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Platelet secretion- release of platelet granule proteins (contain 5-HT, ADP, TXA2, etc.)
Clotting / coagulation cascade Clotting termination Clot removal (fibrinolysis)
COAGULATION TEST Activated Partial Thromboplastin Time (aPTT) • w hen exposed to substances activating the contact - Time it takes plasma to clot when factors which assess intrinsic & common pathways of coagulation (no tissue factor) •
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Prothrombin time (PT) & International Normalized Ratio (INR) - PT: Time it takes plasma to clot when ex posed to tissue factor, which assesses the extrinsic & common pathways of coagulation - INR= [Patient PT ÷ Control PT] ISI
BLEEDING DISORDERS HEMOPHILIA • - A (classic; VIII) - B (Christmas; IX) - C (Rosenthal; XI)
Clotting - important in arresting hemorrhage - 3-6 mins Clot - plugs the opening of the wound Drop / Slide Method - 30 sec interval - from the moment the blood drops to the slide until the formation of the fibrin thread Bleeding Time - refers to the time it takes for a puncture wound to stop bleeding - 1-3 mins - depends upon the depth of the wound and degree of hyperemia in the finger Duke’s Method
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first drop of blood blotted on a filter paper rough surfaces shorten bleeding time appearance of first drop to the time bleeding stops
EXERCISE 29: HYPEREMIA OR CONGESTION Hyperemia Vasodilation in inflammation ❖ Calor- Heat ❖ Rubor-Red ❖ Tumor- Swelling ❖ Dolor- pain Isopropanol: Tissue damage, Increased Blood supply
Congestion ➢ Passive hyperemia - Stagnated deoxygenated blood - Dusky red/ Bluish Color, cold feeling - E.g. Congestive Heart Failure
Hyperemia / Congestion - increase of blood flow or an excess flow of blood in the vessel in a certain area of the body Active Hyperemia Hyperemia - increased blood flow to a specific area / dilation of blood vessels Passive Hyperemia -
slowing down of venous return
EXERCISE 30: 30: CAPILLARY RESISTANCE TEST
Tourniquet Test Test microcapillary fragility • •
Initial screening tool for dengue hemorrhagic fever (not diagnostic)
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Other conditions/ diseases: (not used widely) - Scurvy - Bleeding disorders - Scarlet fever - Endocrine problems (hyperthyroidism) I nflammatory conditions - Medications, Allergy and Inflammatory
Capillary resistance test - measures the ability of the capillary walls to resist r esist pressure - arm : 100 mmHg Thrombocytopenic Thrombocytopenic Purpura Purpura - capillaries will rupture at this pressure Petechiae / Hemorrhages - tiny spots Tourniquet / Rumpel-Leede Rumpel-Leede / Hess Test