Post on 22-Jan-2018
Basics of Haematology
Presented by:
Rayaz Ahmad Bhat
Student:
M.Pharm Pharmacy PracticeM.Pharm Pharmacy PracticeNational Institute Of Pharmaceutical Education And
Research,Guwahati, Assam
Mentor Institute: National Institute Of Biologicals , Noida , Delhi
Mentor:
Dr. Akanksha Bisht
Officer Incharge – HvPI & Scientist Grade-III
Flow Of Presentation
• Properties and Functions
• Components• Blood Indices
Blood: Introduction
• Erythropoiesis• Leukopoiesis• Thrombopoiesis
Haematopoiesis
Blood Grouping Systems
Blood Products: Storage , Shelf-life
Blood
Blood is a connective tissue in fluid form.
Blood is considered as:
fluid of life
fluid of growth
fluid of health
PropertiesColor: Blood is red in color.o Arterial blood is scarle redo Venous blood is purple redo Reaction and pH: Blood is slightly alkaline and its pH
in normal conditions is 7.4.o Specific gravity:
o Specific gravity of total blood : 1.052 to 1.061o Specific gravity of total blood : 1.052 to 1.061o Specific gravity blood cells : 1.092 to 1.101o Specific gravity of plasma : 1.022 to 1.026
o Volume:o Normal adult male: 5 Lo Normal adult female: 4.5 Lo New born baby: 450 ml
o Viscosity: Blood is five times more viscous than water
Functions
o Nutritive function
o Respiratory function
o Excretory function
o Transport of enzymes and hormoneso Transport of enzymes and hormones
o Regulation of water balance
o Regulation of Acid-Base balance
o Regulation of body temperature
o Storage function
o Defensive function
Composition
Blood consists of two major components:
Plasma: Liquid portion
1. Water
2. Organic substances
3. Inorganic substances3. Inorganic substances
4. Gases
Formed Elements:
1. Red Blood Cells or erythrocytes
2. White Blood Cells or leukocytes
3. Platelets or thrombocytes.
Hematocrit
PlasmaPlasma is a straw- colored clear liquid part of blood.
• Normal values:
• Normal values of the plasma proteins are:
• Total proteins : 7.3 g/dL (6.4 to 8.3 g/dL)
• Serum albumin : 4.7 g/dL
• Serum globulin : 2.3 g/dL
• Fibrinogen : 0.3 g/dL• Fibrinogen : 0.3 g/dL
• Albumin/Globulin ratio:
• Ratio between plasma level of albumin and globulin is called albumin/globulin (A/G) ratio.
• It is an important indicator of some diseases involving liver or kidney.
• Normal A/G ratio is 2 : 1
Properties of Plasma Proteins
• Molecular weight• Albumin : 69,000
• Globulin : 1,56,000
• Fibrinogen : 4,00,000
• Thus, the molecular weight of fibrinogen is greater than that of other two proteins.than that of other two proteins.
• Oncotic pressure• Normally, it is about 25 mm Hg . Albumin plays a
major role in exerting oncotic pressure.• Specific gravity: 1.026
• Buffer action: 1/6th total buffering action of blood
Function of Plasma Proteinso Role in coagulation of blood
o Role in defense mechanism of body
o Role in transport mechanism
o Role in maintenance of osmotic pressure in blood
o Role in regulation of acid-base balanceo Role in regulation of acid-base balance
o Role in viscosity of blood
o Role in erythrocyte sedimentation rate
o Role in suspension stability of red blood cells
o Role in production of trephone substances
o Role as reserve proteins
Red Blood Cells(Erythrocytes)
Red blood cells (RBCs) are the non-nucleated,
disc shaped, biconcave formed elements in blood.
Normal Value: 4 to 5.5 milliom/mm3 blood.
Properties of RBC’s
• Rouleaux formation
• Specific gravity of RBC is 1.092 to 1.101
• Packed Cell Volume
• Suspension Stability
Life Span of RBC’s
Average lifespan of RBC is about 120 days. Afterthe lifetime the senile (old) RBCs are destroyed in
reticuloendothelial system.
Fate of RBC’sFate of RBC’s
When the cells become older (120 days), the cell membrane becomes more fragile.The destruction occurs mainly in the capillaries of red pulp of spleen .So, the spleen is called ‘graveyard of RBCs’.
Fate of RBC’s
Function of RBC’s
• Transport of Oxygen from the Lungs to the
tissues
• Transport of Carbon Dioxide from the tissues to• Transport of Carbon Dioxide from the tissues to
the Lungs
• Buffering Action in Blood
• In Blood Group Determination
Variations in RBC count
Physiological• Increase-Polycythemia
• Decrease
• Pathological• Pathological
• Increase
• Primary Polycythemia – Polycythemia Vera
• Secondary Polycythemia
• Decrease
• Anemia
Variations in size
• Microcytes• Microcytes are present in:
• i. Iron-deficiency anaemia
• ii. Prolonged forced breathing
• iii. Increased osmotic pressure in blood.• iii. Increased osmotic pressure in blood.
