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256 Cards in this Set

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clastogen
material that causes breaks in chromosomes via deletion, addition or rearrangement.
caspases
intracellular proteases that fuction as initiators and effectors of apoptosis. They cleave the aspartate-cysteine bonds in proteins.
oncosis
cell death characterised by mitochondrial swelling in ischemic cell death.
MMP
matrix metallo-proteinase (endopeptidase which is a prodeath protein released when TNF / FAS factors bind.
mitotic catastrophe
A type of cell death- abnormal activation of CYCLIN B/CDK1-

Occurs in meta/anaphase - cellular checkpoints ignored / damage = caspases activated. signs are multi and micro nucleation leading to aneuploidy and apoptosis.
TNF
tumour necrosis factor - Induces inflammation, cell death and is activated by macrophages.
Death receptors are..?
FAS / TNF
DIABLO
direct IAP binding protein with low pl

Binds to and inhibits IAP (inhibitor apoptosis protein)
IAP
inhibitor apoptosis protein -> inhibits caspase 3 = no apoptosis
pyknosis
chromatin condenses next to the nuclear membrane in apoptosis

(irreversible)
karyorrhexis
destructive fragmentation of the nucleus, the nuclear membrane becomes permeable.
PARP
"poly - ADP - ribose polymerase" A nuclear protein involved in DNA repair, it can induce AIF (apoptosis inducing factor) to mitochondria to START APOPTOSIS
AIF
apoptosis inducing factor (stimulates mitochondria)
P53
A tumour suppressor protein

conserves the genome & prevents mutations.

Activates DNA repair, induces growth arrest and starts apoptosis
CAD
"caspase activated deoxyribonuclease"

hydrolyses dna to 180-200 bp fragments
APO-1
apoptosis antigen 1 = FAS receptor = TNF receptor

synonyms of each other
Specimen used in apoptosis?
"Caenorhabditis Elegans" a nemutoda, it has 1030 cells and 131 die during development via apoptosis
Specimen used in cytoplasmic streaming?
Elodea.

cyclosis due to actin microfilaments.
Specimen to see flagella?
Euglena.

movement of microtubules
Specimen to see cillary beating?
Paramecium

movment of microtubules
cytochrome c
normal= part of the ETC inner membrane of M

apop= released from M, binds to APAF-1 and ATP to form the apoptosome
APAF-1
"apoptotic protease activating factor" binds to ATP and cytochrome c to form the apoptosome
Apoptosome
cleaves procaspase 9 = caspase cascade! Initiates apoptosis
Annexin V
Is a PHOSPHOLIPID, used to stain phosphatidyl serine with FITC to see protein makers on the cell surface.

FITC = "fluorescein isothiocyanate"
FITC
"flourescein isothiocyanate" flourochrome LABEL used to bind to annexin v
microflourimetry
Quantitative test to tag cell components with flourescent molecules.
microscopy
qualitative test of a specimen under magnification
flourescence
absorption of light by flourochrome, electrons promoted, return to ground state and longer wavelengths of light are emmited with heat.

The emmision energy is flourescence
epi-illumination
light goes through the objective and illuminates the specimen
fluorescein
Direct fluorochrome with unchanged quantum yeild
Acridine orange
A METACHROMATIC direct flurochrome probe with unchanged quantum yield.

DNA = yellow
RNA = orange
Autoflourescence
natural emission of light-

chlorophyll, lipofuscin, Hb, myoglobin, phytochromes, flavins, tryptophan
direct flourescence
flourochromes change QY after binding to cellular structures.

DAPI - DNA BLUE
Hoechst stain
DAPI
4'6'- diamidino-2-phenylindole

A direct flourescent stain.

