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Checkpoint answers for topic 6 Q6.1 Draw up a flow chart that outlines the steps required in DNA amplification and separation to produce a DNA profile. DNA sample obtained DNA fragments created either using restriction enzymes or the polymerase chain reaction. The fragments contain short tandem repeated sequences. Restriction enzymes cut the DNA at specific base sequences on either side of the STR sequence. PCR fragments are created by the binding of primers (DNA complementary to the DNA adjacent to the STR sequence); the action of DNA polymerase and a cycle of temperature changes results in the copying of the STR sequence. Fragments of DNA separated according to their size using gel electrophoresis. Smaller fragment pass more quickly through the gel towards the positive electrode. If restriction enzymes have been used to create the fragments these double stranded fragments are digested to form single strands and transferred to a nitrocellulose or nylon membrane by Southern blotting. The membrane is incubated with a radioactive or fluorescent probe, the probes DNA sequence attaches to the DNA fragment. The probes position on the membrane can be visualised using either an X-ray film or ultraviolet light. If PCR has been used to create the fragments, the primers have fluorescent tags which can be detected using a laser scanner, this is usually highly automated. Q6.2 Produce a bullet-point summary of methods that can be used to determine time of death.  Body temperature: human core body temperature is normally in the range 36.2–37.6 °C, but almost as soon as a person dies their body starts to cool. Using a cooling curve the temperature of the body can be used to estimate the time of death. This assumes that the person’s temperature was normal, 37 °C, at the time of death. The estimate may be somewhat imprecise due to the many factors that can affect post-mortem cooling, such as body size, body position, clothing, air movement and humidity. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 1 of 8  Rigor mortis: after death, muscles usually totally relax and then stiffen; this stiffening is known as rigor mortis. There is a progression in the development of rigor mortis, with smaller muscles stiffening before larger ones. The rigor mortis eventually passes in the same order in which it developed as the muscles start to break down. Most bodies will have complete rigor mortis between 6 and 9 hours after death and the degree of rigor mortis gives an estimate of the length of time since death. However, environmental factors can affect how quickly rigor mortis develops and so the estimate is not likely to be very precise.  Decomposition: the degree of decomposition due to the breakdown of tissues by autolysis can be used to estimate time of death. There is a sequence of characteristic changes that the body goes through during decomposition. The rate of these changes will be affected by the external conditions of the body but if these are known it is possible to estimate for how long the body has been decomposing.  Forensic entomology: the presence of insects allows a forensic entomologist to make a more precise estimate of how much time has elapsed since death. If maggots are found on the body, their stage of development gives an estimate of how long ago the person may have died assuming that the eggs were laid on the body soon after death. If there is more than one species of insect present on the body it is possible to determine the stage of succession that the body has reached. The number of species present increases as succession progresses. The length of each stage in the succession depends on the condition of the body, which in turn depends on environmental conditions. Q6.3 Draw up a table of comparison to allow you to distinguish between the structure of bacteria and that of viruses. Feature Bacteria Viruses Type of cell Prokaryotic cell Size 0.5–5 µm in diameter Structure Contain cytoplasm, a long circular strand of DNA, plasmids, cell surface membrane, free ribosomes, and have a rigid cell wall containing polysaccharide murein. May have mesosomes (infoldings of the cell Viruses are small organic particles with a much simpler structure than bacteria. No proper cell structure. 20–400 nm in length; a wide range of sizes and shapes. A strand of nucleic acid (RNA or DNA) enclosed within a protein coat. May have an outer envelope taken from the host cell surface membrane but containing glycoproteins from the virus. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 2 of 8 Reproduction surface membrane where respiration occurs), flagella and pili. Some have a capsule outside the cell wall. Reproduce asexually by binary fission; after replication of the DNA they divide into identical daughter cells. No sexual reproduction occurs in animals, but cell-to-cell contact through conjugation. Enter the cells of the organisms they infect (the host) and use the host’s metabolic systems to make more viruses. The virus’s genetic material is replicated and then the protein coat synthesised. Newly formed virus particles are released by budding from the cell surface or lysis of the cell. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 3 of 8 Q6.4 Produce a flowchart showing the sequence of events that occurs in the non-specific response at the site of a cut. Damaged basophil white blood cells and mast cells release histamine. Arterioles in the area dilate and venules constrict due to the presence of histamine. Blood flow in the capillaries at the infected site increases. The site of the cut becomes red. It also becomes warm due to increased metabolic activity. Capillaries become leaky as cells in the capillary walls separate slightly. Plasma fluid, white blood cells and antibodies leak from the blood into the tissue causing oedema (swelling). Neutrophils, a type of white blood cell, engulf bacteria. Each neutrophil engulfs between 5 and 20 bacteria before becoming inactive and dying. Macrophages, another type of white blood cell, engulf bacteria, debris from damaged cells and foreign matter. Pus collects at the site of the cut; this thick white liquid is made up of dead white blood cells. The pus gradually breaks down and is absorbed into the surrounding tissues. If viruses have infected cells at the side of the cut, the cells produce the protein interferon. This diffuses to the surrounding cells where it prevents viruses from multiplying by inhibiting viral protein synthesis. