#473 You are required to investigate the effects

 You are required to investigate the effects - Anatomy and Physiology

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As a recent graduate of RMIT University, you have just been employed (3-year contract) by a world-renowned infectious disease institute located in Victoria, Australia. You are required to investigate the effects of the novel small mole clean-viral (DAD030981)to determine the global metabolic changes which take place in the tissue of lungs of monkeys infected bySARS-CoV-2(Infected Group)vs the tissue of lungs of healthy monkeys (Control Group). You have been asked by your laboratory head to design and carry out a “metabolomics experiment.”Q1. What experimental design would do you propose to your lab head? He will first need to approve this and then discuss with you how you are going to prepare samples and analyze them before you begin. What kind of metabolomics approach will you propose to your lab head and why? What considerations with respect to your experimental design should you propose?(5 marks)Q2. Congratulations! Your lab head has approved your project and has now asked you to provide details as to how you are going to prepare samples for your metabolomics experiment. Briefly describe your proposed sample preparation workflow only and provide a justification for each step. What kind of internal standard do you propose to use and why?(5 marks)Q3. Your lab head has now asked you to analyze the polar metabolite extract on the laboratory’s GC-MS. Briefly describe how it may be possible to analyze polar metabolites on the GC-MS. What additional sample should you run with your experiment and why? (5 marks)Q4. Your lab head is quite pleased that you have acquired all data in such a short amount of time and is now eager for you to analyze the data.Briefly explain what and why deconvolution is important in appropriately identifying metabolites. How can your lab head determine which metabolites are up and/or down-regulated?How can your lab head determine which metabolites are statistically significant? (5 marks) BIOL1181 Metabolomics and HPLC Practical Assignment2| P a g eQ5. Your lab head has interpreted your univariate data and noticed that amino acid metabolism appears to be downregulated in the infected group relative to the control group. Your lab head has now asked you to perform a Principal Component Analysis(multivariate analysis). Briefly illustrate or describe how your Principal Component Analysis would look like.What causes your control and infected groups to be separated in three-dimensional space? (5 marks)CONGRATULATIONS! You have carried out to completion a metabolomics workflow and your lab supervisor is exceptionally happy with you!Q6. Your lab head has asked you to quantify 4endogenous steroids present in a patient’s serum sample using an existing HPLC method as described below.

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