DNA = BLUE
Flourescent label
covalently binds to specimen

Phalloidin, DAPI
Flourescent probe
non-covalent binding to specimen

acridine orange
Unchanged quantum yield direct flourescent stains
Acridine orange

Flourescein
Changed quantum yield direct flourescent stains.
DAPI

HOECHST

DiOC6
Phalloidin
labels F actin microfilaments, when conjugated with a flurophore
Flourescence advantages
High sensitivity, speed, safety,

good contrast, fixed specimen
Indirect flourescence
Antibody sandwich, FISH labeled oligonucleotides

another molecule between cellular target and flourescent molecule
FISH
flourescence insitu hybridization
Flourescence disadvantages
Photobleaching, expensive, resolution limit, toxic to cells
BRET
bioluminescence resonance energy transfer

LUCIFERASE railroad worm, squid, bacteria, fireflies (LUCIFERIN)
GFP
Green flourescent protein

Aequorea victoria jelly fish.
Transduces blue chemiluminescence to green flourescent light.
cyropreservation compounds
polyethyleneglycol

glycerol

dimethyl sulfoxide
native stains
DO NOT KILL CELLS

acridine orange, neutral red, janus green
Anti-apoptotic proteins
Bcl-2 (B-cell lymphoma 2)
activates or suppresses based on conc. on the mitochondrion membrane, regulates the release of cytochrome c.

IAP / XIAP (x-linked)

survivin (BIRC5 gene)
Activation genes of A
BAX family
(Bcl-2 associated X protein)

promotes apoptosis
Intrinsic A pathway
Injury to the cell

1. Mitochondria -> cyto C -> APAF-1 activates caspase 3

2. Diablo/smac -> counteract IAP = +caspase 3 & cleavage of PARP

3. ER stress -> mitochondria stress etc...
Extrinsic A pathway
FAS ligand binds to FASr

Fas associated death domain (or TNF)

Procaspase 8 cleavage process ends with CASPASE 3

cleavage of PARP and A
Effects of A on mitochondria
Cyto C released + AIF, -ATP, Ca2+ released,

Activation of endonucleases (caspases) production of O2 radicals, + permeability.
Effects of A on plasma membrane
Externalisation of phosphatidylserine

stain with Annexin v + FITC
Effects of A on cytoskeleton
microfilaments = actin forms blebs

intermediate filaments = cytokeratin fragmentation

microtubules not affected
Regulation of apoptosis?
cytokines - TNF, FAS, GF
necrosis characteristics:
groups of cells

swelling of M, cell, organelles

inflammation! Random & diffuse DNA degeneration

plasma membrane smoothing
apoptosis characteristics:
single cells - shrinkage

blebbed plasma membrane - phosphatidyl serine

clumped and fragmented chromatin (pyknosis)

apoptotic bodies
heat shock proteins
protein chaperons (ubiquitin) aid the correct folding of proteins in response to heat, cold, stress (oxidative or reductive).

examples = HSP90 cardiovascular and HSP70 immunity
Effects of inc temp on cells
+ thermal movement, HSP produced, protein coagulation, cell death, membrane defects and

depolymerisation of the mitotic spindle
Effects of dec temp on cells
- termal movement, HSP produced, metabolic processes retarted, ICICLES = freeze/ thaw. cell death
Activation of apoptosis (4)
DNA damage (PARP -> AIF)
mitochondrial malformation
Caspases
Death ligands (Fas / TNF)
Activation of necrosis
cell membrane damage...
Damage to DNA synthesis possibly results in... (4)
point mutation, chromosomal breaks (addition, deletion, rearrangement), cross links, mitosis defects
Photobleaching effect
irreversible decomposition of fluorochromes due to interactions with molecular oxygen. Light induces the excited state required for the reaction.
human body cells that can autoflouresce?
myoglobin

retinal cells - rhodopsin

haemoglobin
Name 5 components of a flourescence microscope
Light source - UV/Blue/Green
Excitation filter (only excitated light)
objective lens
dichromatic mirror
Emmision filter (only emitted light)
Advantages of laser confocal M over flourescence M (4)
1- more sensitive
2- 3D reconstruction
3- no photobleaching
4- signal to noise ration is better
Principle of Agar diffusion?
substance placed in the middle of an agar plate with cell culture spread evenly on it.

Cell death is proportional to the toxicity of the substance in a circle arrangement as the substance diffuses out and kills the cells
Role of mitochondria in apoptosis?
Bcl-2 proteins on M membrane surface

+ permeability, cytochrome C released, APAF + ATP = apoptosome.
define a gene
a specific length of dna that codes for a protein or rna
explain the term FRAP
"flourescent recover after photobleaching" - proves the fluid mosaic model of the bilipid membrane. Damaged proteins are replaced with fuctional ones via membrane diffusion.
Extrinisc pathway of apoptosis (basic)
Fas ligand -> FADD -> procaspase 8 -> caspase 3 etc...


