Chapter- 4

BLOOD AND CIRCULATORY

SYSTEM
4.1. THE PHYSIOLOGY OF BLOOD
 Learning objectives

 At the end of this lesson learners will be able to:

 Describe the physical characteristics , function and
components blood
 Explain regulatory mechanisms of blood cell production
and its disorder
 Explain the mechanisms of blood clotting
 Identify blood types and its transfusion
 Physical characteristics and volume

 Blood is a sticky, opaque fluid with a metallic taste

 It is a viscus fluid connective tissue, which is heavier and
thicker than water.
 Color varies from scarlet (oxygen-rich) to dark red (oxygen-
poor)
 The pH of blood is 7.35–7.45 and its salt content is 0.9%
 Temperature is 38C, slightly higher than “normal” body
temperature
 Blood accounts for approximately 8% of body weight
 Average volume of blood is 5–6 L for males, and 4–5 L for
females
 Functions of Blood
 Blood performs a number of functions dealing with:
 Substance distribution
 Regulation of blood levels of particular substances
 Protection
 Blood transports:
 Oxygen from the lungs and nutrients from the digestive
tract
 Metabolic wastes from cells to the lungs and to kidneys for
elimination
 Hormones from endocrine glands to target organs
 Functions of Blood…cont’d
 Blood maintains:
 Body temperature by absorbing and distributing heat
 Normal pH in body tissues using buffer systems
(e.g., Hb, HCO3-)
 Adequate fluid volume in the circulatory system
 Blood prevents blood loss by:
 Activating plasma proteins and platelets
 Initiating clot formation when a vessel is broken
 Blood prevents infection by:
 Synthesizing and utilizing antibodies
 Activating WBCs to defend the body against foreign invaders
 Composition of Blood
 Blood is the body’s only fluid connective tissue
 It is composed of -liquid plasma (55%) and
-formed elements (45%)
 Formed elements include:
 Erythrocytes, or red blood cells (RBCs)
 Leukocytes, or white blood cells (WBCs)
 Thrombocytes, or Platelets
 Hematocrit – the percentage or proportion of blood cell to
blood volume
 Components of Whole Blood

Plasma
(55% of whole blood)

Buffy coat:
leukocyctes and platelets
(<1% of whole blood)

Formed
elements
Erythrocytes
1 Withdraw blood and 2 Centrifuge (45% of whole blood)
place in tube
 Plasma
 It is the liquid portion of blood
 It makes up 55% of the blood volume
 Has the osmolality of 300 mosm/l
Composition of plasma
 Blood plasma is composed of
1. Water (90%)
2. Organic constituents (9%)
 Plasma proteins: albumin, globulins, clotting proteins,etc
 Lipids, lipoproteins, phospholipids
 Hormones and enzymes
 Nutrients: CHO, vitamins, amino acids, fats
 Metabolic waste products: urea, creatinine
3. Inorganic constituents: electrolytes (1%)
4. Respiratory gases – oxygen and carbon dioxide
 Plasma…cont’d
 TPP: 7 g/dl
 There are 3 principal plasma proteins
Albumin: 4 g/dl
Globulin: 2.7 g/dl
Fibrinogen: 0.3 g/dl
 Plasma proteins are synthesized by
 hepatocytes
 lymphocytes
 platelets
 endothelial cells
 Plasma…cont’d
Function of plasma proteins
1. Immunologic function: γ-globulins are immunoglobulins
(antibodies)
2. Hemostasis: fibrinogen, prothrombin and most other clotting
factors are plasma proteins
3. Function in starvation
4. Transport of hormones, electrolytes and drugs.
5. pH regulation (buffering function)
6. Maintenance of plasma osmotic pressure (mainly by
albumin)
7. Enzymatic and hormonal function
 Plasma…cont’d
1. Albumin (60%):
 is formed in the liver and constitutes about 60% of the plasma
proteins.
 helps to maintain blood volume, for it can not easily pass
through the capillary membrane.
 During malnourished conditions (deficiency of protein
intake), blood volume decreases due to decreased albumin
content.
 Albumin also serves in transporting nutrients (lipids,hormones,
bilirubin etc
 Plasma…cont’d

