PHSL2101 EXAM 2

● Practicals included
○ Fun with blood
○ Virtual - Membrane Potentials
○ Skeletal Muscle
○ Cardiovascular System
○ Virtual - electrical and mechanical events of cardiac cycle
○ Introduction to CVS
‘Fun with blood’
● 1.0 Experiment 1 - Blood typing
summary of the properties of red blood cells and plasma in each of the
four blood groups.

● 1.1 straight typing

○ "Typing" in blood testing refers to determining the blood group or blood type of an individual.
○ Blood typing is determined by the presence or absence of antigens on the surface of red blood
cells and the presence or absence of antibodies in the blood plasma.
■ E.g. If the blood agglutinates, or clumps, with the anti-A serum, it means the blood has A
antigen on its surface.
■ Antigens on red blood cells, also known as erythrocytes, are glycoproteins and
glycolipids that are present on the surface of these cells. These antigens are responsible
for determining a person's blood type, and they are used in blood typing tests to identify
the blood group of an individual.
■ The ABO antigens are glycoproteins that are found on the surface of red blood cells,
■ The Rh antigens, on the other hand, are a group of five antigens that are found on the
surface of red blood cells. The most important Rh antigen is the D antigen
● antigen on their red blood cells are said to be Rh-positive, while those who do
not have it are Rh-negative.

Blood group Anti-A Anti-B

A agglutinated -

B - agglutinated

AB agglutinated agglutinated

O - -

● 1.2 reverse typing

○ Reverse typing is a blood test that is performed to detect the presence of antibodies in a patient's
blood serum against specific blood group antigens.
○ mixing the patient's serum with known samples of red blood cells that have known antigens on
their surface.
○ The reaction is observed for agglutination or clumping, which indicates the presence of
 antibodies against the specific antigen on the red blood cells.
■ if the patient's serum reacts with red blood cells that have the A antigen on their
surface, this indicates the presence of anti-A antibodies in the patient's blood.

● 2.0 Experiment 2 - Osmosis and Red Blood Cells

○ use human red blood cells (erythrocytes) as a model cell to illustrate principles of osmotic forces,
diffusion and tonicity.

● 2.1 Isotonic (control) solution

○ A drop of whole blood consists of erythrocytes and colourless cells called leucocytes in plasma
■ erythrocytes which contain the protein haemoglobin that is responsible for the transport
of oxygen from the lungs to the tissues
○ study the cells in an isotonic solution that has the same total solute (osmolar) concentration as
the intracellular fluid of the erythrocyte
■ there is no tendency for the water to move in or out of the cell

● 2.2 Hypotonic solutions

● Why is the water now a pinkish colour?
○ As the red blood cells swell and haemoglobin (containing haem that gives the red colour to
erythrocytes) will begin to leak from the cell resulting in the transparent red-pink coloured
solution.
● Do you think all of the red blood cells will lyse?
○ Not all red blood cells will lyse as every blood cell has different numbers of aquaporins that allow
for water to permeate a cell. Hence cells with more aqua-pores would lyse easier.

● 2.3 Hypertonic solutions

● What is the sequence of events that brings about this change?
○ The sequence of events that brings about this change includes:
■ The hypertonic solution has a higher concentration of solutes compared to the inside of
the cell.
■ Water molecules will move out of the cell through the process of osmosis.
■ As water leaves the cell, the cell will shrink and become smaller in size.
■ The water in the molecule moves out faster than it comes through the membran

SUMMARY

Isotonic solution Hypotonic solution Hypertonic solution

What the concentration of solutes a solution with a lower solute the concentration of solutes
is the outside the cell is equal to the concentration than the cell outside the cell is higher than
concentration of solutes inside inside the cell.
conc?
the cell

Result/ there is no net movement of water flows into the cell due to the water moves out of the cell,
obs water across the cell membrane, concentration gradient, causing the leading to shrinkage or
 and the cell maintains its shape cell to swell and potentially burst crenation of the red blood cells
and volume (lyse) if the cell wall is not present

Impact they will retain their In the case of a blood cell, this can concentration of solutes inside
on characteristic shape, which is lead to hemolysis, or the rupturing the cell is higher than outside,
shape? biconcave, and remain intact. of the cell membrane, which can leading to the net movement
release its contents, including of water out of the cell to
This is important for maintaining
hemoglobin and other cellular balance the concentration
the function of red blood cells, components, into the surrounding gradient. In extreme cases,
which is to transport oxygen and fluid. excessive water loss can cause
carbon dioxide throughout the the cell to become dehydrated
body. and lose its function.

● 2.4 Tonicity vs Osmolarity

● Tonicity refers to the effect a solution has on a cell – whether it causes the cell to shrink, stay the same size
or to swell (as seen in the exercises above).
● Osmolarity refers to the concentration of solutes in a solution.

● a) Calculations of osmolarity
○ need to calculate the osmolarity (osmoles/litre) of the glucose
○ solution.
○ Note the osmolarity = n x C x ϕ
■ where n= the number of molecules a solution dissociates into,
■ C is the molar concentration and
■ ϕ is the osmotic co-efcient.

○ For this concentration of glucose ϕ ≈1).

○ Recall that molar concentration is the concentration of a substance expressed in terms of moles
per litre, where a mole is the molecular weight in grams.
■ The molecular weight of glucose is 180 so 180g is 1 mole of glucose.
■ A 5% solution refers to weight (in g, per 100 mls, or 5g/100mls)

Osmolarity of glucose (5%) = n x C x ϕ (n= 1, ϕ = 1)

5 grams per 100 mls = X mols per litre = C

= 50/180
= 0.27778
Is this hyper-osmotic, hypo-osmotic or iso-osmotic with respect to saline (280 momol/l)?

● b) hypothesis:
○ If red blood cells are placed in a 5% glucose solution, will they swell or shrink?

