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PROPERTY OF: JOIE

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FERMENTATION: OPTIMIZE BIO-ETHANOL PRODUCTION

1. As you can see, there are five different phases based on their growth rate. What happens
during the death phase?
More yeast cells die than are created, reducing the total number of yeast cells.
2. Can you determine when the diauxic shift happens?
At 23 hours
3. Is it true that ethanol yield is higher is aerobic fermentation?
No, it’s not
4. Is it true that glucose is consumed faster in anaerobic fermentation?
No, it’s not
5. Is it true that yeast cells grow faster in anaerobic conditions?
Yes, it’s true

WESTERN BLOT TRANSFER: PREPARE FOR PROTEIN DETECTION

1. The electrophoresis that I have already performed separates proteins from size. Where are
the largest proteins found?
Near the top
2. Why is it important to remove air bubbles?
Air bubbles create areas where there is no current
3. In which direction will the proteins migrate?
From the cathode (-) to the anode (+)
4. Have a look at the membrane you are holding. Why do the edges look burnt?
The tank was not cooled
5. Have a look at the membrane you are holding. Why do the bands look blurry?
The layers were not aligned properly
6. That’s your membrane, why do you think it looks like this?
It’s exactly how it’s supposed to look, everything is perfect
7. Have a look at the membrane you are holding. Why does it look like this?
The layers were not properly soaked in transfer buffer
8. Have a look at the membrane you are holding. Why do you think it looks like this?
It’s exactly how it’s supposed to look. Everything is perfect.
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PROPERTY OF: JOIE
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9. Sometimes you can do more than one thing wrong. Why does this membrane look like
this?
The layers were not aligned properly, there was no ice and air bubbles were not
removed
10. Have a look at the membrane you are holding. Why does it look like this?
The gel was facing the anode and the membrane was facing the cathode

EXERCISE PHYSIOLOGY
1. How long is a Wingate sprint?
20-30 s
2. At which level is a Wingate sprint performed?
At supramaximal level
3. Which energy systems contribute to ATP synthesis during a Wingate sprint?
All of them
4. How is the breaking torque calculated?
Torque factor x body weight
5. What does the respiratory exchange ratio (RER) measure exactly?
Oxygen uptake and carbon dioxide release
6. What is the unit in which values in a Wingate power spectrum are presented?
Watt
7. Which molecules causes a hyperosmotic state in muscle cells during the sprint?
Glycolytic intermediates
8. How is hyperosmosis in muscle cells counteracted during a wingate sprint?
By fluid-influx into the cell
9. Which molecule buffers the lower blood pH caused by hydrogen ion release from muscle
cells?
Bicarbonate
10. Why can we observe a continuous increase of RER even after the sprint?
The body tries to get rid of an excess of carbon dioxide
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PROPERTY OF: JOIE
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11. What does the power output measured during the Wingate sprint indicate?
Anaerobic capacity
12. With which response can the acute physiological effects of a Wingate sprint be
associated?
Fight-or-flight
13. Looking at the data, what is the most striking difference between Mike’s first and last
session?
A higher peak power

CARBOHYDRATES: THE SUGARS THAT FEED US

1. In which journal was this article published?
International Journal of Sport Nutrition
2. What is the difference between the high-CHO and the control group?
Carbohydrate intake before trial
3. What do carbohydrates consist of?
Carbon, oxygen, and hydrogen
4. What do the red spheres represent?
Oxygen atoms
5. What is the chemical formula of fructose?
C6H12O6
6. What is the difference between glucose and fructose?
Arrangement of functional groups
7. Which of the following drinks contains a lot of sucrose?
Freshly squeezed orange juice
8. What are the two building blocks of sucrose?
Fructose and glucose
9. By what factor does the surface area increase from the single cube to the 27 cubes on the
right?
3
10. Which of the following sentences is not true?
Enzymes are exclusively involved in digestion
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PROPERTY OF: JOIE
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11. What did your taste buds sense after chewing for a while?
Sweetness
12. Which organ follows your esophagus?
Stomach
13. Where is most of the sugar absorbed?
Small intestine
14. What do the percentages on the right side mean?
Percentage of recommended daily intake, per serving
15. How many grams of starch are contained in one serving?
146 g
16. How much rice do we need to feed our virtual test subject if we want to measure the effect
of 50 g carbohydrates? Make sure you don’t include dietary fiber in the calculation since
this cannot be degraded in the digestive track and is therefore not absorbed.
63 g
17. How long does it take until the blood glucose level peaks?
25 min
18. The glycemic index of carrots is quite high. Why is the glycemic index sometimes
misleading?
It’s relative to the amount of carbohydrate
19. Do you remember the chemical formula of glucose? Which is it?
C6H12O6
20. What is the fasting level of blood glucose?
4mM
21. What could be the reason that person A's blood sugar is acting so differently than that of
person B?
Person A is not capable of producing insulin

