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Blood and its physiology part 2

Basic seminar
Date :- 1-4-2006
Time :- 9.10-10 am
Duration :- 45 mins
 Blood and its physiology
 Introduction
 Composition of blood
 Properties of blood
 Plasma proteins
 Red blood corpuscles
 Leucocytes
 Haemostasis
 Blood groups and blood transfusion
 Conclusion
 References

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 Platelets
 These are small,colourless ,non nucleated and
moderately refractive bodies.
 They are considered to be fragments of the
cytoplasm.
 They have a diameter of 2.5 microns and volume
of 7.5 microns.
 Inactivated platelets are oval /disc shaped and
activated platelets are rod / spherical shaped.

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Stages of platelet formation

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 Morphology and structure.
 They have
1. Cell membrane
2. Microtubules below the cell membrane
3. Cytoplasm.
They do not have DNA or RNA and hence no
protein synthesis occurs. They contain
contractile elements like actin and
myosin.

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 Cell membrane.
 It is made up of 2 layers, outer glycocalyx
layer and inner lipoprotein layer.
 Lipoprotein layer contains lipids like
phospholipids,cholestrol,glycolipids.
 Glycocalyx-layer contains carbohydrates.

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 Microtubules
 They encircle the platelet below the cell
membrane and gives structural support.

 CYTOPLASAM:-it contains
 Golgi bodies, endoplasamic reticulum,
mitochondria, microtubules,
microvessels,filaments and different types
of granules.

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 Alfa granules –when platelets are activated
Alfa granules dissolve and release
PGDF,von willebrands factor,
thrombospondin.
 Dense granules– Non-protein Substances
like serotonin,ATP,ADP,calcium
 Protein Substances like
histamine, ,glycogen, adrenaline.

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 Variation of count
 Normal count –1.5 lakh -4 lakhs /cu mm.
 Physiological variation:-
 Age infants:- 1.5-2 lakhs
3rd month:- normal count
 Sex in females it is reduced during
menstruation
 High altitude increased
 After meals increased

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 Critical count
 Platelet count of less then 40,000 is called critical
count.

 Pathological variation :-
 THROMBOCYTOPENIA
 Acute infection
 Leukemia
 Chickenpox, small pox.
 Splenomegaly,scarlet fever,typhoid

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 Thrombocytosis
 Allergic condition
 Asphyxia
 Haemorrhage
 Bone fracture
 Surgical operation
 Splenectomy
 Trauma

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 Functions of platelets.
 They arrest bleeding by temporary plug
formation
 Help in vasoconstriction of damaged vessels
by releasing thromboxane, serotonin etc
 Repair of ruptured blood vessel-PGDF
 They phagocytose antigen antibody
complexes and thus play a role in defence
mechanism.

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 They have a life span of 10 days and are
destroyed by macrophages and spleen.

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 Haemostasis
 Arrest of bleeding or stoppage of bleeding
is called haemostasis.
 3 basic steps are:-
1. Constriction of bleeding vessel
2. Formation of platelet plug.
3. Coagulation of blood.

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 Applied physiology
 Whenever primary hemostatic mechanisms
fail there will be formation of purpura.
 Haemophilia A and Haemophilia B.

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 Coagulation of blood.
 It is a secondary hemostatic mechanism which
leads to formation of clot.
 Clot is a mesh of thin fibrils entangling the blood
cells.
 These fibrils consist of fibrin which is formed
from fibrinogen.
 There exist 13 clotting factors in human body.
 Factor 1 –FIBRINOGEN

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 Sequence of clotting
mechanisms-3 stages
1. Formation of prothrombin activator
2. Conversion of prothrombin thrombin
3. Conversion of fibrinogen fibrin
Inactive clotting factors are activated and
the enzymes released by them produce the
successive reactions one after another in a
cascading manner.

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 Blood clot
 It consists of fibrin threads which run in all
direction with Rbc’s,Wbc’s and platelets
entrapped in it.
 Clot retraction :-
After the formation, the clot starts
contracting and after about 30-45 mins a
straw colored liquid oozes out.
 Contractile proteins are responsible for this.

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 Health matters
 Clinical methods of hastening clotting
 By applying rough surface such as gauze,
 By applying heat
 By gently squeezing the tissues around the
cut vessel.
 By applying purified thrombin.