If red blood cells are placed in a 5% glucose solution, they will shrink due to the solution being hypertonic
compared to the intracellular fluid of the red blood cells. The concentration of solutes inside the cell is higher than
the concentration of solutes in the surrounding 5% glucose solution, so water will move out of the cell to try to
balance the concentration on both sides. This results in the cell losing water and shrinking.
 ● c) Experiment:
○ Did the 5% weight/volume solution of glucose cause water to diffuse in or out of the red blood
cells?
the red blood cells appeared to shrink and become more crenated, it suggests that water diffused out of the cells,
causing them to lose volume and change shape. Therefore, the 5% glucose solution was hypertonic to the red
blood cells and caused water to move out of the cells through osmosis.

○ Is the 5% weight/volume solution of glucose hypertonic, isotonic or hypotonic?

The 5% weight/volume solution of glucose is hypertonic with respect to normal plasma.

● 3.0 Drugs and fluid movement

● To determine the effect of drugs which alter membrane permeability on fluid movement in red blood cells
by measuring the haematocrit.
● Many antibiotics and other drugs act by altering the permeability of the membrane.
○ When the ability of different ions to cross the cell membrane is altered, there will be subsequent
effects on osmotically-driven fluid movement across the membrane.
● examine how these drugs alter fluid movement by measuring the haematocrit, or the percentage of the
sample occupied by red blood cells (with the remainder predominantly extracellular fluid which for the
blood is plasma).

Control (DMSO) Drug 1: Monensin (in Drug 2: Valinomycin (in

DMSO) DMSO)

haematocrit

Control (Saline) Solution1: 5% Glucose (Iso- Solution 2: 10% Glucose

osmotic & Hypotonic) (Hyperosmotic &
hypertonic)

haematocrit

● Can you predict from the haematocrit results how the drugs have affected the net movement of
water across the erythrocyte plasma membrane?
○ It is difcult to predict how the drugs have affected the net movement of water across the
erythrocyte plasma membrane based solely on the haematocrit results. However, if the
haematocrit value is lower than expected in the tubes with Monensin or Valinomycin, it may
indicate that the drugs have increased membrane permeability, leading to water diffusing out of
the cells and causing them to shrink.
○ Conversely, if the haematocrit value is higher than expected, it may suggest that the drugs have
decreased membrane permeability, leading to water diffusing into the cells and causing them to
swell. However, it is important to note that other factors, such as changes in the red blood cell
 shape or aggregation, can also affect the haematocrit results.

● What gives rise to the osmotic force that causes any such water movement?
○ The osmotic force that causes water movement is generated by the concentration gradient of
solutes between the two sides of the membrane. Water moves from an area of low solute
concentration to an area of high solute concentration, in order to balance the concentration on
both sides. This process is known as osmosis, and the resulting force is called osmotic pressure

● Given that the main extracellular ions are Na+ and Cl- , and the main intracellular ion is K+, how
may each of the drugs have affected the permeability of the membrane to these ions?
○ Monensin is a ionophore that facilitates the diffusion of monovalent cations, including Na+ and
K+, across cell membranes. It is possible that the addition of Monensin increased the
permeability of the erythrocyte membrane to Na+ and K+, leading to an equilibration of ion
concentrations between the intracellular and extracellular environments.
○ Valinomycin is also an ionophore, but it is selective for K+. It forms a complex with K+ ions that
allows for facilitated diffusion of K+ across cell membranes. Therefore, the addition of
Valinomycin likely increased the permeability of the erythrocyte membrane to K+, leading to an
equilibration of K+ ion concentrations between the intracellular and extracellular environments.

● Describe a possible mechanism of action for each of the drugs and solutions.
○ Monensin and valinomycin are both drugs that alter the permeability of the erythrocyte plasma
membrane to ions. Monensin acts as an ionophore and facilitates the transport of monovalent
cations, such as Na+ and K+, across biological membranes. It works by forming a complex with
these ions and shuttling them across the membrane. In erythrocytes, the increased permeability
to Na+ would lead to an influx of this ion, causing an increase in intracellular osmolarity and
resulting in water movement into the cells.

○ Valinomycin, on the other hand, is a potassium-specific ionophore. It forms a complex with K+

ions and transports them across the membrane, leading to an increase in the intracellular K+
concentration. In erythrocytes, this would result in an increase in intracellular osmolarity, leading
to water movement into the cells.

○ The 5% and 10% glucose solutions would both be hypertonic with respect to the erythrocytes.
Glucose is a large, polar molecule that cannot easily cross the erythrocyte plasma membrane.
When these solutions are added to erythrocytes, the high concentration of glucose outside the
cells causes an osmotic gradient that drives water movement out of the cells and into the
extracellular fluid. This results in cell shrinkage.
Virtual - Membrane Potentials

We hope you gain a better understanding of:

● how resting membrane potentials, action potentials and synaptic potentials occur in real cells
● how a combination of ion concentration gradients and membranes that have ion selectivity can
produce different types of membrane potentials
● how the magnitude and sign of these potentials arises from considering electrochemical forces
and membrane permeability.
Skeletal Muscle
● 1.0 Graded stimulation of the sciatic nerve-recruitment of motor units
○ Lowest number means the smallest neurons have been fired as they are the smallest
and easiest to depolarise
○ Recruitment of motor neurons - when all the neurons have been activated

● Your graph should show that a graded mechanical response is obtained when progressively
increasing the intensity of the stimulus applied to the sciatic nerve.
● What Is the mechanism responsible for the increase in force when the stimulus intensity is
increased? Why does the curve rise sharply then reach a plateau?

