Forensics, Disease And Immunity Flashcards
What are fingerprints?
- impressions left by the friction ridges of a human finger
How are fingerprints developed?
- formed between weeks 6-10 of foetal life
- raised portions of the epidermis
- movements of baby in the womb, speed of growth etc. affect the fingerprint pattern
What are the types of fingerprints?
- arch
- tented arch
- whorl
- loop
How are fingerprints left?
- sweat and oil from sebaceous glands cover our fingers
- leaves and impression of the friction ridges when we touch something
Name three ways to make fingerprints visible:
- carbon aluminium or magnetic iron powder (sticks to grease)
- ninhydrin (become purple with AA in sweat)
- superglue vapour
How much of DNA codes for protein?
1%
What is non-coding DNA transcribed into?
- functional non-coding RNA
- transfer RNA, ribosomal RNA, regulatory RNAs
What are the other functions of non-coding DNA?
- transcription functions
- promoters, translation regulation of genes, introns, origins of DNA replication, centromeres and telomeres
What are STRs?
Non-coding DNA contains many short repeated sequences
- 3-7 bases long and repeated from a few to many times
- make up 3% of human genome
- likely involved in chromatin folding and transcription regulation
Describe small tandem repeats?
- STRs occur at the same locus on both homologous chromosomes
- number of STRs at a particular locus can vary on each homologous chromosome
- STRs are inherited like genes
How do STRs help identify people?
- each individual has a large number of STR loci
- this creates a unique DNA profile for each individual depending on the number of STRs at various loci
What is DNA profiling?
DNA fingerprinting
What is DNA profiling used for
- forensic technique used in criminal investigations
- parental testing
- genealogical
- medical research
How is the length of STRs determined?
- using gel electrophoresis
- separates DNA fragments according to size
What is gel electrophoresis
- a method of separating DNA fragments according to size in an agarose gel by applying an electric field
What is step 1 of gel electrophoresis
- mix agarose and buffer
- microwave to melt agarose
- cool, pour into mould
- remove comb when gel set
What is step two in G.E
Loading the gel
- the gel is put into a tank with buffer
- DNA samples are loaded into the wells of the gel with a pipettes
What is step three of G.E?
- electrodes are attached to gel tank
(Cathode near wells, anode on opposite side)
Why do DNA fragments migrate from - to +
Because DNA is negatively charged
Why do small fragments migrate further in gel electrophoresis?
- larger fragmented get caught in the matrix
What are two ways of visualising DNA banding patterns?
- Staining the DNA directly
- Southern blotting
How is DNA stained?
- using ethidium bromide and then visualised under a UV lamp
- ethidium bromide inserts itself between the base pairs in the double helix
- it grows in UV light making DNA visible
What is southern blotting used for?
- to visualise specific DNA fragments
Describe the southern blotting method
- DNA is transferred to nylon or nitrocellulose
- membrane buffer is drawn up through gel
- membrane with bound DNA is removed
- hybridisation
- labelled DNA probe is visualised
Describe hybridisation:
- membrane is incubated with radioactive or fluorescent DNA proble
- (binds with target DNA via complementary base pairing)
What are the four factors to identify time of death
- body temp
- degree of muscle contraction
- decomposition
- entomology and succession
How is body temperature maintained?
- heat released in metabolic reactions (e.g respiration)
How is core body temperature of a dead body measured
- long thermoprobe via rectum or abdominal stab
Describe this graph
A:
- sigmoid curve
- plateau 30-60 mins
- metabolic reactions not fully stopped yet
B:
- linear decline of temperature can be used to estimate time of death (~1.5C/h)
C:
- body temperature reaches ambient temperature
Why does the body cool down when a person dies?
Respiration stops
- no more heat released by cells
Heat lost from body by radiation and evaporation
What affects initial core body temperature?
- fever
- hypothermia
What affects postmortem cooling?
- environmental temperature
- air movement (wind)
- humidity/body found in water
- SA/vol ratio
- fat composition
- body location
- clothing
How does environmental temperature affect cooling?
- the greater the temperature gradient and air movement the more quickly the body loses heat
How does humidity affect cooling?
- the higher the humidity the more slowly the body loses heat by evaporation
How does a body being in water affect body temperature?
- a body in water will lose heat more quickly due to the temperature gradient
How does a body being in water affect body temperature?
- a body in water will lose heat more quickly due to the temperature gradient
Will a small or large body lose heat more quickly?
- small due to a higher SA/Vol ratio
A high fat composition and clothes will…
Insulate the body
What is rigor mortis?
- ‘stiffness of death’
- after death muscles first relax, then stiffen and then relax again
Label the structure of a sarcomere
1 - myosin head
2- thick filament (myosin)
3 - thin filament (actin)
What cause the contraction of a sarcomere?
