INTRO
Pernicious Anaemia (PA) is an autoimmune disease that’s also
known as macrocytic anaemia due to vitamin B deficiency (1). It is
considered an autoimmune disorder as a result of intrinsic factors
and parietal cells both being targeted by gastric autoantibodies.
This also occurs due to the abundance presence of gastric
autoantibodies (2).
-Vitamin B12 deficiency
-Diagnosis
-End stage of chronic atrophic gastritis
-vitamin b12 obtained primarily from diet
-b12 is a fermentation product produced by microorganisms
-b12 bind to secretery product called intrinsic factor
-Intrinisc cells produced by parietal cells found on the lining
of the stomach
-vitamin b12 can only absorbed when binded to intrinsic factor
only occurs with the special receptor that is located in the
terminal ileum (last part of small intenstine)
-Type A gastritis is associated with pernicious anaemia
-Type A gastritis, that leads to pernicious anaemia, involves
the fundus and body areas of the stomach
-Type A chronic atrophic gastritis take approximately 20-30
years to progress to gastric atrophy and pernicious anaemia
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Method
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Result 400 words roughly
Figure legends required too
You need to draw only 2 drawings of the H&E staining, one at
low magnification 100x and one at high magnification 400x (there
are 2 to choose from). The H&E stomach tissue images that you
need to draw from are in prac 2 folder (label the features as
stated in prac manual p. 17)
Immunoperoxidase results need to be place in your report as
images (no drawings). You can select one of each images for -ve,
+ve, P1 and P2 (there are 4 different magnifications for each
sample in PowerPoint presentation - prac 3 folder)
Overall in your report's Result-figures you should have:
· 2 H&E drawings
· 1 WB
· 4 immunoperoxidase images
· Summary table (table is not compulsory to have)
Marker lane has to be labelled with size of protein bands
·
Those 3 bands are 100kda and goes down 75kda and 50kda
(Bio-Rad #161-0318) Marker used
That’s an example result for western blot are under
These 2 require the H&E drawings I have attached the manual
for the prac also a pdf showing what sort of drawing is wanted and
how drawings need to be labelled and have a figure legend
Discussion 800words roughly
References
Appendix 250-300words
a) What is the expected size of the protein being detected
by the sera from pernicious anaemia patients?
b) Which subunit of the proton pump does this correspond
to?
c) Describe another method that could be used to detect
and quantitate the levels of anti- proton pump antibodies in
patient’s sera?
d) The secondary antibody that was used in the Western and
immuno-histochemistry was a sheep anti-human Ig-HRP conjugate. What
is the antigen that this antibody is specific for? What animal was
this antibody raised in? What is the enzyme that is conjugated to
the antibody?