○ The graded mechanical response observed with increasing intensity of the stimulus
applied to the sciatic nerve is due to the recruitment of more motor units within the
muscle. Motor units are composed of a motor neuron and the muscle fibers it
innervates.
○ When a weak stimulus is applied, only a small subset of motor units are activated,
leading to a small amount of force production. As the intensity of the stimulus is
increased, more and more motor units are recruited, resulting in greater force
production.

○ The curve rises sharply initially because the motor units with the lowest activation
thresholds are recruited first. As the stimulus intensity increases, progressively higher-
threshold motor units are recruited, which produces a steeper increase in force output.
○ Eventually, all available motor units are recruited, and the force production plateaus. At
this point, any further increase in stimulus intensity will not result in a further increase in
force output because all motor units are already activated.

○ Overall, the graded mechanical response observed with increasing stimulus intensity is
due to the recruitment of more motor units within the muscle, and the curve rises
sharply then reaches a plateau due to the recruitment of motor units with progressively
higher activation thresholds.

● 2.0 Summation of contraction

● What do you think the mechanism is behind the marked difference in the tension developed
following a single stimulus applied to the nerve and that resulting from repetitive stimulation?
○ The marked difference in tension developed following a single stimulus applied to the
nerve and that resulting from repetitive stimulation is due to the phenomenon of muscle
fatigue.
○ When a single stimulus is applied to the nerve, the resulting muscle contraction is
usually at its maximum force. However, when the muscle is repeatedly stimulated,
fatigue can set in and reduce the force generated by subsequent contractions. This is
because repeated contractions result in a decrease in the availability of energy
substrates and accumulation of metabolic byproducts such as lactate and hydrogen ions
within the muscle fibers. These changes can impair muscle contractile properties, such
as reduced calcium sensitivity, decreased cross-bridge cycling, and altered muscle fiber
membrane excitability.
○ Additionally, during repetitive stimulation, there may be a depletion of the
neurotransmitter acetylcholine (ACh) from the nerve terminals, which can reduce the
amount of ACh available to bind to the muscle receptors and generate an action
potential. This can result in a decreased number of activated motor units and
subsequently reduce force output.

● 3.0 Effect of varying the frequency of trains of supra-maximal stimuli

● Why did the muscle contraction become “smooth” when stimulated at higher frequencies?
● What is the term used to describe this type of tetanus?
○ The muscle contraction becomes "smooth" when stimulated at higher frequencies
because the repeated stimulation causes individual twitches to merge together, resulting
in a sustained contraction without any visible relaxation between twitches. This type of
contraction is called complete tetanus or fused tetanus.

○ During high-frequency stimulation, the time between successive twitches is shortened,

and the muscle fiber does not have enough time to relax fully before the next twitch is
initiated. As a result, the twitches start to fuse together, and the force generated by each
individual twitch is summed together, leading to a sustained contraction.

○ In complete tetanus, the muscle fibers are maximally stimulated and reach their
maximum tension-producing capability. The muscle fibers are unable to relax, and the
muscle remains contracted until the stimulation stops.

○ The smooth sustained contraction observed during complete tetanus is in contrast to the
individual twitches observed during low-frequency stimulation, where there is sufcient
time for the muscle fibre to relax between successive twitches.

● 4.0 . Influence of length on isometric contractions

QUESTIONS:
1. Explain the observations you have made concerning the relationship between the length of the
gastrocnemius muscle and:
● (i) its passive tension?
○ The relationship between the length of the gastrocnemius muscle and its passive tension is
explained by the length-tension relationship of muscle fibers. The passive tension is directly
related to the length of the muscle, and the tension-generating capacity of the muscle decreases
when it is stretched beyond its optimal length.

● (ii) the twitch tension?

○ The twitch tension of the gastrocnemius muscle is related to the degree of overlap between the
actin and myosin filaments within muscle fibers. At an optimal length, the muscle generates
maximal tension with a single twitch, which decreases when the muscle is stretched or shortened
beyond its optimal length.

● (iii) the total tension developed during a response?

○ The total tension developed by the gastrocnemius muscle during a response is the sum of active
and passive tensions. The relationship between muscle length and total tension can be explained
by the length-tension relationship of muscle fibers and the relationship between passive tension
and muscle length. At an optimal length, the active tension generates the greatest total tension,
with passive tension contributing to the total tension at lengths beyond the optimal length.

● (iv) Why is isometric recording essential in investigating these relationships?

○ Isometric recording is necessary to investigate the relationship between muscle length and
tension because it allows for the measurement of tension without changes in muscle length. This
method provides reliable results by eliminating the confounding variable of changes in muscle
length and allows for the determination of the length-tension relationship of muscle fibers and
the contribution of passive tension to the total tension generated by the muscle.

2. Based on all your experimental results and what you have learned in lectures, consider the following
questions related to muscle contractions in vivo.

● How are graded contractions obtained in vivo?

○ Graded contractions in vivo are obtained through the recruitment of motor units within a muscle.
The size principle determines the order of recruitment, with smaller units being recruited first.
Increasing the frequency or intensity of stimulation recruits more motor units, resulting in an
increase in force generated. The length-tension relationship of muscle fibers also contributes to
graded contractions by influencing the degree of overlap between actin and myosin filaments.

● What is the smallest functional unit of contraction in vivo?

○ The sarcomere is the smallest functional unit of contraction in vivo. It consists of overlapping
actin and myosin filaments that generate force when they interact. When a muscle fiber is
stimulated, the sarcomeres within the fiber contract, resulting in the shortening of the fiber and
 the generation of force. The force generated by each sarcomere is transmitted to adjacent
sarcomeres and to the entire muscle fiber, resulting in muscle contraction.

● Which fibre type do you predict would produce a tetanic contraction with a lower frequency of
input (action potentials) from its alpha motor neuron? Why?

○ Slow-twitch (Type I) muscle fibers are predicted to produce a tetanic contraction with a lower
frequency of input from their alpha motor neuron.

