- A.0.15
- B.1.5
- C.15
- D.150
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Thinka Jun 2024 (V2) Cambridge International A Level-Style Mock — Biology (9700)
Paper 12 - Multiple Choice
- A.Both are unbranched polysaccharides containing only \(\beta\)-glucose monomers.
- B.Both contain 1,4-glycosidic bonds formed by condensation reactions.
- C.Both form helical structures stabilized by hydrogen bonds between hydroxyl groups.
- D.Both can be hydrolyzed by the same enzyme to produce maltose.
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Which statement is correct about this inhibitor?
- A.It is a non-competitive inhibitor that binds to an allosteric site on the enzyme.
- B.It is a non-competitive inhibitor that binds to the active site of the enzyme.
- C.It is a competitive inhibitor that binds to an allosteric site on the enzyme.
- D.It is a competitive inhibitor that binds to the active site of the enzyme.
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- Solution P: \(-200\text{ kPa}\)
- Solution Q: \(-400\text{ kPa}\)
- Solution R: \(-600\text{ kPa}\)
- Solution S: \(-800\text{ kPa}\)
In which solution(s) would net movement of water be out of the plant cells, potentially leading to plasmolysis?
- A.P and Q only
- B.S only
- C.P, Q and R
- D.Q and S only
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- A.In the apoplast pathway, water moves through the cytoplasm via plasmodesmata.
- B.In the symplast pathway, water moves through the cellulose cell walls and intercellular spaces.
- C.The Casparian strip in the endodermis blocks the apoplast pathway, forcing water to enter the symplast pathway.
- D.Water movement through the symplast pathway is entirely passive and never crosses any cell membranes.
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- A.It catalyzes the dissociation of carbonic acid into hydrogen ions and hydrogencarbonate ions.
- B.It catalyzes the reaction between carbon dioxide and water to form carbonic acid inside red blood cells.
- C.It binds directly to carbon dioxide to transport it through the plasma as carbaminohaemoglobin.
- D.It actively transports hydrogencarbonate ions out of red blood cells in exchange for chloride ions.
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- A.Antibodies from breast milk: natural passive; Tetanus toxoid injection: artificial active
- B.Antibodies from breast milk: natural active; Tetanus toxoid injection: artificial passive
- C.Antibodies from breast milk: artificial passive; Tetanus toxoid injection: natural active
- D.Antibodies from breast milk: artificial active; Tetanus toxoid injection: natural passive
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- A.glycolysis and the Link reaction
- B.the Link reaction and the Krebs cycle
- C.the Krebs cycle and oxidative phosphorylation
- D.glycolysis, the Link reaction and the Krebs cycle
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Four fractions (1, 2, 3, and 4) were collected:
- Fraction 1 contained the largest, heaviest structures, including nuclei.
- Fraction 2 contained structures containing green pigments.
- Fraction 3 contained structures capable of producing large amounts of ATP under aerobic conditions.
- Fraction 4 contained very small structures consisting only of RNA and proteins.
Which statement about these fractions is correct?
- A.Fraction 2 organelles possess a double membrane, circular DNA, and 70S ribosomes.
- B.Fraction 3 contains organelles that are the primary site of transcription of ribosomal RNA in the cell.
- C.Fraction 4 contains organelles surrounded by a single phospholipid bilayer.
- D.The structures in Fraction 1 contain the only double-stranded DNA in the cell.
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- Fraction 1 contains nuclei (largest/heaviest).
- Fraction 2 contains chloroplasts (possess green chlorophyll pigments).
- Fraction 3 contains mitochondria (produce ATP via aerobic respiration).
- Fraction 4 contains ribosomes (small, non-membrane bound, consisting of rRNA and proteins).
Chloroplasts (Fraction 2) are double-membrane bound organelles containing circular DNA and 70S ribosomes, supporting the endosymbiotic theory. This makes statement A correct.
Statement B is incorrect because ribosomal RNA is primarily transcribed in the nucleolus of the nucleus (Fraction 1).
Statement C is incorrect because ribosomes are not membrane-bound.
Statement D is incorrect because mitochondria (Fraction 3) and chloroplasts (Fraction 2) also contain double-stranded circular DNA.
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During complete acid hydrolysis of this oligosaccharide, how many water molecules are used, and what are the products?
- A.4 water molecules used; producing 3 \(\alpha\)-glucose and 2 \(\beta\)-fructose molecules
- B.4 water molecules used; producing 5 glucose molecules
- C.5 water molecules used; producing 3 \(\alpha\)-glucose and 2 \(\beta\)-fructose molecules
- D.5 water molecules used; producing 5 glucose molecules
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Hydrolysis is the chemical breakdown of a compound due to reaction with water. To completely break down four glycosidic bonds, four molecules of water must be added.
The hydrolysis of these bonds yields the original monomer units: three \(\alpha\)-glucose molecules and two \(\beta\)-fructose molecules.
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1. Enzyme + substrate (control)
2. Enzyme + substrate + Inhibitor X
3. Enzyme + substrate + Inhibitor Y
The experimental results showed:
- For Inhibitor X, the maximum rate of reaction (\(V_{\max}\)) was unchanged, but a higher substrate concentration was required to reach \(\frac{1}{2} V_{\max}\).
- For Inhibitor Y, the maximum rate of reaction (\(V_{\max}\)) was reduced, but the substrate concentration required to reach \(\frac{1}{2} V_{\max}\) was unchanged.
Which statements about the inhibitors are correct?
1. Inhibitor X binds to the active site of the enzyme.
2. Inhibitor Y binds to an allosteric site on the enzyme.
3. The effect of Inhibitor X can be overcome by increasing substrate concentration.
4. Inhibitor Y decreases the affinity of the enzyme for its substrate.
- A.1, 2, 3 and 4
- B.1, 2 and 3 only
- C.1 and 3 only
- D.2 and 4 only
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- Inhibitor X does not alter \(V_{\max}\) but increases \(K_{\mathrm{m}}\) (the substrate concentration at \(\frac{1}{2} V_{\max}\)). This is characteristic of a competitive inhibitor. Competitive inhibitors bind to the active site (Statement 1) and can be outcompeted by high substrate concentrations (Statement 3).
- Inhibitor Y decreases \(V_{\max}\) but leaves \(K_{\mathrm{m}}\) unchanged. This is characteristic of a non-competitive inhibitor. Non-competitive inhibitors bind to an allosteric site away from the active site (Statement 2). Since \(K_{\mathrm{m}}\) is unchanged, the enzyme's affinity for the substrate is not affected, making Statement 4 incorrect.
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The table shows the final mass of the cylinders.
| Cylinder | Sucrose concentration / \(\text{mol dm}^{-3}\) | Final mass / g |
| :---: | :---: | :---: |
| 1 | 0.0 | 5.8 |
| 2 | 0.2 | 5.3 |
| 3 | 0.4 | 4.8 |
| 4 | 0.6 | 4.4 |
Which statement is correct?
- A.The solute potential of the potato cells before the experiment is equivalent to a sucrose solution between 0.0 and 0.2 \(\text{mol dm}^{-3}\).
- B.In cylinder 3, the water potential of the solution was higher than the water potential inside the potato cells.
- C.At a sucrose concentration of approximately 0.32 \(\text{mol dm}^{-3}\), there is no net movement of water into or out of the potato cells.
- D.The water potential of the potato cells in cylinder 4 became less negative during the experiment.
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Looking at the table:
- At 0.2 \(\text{mol dm}^{-3}\), the mass increases to 5.3 g (net water gain of +0.3 g).
- At 0.4 \(\text{mol dm}^{-3}\), the mass decreases to 4.8 g (net water loss of -0.2 g).
The point of zero mass change lies between 0.2 and 0.4 \(\text{mol dm}^{-3}\). By linear interpolation:
\(0.2 + \left(\frac{0.3}{0.3 - (-0.2)}\right) \times (0.4 - 0.2) = 0.2 + 0.12 = 0.32\) \(\text{mol dm}^{-3}\).
Therefore, statement C is correct.
- Statement A is incorrect because the concentration of equivalent solute potential is between 0.2 and 0.4 \(\text{mol dm}^{-3}\).
- Statement B is incorrect because the potato lost mass in cylinder 3, meaning water moved out down a water potential gradient, so the solution water potential was lower (more negative) than inside the cells.
- Statement D is incorrect because losing water concentrates the vacuolar sap, making the water potential more negative.
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- A.xylem vessels \(\rightarrow\) cytoplasm of mesophyll cells \(\rightarrow\) vacuole \(\rightarrow\) stomatal pore
- B.xylem vessels \(\rightarrow\) cell walls of mesophyll cells \(\rightarrow\) evaporation into air spaces \(\rightarrow\) diffusion of water vapour through stomata
- C.xylem vessels \(\rightarrow\) symplast of mesophyll cells \(\rightarrow\) intercellular air spaces as liquid water \(\rightarrow\) diffusion through stomata
- D.xylem vessels \(\rightarrow\) companion cells \(\rightarrow\) phloem sieve tubes \(\rightarrow\) evaporation to air spaces \(\rightarrow\) stomatal pore
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1. Carbonic anhydrase catalyses the hydration of carbon dioxide to form carbonic acid.
2. Hydrogencarbonate ions (\(\text{HCO}_3^-\)) diffuse out of the red blood cells into the plasma.
3. Chloride ions (\(\text{Cl}^-\)) diffuse out of the red blood cells into the plasma to maintain electrical neutrality.
4. Oxyhaemoglobin dissociates more readily because of the decrease in pH.
- A.1, 2, 3 and 4
- B.1, 2 and 4 only
- C.2 and 3 only
- D.1 and 4 only
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1. Carbonic anhydrase catalyses: \(\text{CO}_2 + \text{H}_2\text{O} \rightleftharpoons \text{H}_2\text{CO}_3\) (carbonic acid). (Statement 1 is correct).
2. Carbonic acid dissociates into hydrogen ions (\(\text{H}^+\)) and hydrogencarbonate ions (\(\text{HCO}_3^-\)).
3. \(\text{HCO}_3^-\) ions diffuse out of the red blood cells into the plasma via a transport protein. (Statement 2 is correct).
4. To maintain electrical neutrality, chloride ions (\(\text{Cl}^-\)) diffuse INTO the red blood cells from the plasma (the chloride shift). Thus, statement 3 is incorrect because it says chloride ions diffuse out.
5. The accumulated \(\text{H}^+\) ions bind to haemoglobin (forming haemoglobinic acid), which buffers the cell and lowers the affinity of haemoglobin for oxygen (Bohr effect), causing oxyhaemoglobin to release oxygen more readily in acidic conditions. (Statement 4 is correct).
