Part -1:

Question 1:
Discuss how cell culture can help to understand drug-induced adverse effects observed in a patient. Up to 200 words

Question 2:
How can cellular toxicity be monitored in vitro. Use at least 5 differentand independent parameters (do not list assays!) that could help you to get a detailed understanding of the type of toxicity that is present. Up to 500 words

Question 3:
Discuss the suitability of the WST-1 assay to determine cellular toxicity and list advantages and disadvantages. Also discuss potential artefacts that could affect the results obtained from this assay. Up to 400 words

Question 4:
Discuss the suitability to measure cellular ATP content to determine cellular toxicity and list requirements, advantages and disadvantages. Also discuss potential artefacts that could affect the results obtained from this assay. Up to 400 words

Question 5:
Describe the toxicity (what does it mean) in the following scenario and describe the resulting danger. After exposure of live cells to a novel drug the following is measured in cell culture: no changes in trypan blue assay, increased gammaH2AX signal, no changes in WST-1 assay, no changes in ATP assay, no changes in apoptosis, no changes to protein content/well.
Up to 300 words

Question 6:
Give one example how a viability assay can be used to measure something entirely unrelated. Support your argument by a detailed reaction scheme. Up to 300 words

Question 7:
Describe why many viability assays are standardized on protein content. What is the advantage? Up to 400 words

Part -2:

1. Briefly state the purpose of the experiment - approximately 50 words.

2. Describe the importance of appropriate pipetting and dilution techniques. Please support your description with at least one suitable example - approximately 150 to 200 words.

3. Discuss at least four common mistakes that can occur while aspirating as well as dispensing liquid using micropipettes - approximately 150 to 200 words.

4. Discuss the most appropriate way of aspirating 500 µl of drug solution from an eppendorf tube containing 1000 µl of drug solution. Also, outline the most appropriate way of preparing a 1000 µl of solution (in an empty eppendorf tube) containing 500 µl of aspirated drug and 500 µl of water. Also mention, why do you think that your discussed way is the most appropriate way? - approximately 200 to 250 words.

5. Describe the working principles of UV spectrophotometer and outline at least three pharmaceutical applications of it - approximately 100 to 150 words.

6. Describe the importance of a calibration curve - approximately 50 to 100 words.

7. Outline the methodology you used: a) to prepare caffeine sample using provided stock solution, b) to measure the spectrophotometer UV absorbance of your sample and c) to calculate the concentration of your sample - approximately 200 to 300 words.