- ATP, Ca2+
Describe the mechanism of muscle contraction:
- myosin-ADP binds to actin
- ADP is released resulting powerstroke: muscle contracts
- ATP binds
- myosin ATP is released from actin and muscle relaxes
- ATP hydrolyses to ADP + Pi
Give a summary of rigor mortis development?
- Death: muscle cells become starved of oxygen
- aerobic respiration - Respiration becomes anaerobic
- produces lactic acid - pH falls inhibiting enzymes
- anaerobic respiration stops - ATP is no longer produced
- myosin and actin permanently fixed in contracted state
Why does rigor mortis stop after ~36 hours
- lysosomes break down and release enzyme to break down cell
Apoptosis
Why does rigor mortis start sooner when a person has drowned
- uses a lot of ATP
What affects time of onset of rigor mortis
- fitness -> atp storage
- metabolism
- ambient temperature
Why does rigor mortis start sooner when a person has drowned
- uses lots of ATP
Why is rigor mortis of limited use in determing time of death
- after 36 hours its gone
What is decomposition?
- digestion of cells resulting in breakdown of tissues and release of carbon and nutrients (e.g nitrate and phosphate)
What are the five stages of decomposition?
- Fresh, initial decay
- Bloating, putrefaction
- Active decay
- Advanced decay
- Dry remains
Describe the fresh stage
- 0-3 days after
- autolysis
- anaerobic bacteria in gut start to digest tissues and release gases (start of putrefaction)
Describe bloating/putrefaction
- 3-10 days
- increasing gas production by bacterial activity causes swelling of body and putrid odor
- breakdown of haemoglobin leads to venous marbling of skin + green discoloration of abdomen
Describe active decay
(Black putrefaction)
10-20
- discolouration of skin changes to purple then black
- tissues start to soften and then liquefy
- flesh looks creamy
- loss of fluid and deflation of body
Describe advanced decay
- 20-50 day
- majority of internal tissue lost
- body starts to dry out
Describe dry remains
- 50 to 365days
- soft tissue lost, leaving skin, bone and cartilage
What is autolysis
Self digestion of cells
- when respiration stops, lysosomes release digestive enzymes which digest cell components
- digestive enzymes secreted into gut also contribute to self-digestion
What putrefaction
=putrification
Digestion of proteins and tissues by anaerobic bacteria
Describe the process of putrefaction
- as proteins break down, gases are produced and excreted by anaerobic bacteria (causing a putrid odor)
- gases diffuse to other parts of the body, leading to bloating of torso then limbs
- increased pressure weakens and seperates tissues
What factors effect decomposition rate
- weather, exposure, humidity
= encourages bacteria
How does mummification work?
- remove organs
- dry body with salt for 30 days
- wash body cavity with wine and spices
- wrap body in antrimicrobial bandages
What are plant cell walls made of?
- cellulose (B1-4 glucose)
What is the fungal cell wall
Chitin
What bacterial cell wall
- peptidoglycan
(Polysaccharides held together by oligopeptide crosslinks)
What is oligopeptide
- small peptide 2-20 amino acids
How many types of bacterial cell walls are there and how are they distinguished?
- two
- gram staining
How is gram positive bacteria identified?
- crystal violet CV+ ions stain
- Blue-violet colour
How are gram negative bacteria’s identified
- do not turn blue violet
- absorb the pink counter stain safranin
Label this cell wall
Gram-positive bacterial cell wall
1 - peptidoglycan
2 - teichoic acid
3 - integral protein
4 - transmembrane protein
5 - cell membrane
6 - cell wall
7 - surface protein
Label this bacterial cell wall
1 - lipopolysaccharides
2 - periplasmic space
3 - peptidoglycan
4 - outer membrane proteins
5/6 - membrane proteins
7- cell membrane
8 - cell wall
9 - outer membrane
10- porins
What do porins do?
- allows water and hydrophilic things into the bacteria
Describe the structure of a gram positive bacterial cell
- thick layer of peptidoglycan
- one cell membrane
Describe the structure of a negative bacterial cell wall
- thin layer of peptidoglycan
- cell membrane and another outer cell membrane
Describe the process of bacterial cell walls being stained
- crystal violet ions enter cell walls and gram negative outer membrane
- I- ions bind to and fix CV+ ions
- decolouriser disrupts gram negative outer membrane and so CV+I- is lost
- counter stain adds pink colour which allows us to visualise decolourised cells
Summarise gram+ bacteria
- blue violet stain due to CV
- more susceptible lysozyme and antibodies
- tend to live on skin
Summarise gram negative bacteria
- pink stain due to safranin
- tend to live in wet areas because are susceptible to drying out
How do bacteria reproduce?
- asexual reproduction by binary fission
- this is vertical gene transfer
How can bacteria obtain foreign DNA?
(And therefore obtains genetic variation)
Via horizontal gene transfer
Give the three methods of horizontal gene transfer
- Transformation
- Transduction (via bacteriophage)
- Conjugation (via pillus)