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- A.Sequence of electron transfer: \(\text{water} \rightarrow \text{PSII} \rightarrow \text{electron transport chain} \rightarrow \text{PSI} \rightarrow \text{NADP}\)
Source of electrons to replace those lost by PSII: photolysis of water - B.Sequence of electron transfer: \(\text{water} \rightarrow \text{PSI} \rightarrow \text{electron transport chain} \rightarrow \text{PSII} \rightarrow \text{NADP}\)
Source of electrons to replace those lost by PSII: reduced NADP - C.Sequence of electron transfer: \(\text{PSI} \rightarrow \text{electron transport chain} \rightarrow \text{PSII} \rightarrow \text{NADP}\)
Source of electrons to replace those lost by PSII: photolysis of water - D.Sequence of electron transfer: \(\text{PSII} \rightarrow \text{electron transport chain} \rightarrow \text{PSI} \rightarrow \text{electron transport chain} \rightarrow \text{water}\)
Source of electrons to replace those lost by PSII: reduced NADP
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1. Photolysis of water splits water molecules into oxygen, protons (\(\text{H}^+\)), and electrons: \(2\text{H}_2\text{O} \rightarrow 4\text{H}^+ + 4\text{e}^- + \text{O}_2\). These electrons replace those lost by the reaction centre of PSII (chlorophyll \(a\), P680) when it is photoactivated.
2. Photoactivated electrons from PSII pass along an electron transport chain to PSI.
3. Photoactivated electrons from PSI are used along with protons to reduce \(\text{NADP}^+\) to reduced NADP (NADPH).
Thus, the correct sequence is: \(\text{water} \rightarrow \text{PSII} \rightarrow \text{electron transport chain} \rightarrow \text{PSI} \rightarrow \text{NADP}\), and the electron source for PSII is the photolysis of water.
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- A.Glucose: concentration in glomerular filtrate is the same as in blood plasma; concentration in urine is zero.
- B.Urea: concentration in glomerular filtrate is lower than in blood plasma; concentration in urine is the same as in glomerular filtrate.
- C.Proteins: concentration in glomerular filtrate is the same as in blood plasma; concentration in urine is zero.
- D.Glucose: concentration in glomerular filtrate is lower than in blood plasma; concentration in urine is higher than in glomerular filtrate.
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- **Urea**: Being a small molecule, urea is also freely filtered, so its concentration in the glomerular filtrate is the **same** as in blood plasma. It is not fully reabsorbed, and as water is reabsorbed along the nephron, the concentration of urea in the urine becomes **much higher** than in the glomerular filtrate.
- **Proteins**: Most plasma proteins are too large to pass through the filtration barriers (basement membrane), so their concentration in the glomerular filtrate is **extremely low or absent**.
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- A.Single membrane: lysosome, tonoplast; Double membrane: nucleus, mitochondrion; No membrane: ribosome, centriole
- B.Single membrane: lysosome, ribosome; Double membrane: nucleus, chloroplast; No membrane: microtubule, nucleolus
- C.Single membrane: Golgi body, tonoplast; Double membrane: nucleus, lysosome; No membrane: ribosome, centriole
- D.Single membrane: mitochondrion, tonoplast; Double membrane: nucleus, chloroplast; No membrane: ribosome, nucleolus
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1. Amylose and amylopectin both contain \(\alpha\)-1,4-glycosidic bonds.
2. Glycogen contains more \(\alpha\)-1,6-glycosidic bonds per unit mass than amylopectin.
3. Amylose is a highly branched polysaccharide, whereas amylopectin is an unbranched helical chain.
4. Glycogen and amylopectin are both major storage polysaccharides found within plant starch granules.
Which statements are correct?
- A.1 and 2 only
- B.1 and 3 only
- C.2 and 4 only
- D.3 and 4 only
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In the presence of this inhibitor, the Michaelis-Menten constant (\(K_m\)) of the enzyme increased, but the maximum velocity (\(V_{max}\)) of the reaction remained unchanged.
Which type of inhibitor was used, and what is its effect on the apparent affinity of the enzyme for its substrate?
- A.Competitive inhibitor; decreases the apparent affinity
- B.Competitive inhibitor; increases the apparent affinity
- C.Non-competitive inhibitor; decreases the apparent affinity
- D.Non-competitive inhibitor; has no effect on the apparent affinity
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- Cell P: \(\psi = -600\text{ kPa}\); \(\psi_s = -800\text{ kPa}\)
- Cell Q: \(\psi = -400\text{ kPa}\); \(\psi_s = -700\text{ kPa}\)
- Cell R: \(\psi = -500\text{ kPa}\); \(\psi_s = -900\text{ kPa}\)
- Cell S: \(\psi = -700\text{ kPa}\); \(\psi_s = -1000\text{ kPa}\)
Which option correctly identifies a direction of net water movement between these cells?
- A.from Cell Q to Cell R
- B.from Cell S to Cell P
- C.from Cell P to Cell Q
- D.from Cell R to Cell Q
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- A.Pressure in the left ventricle becomes greater than the pressure in the aorta.
- B.Pressure in the left ventricle becomes greater than the pressure in the left atrium.
- C.Pressure in the left atrium becomes greater than the pressure in the left ventricle.
- D.Pressure in the aorta becomes greater than the pressure in the left ventricle.
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- Type 1: A baby receiving maternal antibodies via breast milk.
- Type 2: An individual receiving an injection of monoclonal antibodies after a venomous snake bite.
- Type 3: A child recovering from an infection of measles.
- Type 4: A person receiving an injection of a weakened pathogen (vaccination).
- A.Type 1: natural passive; Type 2: artificial passive; Type 3: natural active; Type 4: artificial active
- B.Type 1: artificial passive; Type 2: natural passive; Type 3: artificial active; Type 4: natural active
- C.Type 1: natural passive; Type 2: artificial active; Type 3: natural active; Type 4: artificial passive
- D.Type 1: natural active; Type 2: artificial passive; Type 3: natural passive; Type 4: artificial active
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- A.6 \(\text{CO}_2\), 8 reduced NAD, 2 reduced FAD
- B.4 \(\text{CO}_2\), 6 reduced NAD, 2 reduced FAD
- C.6 \(\text{CO}_2\), 6 reduced NAD, 2 reduced FAD
- D.4 \(\text{CO}_2\), 8 reduced NAD, 4 reduced FAD
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- A.The active repressor protein binds; transcription of the structural genes *lacZ* and *lacY* is prevented.
- B.The active repressor protein binds; transcription of the structural genes *lacZ* and *lacY* is activated.
- C.RNA polymerase binds; transcription of the structural genes *lacZ* and *lacY* is prevented.
- D.Lactose binds; transcription of the structural genes *lacZ* and *lacY* is activated.
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- A.The \(K_m\) increases and the \(V_{max}\) remains unchanged.
- B.The \(K_m\) decreases and the \(V_{max}\) remains unchanged.
- C.The \(K_m\) remains unchanged and the \(V_{max}\) decreases.
- D.The \(K_m\) increases and the \(V_{max}\) decreases.
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- Reject B: Competitive inhibitors do not decrease \(K_m\).
- Reject C: This describes non-competitive inhibition.
- Reject D: Competitive inhibitors do not reduce the maximum possible velocity.
Initially, the rate of entry increases linearly with concentration, but then reaches a maximum level (plateaus) at high concentrations. When a respiratory inhibitor is added to the cells, the rate of entry of the solute remains completely unaffected.
Which mechanism of transport is responsible for the entry of this solute?
- A.active transport
- B.facilitated diffusion
- C.simple diffusion
- D.osmosis
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- Reject A: Active transport requires ATP, so its rate would be significantly reduced by a respiratory inhibitor.
- Reject C: Simple diffusion does not involve transport proteins, so its rate would continue to increase linearly without reaching a plateau.
- Reject D: Osmosis refers specifically to the net movement of water molecules, not a polar solute.
- A.root hair cell wall \(\rightarrow\) cell walls of cortex cells \(\rightarrow\) cytoplasm of endodermal cells \(\rightarrow\) xylem vessel elements
- B.root hair cell membrane \(\rightarrow\) cell walls of cortex cells \(\rightarrow\) Casparian strip of endodermis \(\rightarrow\) xylem vessel elements
- C.root hair cell wall \(\rightarrow\) cytoplasm of cortex cells \(\rightarrow\) Casparian strip of endodermis \(\rightarrow\) xylem vessel elements
- D.root hair cell membrane \(\rightarrow\) cytoplasm of cortex cells \(\rightarrow\) cytoplasm of endodermal cells \(\rightarrow\) xylem vessel elements
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- Reject B: Water cannot pass through the hydrophobic Casparian strip in the cell wall.
- Reject C: Movement through cytoplasm represents the symplast pathway, not the apoplast pathway.
- Reject D: This is the symplast pathway entirely, and it starts with crossing the cell membrane.
- A.Carbon dioxide diffuses into the cell and is converted to carbonic acid by carbonic anhydrase; hydrogencarbonate ions then diffuse out of the cell in exchange for chloride ions entering.
- B.Carbon dioxide diffuses into the cell and is converted to carbonic acid by carbonic anhydrase; chloride ions then diffuse out of the cell in exchange for hydrogencarbonate ions entering.
- C.Carbonic acid is formed from carbon dioxide and water in the plasma; hydrogencarbonate ions then diffuse into the cell in exchange for chloride ions leaving.
- D.Oxygen and carbon dioxide compete for the same binding site on the haem group, forming carbaminohemoglobin and oxyhaemoglobin.
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- Reject B: Chloride ions move into the cell, and hydrogencarbonate ions move out of the cell, not vice versa.
- Reject C: Carbonic acid is formed primarily inside red blood cells due to the presence of carbonic anhydrase, not in the plasma.
- Reject D: Carbon dioxide binds to amine groups on the globin polypeptide chains of haemoglobin, not to the haem group where oxygen binds.
- A.To allow the resulting cells to divide repeatedly in culture and secrete a single, specific type of antibody.
- B.To increase the affinity of the antibodies for the target antigen by inducing rapid mutations.
- C.To cause the cancer cells to recognize and bind to the specific antigen.
- D.To enable the plasma cells to synthesize and secrete several different types of antibodies simultaneously.
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- Reject B: Fusion does not aim to induce mutations to increase affinity.
- Reject C: Cancer cells are fused because of their division properties, not because they recognize the antigen.
- Reject D: Monoclonal antibodies are, by definition, identical and specific to a single epitope; hybridomas produce only one type of antibody, not several different types.