KJELDAHL METHOD: ESTIMATE THE PROTEIN CONTENT IN FOOD

1. What form is the nitrogen in the food sample converted to in the acid digestion step?
Ammonium ions
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2. Why is the mixture made basic after the digestion?

To convert the ammonium ions to ammonia, which is volatile and can be distilled
from the mixture
3. Now why is it that you weighed out the milk powder sample very precisely on an analytical
balance?
It’s a quantitative analysis, so we need this for a precise determination of the
protein content
4. Do you recall what happened to the Kjeldahl tube when the digestion unit was done?
I waited 30 min to let it cool
5. So that was the acid digestion. What do you want to do next?
Next step is distillation
6. Before the distillation starts, the unit automatically makes the solution alkaline with
NaOH to convert what?
Ammonium ions into ammonia, which is volatile
7. The moles of acid used in the titration, n(HCl) is found by
CHCl * Vburette
8. The moles of nitrogen present in the original sample, nN, is equivalent to
nNH3 = nHCl
9. Finally, we need to find the percentage of nitrogen in the original sample. Which of these
formulas should we use for this?
((nN*MN) / msample) * 100%
10. What conversion factor should you use for the milk powder samples?
6.38
11. Did you get it? So retention time is an important feature when assessing…..
Compound determination
12. Fragmentation in mass spectrometry is used for…..
Increasing both sensitivity and selectivity
13. Peak area is an important feature when determining…..
Concentration of the compound of interest
14. Are the blanks in fact without any relevant peaks?
There are no peaks in the two blank samples
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15. Does the retention time of the peaks in the recovery controls align with one of the
standards, and do the recovery controls seem to be within the range covered by the
standards?
The retention times align around 3 min, and the recovery controls are within the
range of the standards
16. I have prepared a calibration curve with the standards using linear regression. It has the
following form: Peak Area = a * Concentration + b. How would you use this to calculate the
recovery?
Use the peak area of each recovery control to find its measured concentration, take
the mean value of three, and divide this with the spiked amount
17. What is recovery in a chemical analysis as LC-MS/MS?
It is the amount of analyte that is measured compared to how much was present at
the start
18. What is a control in analytical chemistry?
A sample where the original amount of analyte is known.
19. From the chromatograms, which sample could have a melamine contamination?
Sample 2
20. Would you say the peak in sample 2 is significant when compared to the lowest standard?
Yes, as the peak area of sample 2 is higher than that of the standard
21. Why could the Kjeldahl method not detect the melamine adulteration of sample 2?
The nitrogen in melamine is broken down into ammonium ions during the acid
digestion
22. How do we know that food fraud has been conducted for sample 2?
We detected melamine above its established limit in the sample with a selective
method

PROTEIN SYNTHESIS
1. DNA is the molecule of heredity that contains instructions to build proteins. In 1958,
Francis Crick described the central dogma of molecular biology explaining the process of
gene expression. What do A and B represent in the image?
‘A’ is transcription and ‘B’ is translation
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2. Which statement is true?

