Topic 6 - Infection, Immunity And Forensics Flashcards
What are the uses of DNA profiling?
- Identifying people from remains
- Determining family relationships - father, mother
- Solving crimes - placing people at crime scenes
How many points must be identified for identical fingerprints to match?
16
What is another way to identify individuals other than fingerprints?
Dental records
What are introns?
(Intragenic regions) non-coding blocks
What are exons?
(Expressed regions) Coding regions
What are short tandem repeats (STRs)?
A sequence of repeated bases. A short tandem repeat can contain from 2 to 50 base pairs which can be repeated from 5 to several hundred times.
Where are short tandem repeats found?
Within introns of the DNA.
What is a maternal chromosome?
Chromosome inherited from the mother
What is the paternal chromosome?
Chromosome inherited from the father.
Why are two individuals unlikely to have the same combination of STRs?
Each person has a large number of introns with lots of STR loci. There is a large variation in the number of repeats of each loci.
An STR occurs at the same place on a chromosome of a homologous pair. What’s the difference between them?
The number of times they are repeated on each homologous chromosome can be different.
Describe the process of how DNA is obtained?
1) DNA sample must be obtained from almost all biological tissue (animal or plant) -> check swab/ white blood cells/ bone marrow/ sperm.
2) Tissue sample is physically broken down into a buffer solution that includes salt and a detergent solution to disrupt the the cell membranes.
3) The small suspended particles, including the DNA, are separated from the rest of the cell debris by filtering or centrifuging.
4) Protease enzymes are incubated with the suspension to remove proteins and then cold ethanol is added to precipitate out the DNA.
5) Several stages of washing the DNA in a buffer solution follow.
What are the two ways in which fragments are made?
- Restriction enzymes
- Polymerase chain reaction
Describe how restriction enzymes are used to create fragments?
Restriction enzymes (endonucleases) cut DNA at specific base sequences (usually 4 to 6 base pairs). Restriction sites are either side of STR sequence and fragment of DNA is cut away from the rest of the genome. Restriction enzymes cut a DNA into fragments only where their specific restriction sequence occurs.
What happens if the same restriction enzyme is used to cut two identical DNA samples?
Identical STR fragments are reproduced.
Describe how the polymerase chain reaction (PCR) creates a DNA fragment?
The forensic sample is placed in a reaction tube with DNA polymerase, DNA primers (with fluorescent tags) and nucleotides. The PCR machine goes under a cycle of temperature changes. Firstly, it is 90 degrees Celsius during which the DNA strands separate into two. It then becomes 55 degrees celsius during which the primers attach at the start of the STR sequence. Lastly, it becomes 70 degrees celsius during which the DNA polymerase attaches. The nucleotides are added, extending the DNA from the primer. The STR repeated sequence and DNA adjacent is replicated. As the cycle continues, huge numbers of the target DNA fragments are produced.
What are the DNA samples generally analysed for?
The presence of ten short tandem repeats (STRs/ micro-satellites)
How can you determine the gender from a DNA sample?
A specific primer can be used to determine the gender as it targets the sex chromosome.
Describe the process of gel electrophoresis.
1) DNA is cut into fragments.
2) Fragments of double stranded DNA are loaded into wells of an agarose gel in a tank (provides a stable medium through which fragments can move).
3) Electrodes are connected which produce a potential difference over the gel. The negatively charged DNA moves towards the positive electrode. The fragments separate into invisible bands. They move depending on their size and charge. Smaller fragments end up closer to the positive electrode.
4) DNA is transferred to a nylon or nitrocellulose membrane by solution drawn up through the gel. DNA double strands split and stick to the membrane.
5) Membrane placed in a bag with a DNA probe. Single stranded DNA probe binds to fragments with a complementary sequence.
6) If the DNA probe is radioactive, x-ray film is used to detect the fragments. If the DNA probe is fluorescent it is viewed using UV light as shown above.
What is the benefit of DNA primers with fluorescent tags?
As the DNA fragments move through the gel they pass a laser and the dye in the fluorescent tag is detected, effectively giving a time that it has taken for the fragments to pass through the gel.
What is the size of the fragment determined by?
Number of base pairs it contains.
How does the enzyme used in the polymerase chain reaction not denature?
DNA tacpolymerase is stable because it is from extreme environments (hot springs) and therefore can withstand high temperatures.
What controls the temperature and changes when it is necessary in the Polymerase chain reaction?
A thermocycler
What do DNA primers attach to?
They bind to the complementary base pairs
What are the methods used to determine the time of death?
- Body temperature
- Rigor mortis
- Decomposition
- Forensic entomology
What is the human bodies usual temperature?
36.2 to 37.6 degrees celsius
Why does the body temperature begin to decrease as soon as the person dies?
Absence of heat producing chemical reactions
For how long is temperature measured after the death of a person?