- A.4 \(\text{CO}_2\) and 6 reduced NAD
- B.6 \(\text{CO}_2\) and 6 reduced NAD
- C.6 \(\text{CO}_2\) and 8 reduced NAD
- D.4 \(\text{CO}_2\) and 10 reduced NAD
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1. In the link reaction, each pyruvate is converted to acetyl-CoA, producing 1 \(\text{CO}_2\) and 1 reduced NAD. For two pyruvates, this yields 2 \(\text{CO}_2\) and 2 reduced NAD.
2. In the Krebs cycle, each acetyl-CoA yields 2 \(\text{CO}_2\) and 3 reduced NAD. For two acetyl-CoA molecules, this yields 4 \(\text{CO}_2\) and 6 reduced NAD.
Adding these together:
Total \(\text{CO}_2\) = 2 (link) + 4 (Krebs) = 6 molecules.
Total reduced NAD = 2 (link) + 6 (Krebs) = 8 molecules.
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- Reject A: This omits the products of the link reaction.
- Reject B: This incorrectly states that only 6 reduced NAD are produced in total.
- Reject D: This incorrectly states that only 4 \(\text{CO}_2\) are produced.
A woman with normal colour vision, whose father was red-green colour-blind, has children with a man who has normal colour vision.
What is the probability that their first child will be a red-green colour-blind son?
- A.0.125
- B.0.25
- C.0.50
- D.0.75
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The woman's father was colour-blind (genotype \(X^b Y\)), meaning he must have passed his \(X^b\) chromosome to his daughter. Since she has normal vision, her genotype must be heterozygous: \(X^B X^b\).
The man has normal vision, so his genotype is \(X^B Y\).
Crossing these parents (\(X^B X^b \times X^B Y\)) yields the following offspring possibilities:
1. \(X^B X^B\) (normal-vision female)
2. \(X^B X^b\) (carrier female with normal vision)
3. \(X^B Y\) (normal-vision male)
4. \(X^b Y\) (colour-blind male)
The question asks for the probability that their first child will be a colour-blind son. Out of the 4 equally likely combinations, exactly 1 is a colour-blind son (\(X^b Y\)). Therefore, the probability is 1 in 4, or 0.25.
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- Reject A: 0.125 is incorrect.
- Reject C: 0.50 is the probability of a son being colour-blind given that the child is already known to be male, but the question asks for the overall probability of a first child being a colour-blind son.
- Reject D: 0.75 is incorrect.
- A.During depolarization, sodium ions are actively pumped into the axon; during repolarization, potassium ions are actively pumped out of the axon.
- B.During depolarization, sodium ions diffuse into the axon; during repolarization, potassium ions diffuse out of the axon.
- C.During depolarization, potassium ions diffuse out of the axon; during repolarization, sodium ions diffuse into the axon.
- D.During depolarization, sodium ions diffuse out of the axon; during repolarization, potassium ions diffuse into the axon.
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- Reject A: The movements during depolarization and repolarization are passive diffusion, not active pumping (the sodium-potassium pump is active but does not cause these phases directly).
- Reject C: This reverses the roles of potassium and sodium.
- Reject D: This incorrectly states that sodium diffuses out and potassium diffuses in.
* **Fraction P** contains organelles with a double membrane that are the site of aerobic respiration.
* **Fraction Q** contains organelles bounded by a single membrane that are involved in the modification and packaging of proteins.
* **Fraction R** contains structures with no membrane that are the sites of translation.
* **Fraction S** contains organelles with a double membrane containing DNA and thylakoids.
Which organelles are present in fractions P, Q, R, and S?
- A.P: Mitochondria; Q: Golgi apparatus; R: Ribosomes; S: Chloroplasts
- B.P: Chloroplasts; Q: Lysosomes; R: Nucleoli; S: Mitochondria
- C.P: Mitochondria; Q: Rough endoplasmic reticulum; R: Ribosomes; S: Nuclei
- D.P: Nuclei; Q: Golgi apparatus; R: Microtubules; S: Chloroplasts
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* **Fraction Q** is bounded by a single membrane and modifies and packages proteins, which corresponds to the **Golgi apparatus**.
* **Fraction R** has no membrane and is the site of translation, which corresponds to **ribosomes**.
* **Fraction S** has a double membrane, DNA, and thylakoids, which corresponds to **chloroplasts**.
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- Reject other options because they mismatch organelle characteristics: nucleoli (option B) are sub-nuclear regions with no membrane; nuclei (option D) lack thylakoids; rough endoplasmic reticulum (option C) has a single membrane.
1. Glycogen and amylopectin both contain \(\alpha\)-1,4-glycosidic bonds and \(\alpha\)-1,6-glycosidic bonds.
2. Cellulose contains \(\beta\)-1,4-glycosidic bonds, where alternate glucose molecules are rotated by 180°.
3. Glycogen is more highly branched than amylopectin because it has a higher proportion of \(\alpha\)-1,6-glycosidic bonds.
4. Hydrogen bonds form cross-links between adjacent cellulose chains to form microfibrils.
- A.1, 2, 3 and 4
- B.1, 2 and 3 only
- C.2 and 4 only
- D.1 and 3 only
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1. Both glycogen and amylopectin are branched molecules made of \(\alpha\)-glucose, with linear chains connected by \(\alpha\)-1,4-glycosidic bonds and branches formed by \(\alpha\)-1,6-glycosidic bonds.
2. Cellulose is a linear polymer of \(\beta\)-glucose with \(\beta\)-1,4-glycosidic bonds. To form these bonds, each successive glucose monomer must be rotated 180° relative to its neighbor.
3. Glycogen has branches occurring more frequently (every 8 to 12 glucose units) than in amylopectin (every 24 to 30 glucose units), meaning glycogen contains a higher density of \(\alpha\)-1,6-glycosidic bonds.
4. Parallel cellulose chains run side by side and are held together by numerous hydrogen bonds between hydroxyl groups, forming strong structural units called microfibrils.
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- Reject options B, C, and D as they omit correct statements.
The results are shown in the table below:
| Condition | \(V_{max}\) / arbitrary units | \(K_m\) / mmol dm\(^{-3}\) |
| :--- | :--- | :--- |
| Enzyme alone | 120 | 2.5 |
| Enzyme + Inhibitor X | 120 | 6.8 |
| Enzyme + Inhibitor Y | 45 | 2.5 |
What is the correct explanation of the mode of action of inhibitors X and Y?
- A.Inhibitor X is a competitive inhibitor that binds reversibly to the active site; Inhibitor Y is a non-competitive inhibitor that binds to an allosteric site.
- B.Inhibitor X is a non-competitive inhibitor that decreases the affinity of the enzyme for its substrate; Inhibitor Y is a competitive inhibitor that reduces the maximum velocity of the reaction.
- C.Inhibitor X is a competitive inhibitor that increases the affinity of the enzyme for its substrate; Inhibitor Y is a non-competitive inhibitor that increases the rate of ES complex formation.
- D.Inhibitor X is a non-competitive inhibitor that binds to the active site; Inhibitor Y is a competitive inhibitor that binds reversibly to an allosteric site.
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* **Inhibitor Y** decreases \(V_{max}\) from 120 to 45 arbitrary units, but leaves \(K_m\) unchanged at 2.5 mmol dm\(^{-3}\). This is characteristic of a **non-competitive inhibitor**, which binds to an allosteric site. It reduces the concentration of active enzyme molecules, reducing \(V_{max}\), but does not alter the affinity of the unaffected active sites for the substrate, so \(K_m\) remains unchanged.
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- Reject B, C, and D because competitive inhibitors increase \(K_m\) without changing \(V_{max}\), and non-competitive inhibitors decrease \(V_{max}\) without changing \(K_m\).
* **Mechanism 1**: Movement of a hydrophobic molecule directly through the phospholipid bilayer down its concentration gradient.
* **Mechanism 2**: Movement of ions through a specific membrane channel protein down their electrochemical gradient without the use of ATP.
* **Mechanism 3**: Movement of glucose against its concentration gradient using a membrane protein and energy from a sodium ion gradient established by the active transport of sodium ions.
* **Mechanism 4**: Movement of bulk secretory proteins out of the cell via vesicle fusion with the cell surface membrane.
Which row correctly identifies these transport mechanisms?
- A.1: Simple diffusion, 2: Facilitated diffusion, 3: Secondary active transport, 4: Exocytosis
- B.1: Simple diffusion, 2: Active transport, 3: Facilitated diffusion, 4: Endocytosis
- C.1: Osmosis, 2: Facilitated diffusion, 3: Primary active transport, 4: Exocytosis
- D.1: Facilitated diffusion, 2: Simple diffusion, 3: Secondary active transport, 4: Pinocytosis
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* **Mechanism 2** is **facilitated diffusion** because ions move through a specific membrane channel protein down their gradient without requiring ATP.
* **Mechanism 3** is **secondary active transport** (co-transport) because glucose is transported against its gradient using energy from an ion gradient (established indirectly using ATP).
* **Mechanism 4** is **exocytosis** because bulk materials are released from vesicles fusing with the cell surface membrane.
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- Reject B because mechanism 3 is not simple facilitated diffusion, and bulk release is not endocytosis.
- Reject C because simple diffusion of hydrophobic molecules is not osmosis.
- Reject D because bulk export is not pinocytosis (which is the uptake of fluids).
| Cell | Solute potential (\(\psi_s\)) / kPa | Pressure potential (\(\psi_p\)) / kPa |
| :--- | :---: | :---: |
| **X** | -700 | 200 |
| **Y** | -800 | 400 |
| **Z** | -900 | 300 |
Water potential (\(\psi\)) is calculated as \(\psi = \psi_s + \psi_p\).
In which direction will there be a net movement of water by osmosis between these cells?
- A.from cell X to cell Y, and from cell Y to cell Z
- B.from cell Y to cell X, and from cell Y to cell Z
- C.from cell X to cell Y, and from cell Z to cell Y
- D.from cell Y to cell X, and from cell Z to cell X
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* \(\psi_X = -700 + 200 = -500\text{ kPa}\)
* \(\psi_Y = -800 + 400 = -400\text{ kPa}\)
* \(\psi_Z = -900 + 300 = -600\text{ kPa}\)
Water moves by osmosis down a water potential gradient, from a higher (less negative) water potential to a lower (more negative) water potential.
* Since \(\psi_Y\) (-400 kPa) is higher than \(\psi_X\) (-500 kPa), water moves from **cell Y to cell X**.
* Since \(\psi_Y\) (-400 kPa) is higher than \(\psi_Z\) (-600 kPa), water moves from **cell Y to cell Z**.
Therefore, there is a net movement of water from cell Y to cell X, and from cell Y to cell Z.
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- Reject A, C, and D because they suggest water moving from lower (more negative) water potential to higher (less negative) water potential.
Which row correctly identifies the state of the valves at the point when the pressure in the left ventricle first rises above the pressure in the left atrium, and when the pressure in the left ventricle first falls below the pressure in the aorta?