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 Lysis of blood clot.
 The substance called plasmin or fibrinolysin
is responsible for clot lysis.
 Formation of plasmin:-
After few days of clot formation , some
substances are released from the damaged
tissues and damaged endothelium called
lysosomal enzyme and tissue plasminogen
activator.

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 Human plasminogen
 Human plasminogen consists of 560 amino
acids heavy chain and a 241 amino acid
light chain.
 Glutamate is present at its amino terminal
end, folded into 5 looped structures which
are held together by 3 disulfide bonds.
 These loops are called KRINGLES.

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Human plasminogen

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 Applied physiology
 Human tissue plasminogen activator is
now produced by recombinant DNA
techniques.
 It causes lyses of coronary arteries.
 Streptokinase a bacterial enzyme.

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 Screening tests for Haemostasis
Laboratory test Function measured Associated disorder

1.Bleeding time Platelet function Qualtitative disorder of platelets

von willebrands disease
Vascular integrity Quantitative disorder of
platelets ,acquired vascular
disorders
2.Platelet count Quantification of platelets Thrombocytopenia
Thrombocytosis

3.Prothrombin time Evaluation of extrinsic & Deficiency of factors

common pathway I,II,V,VII,X,DIC, liver disease,
oral anticoagulant therapy
4.Partial thromboplastin time Evaluation of intrinsic & Deficiency of factors
common pathway I,II,V,VIII,IX,X,XI,XII

5.Thrombin time Evaluation of common Dysfibrinogenaemia

pathway Afibrinogenaemia
DIC,Heparin therapy
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 Anticoagulants
 Heparin :- it is produced by mast cells and
basophils.
 It is naturally occurring anti coagulant
which acts in following ways :-
1. It removes thrombin from the circulation
2. It inactivates active form of other clotting
factors.-10,11,12 factors.

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 Uses :

 Surgery
 Dialysis
 Before transfusing blood.

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 Coumarin derivatives
 Ex. Dicoumarol,warfarin.
 They act by inhibiting the action of Vit.K
 Oral anticoagulants.

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 Etheylene diamine tetra acetic
acid.
 Strong anticoagulant.
 It removes calcium and thus prevent
clotting.
 Used in lead poisoning.

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 Blood groups
 It was discovered by Austrian
scientist Karl Landsteiner in 1901.
 When blood from two individuals is mixed
sometimes agglutination occurs which is
due to immunological reaction.
 Antigens are present on Rbc and antibodies
are present in serum.

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 Land Steiner's laws

1. If a particular antigen is present in the red

blood cell the corresponding antibody
must be absent in the serum.
2. If the particular antigen is a absent in Rbc
the corresponding antibody must be
present in the serum.
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 There are more then 20 genetically
determined blood group systems. The two
main blood group systems are ABO and Rh
system.
 Different type of blood group systems are :-
MNS, Duffie ,Lewis ,Kell, Lutheran ,Rh,
Bombay Blood Group System , Punjab D
etc.

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 Antigens
 Normally the Rbc of a person contains
some agglutinogen called antigens ,when
they react with antibodies present in the
serum, they produce antigen antibody
reaction.

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 Antibodies
 There are two types of antibodies:-naturally
occurring and immune / accquired antibodies.
 Naturally occurring antibodies occur without any
previous antigenic stimulus.
 Accquired antibodies are produced in individuals
as a result of previous stimulation of red cell
antigen which is not present in their own red cells.

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 Complement binding antibodies:-the
majority of the antibodies bind to the
complements, with the exception of Rh and
MN antibodies.
 Cold antibodies: most of the antibodies are
IgM type and antigen antibody reaction
occurs at 5 degree-20 degree centigrade.

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 Warm antibodies:-antibodies A,B,Rh are
warm antibodies, as these antigen antibody
reactions do occur at body temp.
 Applied physiology:-As most of the
antibodies are cold antibodies consideration
of ABO and Rh antibodies is sufficient for
blood transfusion.

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 ABO system
 Based on the presence Group Antigen Antibody in
or absence of in Rbc serum
antibodies,blood is A A Anti B
divided into 4 groups,
A,B,AB and O groups. B B Anti A
AB A and B No antibody
O No Anti A and
antigen Anti B

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 Percentage of people having
different blood groups.
Population A B AB O

Europeans 42 9 3 46

Asians 25 25 5 45

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 Determination of the ABO
blood group.
 Principle of blood typing:-
 The blood typing is done on the basis of
agglutination.
 Agglutination is the collection of separate
particles like red blood cells into clumps or
masses.
 It occurs if an antigen is mixed with its
corresponding antibody which is called
isoagglutin.