• Macrocytes• Macrocytes are present in:
• i. Megaloblastic anaemia
• ii. Decreased osmotic pressure in blood
• Anisocytes• Anisocytes occurs in pernicious anemia
Variation in shape
Shape of RBCs is altered in many conditions
including different types of anemia.
• 1. Crenation• 1. Crenation
• 2. Spherocytosis
• 3. Elliptocytosis
• 4. Sickle cell
• 5. Poikilocytosis
Packed Cell Volume• Normal PCV:
• In males = 40% to 45%• In females = 38% to 42%
• PCV increases in:
• 1. Polycythemia• 2. Dehydration• 2. Dehydration• 3. Dengue shock syndrome: Dengue fever (tropical
disease caused by flavivirus transmitted by mosquito Aedes aegypti) of grade III or IV severity
• PCV decreases in:
• 1. Anaemia• 2. Cirrhosis of liver• 3. Pregnancy• 4. Haemorrhage due to ectopic pregnancy.
Significance of PCV
• Determination of PCV helps in:
• 1. Diagnosis and treatment of anemia
• 2. Diagnosis and treatment of polycythemia• 2. Diagnosis and treatment of polycythemia
• 3. Determination of extent of dehydration and
recovery from dehydration after treatment
• 4. Decision of blood transfusion
Blood Indices
• Blood indices are the calculations derived from RBC count, haemoglobin content of blood and PCV. Blood indices help in diagnosis of the type of anaemia.
• Blood indices include:
• 1. Mean corpuscular volume (MCV) .Normal MCV is 90
cu μ (78 to 90 cu μ).cu μ (78 to 90 cu μ).
• 2. Mean corpuscular haemoglobin (MCH) . Normal value of MCH is 30 pg (27 to 32 pg).
• 3. Mean corpuscular haemoglobin concentration
(MCHC) . Normal value of MCHC is 30% (30% to 38%).
• 4. Colour Index : Normal colour index is 1.0 (0.8 to 1.2).
Anaemia
• Anaemia is the blood disorder, characterized by the reduction in:
• 1. Red blood cell (RBC) count
• 2. Haemoglobin content
• 3. Packed cell volume (PVC).3. Packed cell volume (PVC).
• Classification of Anemia
• Anaemia is classified by two methods:• 1. Morphological classification
• 2. Etiological classification
Morphological Classification
• Normocytic Normochromic Anemia
• Macrocytic Normochromic Anemia
• Macrocytic Hypochromic Anemia
• Microcytic Hypochromic Anemia
Etiological Classification
• 1. Hemorrhagic anemia
• 2. Hemolytic anemia
• 3. Nutrition deficiency anemia• 3. Nutrition deficiency anemia
• 4. Aplastic anemia
• 5. Anemia of chronic diseases.
White Blood Cells(Leukocytes)
• White blood cells (WBCs) or leukocytes are the colorless and nucleated formed elements of blood (leuko is derived from Greek word leukos = white).
• Compared to RBCs, the WBCs are larger in size and lesser in number with shorter life span.
• Two Classes:– Granulocytes
– Agranulocytes
Granulocytes
Agranulocytes
• T-Lymphocytes • T-Lymphocytes
• B-Lymphocytes
Size, Lifespan and Normal Values
Total WBC Count: 4000-11000/mm3 of Blood
Diameter and lifespan of WBCs
Differential WBC count
Properties of WBC’s
• Diapedesis
• Ameboid Movement• Ameboid Movement
• Chemotaxis
• Phagocytosis
Functions of WBC’s• NEUTROPHILS
• Neutrophils play an important role in the defense mechanism of the body
• EOSINOPHILS• Eosinophils play an important role in the defense mechanism of the
body against the parasites.
• BASOPHILS• Basophils play an important role in healing processes. So their number
increases during healing process.
• MONOCYTES• Monocytes are the largest cells among the leukocytes.
• Like neutrophils, monocytes also are motile and
phagocytic in nature. These cells wander freely through all tissues of the body
• LYMPHOCYTES• Lymphocytes play an important role in immunity
Pathological Variations In WBC’s
Platelets(Thrombocytes)
• Platelets are small colorless, non-nucleated and moderately refractive bodies of variable shapes.
• Normal platelet count is 2,50,000/cu mm of blood. It ranges between 2,00,000 and 4,00,000/cu mm of blood.4,00,000/cu mm of blood.
