1/21/2021 Physiology – II

2nd Semester (Practical Short Notes)

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Contents

Sr # Practical Title Pg #
Haemopoietic System (Blood)

01 Determination of Bleeding Time 02

02 Determination of Clotting Time 03

04 Determination of ABO Blood Groups 05

Cardiovascular System

04 Measurement of Blood Pressure 07

05 Demonstration of Triple Response 09

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Practical # 01 – Determination of Bleeding Time

Bleeding Time
Time interval between the skin puncture and spontaneous, unassisted stoppage of bleeding.

Bleeding Time Test

 Bleeding time test is a medical test that is a basic assessment of how well your blood
platelets work to form clots.
 Platelets are tiny cell fragments that circulate in your blood. They’re the first cells to react
to a blood vessel injury. They seal off the wound to prevent more blood from escaping.
 A bleeding time test determines how quickly your blood clots to stop bleeding.
 The test involves making small punctures in your skin.

Aim of Bleeding Time Test

 You may need to have a bleeding test if you’ve been experiencing bleeding that won’t
stop, especially from small incisions, punctures, or cuts.
 Doctor can choose from a number of tests to evaluate your platelet function. A bleeding
time test is a common test to screen patients having prolonged bleeding times.
 Abnormal results could also indicate the following conditions:
o A blood vessel defect is any condition that affects how well your blood vessels
transport blood through your body.
o A genetic platelet function defect is a condition present at birth that affects how
well your platelets function. Hemophilia is one example of this type of defect.
o Thrombocythemia is a condition in which your bone marrow creates too many
platelets.
o Thrombocytopenia is a condition that causes your body to produce too few
platelets.
o Von Willebrand’s disease is a hereditary condition that affects how your blood
coagulates (clots).

Procedure of Bleeding Time Test

Apparatus:
 Lancet
 Alcohol swab
 Filter paper
 Stop Watch
Methods to be used: Ivy’s Method & Duke’s Method
Performance of the test using Duke’s method follows these steps:
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1. Clean the pricking/puncture site with an antiseptic (alcohol) to minimize the risk of
infection.
2. Subject is pricked using a lancet, preferably on the fingertip.
3. The prick is about 3-4 mm deep.
4. The patient then swipes the blood every 30 secs on a filter paper.
5. The test ends when the bleeding stops the usual timing is 2-6 minutes.

Result Evaluation
 Learning how long it takes your blood to clot can help your doctor determine if you have
a bleeding disorder, such as hemophilia or von Willebrand’s disease.
 Normal bleeding time is between one and eight minutes. Results outside of that range
could indicate a platelet defect and require further testing.

Practical # 02 – Determination of Clotting Time

Clotting Time
Time interval between the entry of blood into glass capillary tube, or a syringe, and formation of
fibrin threads.
Fibrin: an insoluble protein formed from fibrinogen during the clotting of blood. It forms a
fibrous mesh that impedes the flow of blood.

Clotting Time Test

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A type of coagulation tests, that measure your blood’s ability to clot, and how long it takes to
clot.

Aim of Clotting Time Test

 This test helps to assess the risk of excessive bleeding or developing clots (thrombosis)
somewhere in your blood vessels.
 Conditions that can cause coagulation problems include:
o liver disease
o thrombophilia, which is excessive clotting
o hemophilia, which is an inability to clot normally

Procedure of Clotting Time Test

Apparatus:
 Lancet
 Alcohol swab
 Capillary Tube
 Stop Watch
Method to be used: Wright’s Capillary Glass Tube
Performance of the test includes follows these steps:

1. Clean the pricking/puncture site with an antiseptic (alcohol) to minimize the risk of
infection.
2. Subject is pricked using a lancet, preferably on the fingertip.
3. The prick is about 3-4 mm deep.
4. Note the time when blood appears on the skin.
5. Touch the capillary tube to the drop of blood, and fill it.
6. Break 1 cm of the tube at the end of 2 min, and every 30 secs after that.
7. When fibrin is formed between the two broken pieces of tube, the coagulation/clotting
time is noted.
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Practical # 03 – Determination of ABO Blood Groups

ABO Blood Group System

 The A, B, and O blood groups were first identified
by Austrian immunologist Karl Landsteiner in
1901.
 ABO blood group system, is the classification of
human blood based on the inherited properties of
red blood cells (erythrocytes) as determined by the
presence or absence of the antigens A and B, which
are carried on the surface of the red cells.
 Persons may thus have type A, type B, type O,
or type AB blood.
 Blood containing red cells with type A antigen on
their surface has in its serum (fluid) antibodies against type B red cells. If, in transfusion,
type B blood is injected into persons with type A blood, the red cells in the injected blood
will be destroyed by the antibodies in the recipient’s blood.
 In the same way, type A red cells will be destroyed by anti-A antibodies in type B blood.
 Type O blood can be injected into persons with type A, B, or O blood unless there is
incompatibility with respect to some other blood group system also present.
 Persons with type AB blood can receive type A, B, or O blood.
 Antibodies (agglutinins) for the antigens A and B exist in the plasma and these are
termed anti-A and anti-B. The corresponding antigen and antibody are never found in the
same individual since, when mixed, they form antigen-antibody complexes, effectively
agglutinating the blood.