24hrs
How is the core body temperature recorded?
Via the rectum or an abdominal stab with a long thermometer (sometimes electrical)
When measuring the body temperature what must be noted?
The environmental conditions
Why is it important to note the environmental conditions of where the body was found?
Because the environmental conditions affect the rate at which body temperature changes. In colder environments the faster the core body temperature decreases.
On a graph what does the shape of the temperature follow?
A sigmoid curve (s-shaped)
Why is the original core temperate before death not always 37 degrees celsius?
Their body temperature at the time of death could have been elevated or depressed if they were suffering from fevers or hypothermia.
What are the factors that affect post-mortem cooling?
- Body size
- Body position
- Clothing
- Air movement
- Humidity
- Temperature of surroundings
- If in water will cool more rapidly
What is rigor mortis?
The stiffening of the body after death because of a loss of Adenosine Triphosphate (ATP) from the body’s muscles.
What does the position where the joints become fixed depend on?
Depends on the body position.
Describe the process of rigor mortis.
1) After death, muscle cells become starved of oxygen and oxygen-dependent reactions stop.
2) Respiration in the cells becomes anaerobic and produces lactic acid.
3) The pH of the cells falls, inhibiting enzymes and thus inhibiting anaerobic reactions.
4) The ATP needed for muscle contractions is no longer produced. As a result bonds between the muscle proteins become fixed.
5) The proteins can no longer move over one another to shorten the muscles, fixing the muscles and joints.
Regarding rigor mortis, which muscles stiffen first?
Smaller muscles stiffen before larger ones.
When does rigor mortis occur?
6 to 9 hrs after death
When does rigor mortis set in more early?
Sets in more quickly and lasts for a shorter period of time if the environmental temperature is high or if the person has been physically active before death.
What is decomposition of the body?
Occurs after death when the tissues start to break down.
What is autolysis?
Body’s own enzymes, from the digestive tract and from lysosomes, break down cells.
As well as autolysis, how does decomposition also occur?
Bacteria from the gut and gaseous exchange systems rapidly invade the tissue after death releasing enzymes that result in decomposition.
What are the signs of decomposition?
- Greenish discolouration of the skin and lower abdomen due to the formation of sulfhaemoglobin in the blood. This then spreads across the rest of the body going from a darken to reddish-green and then to a purple-black colour.
- Gas or liquid blisters on the skin.
- Bacteria and gases (hydrogen sulphide, methane, carbon dioxide, ammonia and hydrogen) cause the body to smell and become bloated.
- As the tissue further decomposes, the gas is released and the body deflates. Fluid associated with putrefaction drain away, the soft tissues shrink and the decay rate of the dry body is reduced.
How long does it take until the decolourisation of the body occur?
36 to 72 hrs after death discolouration of the abdominal wall occurs.
How do environmental conditions affect the rate of decomposition?
- Lower temperatures slow decomposition
- Warm temperatures speed it up
- Rate of decomposition is highest between 21 and 38 degrees celsius
- Intense heat denatures enzymes involved in autolysis
What happens if a dead body remains in conditions above 26 degrees?
Gas formation may occur within about 3 days.
What is forensic entomology?
It looks at the presence of insects to make an estimate of how much time has elapsed since death. The forensic entomologists use a predictable sequence of organisms that feed on a decomposing body to determine the time of death.
What do forensic entomologists record?
The information about the location and conditions of the body. They take samples of any insects found on, near or under the body. They record the temperature of the air, ground, body and ‘maggot mass’ are measured.
What do forensic entomologists do with the maggots they find on the body?
Some maggots are killed at the time of collection so that their age can be determined. Some maggots are kept alive where they are taken back to the lab and are fed on meat allowing them to completely develop.
Why do forensic entomologists keep some maggots to let them develop?
It is useful for species identification and to establish when they pupate.
What are the methods in which the ages of maggots are calculated?
- Referring to particular graphs which are made for each species in order to determine the age - you look at the temperature of where the maggot was found as well as its length.
- Identifying the maggot’s stage of development with references to the life cycle of the fly.
- If the maggot is not a C.Vicina and its stage of development is unclear the maggot is allowed to mature which gives the date of pupation. This is calculated by subtracting the date of pupation from the time for the egg to develop.
When using graphs to calculate the age of maggots when does it not work?
When the temperature conditions have changed/fluctuated and has not remained stable.
From the methods used to identify the age of the maggot which is the least accurate? Why?
The method used on maggots which are not C.vicina and works by subtracting the date of pupation from the time for the egg to develop. It is the least accurate because the time of egg laying may give an underestimate of time of death because there is no knowing how long it took the flies to find the body.
How long does it usually take for flies to enter the body?
Normally 1 day
Describe the succession on corpses.