- A.Left ventricular pressure > left atrial pressure: AV valve closes, semilunar valve remains closed. Left ventricular pressure < aortic pressure: AV valve remains closed, semilunar valve closes.
- B.Left ventricular pressure > left atrial pressure: AV valve opens, semilunar valve opens. Left ventricular pressure < aortic pressure: AV valve closes, semilunar valve opens.
- C.Left ventricular pressure > left atrial pressure: AV valve closes, semilunar valve opens. Left ventricular pressure < aortic pressure: AV valve opens, semilunar valve closes.
- D.Left ventricular pressure > left atrial pressure: AV valve remains open, semilunar valve closes. Left ventricular pressure < aortic pressure: AV valve remains open, semilunar valve remains open.
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2. When the pressure in the left ventricle first falls below the pressure in the aorta: this causes blood to start flowing backward towards the ventricle, which instantly catches the pockets of the semilunar (aortic) valve, forcing it to close. At this point, ventricular pressure is still higher than atrial pressure, so the atrioventricular valve remains closed (it only opens once ventricular pressure falls below atrial pressure).
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- Reject options B, C, and D because they mismatch the physiological pressure changes with the corresponding valve actions.
- A.Bronchioles: Cartilage: absent (-), Smooth muscle: present (+), Elastic fibres: present (+). Alveoli: Cartilage: absent (-), Smooth muscle: absent (-), Elastic fibres: present (+).
- B.Bronchioles: Cartilage: present (+), Smooth muscle: present (+), Elastic fibres: present (+). Alveoli: Cartilage: absent (-), Smooth muscle: present (+), Elastic fibres: present (+).
- C.Bronchioles: Cartilage: absent (-), Smooth muscle: absent (-), Elastic fibres: present (+). Alveoli: Cartilage: absent (-), Smooth muscle: absent (-), Elastic fibres: absent (-).
- D.Bronchioles: Cartilage: absent (-), Smooth muscle: present (+), Elastic fibres: absent (-). Alveoli: Cartilage: present (+), Smooth muscle: absent (-), Elastic fibres: present (+).
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* **Alveoli** completely lack both cartilage (-) and smooth muscle (-). However, they contain a rich network of elastic fibres (+) that stretch during inhalation and recoil to push air out during exhalation.
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- Reject B because bronchioles lack cartilage, and alveoli lack smooth muscle.
- Reject C because alveoli contain elastic fibres.
- Reject D because bronchioles contain elastic fibres, and alveoli do not contain cartilage.
What types of immunity are acquired by the person in each of these three situations?
- A.Breast milk: Natural passive; Vaccine: Artificial active; Antivenom: Artificial passive
- B.Breast milk: Natural active; Vaccine: Artificial passive; Antivenom: Artificial active
- C.Breast milk: Natural passive; Vaccine: Natural active; Antivenom: Artificial passive
- D.Breast milk: Artificial passive; Vaccine: Artificial active; Antivenom: Natural passive
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2. **Vaccines**: The introduction of an antigen (attenuated pathogen) medically stimulates the child's own immune response to produce antibodies and memory cells. Thus, it is **artificial active** immunity.
3. **Antivenom**: An injection of pre-formed antibodies (antitoxin) provides immediate protection but does not stimulate the recipient's immune system to make memory cells. Thus, it is **artificial passive** immunity.
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- Reject other options because they mismatch passive vs active or natural vs artificial categories.
Paper 22 - AS Structured Questions
(a) Define the term Michaelis-Menten constant (\(K_m\)) and explain its significance with respect to the affinity of an enzyme for its substrate. [3]
(b) The experiment was repeated in the presence of a reversible competitive inhibitor.
(i) Explain the effect of a competitive inhibitor on the \(V_{\text{max}}\) and \(K_m\) values of the enzyme. [3]
(ii) Suggest how a competitive inhibitor lowers the rate of reaction at low substrate concentrations, and why this inhibition is overcome at very high substrate concentrations. [2]
(c) State two factors, other than the presence of an inhibitor, that can alter the rate of an enzyme-catalysed reaction. [2]
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- Michaelis-Menten constant (\(K_m\)) is defined as the substrate concentration at which the rate of the enzyme-catalysed reaction is half of its maximum velocity (\(\frac{1}{2} V_{\text{max}}\)). [1]
- It is an inverse measure of the affinity of the enzyme for its substrate. [1]
- A low \(K_m\) indicates a high affinity (the enzyme reaches half-maximal velocity at low substrate concentrations), whereas a high \(K_m\) indicates a low affinity. [1]
Part (b):
- (i) A competitive inhibitor increases the \(K_m\) value [1] but has no effect on the \(V_{\text{max}}\) value. [1] This is because the same maximum rate can still be achieved if enough substrate is added to outcompete the inhibitor. [1]
- (ii) At low substrate concentrations, the inhibitor successfully competes with substrate molecules for the active site, reducing the rate of enzyme-substrate (ES) complex formation. [1] At very high substrate concentrations, the probability of substrate molecules binding to the active site is much higher than that of the inhibitor, completely overcoming the inhibition. [1]
Part (c):
- Any two from: Temperature, pH, enzyme concentration, substrate concentration. [2]
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1. \(K_m\) defined as substrate concentration at half-maximum velocity / \(\frac{1}{2} V_{\text{max}}\); [1]
2. \(K_m\) is inversely proportional to enzyme affinity; [1]
3. Low \(K_m\) = high affinity OR high \(K_m\) = low affinity; [1]
Part (b)(i) [3 marks maximum]:
1. \(V_{\text{max}}\) is unchanged / remains constant; [1]
2. \(K_m\) is increased; [1]
3. Idea that additional substrate overcomes the effect of the inhibitor to reach the same maximum velocity; [1]
Part (b)(ii) [2 marks maximum]:
1. Inhibitor binds to the active site, blocking substrate entry / reducing enzyme-substrate (ES) complex formation (at low substrate concentration); [1]
2. High substrate concentration increases probability of substrate binding instead of inhibitor / outcompeting inhibitor; [1]
Part (c) [2 marks maximum]:
1. Temperature; [1]
2. pH; [1]
(Accept: enzyme concentration / substrate concentration if not already specified in the stem)
(a) Describe how sucrose is actively loaded into the companion cells and then enters the sieve tube elements. [5]
(b) Sieve tube elements are highly specialized cells. Explain how the structure of a phloem sieve tube element is adapted to its function of transporting assimilates. [3]
(c) Distinguish between the apoplastic and symplastic pathways of water and solute movement through the plant root cortex. [2]
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- Hydrogen ions (\(\text{H}^+\) / protons) are actively pumped out of the companion cell cytoplasm into the cell wall space using ATP. [1]
- This creates a high concentration gradient of protons in the cell wall compared to the inside of the companion cell. [1]
- Protons diffuse back down their concentration gradient into the companion cell through co-transporter proteins. [1]
- This co-transport protein simultaneously moves sucrose molecules into the companion cell against their concentration gradient. [1]
- Sucrose then diffuses from the companion cell into the phloem sieve tube element through interconnecting plasmodesmata. [1]
Part (b):
- Sieve tube elements have very little cytoplasm, no nucleus, no vacuole, and no ribosomes, which provides a clear pathway with minimal resistance to the flow of sap. [1]
- They have end walls modified into sieve plates with sieve pores, which allow the liquid assimilate to flow from cell to cell easily. [1]
- The cells are elongated and joined end-to-end to form a continuous conduit. [1]
- Thick cellulose walls withstand high hydrostatic pressures. [1] (Max 3)
Part (c):
- Apoplast pathway involves movement through the non-living cell walls and intercellular spaces. [1]
- Symplast pathway involves movement through the living cytoplasm and plasmodesmata of adjacent cells. [1]
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1. Protons / hydrogen ions / \(\text{H}^+\) actively transported / pumped out of companion cells; [1]
2. Active transport requires ATP; [1]
3. Establishes a proton gradient (higher concentration outside in cell wall); [1]
4. Protons diffuse back through co-transporter proteins, bringing sucrose with them; [1]
5. Sucrose enters companion cell against its concentration gradient; [1]
6. Sucrose moves from companion cell to sieve tube element via plasmodesmata; [1]
Part (b) [3 marks maximum]:
1. Little cytoplasm / no nucleus / no vacuole / fewer organelles to minimize resistance to flow / maximize space; [1]
2. Presence of sieve plates / sieve pores to allow mass flow / easy passage of solutes; [1]
3. Elongated cells joined end-to-end to form continuous tube; [1]
4. Strong / thick cellulose walls to resist high hydrostatic pressure; [1]
Part (c) [2 marks maximum]:
1. Apoplast: through cell walls / intercellular spaces / non-living pathway; [1]
2. Symplast: through cytoplasm / plasmodesmata / living pathway; [1]
(a) Describe the structure of a typical IgG antibody molecule, explaining how its structure relates to its function. [4]
(b) Explain the differences between active immunity and passive immunity. [4]
(c) Distinguish between natural active immunity and artificial active immunity. [2]
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- An IgG antibody molecule consists of four polypeptide chains: two identical heavy chains and two identical light chains, arranged in a Y-shape. [1]
- These chains are held together by covalent disulfide bonds. [1]
- It has two identical antigen-binding sites in the variable regions, which have a specific complementary shape to a particular antigen. [1]
- The constant region is identical in all antibodies of the same class and binds to receptors on phagocytes to facilitate phagocytosis. [1]
- A flexible hinge region allows the angle between the arms of the antibody to vary, enabling it to bind to two antigen molecules simultaneously (causing agglutination). [1] (Max 4)
Part (b):
- Active immunity involves the host's own immune system producing antibodies in response to exposure to an antigen, whereas passive immunity involves receiving antibodies produced by another organism. [1]
- Active immunity leads to the production of memory cells, providing long-term immunity, while passive immunity does not produce memory cells and only provides short-term protection. [1]
- Active immunity takes time to develop (lag phase), whereas passive immunity provides immediate protection. [1]
- Active immunity is stimulated by infection or vaccination, while passive immunity occurs naturally via colostrum/placenta or artificially via injection of antitoxins/monoclonal antibodies. [1]
Part (c):
- Natural active immunity occurs when an individual is naturally exposed to a live pathogen, becomes ill, and recovers, developing their own immune response. [1]
- Artificial active immunity is induced by intentional exposure to a weakened or dead pathogen, or its antigens, via vaccination, without causing the disease. [1]
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1. Four polypeptide chains / two heavy and two light chains (forming Y-shape); [1]
2. Chains joined by disulfide bonds / bridges; [1]
3. Variable region forms (two) antigen-binding sites, which are complementary to specific antigen; [1]
4. Constant region binds to phagocytes (to stimulate opsonization / phagocytosis); [1]
5. Hinge region provides flexibility to bind to multiple antigens / agglutination; [1]
Part (b) [4 marks maximum]:
1. Active: host's immune system produces antibodies vs. Passive: external antibodies introduced; [1]
2. Active: memory cells produced vs. Passive: no memory cells; [1]
3. Active: long-term protection vs. Passive: short-term / temporary protection; [1]
4. Active: delay/lag phase in protection vs. Passive: immediate protection; [1]
Part (c) [2 marks maximum]:
1. Natural active: result of infection / pathogen entering body naturally; [1]
2. Artificial active: result of vaccination / injection of weakened or dead pathogen / antigen; [1]
(a) Describe the pathway and role of organelles in the synthesis, modification, and secretion of mucus by a goblet cell, starting from the transcription of the protein component. [6]
(b) Prokaryotic cells, such as bacteria, do not contain the same organelles as eukaryotic cells.