○ This is because slow-twitch fibers have a slower rate of Ca2+ release and uptake by the
sarcoplasmic reticulum compared to fast-twitch (Type II) fibers. This slower rate of Ca2+ cycling
results in a longer duration of Ca2+ binding to troponin, allowing for sustained force production.

○ In addition, slow-twitch fibers have a higher density of mitochondria and oxidative enzymes,
which provide a more efcient supply of ATP for sustained contractions.

○ Therefore, slow-twitch fibers have a greater capacity for sustained force production and fatigue
resistance, which allows them to maintain a tetanic contraction at a lower frequency of input
from their alpha motor neuron compared to fast-twitch fibers.

● During a typical contraction, many of the recruited motor units will not be stimulated strongly
enough to generate a fused-tetanus. What else contributes to making our movements smooth
and fluid?

○ In addition to the recruitment of motor units, the smoothness and fluidity of our movements are
also achieved through the process of motor unit summation, which involves the modulation of
the firing rates of individual motor units.

○ As the force requirements of a given movement increase, the firing rate of individual motor units
can increase, resulting in greater force production. This process allows for a fine degree of control
over movement, as motor units can be activated in a graded manner to generate the precise
amount of force required for a given task.

○ Furthermore, the nervous system can also modulate muscle activation patterns through the use
of reflexes, which provide automatic adjustments in muscle activity based on sensory feedback.

○ Therefore, the smoothness and fluidity of our movements are achieved through a combination of
motor unit recruitment, motor unit summation, and sensory feedback from reflexes.
Cardiovascular System
Introduction
The cardiovascular system comprises the heart, which pumps the blood, and the blood
vessels through which the blood flows.

Figure 1A is a schematic representation of the mammalian

cardiovascular system.
Note that there are two circuits: One in which blood is
pumped from the right ventricle to the lungs and back to
the left atrium. This is the 1.Pulmonary circulation. In the
other circuit, blood is pumped from the 2.left ventricle to
the various organs and tissues, (via their individual
circulations) and then returns to the 3. right atrium via the
venae cavae. This is the systemic circulation.

In Figure 1B
each individual
organ or tissue
system is
supplied by
4.large arteries leaving the aorta. These vessels divide into
smaller vessels, the 5.small arteries and these branch into 6.arterioles and finally 7. capillaries. These
very small vessels then reform into slightly larger vessels, the 8.venules which join to form 9.small veins
and then 10.large veins. These vessels empty into the 11.superior venae cavae or 12. inferior venae
cavae which then enter the right atrium of the heart. As vessels divide into successively smaller vessels
they become more numerous so that the capillaries, which have the smallest diameter of any of the
vessels are also the most common.

THE HEART
The heart is a muscular organ located in the thoracic cavity. It
is covered by a fibrous sac, the pericardium. The walls are
composed of cardiac muscles, the myocardium and the inner
surface is covered by a thin layer of cells, the endocardium.
In Figure 2A note that the heart has a number of chambers
or cavities. How many chambers are there and what are they
called? 13. 4 chambers, L & R atrium and L & R Ventricles.

There is no direct communication between the left and right

sides of the heart in adult
mammals. Note that there are valves between the atria and
the ventricles and between the ventricles and the vessels. The valves between the atria and the
ventricles are the atrio-ventricular (A-V) valves. These permit flow from the atria to the ventricles but
not in the reverse direction. The valve between the left atrium and the left ventricle has two cusps and is
called the 14.bicuspid or 15.mitral
valve. The A-V valve between the right atrium and the right ventricle has three cusps
and is called the 16.tricuspid valve. These valves are connected to the 17.papillary muscles by the
threadlike 18. chordae tendinae which prevent the valves from being sucked into the atria.

The ventricles open to large vessels, the left ventricle to the 19.aorta and the right ventricle to the
20.pulmonary artery. The openings are guarded by the pocket-like 21.semi - lunar valves. There are no
true valves between the atria and the incoming veins, the 22.vena cavae on the right and the
23.pulmonary veins on the left.

The cells constituting the walls of the heart do not exchange nutrients and metabolic end-products with
the blood in the heart chambers. They receive their blood from arteries which arise from the aorta. Look
at Figure 2B. These are the 24.coronary arteries. Note also that the circulation to the heart is shown as a
separate circulation in Figure 1A.

The contraction of the cardiac muscles is initiated by the depolarization of the membranes of certain
cardiac fibres in specialised areas. These cardiac cells are capable of spontaneous, rhythmic self-
excitation and are therefore said to be autorhythmic. Figure 3 shows the electrical conducting system in
the heart. There are 3 main areas of spontaneous activity where contraction may be initiated. The area
with the fastest spontaneous rhythm will be the "pacemaker". This area, where excitation in the heart
usually originates is situated at the opening of the superior vena cava into the right atrium and is called
the 25.SA node. The wave of excitation spreads across the atria to another group of pacemaker cells at
the junction of the right atrium and the ventricles called the 26.AV node and then enters the wall
between the two ventricles via the conducting system fibres termed the 27.Bundle of His. This then
divides into right and left bundle branches and from these 28.purkinje fibres extend and spread
throughout the ventricular myocardium. The electrical events of the heart can be recorded as the
electrocardiogram (ECG or EKG).

The mechanical events of the heart immediately follow the electrical events. Figure 4A shows the heart,
or more specifically the valves, in the heart during the relaxation phase. Note that the 29.atrioventricular
valves are open while the 30.semi - lunar valves are closed so that blood does not flow out. Figure 4B
shows the ventricles in the contraction phase called systole. During this phase blood is pumped from
both ventricles into the circulation. The 31.__semi - lunar__ valves are open to permit this to occur.