2. Globulin (40%) are of 3-types: alpha1&2, beta, and gamma

globulins.
 Globulins are about 40% of the total protein.
 Alpha and gamma globulins are made in the liver.
 Alpha and beta globulins help as carriers to transport lipoproteins,
Fe 2+, hormones, enzymes, nutrients, and other substances in the
body.
 Gamma globulins act as immunoglobulin's (or antibodies) and
help in defending our body against infections.
 Made in Lyphoid tissue
3. Fibrinogen (2-4 %): Fibrinogen is synthesized in the liver and is
mainly involved in blood clotting
 Erythrocytes (Red blood cells)
 Major content of RBCs is Hb (97% )
 Size: Diameter7.5 µm
Thickness 1 and 2 µm
 Shape: Biconcave disk
No nucleus, no organelles
 RBC Count:
 M = 5.2 millions/mm3
 F = 4.6 millions/mm3

 Hematocrit: Percentage of blood cells

M = 42-48%, average-45%
F = 38-43%- average-42%
 Erythrocytes…cont’d
 Function
 Transport of O2 and CO2
 Regulation of acid-base balance.
 Hemoglobin (Hb)
 Male 15 g/dl
 Female 14 g/dl
 Plasma membrane of RBCs is comprised of flexible proteins
 Allow them to change shape as necessary
 ATP is generated anaerobically
 Production of Erythrocytes
 Hematopoiesis – blood cell formation
 Areas of production of RBCs

A. Embryonic life
• RBC are produced in the liver, spleen and lymph
nodes

B. Infants (till 5 years)

• RBC are produced in marrow of all bones
C. Adults (after 20)
– Membranous bones like ribs, sternum, vertebrae, and
pelvic bones, but not in long bones like femur or tibia.
 Production of Erythrocytes…cont’d

Fig. Relative rates of red blood cell production in the bone marrow of
different bones at different ages 17
 Production of Erythrocytes…cont’d

 In adult, RBC and other blood cells are produced in the bone
marrow from a single type of cells called pluripotent
hematopoietic stem cells.
 These cells, then differentiate and form committed stem cells
that produce only specific types of blood cells
 There are Growth inducers (like interleukin 3) that promote
only growth and Differentiation inducers: that initiate only
differentiation of blood cells.
 Factors outside the bone marrow control formation of
growth and differentiation inducers. E.g., erythropoietin
enhances the activity of growth and reproduction inducers
during hypoxia.
Production of Erythrocytes…cont’d
Production of Erythrocytes…cont’d

Proliferation phase Maturation phase

Regulation and Requirements for Erythropoiesis
 Circulating erythrocytes – the number remains constant
(balanced RBC production and destruction)
 Too few red blood cells (Anemia) leads to tissue hypoxia
 Too many red blood cells (Polycethemia) causes increased
blood viscosity

 Erythropoiesis requires diets rich in:

 Proteins, lipids, and carbohydrates
 Iron, and vit. B complex (vit-B12, folic acid)
 Vit-B12 and folic acid are essential for the RBC maturation
(Necessary for the synthesis of DNA of RBC)
 Regulation for Erythropoiesis…cont’d
 Erythropoiesis is hormonally controlled.
 Hormones accelerating erythropoiesis:
 Erythropoietin (EP) from JG-cells and hepatocyte

 GH, T3/T4, Androgens (testosterone)

 Intrinsic factor from parietal cells of the stomach
 Liver plays a critical role in RBC formation as site of globin
syntheis, as a storage area of iron and vit-B12
 Erythropoietin Mechanism

Imb
ala
nce
Start
Normal blood oxygen levels
Stimulus: Hypoxia due to
Imb
a la
decreased RBC count,
nc e
decreased availability of
Increases O2 in blood, or increased
O2-carrying tissue demands for O2
ability of blood
Reduces O2
levels in blood

Erythropoietin
Enhanced Kidney 85% and
stimulates red
erythropoiesis bone marrow liver 15 % releases
increases RBC erythropoietin
count
Erythropoietin Mechanism…cont’d

24
 Relation of Erythropoietin to Ht ratio

 Erythropoietin vs. Hct

 A decrease in Ht ratio in
the blood increases the
concentration of
erythropoietin level
making their relations
inverse.