State three structural differences between a typical prokaryotic cell and a goblet cell. [3]
(c) Name the organelle present in both prokaryotic and eukaryotic cells that is involved in protein synthesis. [1]
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- Transcription of the mucin gene occurs in the nucleus to produce mRNA. [1]
- mRNA leaves the nucleus via a nuclear pore and travels to the rough endoplasmic reticulum (RER). [1]
- Ribosomes on the RER translate the mRNA to synthesize the polypeptide chain. [1]
- The polypeptide enters the lumen of the RER, where it is folded into its tertiary structure and transported. [1]
- Transport vesicles bud off the RER and carry the protein to the Golgi apparatus. [1]
- In the Golgi apparatus, the protein is chemically modified by the addition of carbohydrate chains (glycosylation) to form the glycoprotein (mucin). [1]
- Secretory vesicles containing mucin bud off the Golgi apparatus, move along the cytoskeleton, and fuse with the cell surface membrane to release the mucus via exocytosis. [1] (Max 6)
Part (b):
- Any three from:
1. Prokaryotic cell has circular DNA (not associated with histones / 'naked' DNA), whereas a goblet cell has linear DNA associated with histone proteins. [1]
2. Prokaryotic cell has 70S ribosomes, whereas a goblet cell has 80S ribosomes. [1]
3. Prokaryotic cell has no membrane-bound organelles (e.g. no nucleus, mitochondria, Golgi, RER), whereas a goblet cell has them. [1]
4. Prokaryotic cell has a peptidoglycan cell wall, whereas a goblet cell has no cell wall. [1]
5. Prokaryotes are much smaller in size (typically 1-5 micrometres) compared to eukaryotic goblet cells. [1]
Part (c):
- Ribosome (accept 70S ribosome/80S ribosome). [1]
PastPaper.markingScheme
1. Nucleus: transcription / production of mRNA (from DNA); [1]
2. Nuclear pore: allows mRNA to exit into cytoplasm; [1]
3. Ribosome / RER: site of translation / protein synthesis; [1]
4. RER lumen: folding / transport of polypeptide; [1]
5. Transport vesicles: transport protein from RER to Golgi; [1]
6. Golgi apparatus: modification / addition of carbohydrates (glycosylation) to form glycoprotein; [1]
7. Secretory vesicles: transport mucus to cell surface membrane; [1]
8. Exocytosis / fusion with membrane: to secrete mucus out of cell; [1]
Part (b) [3 marks maximum]:
1. Circular DNA / plasmids vs. linear DNA; [1]
2. Naked DNA vs. histone-associated DNA; [1]
3. 70S ribosomes vs. 80S ribosomes; [1]
4. No membrane-bound organelles / no nucleus / no mitochondria / no Golgi vs. present; [1]
5. Peptidoglycan cell wall vs. no cell wall; [1]
Part (c) [1 mark]:
1. Ribosome; [1]
(a) State the function of each of the following components of the gas exchange system:
(i) cartilage in the walls of the trachea [2]
(ii) elastic fibres in the walls of the alveoli [2]
(iii) smooth muscle in the walls of the bronchioles [2]
(b) Explain how the structure of the alveolar wall is adapted to maximize the rate of diffusion of respiratory gases. [4]
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- (i) Cartilage provides structural support [1] and prevents the trachea from collapsing when the air pressure drops during inhalation. [1]
- (ii) Elastic fibres stretch during inspiration as the lungs fill with air [1] and recoil during expiration to help force the air out of the alveoli. [1]
- (iii) Smooth muscle contracts to narrow the airway lumen (bronchoconstriction) and relaxes to widen it (bronchodilation) [1] to regulate the volume of air reaching the alveoli. [1]
Part (b):
- The alveolar wall consists of a single layer of squamous epithelial cells [1], which provides an extremely thin barrier / short diffusion distance. [1]
- The adjacent capillary walls are also only one endothelial cell thick, making the total diffusion pathway (alveolar wall, basement membrane, capillary wall) very short. [1]
- Alveoli are highly folded, presenting a very large total surface area for diffusion. [1]
- The inner surface of the alveolus is covered by a thin layer of moisture in which oxygen dissolves before diffusing across the cells. [1] (Max 4)
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1. Prevents trachea collapsing / bursting; [1]
2. (Especially) during inhalation / when pressure in trachea decreases; [1]
(Accept: keeps airway open/patent [1])
Part (a)(ii) [2 marks]:
1. Stretch during inspiration / inhalation (preventing bursting); [1]
2. Recoil during expiration / exhalation to force air out; [1]
Part (a)(iii) [2 marks]:
1. Contracts to narrow airway / bronchoconstriction / restricts air flow (e.g., in presence of dust/allergens); [1]
2. Relaxes to widen airway / bronchodilation / allows more air flow (e.g., during exercise); [1]
Part (b) [4 marks maximum]:
1. Squamous epithelium is extremely thin / one cell thick; [1]
2. Capillary wall / endothelium is also one cell thick; [1]
3. Short diffusion distance (total distance is very small / \(\approx 0.5-1.0\ \mu\text{m}\)); [1]
4. Large surface-area-to-volume ratio / highly folded surface; [1]
5. Moist lining / surfactant allows gases to dissolve (speeding up diffusion); [1]
(a) Describe the behaviour of the chromosomes during the following stages of mitosis:
(i) prophase [2]
(ii) metaphase [2]
(iii) anaphase [2]
(b) Explain why it is essential that the DNA is replicated precisely during the S phase of interphase, before mitosis begins. [2]
(c) Suggest the role of telomeres during repeated cycles of DNA replication and cell division. [2]
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- (i) Prophase:
1. Chromatin condenses / coils / shortens and thickens to become visible as distinct chromosomes. [1]
2. Each chromosome is seen to consist of two sister chromatids held together at a centromere. [1]
- (ii) Metaphase:
1. Chromosomes align individually along the equator (metaphase plate) of the spindle. [1]
2. Spindle fibres (microtubules) attach to the centromere (specifically kinetochores) of each chromosome. [1]
- (iii) Anaphase:
1. The centromeres split / divide. [1]
2. Spindle fibres shorten and pull the sister chromatids (now called chromosomes) to opposite poles of the spindle, centromeres leading. [1]
Part (b):
- DNA replication must be precise so that the two new daughter cells receive identical copies of the genome. [1]
- This ensures genetic stability / prevents mutations or loss of genes, allowing the daughter cells to function identically to the parent cell. [1]
Part (c):
- Telomeres act as protective caps at the ends of linear chromosomes. [1]
- Due to the 'end-replication problem', DNA is slightly shortened at each replication; telomeres consist of non-coding repetitive DNA, so they prevent the loss of essential genes during this shortening process. [1]
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1. Chromatin condenses / coils / becomes visible as chromosomes; [1]
2. Chromosomes visible as two sister chromatids joined at the centromere; [1]
Part (a)(ii) [2 marks]:
1. Chromosomes line up along the equator / metaphase plate of spindle; [1]
2. Spindle fibres attach to centromere / kinetochore; [1]
Part (a)(iii) [2 marks]:
1. Centromere splits / divides; [1]
2. Sister chromatids separated / pulled to opposite poles (centromere leading); [1]
Part (b) [2 marks maximum]:
1. To ensure daughter cells are genetically identical / clones of parent cell; [1]
2. To prevent mutations / loss of genes / loss of function; [1]
Part (c) [2 marks maximum]:
1. Prevents the loss of essential / coding genetic information / genes during DNA replication; [1]
2. Consist of non-coding DNA that can be sacrificed as the chromosome ends shorten; [1]
(Accept: prevents chromosome ends from fusing / protects chromosome from degradation [1])
Paper 32 - Advanced Practical Skills
Section instructions: Carry out the experimental procedures as detailed. Record all data and draw biological structures using a sharp pencil.
You are provided with:
- E, a 1.0% amylase solution
- S, a 1.0% starch solution
- I, a 2.0% copper sulfate solution (non-competitive inhibitor)
- W, distilled water
- Iodine, iodine in potassium iodide solution
Part 1: Preparation of Inhibitor Concentrations
You are required to make a two-fold serial dilution of the 2.0% copper sulfate solution, I, to obtain the following concentrations: 2.0%, 1.0%, 0.5%, 0.25%, and 0.125%. The total volume of each solution must be 10 cm³.
(a) Complete Table 1.1 to show how you will prepare these five concentrations of inhibitor using solution I and distilled water W. [3 marks]
Table 1.1
Concentration of inhibitor / %
Volume of solution to be diluted / cm³
Volume of distilled water, W / cm³
2.0
10.0
0.0
1.0
...
...
0.5
...
...
0.25
...
...
0.125
...
...
Part 2: Investigation and Results
1. Label five clean test-tubes with the concentrations of inhibitor: 2.0%, 1.0%, 0.5%, 0.25%, and 0.125%.
2. Add 2 cm³ of the appropriate concentration of inhibitor to each labeled tube.
3. Add 2 cm³ of the amylase solution, E, to each tube. Swirl gently to mix and allow to stand for 3 minutes.
4. During this incubation, add one drop of iodine solution to each well of a clean spotting tile.
5. To the test-tube containing 2.0% inhibitor, add 2 cm³ of starch solution, S. Mix immediately and start a timer.
6. At 30-second intervals, remove a small drop of the mixture using a clean pipette and add it to a well of the spotting tile containing iodine solution.
7. Record the time taken (in seconds) for the blue-black color to first fail to appear. If starch is not completely hydrolyzed after 300 seconds, stop and record ">300".