In the adult person each ventricle pumps 70-80ml/beat (the stroke volume). Since the heart rate is
about 65-75/min, the cardiac output (or volume pumped from either ventricle each minute) must be
about 32.___4.5 to 6_____ litres/min. Note that each ventricle must pump equal volumes of blood per
beat. Therefore, the cardiac output may refer to either ventricle.
SOME BASIC DEFINITIONS AS APPLIED TO THE CARDIOVASCULAR SYSTEM
1. Blood pressure: pressure a volume of fluid (blood) exerts on the wall of its container
(blood vessels). Imparted from cardiac contraction ejecting blood into the container (blood
vessels). Varied within the system by changing the diameter of the vessels and hence
changing the container volume.

2. Cardiac Output (C.O.): volume of blood ejected from the heart per unit time (units = L/min
or ml/min) = Net blood flow (Q) through the vessels since the fluid (blood) is incompressible.

3. Ohm's Law: V = IR
In the cardiovascular system refers to:
. P = QR

Pressure difference = Blood flow x Total Peripheral Resistance

= CO x TPR

Blood Flow simply means the quantity of blood that passes a given point in the circulation in a given
period of time. It is usually expressed in ml/min.

4. Resistance is the impediment to blood flow in a vessel. It is determined by vessel properties:

R = 8 ηL
πr4
Where;
● η = blood viscosity
● L = vessel lengths.
● r = vessel radius

5. Velocity of blood flow (V) = Q

A
Where;
● A = Vessel cross-sectional area.

6. CO = SV x HR
= ml/beat x beats/min
= ml/min
Where;
● SV = stroke volume
= volume blood/beat
● HR = heart rate
= beats/unit time
 7. CO = O2 consumption
[O2] artery - [O2]vein
Where;
● O2 consumption = total body oxygen consumption (ml/min)
● [O2] artery = oxygen content of arterial blood (ml O2/100 ml blood)
● [O2] vein = oxygen content of venous blood (ml O2/100 ml blood)

8. BP = CO x TPR
where;
● TPR = total peripheral resistance
= the total resistance to flow in the periphery

QUESTIONS:
1. Does the palpation method measure systolic arterial pressure or diastolic pressure?
● The palpation method is used to estimate systolic arterial pressure, which is the pressure in the arteries
during the contraction of the heart (systole). The palpation method involves feeling for the pulse in an
artery and using the strength of the pulse to estimate the systolic arterial pressure.

2. Is there a significant difference between the readings obtained by the palpation method and
the corresponding readings obtained by auscultation? If so, what is the explanation of the
difference? Which is the more accurate method?
● Studies have shown that there can be a significant difference between the readings obtained by the
palpation method and the corresponding readings obtained by auscultation, with the palpation method
tending to overestimate systolic blood pressure compared to auscultation.
● The difference between the two methods can be explained by the fact that the palpation method relies on
subjective assessment of the strength of the pulse, which can vary depending on factors such as the
position of the patient, the size of the artery, and the skill of the examiner. In contrast, auscultation
involves the use of a blood pressure cuff and stethoscope to directly measure the sounds of blood flow
through the artery, providing a more objective and accurate measure of blood pressure.
● Therefore, auscultation is generally considered the more accurate method for measuring blood pressure.
3. It is often stated that the mean arterial pressure can be calculated from the readings
obtained by the auscultation method by adding half the pulse pressure to the diastolic
pressure value. Do you consider this a satisfactory method? If not, why not?
● The calculation of mean arterial pressure (MAP) using the equation MAP = diastolic pressure + 1/3 (systolic
pressure - diastolic pressure) is a commonly used and accepted method for estimating MAP based on
blood pressure measurements obtained by auscultation.
● However, the method of adding half the pulse pressure to the diastolic pressure value (MAP = diastolic
pressure + 1/2 pulse pressure) is less accurate than the previous method because it assumes that the
relationship between systolic and diastolic pressures is constant, which is not always the case. In addition,
it does not take into account individual variations in arterial compliance and wave reflection, which can
affect the pulse pressure and thus the accuracy of the calculation.
● Therefore, while the method of adding half the pulse pressure to the diastolic pressure value can provide a
rough estimate of MAP, it is not as accurate as the equation that takes into account the systolic and
diastolic pressures and should not be relied upon as the sole method for calculating MAP.

4. What would be the effect on blood pressure readings of:

a) raising the cuffed arm up above the level of the heart?
● Raising the cuffed arm above the level of the heart during blood pressure measurement
can result in a falsely low blood pressure reading. This is because the hydrostatic
pressure created by raising the arm reduces the pressure in the arteries, which can
cause the pulse sounds to become less audible and result in an underestimation of
systolic blood pressure. Therefore, it is important to keep the arm at heart level during
blood pressure measurement to ensure accurate readings.

b) allowing the cuff pressure to drop too quickly?

● Allowing the cuff pressure to drop too quickly during blood pressure measurement can
result in a falsely low diastolic blood pressure reading. This is because if the cuff pressure
is released too quickly, the sounds of the blood flow may disappear suddenly, resulting
in an underestimation of the diastolic blood pressure. Therefore, it is important to
release the cuff pressure slowly and steadily to ensure accurate blood pressure readings.

c) a very fat arm? How could you avoid this artefact?

● Measuring blood pressure on a very fat arm can result in falsely high blood pressure
readings. This is because the cuff may not fit properly and result in an overestimation of
blood pressure due to the increased volume of the arm. To avoid this artifact, a larger
cuff should be used to ensure a proper fit around the arm. The cuff bladder should also
be centered over the brachial artery, and the arm should be supported at heart level
during the measurement.

5. What sort of calibration is required for the manometers used for these measurements?
 ● Calibration of manometers used for blood pressure measurements is important for accuracy and
reliability. The manometers should be calibrated against a known standard at regular intervals,
and the calibration process should be performed by a qualified technician or service provider.
This involves adjusting the manometer readings to match the known standard and correcting any
deviations from the standard.