Properties and Functions
• Properties of Platelets• Platelets have three important properties (three ‘A’s):
• 1. Adhesiveness
• 2. Aggregation
• 3. Agglutination• 3. Agglutination
• Functions:• Role in blood clotting
• Role in clot retraction
• Role in prevention of
• Role in blood loss (hemostasis)
• Role in repair of ruptured blood vessel
• Role in defense mechanism
A. Inactive platelets. B. Activated platelets
Platelet Disorders
• Thrombocytopenia
• Thrombocytosis
• Thrombocythemia
• Glanzmann’s thrombasthenia
Haemostasis
Blood CoagulationClotting Factors
13 Clotting factorsFactor I - FibrinogenFactor II - ProthrombinFactor III - Thromboplastin(Tissue factor)Factor IV - CalciumFactor V - Labile factor (Factor VI - Presence has not been provedFactor VII - Stable factorFactor VII - Stable factorFactor VIII -AntihemophilicfactorFactor IX - Christmas factorFactor X -Stuart-Prower factorFactor XI- Plasma thromboplastin antecedentFactor XII - Hageman factor (Contact factor)Factor XIII - Fibrin-stabilizing factor (Fibrinase).
AnticoagulantsSubstances which prevent or postpone coagulation of
blood are called anticoagulants.
Anticoagulants are of three types:
1. Anticoagulants used to prevent blood clotting inside1. Anticoagulants used to prevent blood clotting inside
the body, i.e. in vivo.
2. Anticoagulants used to prevent clotting of blood that
is collected from the body, i.e. in vitro.
3. Anticoagulants used to prevent blood clotting both in
vivo and in vitro.
Heparin
• Heparin is a naturally produced anticoagulant in the
body.
• Heparin is a conjugated polysaccharide
• Commercial preparation is available in liquid form or dry
form as sodium, calcium, ammonium or lithium salts.
• Heparin is used as an anticoagulant both in vivo and in
vitro.
• Expensive
How Heparin Acts
• Coumarin Derivatives
• Warfarin and dicoumoral are the derivatives of coumarin
• EDTA
• Ethylenediaminetetraacetic acid (EDTA) is a strong anticoagulant. It is available
in two forms:
• i. Disodium salt (Na2 EDTA).
• ii. Tripotassium salt (K3 EDTA
• OXALATE COMPOUNDS• OXALATE COMPOUNDS
– Oxalate compounds prevent coagulation by forming calcium oxalate, which is
precipitated later.
• CITRATES
– Sodium, ammonium and potassium citrates are used as anticoagulants.
– Citrates are used to store blood in the blood bank as:
– a. Acid citrate dextrose (ACD): 1 part of ACD with 4 parts of blood
– b. Citrate phosphate dextrose (CPD): 1 part of CPD with 4 parts of blood
Tests for Blood Clotting
• Bleeding time: 3-6 minutes, Purpura
• Clotting time: 3-8 minutes, Haemophilia
• Prothrombin time: 12-14 seconds• Prothrombin time: 12-14 seconds
• Partial Prothrombin Time: 30 -45 seconds
• International Normalized Ratio: 1-1.5
• Thrombin time: 12-20 seconds
Bleeding Disorders
• Bleeding disorders are the conditions
characterized by prolonged bleeding time or
clotting time.
• Bleeding disorders are of three types:
• 1. Hemophilia.
• 2. Purpura.
• 3. von Willebrand disease
Haemopoiesis
• After birth and throughout life haemopoiesistakes place in the bone marrow.
• Fetal haemopoiesis occurs mainly in liver and spleen in the early stages.
• The haemopoietic stem cell is the pluripotent• The haemopoietic stem cell is the pluripotentprogenitor cell from which the cells of blood and lymphoid systems are ultimately derived.
• The HSC’s are capable of self-renewal as well as proliferation and differentiation.
• Their proper function depends on haemopoietic
niche.
General Outline
Actual Process of Haemopoiesis
Blood Grouping
Discovery of blood groups by the Austrian ScientistKarl Landsteiner, in 1901. He was honoredKarl Landsteiner, in 1901. He was honoredwith Nobel Prize in 1930 for this discove
Landsteiner Law
Blood Grouping Systems
• At present, 36 blood group systems are listed by the International Society of Blood Transfusion.
• Among these Systems ABO and Rh systems are most important in transfusion and transplantation .most important in transfusion and transplantation .
• According to this system, we have
ABO Blood Typing
Inheritance of ABO Group
Blood Matching
• Matching
Recipients RBC’s + Anti-sera
• Cross Matching
Recipients Serum + Donor RBC’s
Rh System
• Rh factor is an antigen present in RBC’s discovered by Landsteiner and Wiener in Rhesus monkey.
• Many Rh antigens, D more antigenic in humans.