25
 Hemoglobin
 Hb in RBCs is means for transport of respiratory gases
 Normal concentration; 15 g/dl in M and 14 g/dl in F
Composition:
 Composed of a protein globin and heme
 4 heme molecules conjugated with 4 globin molecules to form Hb
 In adult Hb, the globin part has 4 polypeptide chains
2 α-chains, each made up of 141 aa residues
2 β-chains, each made up of 146 aa residues
Types of Hb
1. Adult Hb (HbA-α2 β2)
2. Fetal Hb (HbF -α2 γ2)
3. Sickled Hb (HbS - α2 β2)
 Sickled Hb, in two of the β-chains at position-6 valine is
wrongly substituted for glutamate
Hemoglobin…cont’d
 Hemoglobin…cont’d
 Hb reversibly binds with O2 and most O2 in the blood is
transported in combination with Hb
 Each heme group bears an atom of iron, which can bind to one
O2 molecule
 Each Hb molecule can transport four molecules of oxygen
 Oxyhemoglobin – hemoglobin bound to oxygen
 Oxygen loading takes place in the lungs
 Deoxyhemoglobin – hemoglobin after oxygen diffuses into
tissues
 Carbaminohemoglobin – hemoglobin bound to carbon dioxide
 Carbon dioxide loading takes place in the tissues and is
returned to lungs to be eliminated in expired air
 Destruction of Erythrocytes

 The life span of an erythrocyte is 120 days

 Old erythrocytes become rigid and fragile, and their hemoglobin

begins to degenerate
 Dying erythrocytes are engulfed by macrophages
 Heme and globin are separated and the iron is salvaged for reuse

 The heme part is converted into bilirubin

 Bilirubin is the main component of biliary secretion
 Destruction of Erythrocytes…cont’d
 Jaundice
 Yellow coloration of the skin and sclera

 Due to excessive bilirubin in plasma (>1.5 mg/dl)

 Normal plasma bilirubin level is 0.5 mg/dl
 In case of jaundice, bilirubin level is elevated up to 40 mg/dl

 Types/ causes of jaundice

1. Hemolytic jaundice: ↑RBC destruction

2. Obstructive jaundice:
a. Obstruction of bile ducts by stone
b. Obstruction of bile ducts by head of pancreas
 Destruction of Erythrocytes…cont’d
Steps in the destruction of RBC
1. RBC  Globin + Heme
2. Globin  broken to AA’s  used for protein synthesis
3. Heme  Fe2+ + poryphrine rings
4. Fe 2+  stored in spleen and liver or reused by bone marrow
for new Hb synthesis
5. Pyrol rings  oxidation to green pigment called Biliverdin
and later reduced to bilirubin
a. bilirubin + serum albumin  go and reach liver
b. Bilirubin conjugates with glucuronic acid in liver
c. Liver releases bilirubin as bile to Small intestine
d. Bacterrias change bilirubin into:
Stercobilinogen  stercobilin  feces (brown color)
Uribilinogen  Urobilin  Urine (yellow)
31
Destruction of Erythrocytes…cont’d
Destruction of Erythrocytes…cont’d

33
 Erythrocyte Disorders

 It is a condition char/zed by a decrease in the hemoglobin

level, RBC count or both leading to decrease in the O2
carrying capacity of the blood or increasing RBC count

1. Anemia: means having decreased RBC number and lower

concentration of hemoglobin (Hb) in the circulating blood.
 There is a diminished O2-carrying capacity of the blood.
 Results from excessive loss (bleeding), or destruction
(hemolysis), or lower production of RBC (lack of nutrition) in
the circulation.
 Erythrocyte Disorders…cont’d
 Types of Anemia's include
a. Blood loss anemia
b. Aplastic anemia
c. Megaloblastic anemia &
d. Hemolytic anemia
a. Blood loss anemia:
 During hemorrhage persons loose too much blood.
 The Fe2+ absorbed from the intestine is not enough to produce
adequate Hb to replace the lost Hb that took place during
bleeding
 As the result, the RBC’s produced are smaller in size and
have lower Hb level causing a microcytic, hypocromic anemia
 Erythrocyte Disorders…cont’d
b. A plastic anemia:
 Aplasia means lack of functioning bone marrow.
 It is caused by exposure to excess chemicals, drugs and gamma or
X-ray that destroy the bone marrow.
 Effect is lower RBC production

c. Megaloblastic anemia:
 Maturation failure anemia (pernicious anemia) is caused by
inability to absorb Vitamin-B12 and folic acid from the intestine.
 Absence of intrinsic factor (gastrechtomy) is another cause.
 Effect is that RBC can not proliferate properly, b/s DNA
synthesis does not occur.
 As the result the RBC’s become big, oversized (megaloblastic)
and have fragile membranes that easily rupture and decrease the
number
 Erythrocyte Disorders…cont’d
d. Hemolytic anemia:
 Hereditarily acquired and makes the RBC’s to easily rupture as
they cross through narrow blood vessels like capillaries, spleen
etc. Examples are:
a. Hereditary spherocytosis: spherical shaped RBC (not
biconcave) that rupture easily
b. Sickle cell anemia: is caused by abnormal beta chain in Hb

structure. When exposed to low O2 (hypoxia), the Hb

precipitates forming long crystals and damaging and
rupturing the cell-membrane, their by causing anemia.
c. Erythroblastosis fetalis: Destruction and decrease of
RBC’s number of the fetus by antibodies from Rh- mother.
 Erythrocyte Disorders…cont’d
2. Polycythemia
 Is production of too much RBC in the circulation.
 Has a physiological or a pathological cause:

A. Physiological cause is known as Secondary Polycethemia

 When ever the tissues become hypoxic b/s of low O2-tension)
like in high altitude, the marrow produces excessive RBC (6-7
mill/mm3).

B. Pathological cause is called Polycethemia Vera (erythremia) or

primary polycythemia (about 7-8 million / mm3)
 Is mainly due to genetic causes as in tumerous or cancerous
production of RBC.

38
 Effect of polycethemia on the circulatory system

a. Increased viscosity (Hct) causes sluggish blood flow that

has the effect of decreasing venous return
b. Conversely, the increased Hct increases blood volume. The
increased blood volume in turn increases venous return.
 The effect of the above factors, a & b, mostly compensate
(neutralize ) each other and cause normal CO (cardiac out put)
and blood pressure in polycythemia patients.
c. The color of the skin of polycethemic patients is bluish or
cyanotic (the sluggish blood causes deoxygenation of the Hb-
molecule)
White blood cell (Leukocytes)
 Leukocytes (White blood cell)
 Leukocytes, the only blood components that are complete
cells: Contain nucleus, and other organelles but no Hb
 Make up less than1% of the total blood volume

 Less numerous than RBCs

 Normal WBC count: 4000 – 11000/mm3 (Aver=7000/mm3 )
 When infection occurs, WBC increase in number.

 Are highly mobile and reach tissue fluids

 Function: defense/protection against disease
a. by direct destruction (e.g., Phagocytosis)
b. by producing : i. antibodies ii. sensitized lymphocytes
41
Formation of leukocytes

42
Type of WBC …cont’d

Macrophages

43
 Types of WBC
1.Granulocytes –Polymorphonuclear (i.e.their nuclei have 3-5
lobes)
• Attack pathogen by phagocytosis

a. Neutrophils (~ 62%, 3000-7000/mm3), life span 4-5 days

 Have multilobed nuclei (PMN-cells, 2-5 lobes)
 The 2nd line of defense
 Phagocytic cells (ingest bacteria)
 Their granules have enzymes to digest bacteria

44
 Types of WBC…
b. Eosinophils (~ 2-3%, 100-440/mm3), life span 4-5 days
 Aare bi-lobed and weak phagocytes .
 Have granules in the cytoplasm that have enzymes
 Phagocytize antigen-antibody complexes and destroy them
 Their number increases during asthma and other allergic
attacks

c. Basophiles (~ 0.1-0.4%, 20-50/mm3), life span 4-5 days

 Have granules
 Produce heparin, so act as natural inhibitors of blood clotting.
 They also synthesize & store histamine, bradykinin, and
serotonin

45
 Types of WBC…cont’d
2. Agranulocytes.
a. Lymphocytes(~30%,1500-3500/mm3),life span weeks or months
• Have no granules and the nucleus is not lobed
• Produced in the bone marrow and lymphogenous organs
(spleen, thymus, tonsils , payer’s patches etc) organs
• Are responsible for specific immunity that consists of:
a. Cellular immunity (T lymphocyt)
b. Humoral immunity (B lymphocyte)
b. Monocytes (~5 %, 100-700/mm3), 20 hrs, circulate in blood
and change into macrophages that attach to tissues e.g. alveolar
macrophages in the lung, kuffer cells in the liver etc.
 They are highly phagocytic cells
 They are the largest WBCs
 They leave the circulation, enter tissue, and differentiate into
macrophages 46
 The tissue macrophage system (reticuloendothelial system)

Monocytes are formed in the bone marrow


Enter the circulation

Leave the circulation, enter the tissue,

↑size, ↑lysosomal activities

Become tissue macrophages

Lungs Skin Liver Brain Bone Spleen
     
Alveolar Histocytic Kupffer Microgleal Osteoclasts
Macrophages cells cells Reticular cells
lymph nodes

47
47
Types of Lymphocytic cells

48
Mechanisms of WBC mobility through the tissues

1.Diapedesis:WBC approach the capillary

wall and squeeze out through the
pores(e.g. Neutrophils, Monocytes etc)
2. Amoeboid motion: Produce pseudopodia
and reach the microbes in the tissues.
3. Chemotaxis: WBC are attracted by
chemicals or toxins produced by microbes
or inflamed tissues(Eosinophils)
4. Phagocytosis: engulfing & destroying
the pathogens in the tissues (e.g.,
Neutrophils, macrophages etc.)