6. How could you validate these indirect methods of measuring arterial blood pressure?
● Validation of indirect methods of measuring arterial blood pressure involves comparing the
results to those obtained from a direct measurement, such as an arterial catheter, and comparing
the indirect method to other indirect methods to verify their accuracy and consistency. Validation
studies should be performed on a representative sample of the population and published in peer-
reviewed journals for transparency and reproducibility.

Experiment 2:
QUESTIONS:
1. Compare the thickness of the walls and the size of the cavities of each ventricle.
● The left ventricle of the heart has thicker walls than the right ventricle, as it has to generate enough
pressure to pump oxygen-rich blood to the rest of the body. The right ventricle, on the other hand, pumps
oxygen-poor blood to the lungs, which requires less force and thus has thinner walls. In terms of the size
of the cavities, the left ventricle is also slightly larger than the right ventricle, as it needs to accommodate
a larger volume of blood.

2. Which compartment has the thickest wall?

● The left ventricle of the heart has the thickest wall.

3. Why is this wall the thickest?

● This is because it has to generate enough pressure to pump oxygenated blood to the entire body through
the systemic circulation. The right ventricle, on the other hand, has a thinner wall because it only has to
pump deoxygenated blood to the lungs through the pulmonary circulation. The atria have thinner walls
than the ventricles because they only need to pump blood into the ventricles.

4. What prevents blood flowing back into the atria

(a) on the right side?

● The tricuspid valve, also known as the right atrioventricular valve, prevents the backflow of blood from the
right ventricle into the right atrium during systole (ventricular contraction). The tricuspid valve is a one-
way valve made up of three cusps or leaflets that open during diastole (ventricular relaxation) to allow
blood to flow from the right atrium to the right ventricle, and then close during systole to prevent
backflow. The papillary muscles and chordae tendineae also play a role in the function of the tricuspid
valve by preventing the valve leaflets from prolapsing into the atrium during ventricular contraction.

(b) on the left side?

● The mitral valve, also known as the bicuspid valve or left atrioventricular valve, prevents blood from
flowing back into the left atrium from the left ventricle during systole (ventricular contraction). The mitral
valve is a one-way valve made up of two cusps or leaflets that open during diastole (ventricular relaxation)
 to allow blood to flow from the left atrium to the left ventricle, and then close during systole to prevent
backflow. The papillary muscles and chordae tendineae also play a role in the function of the mitral valve
by preventing the valve leaflets from prolapsing into the atrium during ventricular contraction.

Electrical and Mechanical events of cardiac cycle

Anaesthesia and Surgery
Prior to the procedures, the animal was anaesthetised with an intravenous injection of pentobarbitone – a
barbiturate anaesthetic.

Cannulation of trachea
With the animal lying supine, a midline incision is made in the skin at the font of the neck. Underlying muscles are
then separated, and the trachea identified. The trachea is easily identified by the rings of cartilage in its wall. A
small hole is made in the trachea, the edge of the hole is held and then a plastic cannula is inserted. Through the
tracheal cannula, secretions from the upper respiratory tract can be removed and the animal can be artificially
ventilated if necessary.

Cannulation of the common carotid artery

The common carotid artery is identified in the neck as it lies beside the vagus nerve. It is separated from the nerve,
and a clip is placed on it on the side closest to the heart. A ligature is placed around the carotid artery distal to the
heart and tied into position. A further tie is placed around the carotid artery, but not at this stage tied tightly. A
hole is placed part way through the wall in the carotid artery and a catheter is introduced into the artery through
this hole and advanced towards the clip. The second tie is then used to secure the catheter in place and the clip is
removed. This catheter will be used to measure the animal’s blood pressure.

Cannulation of external jugular vein

The external jugular vein lies on the side of the neck close to the skin. It is identified and separated from
surrounding tissues. As with the carotid artery, a clip is placed on it towards the heart. The distal end of the vessel
is tied and a small hole is cut part way through its wall. The catheter is introduced and advanced past the clip so
that the tip lies in or close to the right atrium. The catheter is secured in place and the clip removed. This catheter
will be used to record the right atrial pressure.

Connection of ECG leads

Electrodes need to be placed via needles through the skin. This is required because the dog’s fur prevents electrical
contact being made with external electrodes as is possible in humans. Three electrodes are used. They can be
combined to produce different ECG leads - leads I, II and III as indicated on Einthoven's triangle below.

Recording carotid arterial pressure

The carotid artery catheter is connected to a blood pressure transducer, which in turn is
connected to a Grass polygraph (chart recorder).

Pen 2 is recording the arterial blood pressure. You can see the pressure rise and fall with
each beat of the heart. Pen 1 is measuring the time in seconds. The record of arterial blood
pressure is calibrated in mmHg. The systolic pressure is about 110 mmHg and the diastolic is
about 90 mmHg.

Recording the electrocardiogram (ECG)

The ECG electrodes are connected to a junction box which is in turn connected to the Grass polygraph. The lead
selector can be used to choose combinations of the electrodes on the animal’s surface.

Lead I combines electrodes on the right arm and left arm.

Lead II combines electrodes on the right arm and left leg,

Lead III combines electrodes on the left arm and left leg.

ECG trace
The recording below (left) shows the ECG trace from Lead II. The P, QRS and T waves are indicated.

The P wave represents atrial depolarisation, the QRS represents ventricular depolarisation and the T wave
represents ventricular repolarisation.

The figure on the right shows two different leads. Although the same waves are seen, their shape is different
because the electrical activity of the heart is being viewed from a different direction.

ECG and carotid arterial pressure

Here both the ECG and carotid pressure are recorded together.

