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