8. Repeat steps 5 to 7 for each of the remaining inhibitor concentrations.
(b) Record your results in a single, appropriately designed table. [6 marks]
(c) Plot a line graph to show the rate of reaction (\(\text{s}^{-1}\)) on the y-axis against the concentration of inhibitor (%) on the x-axis. Calculate the rate as \(\frac{1}{\text{time taken (s)}}\). [4 marks]
(d) Explain the shape of your graph with reference to the mode of action of a non-competitive inhibitor like copper sulfate on amylase. [3 marks]
(e) Identify two sources of error in this experimental procedure that could affect the accuracy of your results, and suggest a specific improvement for each error. [4 marks]
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(a) Dilution Table Completion:
To perform a two-fold serial dilution of 2.0% inhibitor I to obtain 10 cm³ of each concentration:
- 2.0% inhibitor: 10.0 cm³ of 2.0% solution, 0.0 cm³ of W.
- 1.0% inhibitor: Transfer 5.0 cm³ from 2.0% tube, add 5.0 cm³ of W.
- 0.5% inhibitor: Transfer 5.0 cm³ from 1.0% tube, add 5.0 cm³ of W.
- 0.25% inhibitor: Transfer 5.0 cm³ from 0.5% tube, add 5.0 cm³ of W.
- 0.125% inhibitor: Transfer 5.0 cm³ from 0.25% tube, add 5.0 cm³ of W.
Completed table columns (Volume of solution / Volume of W):
- 2.0%: 10.0 cm³ / 0.0 cm³
- 1.0%: 5.0 cm³ of 2.0% / 5.0 cm³
- 0.5%: 5.0 cm³ of 1.0% / 5.0 cm³
- 0.25%: 5.0 cm³ of 0.5% / 5.0 cm³
- 0.125%: 5.0 cm³ of 0.25% / 5.0 cm³
(b) Sample Results Table:
Concentration of inhibitor, I / %
Time taken to reach end-point / s
Rate of reaction, \(1/t\) / \(\text{s}^{-1}\)
2.000
>300 (no reaction)
0.000
1.000
240
0.004
0.500
150
0.007
0.250
90
0.011
0.125
60
0.017
(c) Graph Construction:
- X-axis: Concentration of inhibitor / % (scale from 0 to 2.0% with linear intervals).
- Y-axis: Rate of reaction / \(\text{s}^{-1}\) (scale from 0 to 0.020 \(\text{s}^{-1}\)).
- Plotted Points: Plotted with small 'x' marks. Points show a decrease in rate as concentration of inhibitor increases.
- Line: Points connected by straight ruled lines or a line of best fit.
(d) Explanation:
- Copper sulfate acts as a non-competitive inhibitor because copper ions (\(\text{Cu}^{2+}\)) bind to an allosteric site of the amylase enzyme molecule.
- This binding disrupts ionic/hydrogen bonds, altering the tertiary structure of the protein and therefore the specific shape of the active site.
- The substrate (starch) can no longer fit into the active site, preventing the formation of enzyme-substrate complexes, reducing the rate of starch hydrolysis even at high substrate concentrations.
(e) Errors and Improvements:
- Error 1: The 30-second interval is too large, leading to low precision of the end-point. Improvement: Test the mixture at shorter, more frequent intervals, such as every 10 or 15 seconds.
- Error 2: Fluctuations in room temperature affect enzyme kinetic energy during the experiment. Improvement: Incubate all tubes and carry out the reactions within a thermostatically controlled water bath set to 35°C.
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(a) Dilution table [3 marks maximum]:
- 1 mark for correct volumes of solution and W for all concentrations (5.0 cm³ of previous concentration and 5.0 cm³ of distilled water for all serial dilutions except the starting 2.0% which has 10 cm³ and 0 cm³).
- 1 mark for showing a consistent two-fold dilution pattern (transferring 5.0 cm³ of the preceding solution to the next tube).
- 1 mark for writing all volumes to one decimal place consistently (e.g., 5.0, 10.0, 0.0).
(b) Results table [6 marks maximum]:
- 1 mark for a single table with clear headers including appropriate units (Concentration of inhibitor / %, Time / s, and Rate / \(\text{s}^{-1}\)).
- 1 mark for having no units in the body of the table (only in the column headers).
- 1 mark for recording all five inhibitor concentrations in a logical descending or ascending order.
- 1 mark for recording times as whole numbers (multiples of 30, corresponding to the sampling interval).
- 1 mark for showing a correct trend where the time to end-point increases (or no reaction occurs) as the inhibitor concentration increases.
- 1 mark for calculating rates correctly to an appropriate number of significant figures / decimal places (e.g., 3 decimal places).
(c) Graph plotting [4 marks maximum]:
- 1 mark for correct axes labels with units (x-axis: Concentration of inhibitor / %, y-axis: Rate of reaction / \(\text{s}^{-1}\)).
- 1 mark for a linear scale where the plotted points cover more than half of the grid in both dimensions.
- 1 mark for plotting all points accurately with thin, precise lines/points (e.g., small 'x' or encircled dot within 1 mm of correct coordinate).
- 1 mark for connecting points with a sharp, ruled straight line from point to point, or a smooth line of best fit with no feathering.
(d) Explanation [3 marks maximum]:
- 1 mark for identifying copper sulfate / copper ions as non-competitive inhibitors.
- 1 mark for explaining that the inhibitor binds to an allosteric site (a site other than the active site) of the amylase enzyme.
- 1 mark for stating that this alters the tertiary structure / active site shape, preventing substrate binding / enzyme-substrate complex (ESC) formation.
(e) Errors and Improvements [4 marks maximum]:
- 1 mark for identifying a valid error (e.g., 30-second sampling interval is too wide / subjective end-point determination / lack of temperature control).
- 1 mark for a matching realistic improvement (e.g., sample every 10 or 15 seconds / use a colorimeter to measure absorbance / use a thermostatically controlled water bath).
- Max 2 marks for errors, Max 2 marks for corresponding improvements.
Section instructions: Carry out the experimental procedures as detailed. Record all data and draw biological structures using a sharp pencil.
You are provided with a high-resolution photomicrograph of a transverse section through a dicotyledonous stem, labeled Figure 2.1 (representing a specimen under a light microscope).
[ Figure 2.1: Photomicrograph of a Dicotyledonous Stem TS - Magnification \(\times 120\) ]
Outer boundary: Epidermis & Collenchyma
Cortex parenchyma cells
Ring of Collateral Vascular Bundles
(Xylem vessels on inside, Phloem on outside)
Scale Bar: 10 mm on image = 83.3 \(\mu\)m actual
(a) Draw a large, low-power plan diagram of a sector of the stem shown in Figure 2.1. Your diagram should include at least two vascular bundles to show the tissue organization. Do not draw any individual cells. Label the epidermis and the phloem on your diagram. [6 marks]
(b) Focus on the xylem tissue within one vascular bundle in Figure 2.1. Draw a high-power, detailed diagram of three adjacent xylem vessel elements. Use double lines to represent the cell walls. Label the lumen of one vessel and its cell wall. [5 marks]
(c) You are required to calculate the actual maximum width of one vascular bundle shown in Figure 2.1.
- The magnification of the photomicrograph is \(\times 120\).
- Use a ruler to measure the maximum width of one vascular bundle on Figure 2.1. Give your measurement in millimeters (mm).
- Assume your measurement of the vascular bundle on the diagram is 48 mm.
Calculate the actual width of this vascular bundle in micrometers (\(\mu\text{m}\)). Show your working clearly. [5 marks]
(d) Prepare a table to compare the observable structural features of the xylem vessels with those of the phloem sieve tube elements, based on Figure 2.1 and your biological knowledge. [4 marks]
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(a) Low-Power Plan Diagram:
- The diagram should depict a wedge-shaped sector of the stem.
- Outermost layer: Epidermis drawn as a single, thin, continuous layer of uniform width.
- Cortex: A layer of parenchyma tissue between the epidermis and the vascular bundles.
- Vascular bundles: Wedge-shaped or oval bundles arranged in a ring. Within each bundle, a distinct line or boundary should separate the outer phloem from the inner xylem.
- Pith/Medulla: Central region of the stem.
- No individual cells should be drawn in any tissue layer.
- Clear label lines pointing to the outermost cell layer (epidermis) and the tissue located outer/above the xylem within the vascular bundle (phloem).
(b) High-Power Drawing:
- Drawing should contain exactly three adjacent cells.
- The cell walls must be drawn as double lines to represent their thickness, with a clear middle lamella boundary if applicable, and no gaps between adjacent cells (shared walls).
- Cell shape should be angular / polygonal (typical of xylem vessels in cross-section).
- Lumens must be completely empty (no nuclei, cytoplasm, or organelles).
- Labels: Cell wall (pointing to the double boundary) and lumen (pointing to the empty center of a cell).
(c) Calculation:
- Measured image width of vascular bundle = 48 mm.
- Convert mm to micrometers (\(\mu\text{m}\)):
\(48\text{ mm} \times 1000 = 48000\text{ }\mu\text{m}\).
- Magnification = \(\times 120\).
- Using the formula:
\(\text{Actual size} = \frac{\text{Image size}}{\text{Magnification}}\)
\(\text{Actual size} = \frac{48000\text{ }\mu\text{m}}{120} = 400\text{ }\mu\text{m}\).
- Therefore, the actual width of the vascular bundle is 400 \(\mu\text{m}\).
(d) Comparison Table:
Feature
Xylem Vessels
Phloem Sieve Tube Elements
Thickness of cell walls
Thick, lignified secondary cell walls
Thin, non-lignified primary cellulose walls
Cell contents / lumen
Completely hollow lumen, no cytoplasm or organelles (dead cells)
Peripheral cytoplasm, no nucleus or vacuole (living cells)
Relative cell diameter
Larger/wider diameter
Smaller/narrower diameter
End walls
Completely absent / perforated to form continuous tubes
Sieve plates present with sieve pores
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(a) Low-power plan diagram [6 marks maximum]:
- 1 mark for clean, continuous, sharp lines drawn with a sharp pencil (no feathering, no shading).
- 1 mark for drawing a sector of the stem encompassing at least two vascular bundles.
- 1 mark for drawing tissues in correct proportions (e.g., width of cortex proportional to the size of the vascular bundle).
- 1 mark for representing vascular bundles as distinct wedge-shaped or oval areas with a clear boundary line separating xylem and phloem.
- 1 mark for ensuring NO individual cells are drawn anywhere in the entire plan diagram.
- 1 mark for correct labels of 'epidermis' (outermost layer) and 'phloem' (outer region of vascular bundle) with straight, uncrossed label lines.
(b) High-power cell drawing [5 marks maximum]:
- 1 mark for drawing exactly three adjacent xylem vessel cells with no gaps between them.
- 1 mark for drawing all cell walls as clear double lines to represent thickness.
- 1 mark for showing correct polygonal/angular shapes with lumens that are completely empty of any contents.