49
Mechanism of Phagocytosis
1. Opsonization
2. Attachment
3. Engulfment
4. Intracellular killing by
producing:
 Lysosomes, and
 oxidizing agents like lipases,
peroxisomes, H2O2, OH- ions
are produced from
macrophages that are lethal
for bacterial cell membranes.

50
 Activation of T-Lymphocytes
The different types of T-cells include:
a. Killer (cytotoxic) T-cells: Kill pathogens directly and stimulate
other macrophages for phagocytosis
b. T-memory cells: Remain as reserve in the lymph node and
protect when the same type pathogen attacks the body again
c. Helper T- cells: support or help the B-cells by producing
chemicals known as Lymphokines or (cytokines
 HIV, the virus that causes AIDS normally infects helper T-cells
(and other immune cells) and inactivates the total immune
response
d. Suppressor T-Cells: Regulate the immunological response of
T-and B-cells
 Immunoglobulins
 B-cells that bind with an antigen will
subsequently differentiate into Plasma cells
& Memory cells
 Plasma cells - begin to produce
antibodies (up to 2,000 per second)
 Memory cells - remain dormant until a
person is again exposed to the same
antigen
 There are 5-classes of antibodies

 IgA, IgE, IgD, IgG and IgM

 Five classes of Immunoglobulin
 IgM (10%) = are pentameters and is 1st or early antibody that
appears in circulation after infection.
• It forms the natural antibodies of the ABO blood antigens
 IgG(75%) - crosses' placenta and provides passive defense of
the fetus
 IgE (0.004%) = increased in allergic reactions and parasite
infections.
• It can bind to mast cells and induce the liberation of histamine
(Hay fever ,asthma & hives (Urticaria) , Anaphylaxis)
 IgA(15%) - found in secretions of digestive (saliva), respiratory
(bronchus), urinary, & reproductive systems, as well as in breast
milk and in tears
 IgD (0.2%) = Are found on surfaces of B-cells and help to
recognize the antigen, otherwise their other function is not known
 Mechanism of action of antibodies
A. By direct attack: inactivate the invaders by the following
methods
1. Agglutination: Involves the clumping binding of bacterial
antigens to each other, so that they become dysfunctional.
2. Precipitation : The complex antigen-antibody reaction is
made insoluble and precipitates down
3. Neutralization: The antibodies cover the active sites of the
invader and inactivate the toxic sites of the antigens
4. Lyses: Antibodies directly attack membranes of virulent
agents cause their rupturing.
 Mechanism of action of antibodies...
B. Complement system
 The complement system consists of a series of proteins that work
to "complement" the work of antibodies in destroying bacteria. 
 Complement proteins circulate in the blood in an inactive form.
 The so-called "complement cascade" is set off when the first
complement molecule, C1, encounters antibody bound to antigen
in an antigen-antibody complex.
 Each of the complement proteins performs its specialized job in
turn, acting on the molecule next in line.
 The end product is a cylinder that punctures the cell membrane
and, by allowing fluids and molecules to flow in and out, dooms
the target cell.
Complement system…cont’d

56
 Complement system…cont’d
 The complement system simply amplifies the following action

1. Opsonization and phagocytosis

2. Lysis
3. Agglutination
4. Neutralization
5. Chemotaxis
6. Activation of mast cells and basophiles: to release histamine
and heparin to cause vasodilatation and increased local
blood flow

57
 Mechanism of self tolerance
 Self tolerance is a condition of self-recognition of the bodies
antigens, i.e, the immunity system does not form antibodies or
sensitized lymphocytes against its own antigens.
 Normally, self cells have a specific antigen called human
leukocyte antigen (HLA) which stands for each individual.
 This HLA is expressed by a group of genes called major
histocompatability complex (MHC).
 The importance of MHC proteins is that they allow T-cells to
distinguish self from non-self
 All clones of lymphocytes that are specific and sensitive to the
bodies own antigens are deleted or destroyed during the fetal
period while being pre-processed in the thymus and in the pre-
processing area of B-cells (Liver, spleen, or bone marrow)