The RH System
 Rh antigens, named for the rhesus
monkey in which they were first
discovered, are also surface antigens
expressed on red blood cells. There are
a few Rh antigens (common one is
called D). Red cells expressing the Rh
antigens are called Rh positive. Red
cells which do not express this surface
antigen are Rh negative (about 15% of
the human population is Rh negative).
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Procedure of Determining ABO Blood Group System

1. Obtain clean microscope slides. Using a glass marking pencil, mark A, B and D on it.
2. Prick the tip of the finger under sterile conditions to obtain blood and place one drop of
blood on each marked spots.
3. Add one drop of anti-A serum to side marked A. Add one drop of anti-B serum to the
side marked B. And add anti-D serum to side marked D.
4. Mix the antiserum and blood on each side with a tooth pick, using always a new stick for
each side. Gently move the slide back and forth and observe for any agglutination of red
blood cells.
5. If agglutination occurs on side ‘A’ only then blood type is group ‘A’. And if
agglutination occurs on side ‘B’ only then blood group is ‘B’. If agglutination occurs on
both sides, the blood group is ‘AB’. If no agglutination occurs on either side, the blood
group is ‘O’. Sometimes it is necessary to observe the cells under the microscope to
ascertain the agglutination in doubtful cases. Agglutination generally looks like red
pepper granules.

Precautions
 Read the labels for expiry on bottles of antisera.
 The slides should be clean and dry.
 The mixing of blood and serum should be quick to avoid coagulation.
 Make sure that you do not mix the anti-A and anti-B antisera.

Interpretation of Results

Glass slide

A B D
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Practical # 04 – Measurement of Blood Pressure

Blood Pressure
The pressure of the blood in the circulatory system, often measured for diagnosis since it is
closely related to the force and rate of the heartbeat and the diameter and elasticity of the arterial
walls.
Systolic blood pressure is the pressure when the heart beats – while the heart muscle is
contracting (squeezing) and pumping oxygen-rich blood into the blood vessels.
Diastolic blood pressure is the pressure on the blood vessels when the heart muscle relaxes. The
diastolic pressure is always lower than the systolic pressure.

Measuring Blood Pressure

 Blood pressure is measured in units of millimeters of mercury (mmHg). The readings
are always given in pairs, with the upper (systolic) value first, followed by the lower
(diastolic) value.
 Normal systolic blood pressure is 120 mmHg.
 Normal diastolic blood pressure is 80 mmHg.
 Necessary for diagnosing and evaluating heart diseases.

Procedure of Blood Pressure Measurement

Apparatus:
 Sphygmomanometer
 Riva Rocci rubber cuff
 Stethoscope
Riva Rocci Sphygmomanometer, an instrument for measuring blood pressure, typically
consisting of an inflatable rubber cuff which is applied to the arm and connected to a column of
mercury next to a graduated scale, enabling the determination of systolic and diastolic blood
pressure by increasing and gradually releasing the pressure in the cuff.
Method to be used: Ascultatory Method
Performance includes follows these steps:
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When measuring blood pressure using the auscultation method, turbulent blood flow will occur
when the cuff pressure is greater than the diastolic pressure and less than the systolic pressure.
The "tapping" sounds associated with the turbulent flow are known as Korotkoff sounds.
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Practical # 05 – Demonstration of Triple Response

Triple Response
 The three cardinal circulatory responses of the skin (i.e. reddening, flare formation or
flushing of adjacent skin, and wheal formation or swelling) to any form of injury, such
as pricks, scratches, burns, exposure to corrosive chemical agents, and so on.
 This response was first described by Lewis in 1927. That’s why this response is also
called Lewis’ Response.
 With light injury, only the ‘white line’ is seen, while with a stronger stimulus, all the
three stages of ‘triple response’ can be seen.

White Line (White Reaction)

 Seat the subject on a stool with his forearm resting on the table.
 Draw a blunt pointed object like edge of the key or a blunt pencil, lightly on the skin of
the ventral surface of forearm.
 The response which appears in 8 – 10 seconds, is a pale or white line along the track of
the stimulus, the mechanical stimulus, causes contraction of the precapillary sphincters,
squeezes out the blood from the capillaries and small venules, leaving behind white line.

Triple Response
After the white line disappears in about a minute, use a stronger stimulus with a sharp pencil, the
response might vary from person to person. A full-fledged triple response, especially in sensitive
skins, consists of the following 3 stages.
1. The Red Line (Red Reaction): It appears in about 10 seconds. Red line appears due to
vasodilation of capillaries and venules in response to injury.
2. The Flare: It is an irregular zone or flush surrounding the red line, that develops due to
arteriolar dilation.
3. The Wheal: It is a zone of edema, due to fluid exudation.