Q1. (A) Describe and Explain How Cell Fractionation and Ultracentrifugation Can Be Used To Isolate
Q1. (A) Describe and Explain How Cell Fractionation and Ultracentrifugation Can Be Used To Isolate
Q1.(a)     Describe and explain how cell fractionation and ultracentrifugation can be used to isolate
            mitochondria from a suspension of animal cells.
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      (b)     Describe the principles and the limitations of using a transmission electron microscope to
             investigate cell structure.
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Q2.          (a)     Small samples of plant tissue were placed in a cold, isotonic solution and then treated
               to break open the cells to release the organelles. The different organelles were then
               separated. Describe a technique that could be used to
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      (b)     One group of organelles was placed in a hypotonic solution. The diagram shows one of
             these organelles seen under an electron microscope before and after it was placed in the
             hypotonic solution.
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(Total 3 marks)
            In a human, there are over 200 different types of cell clearly distinguishable from each other.
            What is more, many of these types include a number of different varieties. White blood cells,
            for example, include lymphocytes and granulocytes.
              Although different animal cells have many features in common, each type has adaptations.
      5       associated with its function in the organism. As an example, most cells contain the same
              organelles, but the number may differ from one type of cell to another. Muscle cells contain
              many mitochondria, while enzyme-secreting cells from salivary glands have particularly large
              amounts of rough endoplasmic reticulum.
              The number of a particular kind of organelle may change during the life of the cell. An
      10      example of this change is provided by cells in the tail of a tadpole. As a tadpole matures into
              a frog, its tail is gradually absorbed until it disappears completely. Absorption is associated
              with an increase in the number of lysosomes in the cells of the tail.
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Use information from the passage and your own knowledge to answer the following questions.
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           (ii)     rough endoplasmic reticulum and enzyme-secreting cells from salivary glands
                   (lines 7 - 8).
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    (b)     Use information in the passage to explain how a tadpole’s tail is absorbed as a tadpole
           changes into a frog.
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    (c)     Starting with some lettuce leaves, describe how you would obtain a sample of
           undamaged chloroplasts. Use your knowledge of cell fractionation and ultracentrifugation
           to answer this question.
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                                                                                                                                     (Total 13 marks)
 Q4.          Liver was ground to produce a homogenate. The diagram shows how fractions containing
      different cell organelles were produced from the filtered homogenate.
(a) Explain why the homogenate was filtered before spinning at low speed in the centrifuge.
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      (b)     The main organelles present in sediment B were mitochondria. Suggest the main
             organelles present in
(i) sediment A;
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(ii) sediment C.
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(c) What property of cell organelles allows them to be separated in this way?
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      (d)     Explain why the organelles in sediment C could be seen with a transmission electron
             microscope but not with an optical microscope.
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Q5.(a)     Describe how you could make a temporary mount of a piece of plant tissue to observe the
            position of starch grains in the cells when using an optical (light) microscope.
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                                                                             © Science Photo Library
(b) Give the name and function of the structures labelled W and Z.
Name of W .......................................................................................................
Function of W ...................................................................................................
Name of Z ........................................................................................................
         Function of Z ....................................................................................................
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    (c)     A transmission electron microscope was used to produce the image in the figure above.
           Explain why.
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(d) Calculate the magnification of the image shown in the figure in part (a).
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Q6. The diagram shows how some organelles may be distinguished from each other.
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(i) is a ribosome;
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      (c)     Some liver tissue was ground, filtered and centrifuged to make a suspension of
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            organelle D.
(i) Explain why the solution in which the liver tissue was ground should be ice-cold.
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             (ii)     The ground liver was centrifuged at low speed. The pellet that formed at the bottom
                     of the centrifuge tube was thrown away and the supernatant centrifuged again at
                     higher speed. Explain why it was necessary to first centrifuge the ground liver at low
                     speed in order to obtain a suspension of organelle D.
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Q7. (a) Name the type of bond that joins amino acids together in a polypeptide.
