Mycobacterium tuberculosis (Mtb) is the bacterium that causes tuberculosis(TB). Mtb can cause

BIOL22111/BIOL24413 Immunoopharmacology practical (BMS and DA only) (Term 1) Session 2022-2023. The effect of toll-like receptor (TLR) activation on cell viability and nitrite production in J774 macrophage cells: the effect of Dexamethasone.

In what ways do the plant cells in your sample differ from one another? Describe some features that vary among the cells that you're looking at. b).

B. Animal Cells: You will be looking at a prepared slide of epithelial cells and microorganisms from the cheek tissue inside the mouth. Under the microscope, these cells look a bit like fried eggs. The cells on this slide have been stained, and you will see that some parts of the cells stain more strongly than other parts. Examine this sample with your microscope, starting at the lowest magnification and working up to the highest. i. Create a detailed, neatly drawn sketch of 2 to 3 cells, labelling key features of these cells. Be sure to include a figure caption (see Appendix C). Use the ocular micrometer on your microscope to measure the length of two of the cheek cells that you chose to draw and record those two length values in your figure caption and on the class spreadsheet. ii. Now, describe those features that make it clear what type of cell you are looking at. That is, how do

C. Prokaryotic Cells: Use a toothpick to drop some yogurt onto a microscope slide. Cover the sample with a coverslip and observe right away before the yogurt dries out. Two of the main bacterial types used in making yogurt are Streptococcus thermophilus (chains of spheres) and Lactobacillus bulgaricus (rod- shaped). The bacteria are barely visible using the 40X objective, and most of what you will see on the slide are clumps of casein (milk protein). Starting with the lowest power objective, focus on the casein clumps, then switch to the next highest power objective, and refocus. When you get to 400X total magnification, look for an open space between two thick areas of casein, and look for the tiny bacteria streaming through this space. You might notice the bacteria vibrating a bit as well. It may help to decrease your microscope illumination slightly to see these cells.

A 77-year old male is presented to hospital with chest pain and neck pain that reflects to his left arm. The onset of his pain was 24 hours prior to admission. The patient has developed electro cardiograph (ECG) changes that are interpreted as

A Biomedical Scientist has been asked to prepare DNA samples from cells isolated from a tumour biopsy. After following the standard protocol for the DNA extraction in the laboratory, the scientists eluted the final DNA sample in 20 μl of elution buffer and measured absorbance at 260 nm and 280 nm. Values obtained are as follows: A260=1.446 A280 = 0.784 Based on the data above, A) Briefly describe the protocol for DNA isolation. B) Comment on the purity of the DNA sample obtained. Provide appropriate calculations to back up your comment. C) Assuming a pure solution of double-stranded DNA has a concentration of 50 µg/ ml and A260 of 1.0, calculate the concentration of DNA in the solution obtained by the scientist and give the total DNA yield. D) How can the scientist assess DNA breakage in the sample obtained?

CULMINATING PERFORMANCE TASK Culminating Activity Rationale: In this project, you will analyze a device/technology related to a topic learned in this course, introduce a scientist, and explore a career. Overall Expectations: A1.Describe the structure and function of the macro molecules necessary for the normal metabolic functions of all living things,and the role of enzymes in maintaining normal metabolic functions. B1.Investigate and explain the relationship between metabolism and the structure of bio molecules C1. Determine the similarities and differences between cellular respiration and Photosynthesis D1.Investigate the effects of genetic manipulation on an organism by observing of survival and growth in different environment. E1. Describe and explain mechanism involved in the maintenance of homeostasis F1. analyze the components of population growth, and explain factors that affect

Experiment 1: Study the effect of temperature on membrane permeability using colorimetry 1. Propose an aim and hypothesis based on the theoretical context; when temperature rises will membrane permeability increase or decrease? (AC 1.1) 2. Devise a methodology to investigate your hypothesis, identifying any risks and outlining steps taken to minimise risks (AC 1.2 and AC 1.3) 3. Carry out the proposed practical investigation (AC 1.3 and AC 1.4) 4. Record and present data in a format suitable for analysis and to verify or discredit the predictions made in the hypothesis (AC 2.1 and AC 2.2) 5. Evaluate the validity of your hypothesis (AC 2.3) 6. Review the practical investigation for any anomalies, errors or limitations linked to equipment or methodology and suggest proposed improvements (AC 3.1, AC 3.2, AC 3.3 and AC 3.4) 7. Ensure that the final lab report follows a format agreed with the tutor (AC 4.1)

Experiment 2: Experiment to investigate the effect of different concentrations of catalase enzyme on the breakdown of hydrogen peroxide 1. Propose an aim and hypothesis based on the theoretical context; when enzyme concentration increases, will rate of hydrogen peroxide decomposition increase? (AC 1.1) 2. Devise a methodology to investigate your hypothesis, identifying any risks and outlining steps taken to minimise risks (AC 1.2 and AC 1.3) 3. Carry out the proposed practical investigation (AC 1.3 and AC 1.4) 4. Record and present data in a format suitable for analysis and to verify or discredit the predictions made in the hypothesis (AC 2.1 and AC 2.2) 5. Evaluate the validity of your hypothesis (AC 2.3) 6. Review the practical investigation for any anomalies, errors or limitations linked to equipment or methodology and suggest proposed improvements (AC 3.1, AC 3.2, AC 3.3 and AC 3.4) 7. Ensure that the final lab report follows a format agreed with the tutor (AC 4.1)

Discuss the general functions and mechanisms of molecular chaperones. Give THREE examples of different chaperone systems and explain how they are thought to carry out their function.

i. To examine the main blood vessels of the heart and the coronary artery. ii. To carry out a detailed examination of the differences between the heart chambers and the walls between them. iii. To produce scientific annotated drawings of the dissected human heart. iv. Name the structure and the function of the animal’s heart organs. v. Understand the anatomy and physiology of a sheep’s heart.