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 4 of 8 Q6.5 Sketch a diagram or use downloaded figures from the mediabank book artwork to show how Figures 6.36, 6.38A, 6.38B and 6.39 are interconnected. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 5 of 8 Q6.6 Produce a flowchart which summarises the course of disease for TB and for AIDS. TB Tuberculosis (TB) is a contagious disease caused by the bacterium Mycobacterium tuberculosis. Respiratory or pulmonary TB is the most common form. Infection may occur when M. tuberculosis bacteria are inhaled and lodge in the lungs. Here they start to multiply. The first phase (primary infection) can last for several months; it may have no symptoms. An inflammatory response by the host’s immune system occurs. Macrophages engulf the bacteria. Anaerobic tissue masses known as a granuloma or tubercules form in response to infection. They contain dead bacteria and macrophages at their centre. After 3–8 weeks, the infection is controlled and the infected region of the lung heals. Some M. tuberculosis bacteria may survive inside macrophages. The bacteria have very thick waxy cell walls, making destruction inside the macrophages very difficult. The bacteria can lie dormant for years. The second phase (active tuberculosis) occurs if there are too many bacteria for the immune response to deal with, or if an old infection breaks out because the immune system is not working properly. The bacteria multiply rapidly and destroy the lung tissue, creating holes or cavities. The lung damage will eventually kill the sufferer if they are not treated with an appropriate antibiotic. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 6 of 8 AIDS AIDS, acquired immune deficiency syndrome, is caused by infection with the human immunodeficiency virus, HIV. HIV infection occurs when the body fluid (blood, vaginal secretions and semen, but not saliva or urine) of an infected person is transferred directly into the body of an uninfected person. This can occur through unprotected sex. This can occur when sharing needles, whether used illegally or legally. This can occur with direct blood-to-blood transfer through cuts and grazes. This can occur from mother to child across the placenta or in breast milk. HIV invades T helper cells and macrophages. The HIV gp120 molecules attach to their CD4 receptors allowing the virus envelope to fuse with the host cell surface membrane, enabling the viral RNA to enter the cell. Once inside, the virus uses reverse transcriptase to produce DNA from its RNA. The DNA is integrated into the host’s DNA by another HIV enzyme, integrase. The viral DNA is transcribed and translated to produce new viral proteins and assemble new viruses. The new virus particles bud out of the T cell, taking some of the surface membrane with them as their envelope, and killing the cell as they leave. When a person is first infected by HIV, there is an acute phase of infection. There is rapid replication of the virus and loss of T helper cells. As the number of viruses increases, the number of host T helper cells decreases. Macrophages, B cells and T killer cells are not activated and the infected person’s immune system becomes deficient. The infected person may experience symptoms such as fever, sweats, headache, sore throat and swollen lymph nodes, or they may have no symptoms. The virus continues to reproduce rapidly, but the numbers are kept in check by the immune system. T killer cells recognise the infected T helper cells and destroy them. There may be no symptoms during this chronic phase, but there can be an increasing tendency to suffer various infections which are slow to go away. Dormant diseases such as TB and shingles can reactivate. The chronic phase can last for years, especially if combined with drug treatment. An increased number of viruses in circulation (viral load) and a declining number of T helper cells indicate the onset of AIDS, the disease phase. The weakened immune system makes the patient more prone to opportunistic infections such as pneumonia and TB. There may also be significant weight loss, dementia (memory and intellect loss) and the cancer Kaposi’s sarcoma. AIDS is usually fatal. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. Page 7 of 8 Q6.7 Write a definition for each of the four types of immunity: passive natural immunity, active natural immunity, active artificial immunity and passive artificial immunity. Passive natural immunity occurs when antibodies pass from a mother to baby either across the placenta before birth, or via breast milk after birth. This helps protect the baby for a short time. Active immunity occurs when the body makes its own antibodies in response to an antigen. Active natural immunity develops following an infection. The ‘specific immune response’ to the foreign antigens produces a supply of antibodies and B memory and T memory cells that will respond quickly if the body is reinfected with the same pathogen. Active artificial immunity develops following immunisation. Antigens in the vaccine trigger a ‘specific immune response’ by the body’s B and T cells and antibodies are produced. Vaccines usually contain weakened (attenuated) forms of the virus or bacterium. These stimulate the immune system to produce the antibodies but do not cause the disease. For some infections dead pathogens or antigen fragments may be used to trigger the response. Passive artificial immunity develops when a person is given ready-made antibodies in a medical procedure. This method provides immediate protection in emergency situations but is short lived. The protein antibodies are broken down after a few days or weeks. Salters-Nuffield Advanced Biology, Pearson Education Ltd 2009. ©University of York Science Education Group. This sheet may have been altered from the original. 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