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 These antigens are carbohydrates attached
to a precursor backbone and are secreted
into plasma and body fluids as
glycoproteins.
 H substance is the immediate precursor
upon which A and B antigens are added.

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 Applied physiology
 Rare individuals lack the H gene and cannot form
H substance.
 These individuals are homozygous for the silent
allele (hh) and have Bombay phenotype (Oh)
 Individuals with Bombay phenotype produce
antibodies to H substance as well as both A and B
antigens and are therefore compatible only with
other hh donors.

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 Applied physiology
 Non secretors of A and B antigens are
susceptible to a variety of infections as
many organisms bind to polysaccharides on
the cells.

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 Requisites for blood typing
 To determine the blood group of a person, a
suspension of his red blood cell and testing
antisera are required.
 Suspension of red blood cells is made by
adding blood drops in isotonic saline(0.9%).
 The test sera are :Antiserum A and
Antiserum B.

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Procedure :-

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 Importance of ABO groups in
blood transfusion:-
 Donor
 Recipient
 During blood transfusion antigen of the
donor and the antibody of the recipient are
considered.
 Universal donor O group
 Universal recipient AB group.

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 Cross matching
 It is done by mixing the serum of the
recipient and the red blood cell of the
donor.
 It is always done before blood transfusion.

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 Inheritance of ABO
agglutinogens and agglutinins
 A and B antigens appear in the 6 th week of
fetal life.
 Their concentration at birth is 1/5 th the
concentration of adult and this rises to adult
concentration at puberty.
 These antigens are inherited as Mendelian
characters.

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 The blood group of an individual depends
upon genes received from each parent.
 Gene A and B are dominant whereas gene
O is recessive.

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 Inheritance of ABO blood
group
Gene received Group of the off Genotype
from parent spring
A+A A AA or
A+O AO
B+B B BB or
B+O BO
A+B AB AB
O+O O OO
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 Rh factor
 It is an antigen present in the red blood cell.
 There are many types of Rh antigen but
only D antigen is more antigenic.
 Among Asians 85% of people are Rh +ve
whereas 15% are Rh –ve.
 There is no natural antibody to Rh antigen.

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 However if a Rh -ve person receives Rh
+ve blood for the first time he develops Rh
antibody.
 Rh factor is an inherited as a dominant
factor.
 Rh positive can be DD or Dd, Rh negative
can occur only with dd (homozygous
antigen.)

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 Hemolytic disease of the
newborn.
 When the father is Rh +ve , mother is
Rh –ve and the baby is Rh +ve, usually the
first child escapes the complications of Rh
incompatibility.
 Rh antigen cannot pass through the
placental barrier and enter the fetal
circulation.

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 When the mother conceives for the second
time and if the fetus is Rh positive, then it
can develop erythroblastosis foxtails.

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Transfusions reactions due to
Rh incompatibility.

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 Hydrops fetalis

 If hemolysis is severe then it can lead to

intrauterine death of the fetus.
 The death mainly occurs due to
development of edema, enlargement of liver
and spleen and cardiac failure.

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 Kernicterus
 If the child survives the anemia of
erythroblastosis fetalis then child is affected
by neurological syndromes.
 The child is more prone to develop
disturbance of motor
activities,deafness,chorea and spasticity.

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 Blood Transfusion
 When blood collected from an individual is given
to another person through vein (intravenously ), it
is called blood transfusion.
 Auto transfusion
 Indications for blood transfusion
 Haemorrhage
 Trauma
 Burns
 Anemia

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 Precautions to be taken before
blood transfusion
 Donors must be healthy without any
diseases like syphilis, AIDS, hepatitis etc.
 Only compatible blood must be transfused.
 Both matching and cross matching of
recipient blood and donor must be done.
 Rh compatibility must be confirmed.

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 Precautions to be taken while
transfusing blood.
 Apparatus used for this must be sterile.
 Temperature of blood must be same as body
temperature.
 Transfusion rate must be slow.

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 Collection and storage of
blood.
 Blood is mixed with commonly used
anticoagulant ACD.(Acid citrate dextrose)
 100 ml of ACD is sufficient for 500 ml of
blood.
 It is stored at 4 deg.centigrade. for about 21
days.