On the ECG trace, the QRS complex represents depolarisation of the ventricle or the commencement of ventricular
systole (contraction). The upstroke of the pressure wave in the carotid artery also represents the commencement
of systole. However, it can be seen that the QRS complex precedes the upstroke of pressure in the carotid artery.
Why is there a delay between the onset of the QRS and the onset of the rise in arterial pressure in the
carotid artery?
Three factors contribute:

1. The time taken for excitation-contraction coupling to occur. It takes time for the action potential to travel
down the T-tubules, release calcium ions etc.
2. The time of isovolumetric contraction. This lasts about 0.05 sec until the pressure in the left ventricle
exceeds aortic pressure and the aortic valves open.
3. Time taken for the pulse wave to travel along the wall of the arteries to the point of recording in the
carotid. Note that because of the elasticity of the walls of the arteries, the pulse wave travels slower in
these vessels than in a rigid catheter. In a young adult, this wave travels at about 4 m/sec in the aorta, and
8 m/sec in large arteries. Note that this pressure wave moves at a faster velocity than the rate of blood
flow.

ECG and femoral arterial pressure

Now blood pressure is measured from the femoral artery in the thigh (in the same way as the carotid arterial
pressure was recorded).

Simultaneous recordings of the ECG and femoral arterial blood pressure are shown. For comparison, the carotid
trace is also shown. Note that typically the femoral arterial pulse has a high systolic peak and a secondary diastolic
wave. The delay between the QRS complex and the commencement of the rise in arterial pressure is quite long.

Changes in the arterial pressure wave with recording site

As the recording site moves further from the heart the following
changes occur:

1. There is a longer delay to the onset of the initial rise in

pressure.
2. There is an increase in the amplitude of the systolic peak.
 3. The pulse pressure (difference between systolic and diastolic pressure) becomes greater.
4. The systolic portion narrows and the time to peak pressure is reduced.
5. High frequency components like the incisura (the notch at the end of ventricular ejection) are damped out
and disappear.
6. A hump or secondary wave may become prominent on the diastolic portion of the wave.

What causes these changes in arterial pressure wave shape?

A number of factors contribute including the resonance of the arterial tree (which enhances certain frequencies
and dampens others), tapering of the vessel (as the vessel progressively narrows the pressure wave is amplified)
and reflection of the wave at branch points or when the arterial dimension changes.

Note that when the arteries are less compliant the shape of the pressure wave changes. A good example of this is
arteriosclerosis (“hardening of the arteries”). (See fig)

Left ventricular pressure

This pressure recording is made through the same catheter that was previously used to record carotid arterial
pressure, but the catheter tip is advanced so that it lies within the left ventricle in the thorax.

Simultaneous recordings of the ECG and left ventricular pressure are shown. Contraction of the left ventricle cause
a wide pressure pulse. The diastolic pressure is close to zero and the systolic peak is around 110 mmHg (ie the
same as the carotid arterial pressure).

The large ventricular pressure pulse is preceded by a

small notch of pressure caused by the atrial contraction.

Left ventricular and aortic pressure

Here the catheter is gently withdrawn from the left ventricle into the aorta. Note that the systolic pressure remains
the same but the diastolic pressure is much higher. On advancing the catheter tip back into the ventricle, the wide
pulse pressure is again seen. The catheter tip lying within the ventricle irritates the ventricle and causes occasional
ectopic beats.
Carotid sinus reflex
This reflex is seen when the common carotid artery is occluded with a clip. This causes a drop in blood pressure in
branches upstream of the occlusion.

The trace shows heart rate, ECG, and arterial blood pressure. The marker pen indicates the period of occlusion.
When the carotid is occluded, the heart rate rises and the blood pressure rises. These changes are maintained for
the period of the occlusion. When the occlusion is removed,
the heart rate and blood pressure return to normal.

Note that the paper speed is quite slow, so it is not possible to

see the individual components of the ECG and pressure waves.

Mechanism of the carotid sinus reflex

The carotid sinus is a dilation of the internal carotid artery just above the bifurcation of the common
carotid artery. The baroreceptors are branched, coiled bare ends of sensory nerve fibres located in
the adventitia of the carotid sinus. They act as stretch receptors, detecting stretch of the wall due to
changes in arterial pressure. Their output is transmitted via afferents in the carotid sinus nerve to the
glossopharyngeal nerve (Cranial Nerve IX) and then to the nucleus tractus solitarius (NTS) in the
medulla.

When blood pressure rises, afferent impulses increase and the efferent response is to decrease the
sympathetic discharge to the heart and vasculature and to increase in vagal discharge to the heart.
This causes peripheral vasodilation, venodilation, a drop in blood pressure, a fall in heart rate and a
drop in cardiac output. This returns blood pressure to normal.

Occlusion of the carotid below the sinus simulates a fall in blood pressure. There is a reduction in stretch of the
carotid sinus, so afferent impulses are reduced. This results in a compensatory increase in sympathetic discharge to
the heart and blood vessels and a decrease in vagal discharge. Therefore, the blood pressure and heart rate rise.

Note: there are also baroreceptors in the arch of the aorta which project to the NTS via the vagus nerve. If these
baroreceptors had been blocked the response to carotid occlusion would have been greater.
Recording right atrial pressure
Right atrial pressure is measured via the catheter in the external jugular which was advanced so that its tip was in
the right atrium. There is a periodic fall in right atrial pressure. This occurs each time the animal inspires. When the
catheter is open to the atmosphere to show zero pressure, it indicates that right atrial pressure is below
atmospheric pressure. This is because the thin walled atrium is affected by intrapleural pressure which is usually
negative.

Right atrial pressure and tracheal occlusion

Here the tracheal cannula is occluded when the animal attempts to breath in. Right atrial pressure falls much more
when the attempt to draw air into the lungs is impeded.

Arterial pressure and right atrial pressure

Simultaneous recordings of arterial blood pressure and right atrial blood pressure are shown. When the trachea is
occluded during the animal’s inspiration, the fall in right atrial pressure is associated with a corresponding rise in
arterial blood pressure. This sequence is shown three times.