- 1 mark for drawing cells to a large scale (at least 4 cm across for each cell).
- 1 mark for correct, clear label lines pointing to 'lumen' and 'cell wall'.
(c) Actual size calculation [5 marks maximum]:
- 1 mark for stating the correct relationship: \(\text{Actual size} = \frac{\text{Image size}}{\text{Magnification}}\).
- 1 mark for converting the measured image size from millimeters to micrometers (e.g., \(48\text{ mm} \times 1000 = 48000\text{ }\mu\text{m}\)).
- 1 mark for correct substitution of values: \(\frac{48000}{120}\).
- 1 mark for the correct final numerical answer of 400 (or equivalent based on measurement if slightly different, e.g., 48 mm gives exactly 400).
- 1 mark for expressing the final answer with correct units (\(\mu\text{m}\)) and to appropriate significant figures (2 or 3 significant figures, e.g., 400 or 400.0 is acceptable, reject incorrect units like mm or cm).
(d) Comparison table [4 marks maximum]:
- 1 mark for drawing a structured table with appropriate column/row headers ('Feature', 'Xylem Vessels', 'Phloem Sieve Tube Elements').
- 1 mark for a correct comparison of cell wall thickness (thick/lignified vs thin/cellulose).
- 1 mark for a correct comparison of cell contents or lumen (empty lumen/no cytoplasm vs peripheral cytoplasm present/sieve plates).
- 1 mark for a correct comparison of cell size/diameter (larger/wider lumen vs smaller/narrower lumen).
Paper 42 - A Level Structured Questions
(b) At high temperatures, the rate of photorespiration in C3 plants increases significantly. Explain the biochemical basis of photorespiration and why it reduces the efficiency of photosynthesis in C3 plants. [4]
(c) Some plants, known as C4 plants, have evolved mechanisms to minimize photorespiration. Explain how the leaf anatomy of C4 plants is adapted to reduce photorespiration. [3]
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(b) Describe the detailed intracellular mechanism of action of ADH on the cells of the collecting duct of the kidney nephron. [5]
(c) Explain the effect of ADH on the volume and concentration of urine excreted. [2]
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(b) Brown adipose tissue contains a high concentration of mitochondria that express uncoupling protein 1 (UCP1) in their inner mitochondrial membranes. UCP1 acts as a proton channel, allowing protons to diffuse back into the mitochondrial matrix down their electrochemical gradient.
(i) Explain the effect of UCP1 activity on the rate of ATP synthesis by oxidative phosphorylation. [2]
(ii) Explain how the activity of UCP1 contributes to the thermoregulatory needs of newborn mammals. [3]
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(a) Describe how single-stranded cDNA is prepared from the mRNA of both cancer and normal cells for use in a microarray. [3]
(b) Outline the steps involved in using a microarray chip to compare the gene expression profiles of these two cell types. [5]
(c) Explain how the data obtained from a microarray can be useful in the diagnosis and personalized treatment of cancer. [2]
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(b) Describe the sequence of events that occurs at a cholinergic synapse following the arrival of an action potential at the presynaptic membrane, up to the generation of a new action potential in the postsynaptic neurone. [5]
(c) Outline the role of acetylcholinesterase at a cholinergic synapse and explain the consequences of inhibiting this enzyme. [2]
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(b) Explain how natural selection can lead to the formation of two distinct species of cichlid fish inhabiting different depth zones within the same freshwater lake (sympatric speciation). [5]
(c) State two pre-zygotic isolating mechanisms, other than ecological isolation, that prevent hybridisation between closely related species. [2]
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(a) Define the term *operon* and describe the structure of the *lac* operon, naming its regulatory and structural components. [4]
(b) Explain how the transcription of the structural genes of the *lac* operon is regulated:
(i) when lactose is absent from the growth medium. [3]
(ii) when lactose is present but glucose is absent. [3]
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(i) Explain how auxin (IAA) stimulates cell elongation in plant stems according to the acid growth hypothesis. [5]
(ii) Outline how gibberellin (GA) stimulates the production of amylase during the germination of barley seeds. [3]
(b) State the role of the endosperm and aleurone layer in seed germination. [2]
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(a) Describe how the arrival of an action potential at the presynaptic membrane of a cholinergic synapse results in the release of acetylcholine into the synaptic cleft. [4]
(b) Fasciculin-X is a toxin produced by some species of snakes. It acts as a highly specific, reversible inhibitor of the enzyme acetylcholinesterase.
Explain the effects of Fasciculin-X on the transmission of impulses across the cholinergic synapse. [4]
(c) Compare the structural features of a sensory neurone with those of a motor neurone. [2]
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(b) Fasciculin-X inhibits acetylcholinesterase, the enzyme responsible for hydrolysing acetylcholine into choline and ethanoic acid. As a result, acetylcholine is not broken down and remains in the synaptic cleft. It continuously binds to ligand-gated sodium channels on the postsynaptic membrane, keeping them open. This leads to a continuous influx of sodium ions (\(Na^{+}\)), keeping the postsynaptic membrane permanently depolarised. This prevents the membrane from repolarising, meaning no new action potentials can be generated/transmitted, leading to synaptic transmission block and muscle paralysis or continuous spasm.
(c) A sensory neurone has its cell body situated along the axon (in the dorsal root ganglion), whereas a motor neurone has its cell body at one end of the neurone (within the central nervous system/grey matter). Furthermore, a sensory neurone has a long dendron and a short axon, whereas a motor neurone has short dendrites and a long axon.
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1. Depolarisation of presynaptic membrane causes voltage-gated calcium channels to open. [1]
2. Calcium ions / \(Ca^{2+}\) diffuse into the presynaptic knob/neurone. [1]
3. Calcium ions cause synaptic vesicles (containing acetylcholine) to move towards / fuse with the presynaptic membrane. [1]
4. Acetylcholine is released into the cleft by exocytosis. [1]
Part (b) [Max 4 marks]:
1. Acetylcholine is not hydrolysed / broken down (into choline and ethanoic acid). [1]
2. Acetylcholine remains bound to receptors / ligand-gated sodium channels on the postsynaptic membrane. [1]
3. Sodium channels remain open, resulting in continuous influx of sodium ions / \(Na^{+}\). [1]
4. Postsynaptic membrane remains depolarised / cannot repolarise. [1]
5. No discrete/new action potentials can be generated (causing permanent muscle contraction / spasm / paralysis). [1]
Part (c) [Max 2 marks]:
1. Cell body location: sensory neurone cell body is in the dorsal root ganglion / middle of the neurone, whereas motor neurone cell body is in the CNS / grey matter / at the end of the neurone. [1]
2. Length of fibres: sensory neurone has a long dendron and short axon, whereas motor neurone has short dendrites and a long axon. [1]
(a) Explain the roles of reduced NADP and ATP in the conversion of glycerate 3-phosphate (GP) to triose phosphate (TP) in the Calvin cycle. [3]
(b) Rubisco (ribulose bisphosphate carboxylase-oxygenase) is the enzyme responsible for fixing carbon dioxide in the Calvin cycle. However, Rubisco can also catalyse a reaction with oxygen instead of carbon dioxide, in a process known as photorespiration. This occurs more frequently under high temperatures when stomata are partially closed.
(i) Describe the normal reaction catalysed by Rubisco during carbon fixation. [2]
(ii) Explain why an increase in photorespiration leads to a decrease in the rate of light-independent photosynthesis and a reduced crop yield. [3]
(c) The thylakoid membranes within chloroplasts are organised into grana.
Describe how the structure of grana is adapted to its function in the light-dependent stage of photosynthesis. [2]
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(b) (i) Rubisco catalyses the carboxylation of ribulose bisphosphate (RuBP), which is a 5-carbon sugar. RuBP reacts with carbon dioxide (\(CO_2\)) to form an unstable 6-carbon intermediate, which immediately splits into two molecules of the 3-carbon compound, glycerate 3-phosphate (GP).
(ii) In photorespiration, oxygen acts as a competitive inhibitor and binds to the active site of Rubisco instead of \(CO_2\). This means less RuBP is carboxylated, resulting in a lower production of GP and subsequently less TP. Consequently, there is less TP available to regenerate RuBP and less TP available to synthesise organic macromolecules such as glucose, sucrose, and starch. Additionally, ATP and reduced NADP are consumed in photorespiration without producing sugars, making photosynthesis less efficient and reducing the overall biomass and crop yield.
(c) The thylakoid membranes are stacked into grana to provide a very large surface area. This large surface area accommodates a high density of photosystems (containing chlorophyll and accessory pigments) for maximum light absorption. The membranes also contain components of the electron transport chain and ATP synthase complexes, and they act as a barrier to maintain a proton gradient necessary for chemiosmosis / photophosphorylation.
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1. ATP hydrolysed to release energy / provides energy (for the conversion). [1]
2. Reduced NADP provides hydrogen / electrons / acts as a reducing agent. [1]
3. To reduce glycerate 3-phosphate (GP) to triose phosphate (TP). [1]
Part (b)(i) [Max 2 marks]:
1. Ribulose bisphosphate (RuBP) / 5C compound combines with carbon dioxide / \(CO_2\). [1]
2. To produce two molecules of glycerate 3-phosphate (GP) / 3C compound (via an unstable 6C intermediate). [1]
Part (b)(ii) [Max 3 marks]:
1. Oxygen acts as a competitive inhibitor / competes with \(CO_2\) for the active site of Rubisco. [1]
2. Less RuBP reacts with carbon dioxide / less GP is produced. [1]
3. Less triose phosphate (TP) is formed to regenerate RuBP / less TP to make glucose / starch / organic molecules. [1]
4. Energy (ATP) and reducing power (reduced NADP) are wasted / used inefficiently, lowering crop biomass. [1]
Part (c) [Max 2 marks]:
1. Grana / stacked thylakoids provide a large surface area for the attachment of photosystems / light-harvesting complexes / photosynthetic pigments. [1]
2. Large surface area holds many electron carriers / electron transport chains / ATP synthase molecules. [1]
3. Thylakoid membranes are impermeable to protons, allowing a proton gradient to be established between the thylakoid lumen and the stroma. [1]
Paper 52 - Planning, Analysis and Evaluation
(a) Describe a method the student could use to carry out this investigation. Your description should include:
- how the independent variable is varied
- how key confounding variables are controlled
- how the mitochondria are isolated and kept active
- how the rate of oxygen consumption is measured
- how safety and reliability are ensured. [10]
(b) Outline how the student could extend this investigation to determine whether sodium malonate acts as a competitive or a non-competitive inhibitor of succinate dehydrogenase. [3]
(c) Predict and explain the effect of adding an excess concentration of succinate on the rate of oxygen consumption in the presence of sodium malonate, assuming malonate is a competitive inhibitor. [2]
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1. Independent variable: Prepare at least five different concentrations of sodium malonate (e.g., 0.0, 0.2, 0.4, 0.6, 0.8, and 1.0 mol dm^{-3}) by performing simple dilution of a 1.0 mol dm^{-3} stock solution with distilled water.