FADD = Fas associated death domain (intracellular process)
what is REACH?
registration, evaluation, authorisation and restriction of chemicals

aim is to improve protection of human health and environment. EU chemicals industry 2006
what is a terminator?
a dna sequence at the end of a gene operon used to terminate transcription.

Rhoe factor or intrinsic hairpin loop
What kind of dna is in mitochondria?
circular, not associated with histones, maternal origin in humans.

mtDNA though to derive from bacteria which were englufed by eukaryotic cells.
what is the osmolarity of human plasma?
290 mOsm/kg

mili-osmoles per kg
three phases of cell culturing?
lag, log, plateau phase
three chemicals in cyropreservation
glycerol

polyethyleneglycol

dimethyl sulfoxide
what is a cell strain?
it is a subline or clone of the original cell line.
what is HeLa?
Henretta lacks cervical cancer, continuous cell line.
primary culture?
dissaggregation of cells and placed on a cultivation surface (Eagles MEM)
Enzymatic disaggregation of cells?
Trypsin, EDTA, collagenase

ethylenediamine tetraacetic acid
secondary culture?
PC dispersed in cell suspension and reseeded into fresh vessels = passage

Cell line produced.
Fate of cell lines?
normal= growth and death (lag, log, plateau)

Cell strain= selected by cloning, subline of original cell line produced

Continuous cell line = transformation invitro
What is a stem cell line?
A continous cell line (SC transform invitro) with stem cell properties to still differentiate.
Explain histiotypic culture...
Cells are reaggregated in suspension then infiltrated onto a 3D tissue like structure matrix.
Explain organotypic culture
reassociation of cells from DIFFERENT lineage. (many cells make up a tissue, many tissues make up an organ)
What is a hybridoma?
a hybrid cell resulting from the fusion of a lymphocyte and a tumor cell; used to culture a specific monoclonal antibody.
advantages of tissue culture?
precise control of physiologic conditions
better biological standardisation of experiements
cyropreservation = years of work on the same cell line
invitro culture = cheaper than animals
no ethical and legal problems
disadvantages of cell tissue cultures
cells dissociated from 3D geometry and ECM = loss of cell interactions

lack homeostatic mechanisms that you would find invivo.

PCC are not stable due to overgrowth or dedifferentiation.
five types of bio-modelling?
computer programs
3D models
cell cultures
laboratory animals
human volunteers
Explain FRAME
fund for the replacement of animals in medical experiments
what is a xenobiotic?
a chemical found in an organism which is not normally produced or expected to be present.
How did we induce cell death (necrosis) in paramecium?
eosin
cell culture environment includes what? (4)
substrate, culture medium, gas phase, temperature
what is hayflick's limit?
is the number of times a normal cell population will divide before it stops, presumably because the telomeres shorten to a critical length.
Two main constitutes of culitvation medium?
Eagles MEM (minimum essential medium)

Bovine/calf SERA (serumalbumin) 10%: contains a wide range of minor components which effect cell growth
Explain conditions for the GAS PHASE of culture experiments
5% CO2 with Ph at 7.4, in equilibrium with sodium bicarbonate in the medium

Organ culutures require 95% O2
What signs of a cell culture show it needs a medium change?
A Ph drop below 6.5

Morphological deterioration - nucleus granulates, vacuolisation of the cytoplasm, rounding up of cells.
what is the 3R concept?
Refinement, reduction and replacement

of laboratory animals with other toxicological testing methods. = invitro mammalian cells.
Name three "targets" of measurement to asses proliferation in a cytotoxicity test.
Cell protein count (coomasie blue test)

DNA count

ATP / ADP ratio (best one) - ratio is constant in proliferating / live cells
Give an example of a metabolic activity - cytotoxic test
Viable cells reduce TETRAZOLIUM salts (MTS/MTT) to coloured FORMAZEN compounds.

Focus is on bio functions inside intact cells.
Give an example of four cytotoxicity methods.
Viability - Trypan blue

Proliferation - ATP/ADP ratio

Metabolic assay - tetrazolium -> formazen

DNA chips / microassays- gene expression level determined
How do DNA chips / microassays work?
DNA sequences are applied to a slide / nitrocellulose membrane.