 Moreover, T-suppressor cells destroy or suppress any reaction

that comes against self antigens 58
 Disturbance of WBC
1. Leukemia - increased WBC No. or cancerous production of
WBC.
• Their increased production takes the space of platelets & RBC
causing anemia + impaired blood clotting
 Increased WBC causes
A. Metabolic starvation:
• WBC consume too much metabolic substrates. Thus energy
source is depleted and too much use of a.a by cancerous cells
causes rapid deterioration of other body tissues.
B. Anemia and increased bleeding tendency can occur.

2. Leucopenia - decreased production of WBC.

• Bone marrow stops producing them.
• It can be caused by: - Drug poison, X-rays
59
Platelets (Thrombocytes)

60
 Platelet
 Platelets are small disk-shaped cell fragments that emerge from
megakaryocytes in red bone marrow (2-4 microns in diameter)
 Life span : 4-12 days
 Platelet count : 250x109/L of blood
 Cytoplasm: have no nucleus and thus can not reproduce
 They release some chemicals in their cytoplasm
(prostaglandins, thromboxane, fibrin stabilizing factor,
phospholipids, minerals and growth factor etc).
 Platelets function in the blood clotting mechanism by forming a
temporary plug that helps seal breaks in blood vessels

61
 Genesis of Platelets
 The stem cell for platelets is the hemocytoblast

 The sequential developmental pathway is hemocytoblast 

megakaryoblast  promegakaryocyte  megakaryocyte  platelets

Figure62
17.12
 Properties of platelets

1. Adhesiveness : platelets stick when they come in contact with

wet and rough surfaces.
 Normally, their glycoprotein (structure in their cell membranes)
prevent adhesiveness to the normal endothelium)
2. Aggregation: When platelets are activated, they usually group
together
 Their aggregation and stickiness is mainly due to ADP and
Thromboxane A2 found in their cytoplasm
3. Agglutination: Platelets clump together and form clots
63
 Hemostasis / blood clotting
• Hemostasis means arrest or prevention of bleeding or series of
reactions designed for stoppage of bleeding

• Platelets: are responsible for clotting b/s the cytoplasm of

platelets synthesizes chemicals which enhance clotting. These are:
a. Thromboxane -A2,
b. Serotonin,
c. Histamine,
d.Von Willbrand’s factor (important for adherence of platelets)
e. Minerals like Cu2+ and Fe2+ that are involved in clotting
processes.

64
 Hemostasis…cont’d
 During hemostasis, five phases occur in rapid sequence
1. Vascular spasm – immediate vasoconstriction in response
to injury
2. Platelet plug formation
3. Coagulation (blood clotting)
4. Fibrin mesh development (clot retraction)
5. Clot dissolution

65
1. Vascular spasm
 Local vasoconstriction that is caused by:
 Reflex sympathetic discharge
 Local myogenic contraction of the vascular wall
 Local metabolites: The vasoconstrictor substances released
include serotonin, histamine, Thromboxane A2 etc.

 In effect both reaction minimize rate of bleeding by narrowing

the diameter of the hole (rupture) on the wall of the blood
vessels

66
 2. Platelet Plug Formation
 Normally, platelets do not stick to each other or to the endothelial
lining of blood vessels
 Upon damage to blood vessel endothelium (which exposes
collagen) platelets:
 Adhere and Stick to exposed collagen to form platelet plug:
facilitated by Von Willebrands factor released in plasma
 Release serotonin, thromboxane A2 and ADP, which attract
additional platelets
 The platelet plug is limited to the immediate area of injury

67
 3. Blood Clot formation (Coagulation)
 A set of reactions in which blood is transformed from a liquid
to a gel (clot)
 It is slow but has long lasting effect to stop bleeding
 Coagulation follows intrinsic and extrinsic pathways
 Coagulation undergoes three steps of reactions:
1. Formation of prothrombin activators through
 The intrinsic pathway
 The extrinsic pathways
2. Conversion of prothrombin into thrombin by the action of
prothrombin activators (prothrombinase)
3. Conversion of fibrinogen into fibrin thread by the action of
thrombin