What causes this rise in arterial blood pressure? When the intrathoracic pressure becomes more negative (due to
the attempts to breath in with an occluded trachea), this causes an increase in venous return to the right side of
the heart. This causes is a corresponding rise in right ventricular output, which increases venous return to the left
side of the heart, and in turn increases left ventricular output and blood pressure.

Although this response is marked in the dog, in man the

increase in blood pressure may be less apparent. This is
because there is also a tendency for blood to pool in the lungs
when intrathoracic pressure is reduced. This means that the
increase in venous return to the left side of the heart is not as
great and so there is less increase in left ventricular output and
blood pressure.

Effect of positive pressure

A positive pressure can be produced by attaching a rubber tube to the tracheal cannula and blowing into the tube.
This situation is similar to mouth-to-mouth ventilation or positive pressure ventilation in a hospital situation.
During the period of blowing the right atrial pressure rises and there is a simultaneous fall in arterial blood
pressure. The positive pressure applied at the trachea impedes the venous return to the heart by increasing the
right atrial pressure. This means that right ventricular output is reduced, so venous return to the left side of the
heart is reduced, as are left ventricular output and arterial blood pressure.

Components of the right atrial pressure wave

When the recording is made at a fast paper speed, the individual waves of the atrial pressure trace become
apparent. These waves are called the a, c and v waves.

Here a simultaneous recording of ECG, arterial pressure and right atrial pressure at a fast paper speed is shown.
Individual waves of the atrial pressure can be identified by comparing them with the electrocardiogram. This is
shown more clearly on the next slide.

●
● The P wave of the ECG represents atrial depolarisation and is associated
with the a wave of atrial contraction in the atrial pressure recording.

● The QRS wave represents ventricular depolarisation and is followed by the

small c wave in the atrial pressure recording which represents bulging of
the tricuspid valves when the ventricle contracts.
 ● The v wave represents venous return to the atrium while the tricuspid valve is closed.

Stimulation of the vagus nerve

Electrical stimuli are obtained from a Grass stimulator and applied through a pair of electrodes which are placed on
the vagus nerve. When the nerve is stimulated, a train of action potentials passs along the nerve to the heart.

The marker pen indicates when the stimulation occurs. When the nerve is stimulated at 5 impulses per second, the
ECG shows a small amount of slowing. When the stimuli are delivered more frequently, 10 per second, the slowing
of the heart is more pronounced. At 15 per second the slowing is profound. Thus, stimulation of the vagus slows
the heart and the greater the rate of stimulation, the more pronounced the slowing. As the heart slows, the blood
pressure also falls.

Vagal escape
When stimulated at 20 Hz, the heart stops completely and then resumes beating at a slower rate. Even though the
vagal stimulation is continuing, the heart escapes from its influence.

Consider what causes this vagal escape? Then look at the next slide.

Mechanism of vagal escape (a.k.a. ventricular escape)

The various parts of the conduction system of the heart (see fig) are capable of spontaneous discharge. Normally
the Sinoatrial (SA) node is the pacemaker because it discharges most rapidly. When it depolarises, the
depolarisation spreads to the other regions of the heart before they discharge spontaneously.

The vagus nerves are distributed to the SA and AV nodes, and to a lesser extent to the atrial muscle. Although there
is sympathetic innervation to all parts of the heart, the ventricles receive little vagal innervation. Stimulation of the
vagus nerve both decreases the rhythm of the SA node and decreases the excitability of the A-V junctional fibres
between the atrium and the AV node. When the vagus nerve is intensely stimulated, spontaneous discharge of the
SA node is abolished and/or transmission of the cardiac impulse from the atria to the ventricles is abolished. This
allows a different part of the conduction pathway with a slower rate to take over as the pacemaker.

Typical spontaneous discharge rates are SA node – 70-80 times/min; AV node 40-60 times/min; Purkinje fibres 15-
40 times/min.

Termination of the experiment - Overdose of barbiturate anaesthetic

Simultaneous recordings of ECG, arterial pressure and right atrial pressure are shown. The right atrial pressure
trace can be used to indicate respiration. When the overdose of barbiturate anaesthetic is given, the blood
pressure commences falling. At first the animal breathes a little more, but then the breathing becomes less and the
blood pressure fall continues. Barbiturate anaesthetics are very powerful depressants of respiration. The dose of
barbiturate was lethal. The blood pressure has fallen to such a low level that the vital organs can no longer be kept
alive.

Post lab questions

Introduction to CVS
1. Tactile Sensation
a) Detection of punctate tactile stimuli.
Is there any difference in the percentage of failures in the three areas? If so what is the likely
explanation?
● It is easier to feel pricks in the arm and fingertips compared to being pricked on the back due to the
differences in the distribution of sensory receptors. The skin in the arm and fingertips has a higher density
of sensory receptors, including mechanoreceptors and nociceptors, compared to the skin on the back. This
means that there are more nerve endings per unit area in these regions, which makes them more sensitive
 to touch, pressure, and pain. In addition, the skin on the back is thicker and has a larger area of
innervation, which further reduces the sensitivity to touch and pain.

b) Two-point discrimination.
How does this data relate to those from part 1a?
● The ability to discriminate between two points on the skin's surface is known as two-point discrimination.
It varies in different areas of the body, with greater sensitivity in areas with a high density of touch
receptors, such as the fingertips. This is due to the smaller receptive fields of touch receptors in these
areas, which allow for more precise localization of the stimulus. In contrast, areas with larger receptive
fields, such as the back, have less two-point discrimination ability, making it more difcult to distinguish
between two closely spaced stimuli. The ability to feel pricks in the arm and fingertips more easily than in
the back may reflect these differences in two-point discrimination ability.