One group of organisms feed on a decomposing body. This causes the conditions on the corpse to change and it becomes attractive to other groups of organisms. As the corpse continues to change, different organisms are attracted to the conditions and this continues until there is only the skeleton left.
Are all of the insects found on the corpse attracted to it?
No some are attracted to the insects on the corpse rather than the decaying corpse itself.
What influences the species present found on a decomposing body?
- Season
- Weather
- Size
- Location of the body
What is the difference between plant and corpse succession?
Unlike plant succession, where many of the early species are replaced as conditions change, most of the early insects remain on the body until the advanced stages of decay.
What can insects also help to determine regarding forensic entomology? How?
Insects can help to determine whether a body has been moved. Species of insects found on a body that are not from that location shows the body has been moved.
What are the other decomposers that decompose a body?
Bacteria and fungi
Where is the bacteria from that helps decompose the body?
- Bacteria from the gut quickly invades the tissue after death.
- Bacteria and fungi from the surroundings colonise the corpse.
What is the sequence in which the bacteria and fungi colonise?
There is no particular sequence in which they colonise.
What do corpses provide to decomposers?
- Organic carbohydrates
- Proteins
- Fats
- Nucleic acids
- Energy to grow and multiply
What is released into the atmosphere by decomposers?
Carbon dioxide is released into the atmosphere. This can be recycled back into a form which can be used for photosynthesis by plants to synthesise more organic molecules.
What are viruses?
Small organic particles with a structure that is quite different from that of bacteria and very much simpler. They consist of a strand of nucleic acid (RNA or DNA) enclosed within a protein coat.
Viral DNA can be both … and … …
Single and double stranded.
What do you some viruses have on their outer surface?
Some viruses have an outer envelope taken from the hosts cell-surface membrane. This contains lipids, proteins and glycoproteins.
What is the importance of the envelope surrounding the virus?
The envelope is important as it helps the virus attach to the cell and penetrate the cell membrane.
Describe the replication of viruses.
1) The virus attaches to the host cell.
2) The virus inserts it’s nucleic acid.
3) The viral nucleic acid replicates.
4) The viral protein coat synthesised.
5) New virus particles are formed.
6) The virus particles are released due to cell lysis.
What type of cell is bacteria?
It is a prokaryotic cell.
Which are smaller bacteria or viruses?
Bacteria are larger than viruses normally between 0.5 to 5 micrometers whereas viruses are usually only 20 to 400 nm.
What is the structure of bacterial cells?
- Capsule
- Cell wall
- Cell membrane
- Main circular DNA
- Plasmids
- Mesosomes
- Flagellum
- Pilus
- Ribosomes
What is the structure of viruses?
- Protein coat
- Nucleic acid (DNA or RNA depending on virus)
How do bacteria reproduce?
They reproduce by form of asexual reproduction known as binary fission which divides the bacterial cell into two identical daughter cells.
How can bacteria exchange genetic material?
Pili can attach to other bacteria and exchange genetic material.
Can viruses exist outside a living body?
Viruses cannot exist outside of living body this is because they must get inside the cell in order to make more virus particles as they do not have the necessary organelles needed for reproduction.
Are all viruses and bacteria pathogens?
All viruses of pathogens however not all bacteria are pathogens.
When is a post-mortem examination performed?
The post-mortem examination is performed if there is a sudden or an unexpected death, or if the cause of death is unknown.
What is the first part of the post-mortem examination?
The external examination is the first part made by a pathologist.
What follows the external examination part of the post-mortem?
The internal examination.
What does the internal examination include?
It involves an incision being made down the front of the body so that the organs can be taken out for a detailed examination. The state of the organs may allow conclusions to be made about the health of the person and the illnesses suffered.
Blood and tissue samples may also be taken during a post-mortem examination. What are these tested for?
They are tested for toxins, infections or tumours.
What is the Capsule?
A protective mucus layer which prevents dehydration and also allows bacteria to form colonies.
Describe the cell wall properties of bacteria.
It does not contain cellulose it is made up of a peptidoglycan: a polysaccharide cross-linked by peptide chains.
What is the function of the ribosomes within the bacteria?
They are the site of protein synthesis and they occur free in the cytoplasm.
What are pili?
Protein tubes that allow bacteria to attach to surfaces and are involved in cell to cell attachment.
What are plasmids?
Small circles of DNA.
What is the flagellum?
It is used for cell movement.
What are the two types of bacterial cell walls?
The gram-positive bacteria cell walls which are thickened with additional polysaccharides and proteins. The gram negative bacteria walls have thinner walls but have a surface layer of lipids for protection.
How is TB carried?
It is carried in droplets of mucus and saliva.
How does TB spread?
When it is released into the air for example when an infected person talks, coughs or sneezes.
How to others develop TB?
When others inhale the droplets through droplet infections.