68
 Blood clotting factors
1. Factor-I: Fibrinogen
2. Factor-II: Prothrombin, α1- globulin
3. Factor-III: Tissue factor, tissue thromboplastin, a
phospholipoprotein
4. Factor-IV: Calcium ion, essential for clotting
5. Factor-V: Labile factor, β-globulin. Defiency leads
to parahemophelia
6. Factor-VII: Stable factor, α1-glubulin
7. Factor -VIII: Antihemophilic factor-A, β2-globulin
8. Factor -IX:Christmas factor, AHF-B, α1- globulin
9. Factor -X: Stauter-power factor
10. Factor-XI: AHF-C, plasma thromboplastin anticident
γ-globulin
11. Factor-XII: Contact factor, Hagen factor, glass factor
12. Factor-XIII: Fibrin stabilizing factor, β-globulin,
polymerizes fibrin threads 69
 Additional clotting factors
 Prekallikrin: Fletcher factor, β-globulin
 Kininogen: α-globulin
 Van Willebrand factor: For platelet adhesion

Vitamin-K and blood clotting

 Vitamin-K is required for the synthesis of clotting factors: II,
VII, IX and X
 Vitamin-K deficiency occurs in case of obstructive jaundice,
chronic diarrhea, liver disease (hepatitis, cirrhosis, malignancy)
 Blood clotting factors are produced by
 Liver
 Platelets
 Endothelial cells

70
 Stages of Blood Coagulation...cont’d

 There are more than 50 different types of anticoagulants and

pro-coagulants circulating in the blood.
 Anti-coagulants dominate a little over procagulants when
blood circulates in the circulatory system.
 During tear or injury, procagulants dominate near the site of
injury, otherwise they remain more or less in balance.
 Be it through intrinsic (damage to the blood itself) or extrinsic
(rupture of the vessel) blood clotting mechanism, both of
them have a common ending.
 That is, they end in producing a substance called
Protrombin activator that activates Protrombin to change to
thrombin

71
72
 Stages of Blood Coagulation...cont’d
Extrinsic pathway:

Begins with traumatized (injured) tissues that release a

substance called tissue thromboplastin and together with factor
VII activates factor X to its active form Xa.
 Activated Factor Xa with Ca2+ and Factor V forms a
Prothrombin activator that in turn activates and converts
Prothrombin ----------- Thrombin
 Thrombin together with Ca2+ converts

 Fibrinogen ------------- to Fibrin (loose solid clot)

73
Stages of Blood Coagulation...cont’d

Extrinsic pathway
 Stages of Blood Coagulation...cont’d
Intrinsic pathway:
 Begins with trauma to the blood itself or exposure of the blood to
collagen at injured blood vessel wall.
 The blood then activates factor XII to its active form XIIa
 Then cascade of reactions continues until the formation of
activated Factor Xa .
 Factor VIII + IX + platelet phospholipids (tissue
thromboplastin) are involved in the activation
 The absence of Factor VIII causes classic hemophilia and
deficiency in platelet number causes the bleeding disease
thrombocytopenia.

75
77
 Fibrinolysis: dissolution of clot
Steps:
Thrombin
+Thrombomodulin
Protein-C Activated Protein-C

F-VIIIa VIII Va V

Activates tPA
Plasminogen Plasmin (fibrolysin)

Degradation of Fibrin
78
 Prevention of unwanted intravascular clotting
1. Smoothness of the endothelial cell surface. prevents platelet
adhesion and contact activation of the intrinsic clotting system
2. A glycocalyx (a polysaccharide on endothelial cell surfaces)
repels clotting factors and platelets thereby preventing
activation of clotting factors.
 So, when the endothelia wall is traumatized, the smoothness
and the glycocalyx layer are lost or injured.
3. The presence of intravascular anticoagulants
(thrombomodulin ) produced by endothelial cells, platelets
and WBCsA
 Thrombomodulin:
 inactivates thrombin
 Initiates the formation of protein-C
 Protein-C inactivates F-V and F-VIII
 Over all, prevents clotting.
79
Hemostasis Disorders:Thrombo-embolytic Conditions
 Thrombus – a clot that develops and persists in blood vessel
 Thrombi can block circulation, resulting in tissue death and
organ failure
 Embolus – a thrombus (blood clot) freely floating in the blood
 Pulmonary emboli can impair the ability of the body to obtain
oxygen
 Cerebral emboli can cause strokes

80
 Bleeding disorders
Excessive bleeding can be caused due to 3-major factors:
1. Hemophilia:
- It is due to insufficiency of Factor VIII and IX
- bleeding occurs at big blood vessels
- is an “X” linked inherited recessive bleeding disorder
(i.e., the person does not have enough clotting factors)