DNA's are used to probe target sequences. the effects of a toxicant increases or decreases the level of gene expression.
what were we testing when we used Iodonal B? What is its active ingredient?
cell viability and the compound was iodophor
Which three things are most desirable in microscopy?
Magnification - magnified image of the specimen

Resolution - separate the details

Contrast - to make the details visible
Focus length and resolution of human eye?
10cm and 0.1mm
Microscope magnification & resolution?
1000x and 0.2um (micrometers)
Why cant you increase the magnification of a LM beyond 1000x?
The resolution would not improve rendering the image blurry
Why is oil immersion sometimes used.
It creates a continuous film between the coverslip and the lens so light rays do not refract away.
What kind of microscope do we use?
NIKON SE COMPOUND LIGHT MICROSCOPE
what is the numerical aperture at 10x?
10X = 0.25 NA
what is the numerical aperture at 40x?
0.65 NA
what is the numerical aperture at 100x?
1.30 NA
Describe bright field microscopy
Sample illumination is transmitted, via objective to be viewed from above
several objective lenses
limited resolution
dark image on clear background
What does "parafocal principle" mean?
When an object is in focus with the course adjustment knob it will still be in focus at all other magnifications.

corresponding focal points in the same plane
what does "paracentral principle" mean?
the object must be in the center of the field of view before increasing the magnification, or it will be left out.
what is the "field of view"?
the circle of light you see when you look into the microscope. Inc Mag = smaller FOV
What is the condenser and how does it affect resolution and contrast?
condenser ensures that all parts of the specimen are evenly lit via the iris diaphragm.

inc light = + resolution / poor contrast

dec light = + contrast / poor resolution
what are the "vernier scales"?
the vertical and horizontal readings on the mechanical stage of the M.

used to find the same spot on a slide.
If at 1000x, 10 cells fit in the FOV what size are they?
18um
If 30 cells fit in your FOV at 400x what is the cell diamter?
15um
FOV diameter at 100x?
1.8mm
FOV diameter at 1000x?
180um
FOV diameter at 40x?
450um
size estimation is what?
field of view (mm or mm)

divided by

estimated no of cells in transect of FOV.
Explain what "phase contrast" means
The PC-M exploits interference effects that take place when two types of light recombine.
What is numerica aperture?
it is a dimensionless number characterising the number of angles of which a system can accept or emit light. inc mag = + NA
what is Dark Field microscopy?

Differential / Normaski M uses the same principle
DF-M is a technique used to increase the contrast of unstained transparent specimens.
Phase contrast other details
the phase annulus (clear) must match the phase plate (darker)

phase differences of light passing through an object are converted to the changes in intensity (amplitude)
How is the best phase contrast achieved?
one has to shift the phase of light by 1/4 lambda in the +/- sense.

high refractive index = darker
What kind of light is used in phase contrast microscopy
monochromatic yellow-green light which is most sensitive to our eyes.
what is a telomere?
it is a length of repetitive DNA at the 3' end of the chromosome with a TTAGGG sequence.

It protects coding genes from shortening which occurs during mitosis
what is CAAT?
the "center for alternatives to animal experimentation"
what is overtons rule?
Increases lipid solubility of a molecule increases its diffusion rate across a plasma membrane.
when does haemolysis occur?
when red blood cells are placed in a hypotonic solution.

cell swelling and burst.
what makes the fluid mosaic model unique?

singer & nicolson 1972
the dynamic interspersed relationship between the phospholipid bilayer and membrane proteins (integral and peripheral)
which part of the plasma membrane confers antigenic specificity to the cell
carbohydrate chains from integral glycoproteins.
what is the thickness / are the basic functions of the plasmamembrane?
7.5nm (nanometers).