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 Preservation injuries
 Reduced 2,3 DPG
 Raised potassium ,sodium concentration.
 Reduced viability of Rbc’s so they cannot
work for longer time due to decreased ATP
in them.
 Reduced platelet count.

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 Hazards of blood transfusion
Hazards

Due to Due to
mismatched matched
blood transfusion

Immediate –Hemolysis Immediate –pyrogenic

Delayed- jaundice reaction, Shock, Delayed –transmission
Shock ,
Overload Of disease,
Renal failure
Hyperkalemia Hemosiderosis
Hypocalcemia

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 Incompatibility
 Severe hemolytic reaction is characterized
by 4 phases.

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 Hemolytic Phase
 Symptoms may occur immediately or may be
delayed by 2 hrs.
 Manifestations:-aching pain in lumbar region,
flushing of face, throbbing in the
head,anxiety,breathlessness,nausea,vomiting,hypo
tension.
 Hemorrhagic diathesis manifestations may be
masked in patients under general anesthesia.

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 The post shock phase
 Two main features are haemoglobinuria and
jaundice. Jaundice is evident after 12 hrs and
persists for several days, commonly deepest on the
day after transfusion.
 Haemoglobin value falls in proportion to the
amount of blood destroyed.
 Red cell agglutination may be present on the
blood film, and a moderate leucocytosis may also
be seen.

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 The Oliguric Phase
 It’s the first sign of renal failure, thus an
accurate record of fluid intake and urinary
output should be commenced immediately,
as fluid balance is critical in this situation.

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 The diuretic phase
 End of oliguric phase is marked by diuretic
phase
 Here there is excess loss of sodium,
potassium and water which if uncorrected
may cause death of the patient.

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 Management of immediate
phases are
 Cessation of transfusion
 Administration of 80-120 mg frusemide iv.
 Transfusion of compatible red cells and
plasma volume expander.
 IV hydrocortisone
 When abnormal bleeding occurs measures
to treat disseminated intravascular
coagulation.

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 Management of oliguric and diuretic phases
is that of renal failure.
 Fluid balance is critical
 Peritoneal or haemodialysis is needed in
severe cases.

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 Febrile Reactions
 They are relatively common in multiparous
women, people with repeated transfusions,.
 They have developed antibodies to
leucocytes or platelets.

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 Allergic Reactions
 They can range from small wheal to
circulatory collapse.
 Headache, nausea, vomiting,
dyspnea,edema, clinical picture of shock.
 Reduced serum antibody IgG is usually
seen.

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 Reactions due to infected blood.
 Thrombophlebitis.
 Transfusion hemosiderosis

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 Recent advances in health
matters
 Food and drug administration has
recommended that all blood used for
(homologous) transfusion be subjected to
leukoreduction.
 Efforts to increase the safety of blood used
in transfusions are ongoing like a new
screening procedure called nucleic acid test
(NAT)

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 Artificial blood:-researches are going on to
develop a duplicate blood that could carry out one
or more functions of blood.
 These intravenous solutions called oxygen
therapeutics began with a flurorchemical oxygen
carrying compound called fluosol.
 Polyheme is produced from chemically treated
haemoglobin obtained from outdated human
blood.Hemopure is produced from bovine
haemoglobin.

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 Conclusion
 Though blood can be considered as a
gift of life , but it can turn into death trap if
handled carelessly.
 Knowledge of Blood groups is essential:-
medically, socially and judicially.
 Hence basic knowledge of blood grouping
and transfusion reactions are of immense
help in saving patients lives.

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 References
 Review of medical physiology. William F.
Ganong.19th ed. Pg 493-520
 Human physiology –Andrew
Davies,Asa.G.H,Blakeley,Cecil Kidd.
Pg:-530-562
 Text Book Of Physiology –
Robert.M.Berne,Matthhew, Bruce,Satanton.
5 th ed. Pg:-265-274.
 Anthony’s Text Book Of Anatomy And
Physiology.17th ed. Pg:-25-38

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 De’gruchy’s text book of clinical hematology.
Pg.475-488
 Essentials of medical physioloy.
Prema Sembulingam, K.Sembulingam.
3rd ed. Pg 33-97
 Essentials of medical physiology-Anil Baran
Singh Mahapatra. Pg19-64
 Concised Medical Physiology-Chowdhary.
4th ed. Pg-19-65.

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