2. Vit. K deficiency: (liver diseases, hepatitis)

- important to synthesize Prothrombin in the liver

3. Thrombocytopenia (platelet deficiency)

- Bleeding at small vessels, capillaries and Venules.
-Platelets usually plug small blood vessels, so their absence
causes thrombocytopenic purpura ( small purplish blotches )
81
 Human Blood Groups
 Landsteiner (Austrian): is the 1st scientist to identify blood
group antigen.
 Blood group antigens are located at the cell membrane surface
of Erythrocytes (RBC’s).
Why blood typing?
 Blood typing is important because during transfusion of blood
mismatching can take place and cause unwanted complications
like hemolysis, renal failure, and sometimes death.
 In blood transfusion, because antibodies in plasma react with
antigens on red cells, it is important to identify which blood
antibodies combine with which blood antigens to produce
compatible blood types

82
 Major blood groups

 Over 14 blood groups and over 100 antigens are identified

from erythrocytes
 Major blood groups are
 ABO blood group
 Rh system
 Other blood groups (medically less important)

83
 ABO Blood Group
Based on the ± of two agglutinogen (A and B), the ABO blood group
can be classified into four types of blood
1. When agglutinogen A is present on RBCs,it has anti-B antibody
(agglutinin) in his serum  the blood is type-A
2. When agglutinogen B is present on RBCs, Anti-A antibody is
present in its serum  the blood is type-B
3. When both agglutinogens A & B exist together, then it has no
antibody  type-AB
4. When neither agglutinogen A nor B are present, has both anti A
and B antibody in its serum  type-O

84
ABO Blood groups

85
86
 Method of blood typing
 On a slide, drops of anti-A antibody and Anti-B antibody are
added at opposite sides.
 2-3 drops of blood (RBC’s) are added on the prepared antibodies
and agglutination reactions are observed after a few minutes.
 If agglutination occurs on anti-A antibody (sera), then the blood is
group A.
 If agglutination occurs on anti-B antibody , the blood is blood
group B.
 If no agglutination occurs, then it is blood group O.
 If there is agglutination in both anti-A and anti-B antibodies, then
it is blood group AB.

02/23/2022 by H.F. 87
88
 ABO Blood Typing
To determine a blood type, a drop of blood sample is mixed with a
known prepared antibodies
Known agglutinins
Blood
samples Anti-A
Anti-B Blood type
antibody
antibody

1 + ‒ A

2 ‒ + B
3 + + AB
4 ‒ ‒ O
(+) = Agglutination reaction
(‒) = No agglutination reaction
89
 ABO blood group cross matching
Cross matching is a means to determine the compatability
of the blood of donors with that of recipients for transfusion

Recipients blood types

Donors
Blood
types A B AB O

A ‒ + ‒ +
B + _ ‒ +
AB + + ‒ +
O (+) = Agglutination
‒ ‒
reaction
‒ ‒
(‒) = No agglutination reaction 90
 Rh Factor

 It was first worked out on Rhesus monkey to the name Rh-

system
 Grouped as Rh+ and Rh- based on the presence or absence of
agglutinogen D on the surface of RBCs
 The presence of agglutinogen on RBCs: Rh+
 Missing agglutinogen RBCs: Rh-
 Agglutinin (anti-D antibodies) are not normally present in the
serum, produced secondary to exposure of an Rh- blood to Rh+
blood (antigen-D)

91
 Erythroblastosis Fetalis
 Occurs when an Rh- mother marries an Rh+ father and conceives
an Rh+ fetus
 During delivery, there could be a leakage of Rh+ blood from the
fetus to the circulation of the mother.
 Rh+ blood induces production of anti-D antibodies in the
circulation of the mother.
 During 2nd conception of Rh+ fetus, anti-D antibodies cross the
placenta and attack RBCs of the fetus and agglutinate or
hemolyze the RBC’s the fetus.
 This type of hemolytic disease is called Erythroblastosis fetalis.
92
 Blood Transfusions
 Whole blood transfusions are used:
 When blood loss is substantial
 In treating thrombocytopenia
 Packed red cells (cells with plasma removed) are used to treat
anemia
 Transfusion of whole blood or its parts (plasma, or red blood
cell concentrates or other specific clotting factors is supplied
in order to:
A. Restore blood volume
B. Improve O2 transport to tissues
C. Correct bleeding abnormalities etc.

93
Thanks!!!

94