DNA has no hydroxyl group on the second carbon
3. The mRNA strand is synthesized in the 5’ to 3’ direction. The circles in the image dictate
the chemical group in which the next base is attached. Which image has the circle in the
correct position?
C
4. DNA and mRNA synthesis proceeds in the 5’ to 3’ direction. What does 5’ and 3’ mean?
The position of a carbon atom in the sugar backbone
5. Protein synthesis is performed by an RNA-protein complex called the ribosome. As you
can see in the image, a ribosome is composed of 3 sites called E, P, and A. To which site
does tRNA add its amino acid to the polypeptide chain?
P
6. Where do eukaryotic cells synthesize proteins and which number represents this organelle
in the image?
Ribosomes, 2
7. What is the name of each sequence of three nitrogen-containing bases of RNA that codes
for one amino acid?
Codon
8. The biosafety cabinet is important for physical containment. In this lab we will use a
biosafety cabinet class II. Who or what does a level II biosafety cabinet protect?
The personnel, the experiment, and the environment
9. The tRNA shown in the diagram will be able to form a base-pair interaction with which
mRNA?
5’-AGU
10. Three types of RNA cooperate to perform translation.
A: Read codons and deliver the amino acids
B: Carry the genetic information from DNA
C: Catalyze the assembly of polypeptide chains
Which type of RNA matches the A, B, and C descriptions respectively?
tRNA, mRNA, rRNA
11. Which strand is the template strand?
Bottom
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12. Which is the correct mRNA sequence resulting from the EPO gene transcription process?
5’- AUU ACA AUG GGG GUG CAC GAA UGU CCU -3’
13. What is the correct amino acid translated from the resulting mRNA?
MGVHECP
14. The erythropoietin protein is 193 amino acids long. With this information, how many
nucleotides are there in the mRNA including the initiator and terminator codons?
582
15. Downstream means near the 3’ end, so notice that the first codon is not methionine. What
is the correct amino acid sequence?
TGDR
16. Amino acids are linked by a specific type of covalent bond. The reaction of this bond
produces one water molecule. This bond links multiple amino acids together, what is it
called?
Peptide bonds
17. A sequence of amino acids, or a polypeptide, grows from the N-terminus to the
C-terminus. This polypeptide chain represents the primary structure of a protein. The
linear sequence of amino acids within a protein is considered the ….. structure of the
protein.
Primary
18. Which level of protein structure is formed from weak bonds between oxygen and hydrogen
atoms within the polypeptide backbone?
Secondary structure
19. The main two secondary structures of proteins are alpha helix and beta sheet which are
represented respectively by which two letters in the image?
B and A
20. Based on the mass spectrometer result, what is the mass difference between rhEPO
expressed in CHO and E.coli?
12 kDa
21. Glycosylation sites are marked with a diagram of glycan. Based on the mass spectrometry
results, rhEPO is glycosylated in which expression system?
CHO
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22. Based on the mass spectrometry results, how many glycosylation sites exist in
rhEPO-CHO?
4
23. Bacterial cells often cannot produce recombinant proteins that are identical to their wild
type. Why is this?
Bacteria lack the machinery for making post-translational modifications
24. Recombinant proteins produced by E.coli cells are not glycosylated. This is because E.coli
cells do not possess which organelle?
Golgi apparatus
25. Based on the mass spectrometry result, the masses of rhEPO expressed in CHO and E.coli
are 30 kDa and 18 kDa, respectively. We also know that the EPO proteins expressed in
mammalian cells are glycosylated. Therefore, we can say that what weight percentage of
rhEPO expressed in CHO are polysaccharides.
40%
26. If hypothesis B was picked: Do your results confirm the initial hypothesis that bacterial
systems can produce biologically active and stable erythropoietin?
No, erythropoietin is best expressed in mammalian cells
If hypothesis A was picked: Are mammalian expression systems better than bacterial
systems for the production of biologically active and stable erythropoietin?
Yes, erythropoietin is best expressed in mammalian cells
27. Mass spectrometry identifies molecules based on what?
Mass and charge
28. MALDI is an abbreviation for what?
Matrix-assisted laser desorption/ionization
29. The macromolecule ionization method, such as MALDI MS, allows us to obtain a peptide
mass fingerprint, or PMF, that can be used to do what?
Predict the identity of an unknown protein
30. If X, Y, and Z refer to the parts of a mass spectrometer that perform different processes
such that:
X detects the number of different ions
Y is where ion production from the sample takes place
Z separates ions with different mass-to-charge ratios
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In which order do X, Y, and Z occur?

Y, Z, then X
31. Protein samples are mixed with a matrix prior to analysis. What is the function of a matrix
in MALDI MS analysis?
Ensures ionization of protein molecules
32. In mass spectrometry protein molecules are separated based on their mass to charge ratio.
The data recorded in a histogram for analysis. Which of these histograms are NOT
coherent with the mass spectrometry principle?
II, III, IV
33. Compare the sample results from each athlete with the control results. There are two
controls:
1. Positive control: contains rhEPO only
2. Negative control: only includes urine. No rhEPO in this control.
Which athlete is using rhEPO?
C (pabago bago naman ata to 🧚goodluck nalang)
34. EPO can increase production of energy (ATP) by increasing production of what?
Red blood cells