surrounds protoplasm and regulates which substances enter and exit the cell.
define osmosis
the diffusion of water across a membrane in response to a concentration gradient
what will be the result if a plant cell is placed in a hypertonic solution?
plasmolysis - water leaves the cell by osmosis, plasma membrane shrinks away from the cell wall.
define hypertonic solution
The concentration of solute in an external solution is greater than that of inside a cell.
define a hypotonic solution
The concentraion of solute in an external solution is lower than that of inside a cell.
Four characteristics of continuous cell lines.
- loss of contact inhibition
- "imortal"
- transformed invitro / tumour derived
- dedifferentiation.
Four characteristics of primary cell lines.
- diploid karyotype
- terminal differentiation
- contact inhibition of growth & proliferation
- senesence and death
Structure and function of aquaporins
six transmembrane alpha helices in circular fomation create a selective water pore which regulates water inflow to a cell.
what happens if you place a red blood cell in a hypertonic solution?
crenation / plasmolysis as water moves out of the cell by osmosis.
give six examples of INTRACELLULAR movement
-cytoplasmic streaming (cyclosis due to actin microfilaments)
-vesicle trafficing between vesicles and cell surface
- cell division for cytokinesis
- movement of chromosomes in mitosis
- pigment movement for colouration
- discharge of vesicle content
Genetic microtubule defects (3)
anosmia (loss of olfaction
hyrdocephalus
kartagner syndome (primary cillary dyskinesis)
aquired microtubule defects (3)
-hydrocephalus (lack of vit D)
- cillary beat arrest (toxins, bacteria)
- cancer
MOA of Taxol
prevents dissasembly of microtubules

= mitotic arrest
MOA of nocodazole
depolymerises the mitotic spindle
MOA of colchicine
inhibits microtubule polymerisation
MOA of vinblastine, vincristine
binds to tubulin preventing dynamic instability at both ends
New sea sponge derived microtubule inhibitors
Eribulin - prevents dynamic instability

Discodermolide - prevents depolymerization in the G2/M phase
what is kinesin?
motorprotein which caries vesicles towards the + end of the microtubule.
what is dynein?
motorprotein which caries vesicles towards the - end of the microtubule
what is a topoisomerase?
an isomerase enzyme that acts on the unwinding or overwinding of DNA in 3D space.
what are topoisomerase inhibitors?
drugs which prevent unwinding of DNA during replication = damaged replication (quinolones)
what does permeability of a molecule depend on throug a PM?
size, charge, concentration, solubility
name 5 online information sources
EBESCO
pubmed
nature.com
science direct
medscape
during A mitochondira release
Diablo / smac
AIF
cytochrome C
endopeptidase G
what is the photochemical effect?
When light hits phototoxic compounds inside the cell

eosin / porphyrin
what is porphyria?
it is a rare genetic disease characterised by dysfunction of heme metabolism and deposition of porphyrins in teeth, skin and bones.
Two ways of DNA isolation?
phenol based

phenol free
what is PDT?
PHOTODYNAMIC THERAPY

- a photosensitizer is exposed to a group of cells.
- when light is shone on the area, the cells react with molecular oxygen and initiate apoptosis/ necrosis.

cancer: light wavelengths and PS are specific to each other!
basically describe the phenol based extraction of DNA
transfer of nucleic acid into a water phase of a solution. Removal of acid associated proteins and lipids in phase separation.
How would you dissociate DNA/RNA in a cell?
Degredation via mechanical force- chemicals- then enzymes:
Proteinase K or PRONASE

or detergent solutions.
What are the best conditions for enzyme digestion of cells to extract DNA?
37*C with proteinase k or pronase
left over night.
what does SDS stand for?
sodium dodecyl sulfate - emulsifies lipids and enables the dissociation of DNA and proteins (the histone complex)
What does EDTA stand for and what does it do?
ethylenediaminetetraacetic acid

deactivates metal dependant enzymes and DNA associated ions by chelating the metal ions.
what is the ideal PH for DNA extraction?
pH 8
which buffer is used to keep the pH (8) stable during extraction.
TRIS - buffer capacity of 6.5 -> 9.7

trishydroxymethylaminomethane
What is PCR?
polymerase chain reaction, used to exponentially amplify DNA with the use of primers and polymerase enzymes. (in vitro method)
what is a DNA dipstick?
a kit used for measuing nucleic acid concentration and yield
what is NONIDET solution?
it is a solution used in DNA extraction to breakdown membranosus structures
what is IF?
impact factor -
it is a measure of citations to science journals, a proxy for the importance of a journal to its field.
Give an example of:

positive taxis

Negative taxis
plants growing towards light

protective reflex (hand withdrawl)
define kinesis
Reflex - LIKE - activity.

stimulus = change in speed/direction away from noxious stimulus.
when a cell exhibits ameboid movement name the four changes of actin-PM interactions.
stress fibers, ruffling membrane, lamellipodia, filopodia
Describe the motility seen in Euglena
positive phototaxis as a result of photoreceptors at the base of the flagellum (microtubule movement)
What is depth of field in microscopy?
The area infront and behind of the specimen that will be in the acceptable area
Define the mechanism and occurance of bioluminescence
A biochemical oxidative process that results in the release of energy as emitted light.