2. Mitochondrial isolation: Homogenize germinating mung beans in ice-cold, isotonic phosphate buffer (pH 7.4). The cold temperature prevents enzyme denaturation, the buffer maintains pH, and the isotonic medium prevents the osmotic lysis or shrinkage of mitochondria. Filter the homogenate through muslin to remove intact cells and cell walls. Centrifuge the filtrate at a low speed (e.g., 1000g) for 5 minutes, discard the pellet (nuclei and heavy debris), then centrifuge the supernatant at a high speed (e.g., 10000g) for 15 minutes to pellet the mitochondria. Resuspend this mitochondrial pellet in fresh ice-cold isotonic buffer and store on ice.
3. Control of variables: Keep temperature constant (e.g., 25\u00b0C) using a thermostatically controlled water bath. Keep the volume and concentration of the respiratory substrate (succinate) constant in each test (e.g., 1.0 cm^{3} of 0.1 mol dm^{-3} succinate).
4. Measurement: Set up a sealed reaction vessel (to prevent atmospheric oxygen from dissolving in the mixture). Add standard volumes of mitochondrial suspension, buffer, succinate, and the selected concentration of sodium malonate. Insert the dissolved oxygen probe connected to a data logger. Record the oxygen concentration (%) at regular intervals (e.g., every 10 seconds) for 5 minutes. Calculate the initial rate of oxygen consumption from the linear gradient of the oxygen concentration against time graph.
5. Reliability & Safety: Repeat the measurement at least three times for each concentration of sodium malonate, identify any anomalous results, and calculate a mean rate. Wear eye protection, gloves, and a lab coat as sodium malonate is an irritant, and keep electrical equipment (data loggers/probes) away from water baths.
(b) Extension to determine inhibition type:
1. Measure the rate of oxygen consumption at a range of different succinate (substrate) concentrations (e.g., 0.02 to 0.2 mol dm^{-3}).
2. Carry out this process twice: once in the absence of sodium malonate, and once in the presence of a fixed, constant concentration of sodium malonate.
3. Plot a graph of the rate of oxygen consumption against succinate concentration for both treatments. If the maximum rate (Vmax) is the same in both treatments (achieved at higher succinate concentrations), the inhibition is competitive. If the maximum rate (Vmax) is significantly lower in the presence of the inhibitor, the inhibition is non-competitive.
(c) Prediction and explanation:
- Prediction: The rate of oxygen consumption will increase and eventually return to the maximum uninhibited rate (Vmax).
- Explanation: Since sodium malonate is a competitive inhibitor, it binds reversibly to the active site of succinate dehydrogenase. An excess concentration of succinate greatly increases the probability of a succinate molecule binding to the active site rather than a malonate molecule, successfully outcompeting the inhibitor.
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- 1. Dilution: Describes simple or serial dilution of stock sodium malonate to produce at least 5 different concentrations (must state example concentrations or volumes).
- 2. Homogenisation: Homogenise mung beans in ice-cold, isotonic, and buffered solution.
- 3. Isolation Role: Explains that ice-cold temperature prevents denaturation / isotonicity prevents osmotic lysis of mitochondria / buffer maintains optimum pH for enzymes.
- 4. Centrifugation: Centrifuge homogenate at low speed first to remove cell debris/nuclei, then supernatant at high speed to pellet mitochondria.
- 5. Standardisation (Temp): Keep temperature constant using a water bath.
- 6. Standardisation (Substrate): Keep the concentration and volume of succinate constant.
- 7. Sealed Vessel: Use a sealed reaction chamber to prevent oxygen entering from the air.
- 8. Detection: Use a dissolved oxygen probe and data logger to record oxygen concentration.
- 9. Rate Calculation: Find the rate by calculating the initial gradient of the oxygen concentration vs. time curve.
- 10. Reliability: Repeat each concentration at least 3 times and calculate the mean / identify anomalies.
- 11. Safety: Goggles / gloves / lab coat because sodium malonate is an irritant / safety precaution with electrical equipment near water.
(b) [Max 3 marks]
- 1. Measure the rate of oxygen consumption across a range of different succinate (substrate) concentrations.
- 2. Repeat this in the presence of a constant concentration of sodium malonate and in its absence (control).
- 3. Compare the maximum rate (Vmax) reached: if Vmax is unchanged at high substrate concentrations, it is competitive; if Vmax is reduced, it is non-competitive.
(c) [Max 2 marks]
- 1. (Prediction) The rate of oxygen consumption increases / returns to maximum / Vmax is restored.
- 2. (Explanation) Succinate and malonate compete for the same active site on succinate dehydrogenase; high substrate concentration increases the frequency of substrate-active site collisions, outcompeting the inhibitor.
At each site, the ecologist collected 15 leaves from 15 different plants of *P. maritima*.
The stomatal density (number of stomata per \( \text{mm}^2 \)) was determined for each leaf using clear nail varnish impressions viewed under a light microscope. The results are summarized below:
- Site A (sheltered): Mean = \( 124.6 \text{ stomata mm}^{-2} \), Standard Deviation (\( s_1 \)) = \( 12.3 \), \( n_1 = 15 \)
- Site B (exposed): Mean = \( 141.2 \text{ stomata mm}^{-2} \), Standard Deviation (\( s_2 \)) = \( 16.8 \), \( n_2 = 15 \)
(a) Describe a method the ecologist could use to ensure that the selection of leaves at each site was random and representative. [3]
(b) State a null hypothesis for this investigation. [1]
(c) Calculate the value of the Student's t-test using the formula:
\( t = \frac{|\bar{x}_1 - \bar{x}_2|}{\sqrt{\frac{s_1^2}{n_1} + \frac{s_2^2}{n_2}}} \)
Show your working and give your answer to two decimal places. [3]
(d) Calculate the degrees of freedom for this test. Using the critical values table below, state whether the difference in means is significant at the \( p < 0.05 \) probability level, and explain how you reached your conclusion. [5]
Critical values table for Student's t-test at \( p = 0.05 \):
- \( \text{df} = 26 \): 2.056
- \( \text{df} = 28 \): 2.048
- \( \text{df} = 30 \): 2.042
(e) Suggest one physiological reason why a higher stomatal density might be advantageous to *P. maritima* in an exposed coastal environment. [3]
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1. Establish a sampling grid at each site using two long tape measures laid out at right angles to each other (e.g., 20 m x 20 m grid).
2. Generate random coordinates using a computer random number generator or calculator to remove investigator bias. Avoid 'throwing' quadrats.
3. To ensure representativeness, standardise the leaf selected from each plant located at the coordinates (e.g., always select the third fully expanded leaf from the base of the plant of a similar size/age/height).
(b) Null hypothesis:
There is no significant difference between the mean stomatal density of *Plantago maritima* leaves at Site A (sheltered) and Site B (exposed).
(c) Calculations:
1. Difference in means: \( |\bar{x}_1 - \bar{x}_2| = |124.6 - 141.2| = 16.6 \)
2. Variance terms:
\( s_1^2 = 12.3^2 = 151.29 \) and \( \frac{s_1^2}{n_1} = \frac{151.29}{15} = 10.086 \)
\( s_2^2 = 16.8^2 = 282.24 \) and \( \frac{s_2^2}{n_2} = \frac{282.24}{15} = 18.816 \)
3. Sum of variances: \( 10.086 + 18.816 = 28.902 \)
4. Square root of sum: \( \sqrt{28.902} \approx 5.376 \)
5. Calculate t-value: \( t = \frac{16.6}{5.376} = 3.0878 \approx 3.09 \)
(d) Degrees of freedom & Significance:
1. Degrees of freedom: \( \text{df} = (n_1 - 1) + (n_2 - 1) = (15 - 1) + (15 - 1) = 28 \).
2. Critical value: The critical value at \( \text{df} = 28 \) for \( p = 0.05 \) is 2.048.
3. Comparison: The calculated t-value (3.09) is greater than the critical value (2.048).
4. Significance: Because the calculated t-value exceeds the critical value, the difference in mean stomatal density between the two populations is statistically significant.
5. Conclusion: There is a less than 5% probability (\( p < 0.05 \)) that this difference is due to chance alone. The null hypothesis is rejected.
(e) Physiological advantage of higher stomatal density:
In an exposed coastal environment, wind speeds are high, which rapidly removes the boundary layer of water vapour from around the leaves, potentially causing high transpiration rates but also maintaining a steep diffusion gradient for carbon dioxide. A higher stomatal density allows the plant to rapidly take in carbon dioxide for photosynthesis during brief periods when conditions are favorable (e.g., when the air is humid or when the plant has sufficient water to keep stomata fully open). Rapid photosynthesis allows the plant to synthesize organic compounds quickly to withstand stress, and the stomata can be shut tightly during dry, highly windy periods to prevent excessive water loss.
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- 1. Method of randomisation: Use of tape measures at right angles to construct a grid and generating random coordinates using a random number generator.
- 2. Location: Select the plant closest to the generated coordinates.
- 3. Standardisation: Select leaves of the same age / position on the plant / leaf size to ensure the samples are representative.
(b) [1 mark]
- 1. States that there is no significant difference between the mean stomatal density of *Plantago maritima* at Site A and Site B / sheltered and exposed sites.
(c) [Max 3 marks]
- 1. Calculates the numerator correctly as 16.6.
- 2. Calculates the denominator correctly as 5.38 (or \( \sqrt{28.90} \)).
- 3. Final calculated t-value of 3.09 (accept 3.08 to 3.10 based on rounding steps, but must be to 2 decimal places as requested).
(d) [Max 5 marks]
- 1. States degrees of freedom = 28.
- 2. Correctly identifies the critical value of 2.048.
- 3. States that the calculated t-value is greater than the critical value.
- 4. Concludes that the difference is statistically significant (or that the probability of the difference occurring by chance is less than 0.05 / 5%).
- 5. States that the null hypothesis is rejected.
(e) [Max 3 marks]
- 1. Wind removes the boundary layer of water vapour, promoting high diffusion rates.
- 2. A higher stomatal density maximizes carbon dioxide uptake for photosynthesis when stomata are open / when water is available.
- 3. Enables high transpiration pull to transport essential mineral ions from nutrient-poor / saline coastal soils.
- 4. Facilitates rapid gas exchange during brief periods of stomatal opening, allowing quick closing to prevent dehydration in high winds.
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