(a)Describe the methods used to study protein structure (give a list of methods mentioning the achievable resolution and sample requirements for each method). (b)You are given 100mg of a stable, soluble and pure 500 kDa protein. Which low- and high-resolution techniques would you use to obtain structural information of the protein? Which methods would you not use and why?Discuss the advantages and disadvantages of each method. (c)Why do some proteins crystallize easily while others do not? Focus on the properties of different proteins. (d)Discuss the differences between different amino acids (discuss amino acidgroups and list amino acids belonging to each group). Draw the structures ofarginine and alanine and compare these 2 amino acids.

(a)List four properties of proteins that are exploited in their purification. For each give an example of a protein purification procedure that takes advantage of that property.(2 marke) (b)Name two procedures that can be used to break open bacterial cells? In your answer, briefly state the principles behind how each method achieves this. 흐What conditions/reagents can be used to preserve the biological integrity of proteins once cells have been broken open? You must provide at least three examples and explain how the condition/reagent works to preserve the biological integrity of the proteins. (d)Use the table below to calculate the specific activity, fold purification and percent recovery for the last three stages of a purification procedure. Please show at least one full calculation for each column and where appropriate include the correct units. (e)Explain what is meant by a crude extract (1 mark) and why ammoniumsulphate is often used as the first step in the purification procedure afterproducing a crude extract (2 marks). (f)Explain the effectiveness of using a CM-cellulose ion exchange chromatography for the purification of a protein with a pl of 6.7, when using a buffer with pH 7 42 6A small sample of the purified protein was run on an isoelectric focussing gel.Briefly, explain the technique and how an isoelectric focussing gel is set up.The answer should also include an explanation to the principle of isoelectric focussing.(2 marks)

(a) What are the names of the enzymes that carry out the following reactions, and what are their EC numbers based on systematic classification? (4 marks) (i) Tyrosine + a-ketoglutarate D p-Hydroxyphenylpyruvate + Glutamate Glucose + ATP → Glucose-6-phosphate + ADP (iii) Succinate + CoA +GTP D Succinyl-CoA + GDP + Pi (iv) Glutamine + H₂O → Glutamate + NH4+ The enzyme that carries out the reaction Tyrosine + a-ketoglutarate D p-Hydroxyphenylpyruvate + Glutamate is not solely composed of protein, there is an accessory organic molecule at the active site. What is this name of this molecule, and what is its function in the mechanism of the enzyme?(3 marks) (c) Briefly describe three mechanisms that contribute to the decrease of activation(3 marks)energy at an enzymes active site. (d) In developing their mathematical model for invertase, Michaelis and Mentenmade a number of assumptions to develop the model upon which their equation to describe enzyme kinetics is based. Explain what is meant by the'steady-state' assumption.(2 marks) (e) An enzyme that catalyses the conversion of molecule A into molecule B with aKm value of 2.4mM for molecule A and a keat value of 54 s-¹ (i)At what concentration of molecule A would the enzyme operate at 25% of the maximum rate?(2 marks) (ii)Determine the fraction of Vmax that is achieved when the [A] is equal to each-of the following multiples of Km: 0.33.Km, 5.Km, 40.Km and 100.Km (give values to 2d.p.).(3 marks) (iii)If the rate of reaction observed at 1.2mM A is 4.1 x 10-7 moles/L/min, what is the enzyme concentration in the reaction? Express your answer in nM.(3 marks)

4. Place the following steps of glycosylation in order: a. The carbohydrate is further modified by addition, subtraction or post-transnational modification (such as phosphorylation) b. Dolichol-phosphate is inserted into the membrane c. The core glycosylation is transferred to an asparagine residue on the protein d. A core carbohydrate made up of 2 N-acetyl glucosamine, 4 mannose and 3 glucose molecules is built Where in the cell do each of these steps (a-d) occur?

DNA ischarged due theelectrode.backbone and hence will move towards

6. In gel electrophoresis DNA molecules are separated based on

3. Directions: Identify the restriction sites for each of the examples given. Show the cuts, mention if they are sticky ends or blunt, 5' overhangs or 3 'overhangs and the number of DNA fragments produced and the number of base pairs in each (count the top row).Remember: Analyze in the 5'→ 3' direction 3

3. You create a fusion protein that adds a Green Fluorescent protein domain to thesoluble cargo protein in the diagram above. Where does the cargo proteinaccumulate if :a. Rab is mutated and does not function?b. The transmembrane cargo protein is missing?c. The coat protein is missing?d. The v-snare is mutated and does not function?e.The t-snare is mutated and does not function?f. The pH at the final destination is the same as the pH of the transportvesicle?