Expt 26

2. How does one develop Rh antibodies?

When Rh-positive red blood cells from the fetus enter the blood of an Rh-negative woman through
a tear in the placenta, the mother is sensitized to the Rh antigen and produces anti-Rh antibodies
before or during delivery.

3. What is HDN? Explain how HDN is acquired.

During a subsequent pregnancy of an Rh-negative mother with an Rh-positive fetus, the Rh-
positive red blood cells once again cross the maternal circulation where it can stimulate the mother
to produce antibodies against the Rh antigen in a rapid antibody production due to initial Rh
sensitization. The anti-Rh antibodies from the mother cross the placenta, causing agglutination
and hemolysis of fetal red blood cells, and hemolytic disease of the newborn (HDN) develops.

Expt 27
1. What causes the absence of intravascular clotting?
Clots are dissolved by a process called fibrinolysis where an inactive plasma protein called
plasminogen is converted to its active form, plasmin. Thrombin, other clotting factors activated
during clot formation, and tissue plasminogen activator (t-PA) released from surrounding tissues
can stimulate the conversion of plasminogen to plasmin, which will slowly break down the fibrin
in a blood clot.

2. What is the importance of determining clotting time?

The time taken for blood to clot mainly reflects the time required for the generation of thrombin. If
the plasma concentration of prothrombin or of some of the other factors is low (or if the factor is
absent or functionally inactive), clotting time will be prolonged.Clotting time is done in 2 main
ways: prothrombin time (PT) or partial thromboplastin time (pTT). This is important to determine
if the patient doesn’t have any clotting disorders such as hemophilia or liver disease, to monitor
anticoagulant therapy such as warfarin or heparin, or to check whether the patient may need
transfusions of blood products after having a large hemorrhage.

3. Discuss the mechanism of hemostasis.

After inactive clotting factors are activated by exposure to connective tissue or by chemicals from
tissues, these clotting factors form prothrombinase which converts prothrombin to thrombin, which
further convers fibrinogen to fibrin and forming the clot. The formation of this clot is controlled to
prevent spread from the point of initiation throughout the blood vessels. Anticoagulants prevent
clotting factors from forming clots under normal conditions. For example, antithrombin and heparin
inactivate thrombin. Without thrombin, fibrinogen is not convrted to fibrin, and no clot forms. At an
injury site, however, the activation of clotting factors is very rapid. Enough clotting factors are
activated that the anticoagulants can no longer prevent a clot from forming. Away from the injury
site, there are enough anticoagulants to prevent clot formation from spreading.

Expt 28
1. Can there be prolonged bleeding time with normal clotting time or vice versa? Explain your
answer.
There can b a prlonged bleeding time with normal clotting time or a shortened bleeding time with
abnormal clotting time. This is because bleeding time depends upon the depth of the wound and
the degree of hyperemia in the body part influenced by tissue fluids, the elasticity of the
surrounding tissues, and the chemical effects of the destroyed platelets, and not exactly pertaining
ot the ablity of the blood to clot (which is measured by the clotting time).
2. Enumerate the advantages and disadvantages of using the earlobe as puncture site for
determining bleeding time.
Advantages of skin puncture using the earlobe:
1. It is less painful due to lesser nerve endings.
2. There is more free flow of blood due to thinner skin
3. There is less tissue juice contamination of blood due to lesser tissue and muscles in the
earlobe
4. It is ideal when searching for abnormal cells
Disadvantages of skin puncture using the earlobe:
1. Poor reproducibility and unreliable test results due to variability of earlobe thickness and
variability of puncture depth
2. Outdated and rarely used site for puncture as new methods use the forearm for an
improved sensitivity and reproducibility
Use of the earlobe to determine bleeding time was developed by Dr. William W. Duke in 1910. It
was replaced by the Ivy Bleeding Time with the following advantages:
 "Surgical" incision more closely approximated patient's
 hemostatic response to surgery
 Large surface area of template (longer incision) minimized
 skin displacement
 Depth of incision was controlled

Clotting Time
In order for blood to clot, the enzyme thrombin must be generated from the plasma precursor
prothrombin. Thrombin then converts soluble fibrinogen into insoluble fibrin. Generation of
thrombin involves the sequential activation of a number of other plasma clotting factor, this
process is also being assisted by Ca++ and by factors released by platelets and damaged tissues
. The time taken for blood to clot mainly reflects the time required for the generation of thrombin
in this manner. If the plasma concentration of prothrombin or of some of the other factors is low
(or if the factor is absent, or functionally inactive), clotting time will be prolonged. The expected
range for clotting time is 4-10 mins.