Look at the following dot plot from flow cytometric analysis of lymphocytes that have been stained with fluorescently labelled antibodies to CD3 (CD3 PE) and CD19 (CD19 FITC).  The fluorophores are phycoerythrin (PE) and fluorescein isothiocyanate (FITC).

Briefly explain what the dot plot tells you about this population of lymphocytes.

Use the following image of immune cells to help you answer this question:

Question:

Look at the following dot plot from flow cytometric analysis of lymphocytes that have been stained with fluorescently labelled antibodies to CD3 (CD3 PE) and CD19 (CD19 FITC).  The fluorophores are phycoerythrin (PE) and fluorescein isothiocyanate (FITC).

What percentage of this population are T-cells? [Your answer is to be given as a number only]

Look at the immunofluorescence micrograph below.  It shows a group of human PBMCs (peripheral blood mononuclear cells), as the name suggests, these cells are isolated from peripheral blood and contain cells with large, round nuclei, such as lymphocytes, monocytes and macrophages).

The cells below have been stained with two different types of agents.  The first is called DAPI (4', 6-diamidino-2-phenylindole), this is a fluorescent dye that binds strongly to adenine/thymine-rich regions in DNA.  DAPI appears blue under fluorescent light.  The second is an antibody linked to a green fluorophore.  The antibody is against the human glycoprotein CD4, this antibody will bind to the surface of cells that express the CD4 glycoprotein on their surface.

(A)  Which region of the cells is visible due to staining with the DAPI stain?     (1 mark)

(B)  The primary antibody used to bind to the human CD4 glycoproteins on the cells is a monoclonal mouse antibody.  Its full description is a mouse anti-human CD4 IgG1 monoclonal antibody.

This primary antibody does not contain a fluorophore (nothing that will fluoresce under UV light).  Therefore a secondary antibody linked to a green fluorophore (called FITC - fluorescein isothiocyanate) has been used.

(i)  What is the secondary antibody raised against?     (1 mark)

(ii) Which species (e.g. mouse, human, goat, rabbit, donkey... etc) would the secondary antibody have been raised in?     (1 mark)

You have expressed human insulin in Escherichia coli and then lysed the cells.  You are going to purify the insulin from the bacterial proteins by passing the protein lysate over an immunoaffinity column with a Bio-Gel® support (beads).  What needs to be adsorbed to the support in order for your purification to be successful?

You wish to purify acetylcholinesterase from a sample using immunoaffinity chromatography with a column that contains Reacti-Gel® as a support (beads). You have appropriately set up your column to purify the acetylcholinesterase. You pass your sample over the column and are confident the acetylcholinesterase has been retained on the support. 

Which one of the following statements best describes the interaction between acetylcholinesterase and the support at this point in the assay?

I agree to participate in the BPS3022 Final Assessment Part 1 (Lab-based exam) for Monash University (Parkville Campus) at my allocated time.

I understand that the exam questions presented to me in this assessment are the property of the Faculty of Pharmacy and Pharmaceutical Sciences.

Student's statement: 

1. I will not communicate or divulge any information regarding my exam questions to any other parties (with the exception of Faculty staff at Monash University).
2. I will not discuss the cases with any other students of Monash University.
3. I acknowledge that breaking the above conditions is a breach of academic integrity and there will be consequences for such a breach.  

The major significance of Koch’s work was the demonstration that:

A.   Microorganisms are present in a diseased animal

B.   Microorganisms can be cultured

C.   Microorganisms cause disease

D.   Microorganisms are the result of disease

(i)   Which response (A-D) is the best choice? (1 mark)

(ii)  Briefly explain your reasoning for your choice (2 marks)

Hyperbaric oxygen therapy can be used to increase oxygen concentrations in the tissues of people with certain infections.  Which one of the following types of disease-causing microbes would be targeted with this type of therapy?

A.   Strict aerobes

B.   Facultative anaerobes

C.   Aerotolerant anaerobes

D.   Strict anaerobes

(i)   Which response (A-D) is the best choice? (1 mark)

(ii)  Briefly explain your reasoning for your choice (2 marks)

Which one of the following techniques is NOT a good choice for sterilising a product?

A.   Pasteurisation

B.   Dry heat (170°C for 2 hours)

C.   Autoclaving (121°C for 20 minutes)

D.   Ionising radiation

(i)  Which response (A-D) is the best choice? (1 mark)

(ii ) Briefly explain your reasoning for your choice (2 marks)

  When comparing the metabolism of glucose in bacterial and mammalian cells:

A.  Glycolysis is highly conserved; the citric acid cycle and oxidative phosphorylation differs in some bacteria compared with mammalian cells.

B.   Bacteria can only use glucose as an energy source whereas mammalian cells can use alternative energy sources.

C.   Bacteria only use glycolysis for the formation of ATP; they do not possess ATPase and do not use chemiosmosis.

D.   Bacteria do not respire, they only ferment.

(i)   Which response (A-D) is the best choice? (1 mark)

(ii)  Briefly explain your reasoning for your choice (2 marks)