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      (b)     The cytoplasm at F contains amino acids. These amino acids are used to make proteins
             which are secreted from the cell.
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         Place the appropriate letters in the correct order to show the passage of an amino acid
         from the cytoplasm at F until it is secreted from the cell as a protein at K.
 
F K
(2)
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    (d)     A group of scientists homogenised pancreatic tissue before carrying out cell fractionation
           to isolate organelle G.
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                                                                                                                                   (Total 10 marks)
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Q8.A stomach ulcer is caused by damage to the cells of the stomach lining. People with stomach
     ulcers often have the bacterium Helicobacter pylori in their stomachs.
         A group of scientists was interested in trying to determine how infection by H. pylori results in
         the formation of stomach ulcers.
         The table below shows the substances released by each of these strains.
 
                                       Substances released by the H. pylori cells
             H. pylori strain
                                         Toxin            Enzyme that neutralises acid
A
B
C
         The scientists centrifuged the cultures of each strain to obtain cell-free liquids. They added each
         liquid to a culture of human cells. They then recorded the amount of damage to the human
         cells.
Their results are shown below. The error bars show ± 1 standard deviation.
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    (a)     Describe and explain how centrifuging the culture allowed the scientists to obtain a cell-
           free liquid.
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    (b)     The scientists measured cell damage by measuring the activity of lysosomes.
           Give one function of lysosomes.
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    (d)     What do these data suggest about the damage caused to human cells by the toxin and
           by the enzyme that neutralises acid?
           Explain your answer.
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    (e)     The scientists carried out a further investigation. They treated the liquid from strain A with
            a protein-digesting enzyme before adding it to a culture of human cells. No cell damage
            was recorded.
            Suggest why there was no damage to the cells.
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                                                                                                                                      (Total 12 marks)
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            5.      Centrifuge (at lower speed / 1000 g) to separate nuclei / cell fragments / heavy
                    organelles;
                          5. Ignore incorrect numerical values.
      (b)     Principles:
              1.      Electrons pass through / enter (thin) specimen;
              2.      Denser parts absorb more electrons;
              3.      (So) denser parts appear darker;
              4.      Electrons have short wavelength so give high resolution;
                            Principles:
                            Allow maximum of 3 marks
            Limitations:
            5.      Cannot look at living material / Must be in a vacuum;
            6.      Specimen must be (very) thin;
            7.      Artefacts present;
            8.      Complex staining method / complex / long preparation time;
            9.      Image not in 3D / only 2D images produced.
                          Limitations:
                          Context of limitation must be clear, not simply explaining how TEM
                          works
                          E.g “allows you to see organelles as a thin section is used” is not
                          a limitation
                          Allow maximum of 3 marks
                          Ignore ref to colour
                                                                                                       5 max
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       (b)     (i)      nuclei;
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(ii) Photosynthesis;
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                     Uses light (energy);
      (b)     (i)      A;
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             (ii)     C;
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             Release ATP / energy for active transport / transport against the concentration
             gradient / protein synthesis / exocytosis;
                         Q Reject: anaerobic respiration
                         Q Reject: produces / makes energy
                         Accept: produces ATP for energy
                         Reject: produces ATP for respiration
                         Neutral: protein secretion
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      (d)     (i)      Breaks open cells / disrupts cell membrane / releases cell contents / releases
                       organelles / break up cells;
                            Reject: breaks down cell wall
                            Neutral: separates the cells
                            Reject: breaks up cells so they can be separated
                            Reject: breaks up / separates organelles
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            (iv)    Prevents osmosis / no (net) movement of water / water does not enter
                    organelle / water does not leave organelle;
                  So organelle / named organelle is not damaged / does not burst / does not
                  shrivel;
                        Neutral: ref. to water potential
                        Q Ref. to cells rather than organelles negates the second mark
                        only
                        Reject: ref. to turgid / flaccid for second mark
                        Reject: organelle ‘explodes’ for second mark
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          2.      Some cell damage when either there on their own / some cell damage in B
                  and C;
                       MP1 and MP2 − figures given from the graph are insufficient.
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