• Among Indian population, 85% of people are Rhpositive and 15% are Rh negative. Percentage of Rh positive people is more among black people.
Inheritance of Rh Antigen
Rh Incompatibility
Other Blood Groups
• Lewis group
• MNS group
• Auberger groups
• Diego group• Diego group
• Bombay group
• Duffy group
• Lutheran group
Blood Components
Cellular components
• Red cell concentrate
• Leucocytes-reduced red
cells
• Platelet concentrates
Plasma Components• Fresh frozen plasma
• Single donor plasma
• Cryoprecipitate
• Cryo-poor plasma• Platelet concentrates
• Leucocytes-reduced
platelet concentrates
• Platelet Apheresis
• Granulocytes, Apheresis
Plasma derivatives• Albumin 5% & 25%
• Plasma protein fractions
• Factor viii concentrate
• Immunoglobulin
• Fibrinogen
• Other coagulation factors
Blood components and Products
• Rule 122-EA, Part X-B, D&C Rules 1945 Item ‘e’
“ BloodComponent” means a drug prepared, obtained, derived or seperated from a unit of blood drawn from a donar.blood drawn from a donar.
• Rule 122-EA, Part X-B , D&C Rules,1945, Item ‘f’
“Blood product” means a drug manufactured or obtained from pooled plasma of blood by fractionation, drawn from donors.
Categories Of Blood Components
• Part XII-B ,Section III, Rule E of D&C Rules,1945
• Concentrated Human Red Blood Corpuscles
• Platelet Concentrates• Platelet Concentrates
• Granulocyte Concentrates
• Fresh Frozen Plasma
• Cryoprecipitate
Indications
• Anaemia
• Major surgical operations
• Accidents
• Cancer patients• Cancer patients
• Women's in child birth in some cases
• Patients of hereditary diseases like Haemophilia, Thalassaemia, Sickle cell disease
• Severe burn victims
Legal Framework
• Conditions for Donor-D&C Rules 1945,Part XII-
B, Rule-H
• Anticoagulants- Rule I, 4 (b)• Anticoagulants- Rule I, 4 (b)
CPDA 14ml for 100ml of blood
ACD 15ml for 100 ml of blood
• Additives used: Rule I , item 4b-iii– SAGM,
ADSOL, NUTRICEL
Whole Human Blood
• 122-EA, c, D&C Rules 1945
Means Blood + Anticoagulant
Storage Conditions :Volume Storage Shelf-lifeVolume Storage Shelf-life
45o ml blood + 63 ml CPD
2-6 oC 42 Days unopened
359 ml blood + 49 mlCPD
2-6 oC 42 Days unopened
45o ml blood + 63 ml CPDA-1
2-6 oC 35 Days unopened
359 ml blood + 49 mlCPDA-1
2-6 oC 35 days unopened
• Indications
Red Cell replacement in acute blood loss
Exchange transfusion
Note: Transfusion should be started within 30 minutes after blood is taken out of storage.
Platelet Concentrates
• Storage:
20- 24 o C for 5 days
Indications:Indications:
Bleeding Disorders due to
• Thrombocytopenia
• Platelet function defects
• Granulocyte Concentrate Storage:20 – 24 o C for 24 hours
Indications:Indications:• Chronic Granulomatous Disease(Congenital
Neutrophil defects)• Chemotherapy induced Neutropenia
• Treatment of infection• Infection prophylaxis
• Aplastic Anaemia• Neonatal Sepsis
Plasma Components
• Fresh Frozen Plasma:– Plasma frozen within 6 hours of being collected,
containing all coagulations factors.
Storage:Minus 30 o CMinus 30 o C
Shelf-life : 1 year
Indications:Multiple coagulation factor deficiencies
Liver diseases
Warfarin overdose
Cryoprecipitate
• Concentrate of plasma rich in anti-haemophilliac factor and fibrinogen , prepared by thawing of FFP.
• Storage: Minus 30 o C• Storage: Minus 30 o C
• Shelf-life: 1 year
• Indications:– Inherited deficiency of factor Viii
– Haemophilia
Mandatory screening tests for donated blood
• ELISA for HIV I & II
• Test for Hepatitis B surface antigen
• ELISA for Hepatitis C Antibody
• Test for Malarial parasite
• VDRL/RPR for Syphills
References
K Sembulinga, Prema Sembulingam;Essentialsof Medical Physiology; Section 2;Body Fluids;Page:51-139
The Drugs and Cosmetics Rules,1945 as amended by The Drugs and Cosmetics ( Second
The Drugs and Cosmetics Rules,1945 as amended by The Drugs and Cosmetics ( Second Amendment)Rules,2016 (23 0f 1940)
UpToDate.com: Granulocyte transfusionsIntenational Society of Blood Transfusions webHemogenomics blog: Hemogenomics.com