Luciferin + molecular oxygen catylased by luciferase.
In the DNA isolation practical which two solutions were used?
EDTA

Sodium dodecyl sulphate
Which of the following represent molecular motor proteins?
A. Gelsolin
B. Myosin +
C. Kinesin +
D. Actin
E. Dynamin +
What is gelsolin?
Gelsolin is an actin-binding protein that is a key regulator of actin filament assembly and disassembly.
What is wiskott-aldrich syndrome?
An X-linked recessive genetic disease characterised by thrombocytopenia (lack of platelets)
What is a chaperone protein?
Chaperone proteins aid in the non-convalent folding or unfolding of other macromolecules
What is YFP?
yellow flourescent protein
Cyclosis is an example of movement of...

Which is based on...

we observed the phenomenon in....
cyctoplasmic streaming

actin microfilaments

Elodea
Monoclonal antibodies are formed from?
hybridomas
Hybridomas arise from?
B lymphocytes fused with tumour cells
Applications of monoclonal antibodies?
immunohistochemistry diagnostic tests

cancer treatment, for specific cancer antigens
Transfusion of pt occured with DISTILLED WATER (in a rush) what is the main consequence?
RBC's are hypertonic relative to pure water and will burst undergoing haemolysis

serious possibly fatal
FRAP means and is used for?
flourescence recovery after photobleaching. It is used to study the fluid mosaic model of plasma membranes with integral and peripheral membrane proteins
GIve three examples of human cells which use locomotion
macrophages - ameboid movement
neurons - growth cones
mesenchymal cells - cell crawling (mesenchymal movement) with MT help (4 things there)
The most famous model organism for apoptotic studies is..
C. Elegans 1030 cells and 131 undergo apoptosis.
what are stress fibers?
high order structures in the cell containing actin fillaments, myosin motor proteins and crosslinking proteins. Support the cell and have a role in movement
What is PEG
polyethylene glycol

A polyether used in cyropreservation
Define totipotency
Cells which are totipotent can differentiate into any cell of the body including the extraembryonic tissues. The zygote is the only source of totipotent cells.

all plant cells are totipotent
FAS
is a transmembrane protein related to the Tumour Necrosis Factor family. (it is a receptor) Binding results in the extrinsic pathway of apoptosis.
In agar diffusion testing of cell viability what is the dye used called?
Neutral Red
MOA of vinblastine
prevents depolymerisation of the microtubules
What does stoke shift mean?
It is the difference in wavelength between excitation wl and emission wl.

Emission is always longer than the excitation (energy lost...)
Rough emission spectra wavelenths for colours

blue
green
red
B: 420-500

G: 510-560

R: 590- 650
promidium iodide stains what?
intercalates with DNA - red
ethidium bromide stains what?
intercalates with DNA - orange
Clinical dissorders associated with aquaporins?
diabetes insipidus, cataracts of the cornea, cirrhosis (liver inc AQ)
what is a bibliographic record?
A record of an article or book in a database, it includes info about authors, sources and an abstract
What is botulinum toxin?
It is a microtubular poison preventing the transmission of signals at the neuromuscular junction. Produced by a bacterium
what is crenation?
osmotic responce to RBC's being placed in a hypertonic solution - cell shrinkage
As magnification increases....
Numerical aperture increases

Field of view decreases

Depth of field decreases
Give an example of a light source for flourescence microscopy...
xenon arc lamp
how does a dichromatic mirror work?
It reflects wavelenghts shorther than a specific wavelenght and passes light longer than a specific wavelength
where does the word caspase come from?
Caspase cleaves cysteine/aspartate amide bonds = C-ASPA -ase = enzyme..
what is ex vivo?
Tissue taken from invivo, processed in a lab and then pub back invivo
name THREE cultivation mediums
fibroblast growth medium
melanocyte medium
CADMEX growth medium
EAGLES MEM growth medium
Name five essentials for cell culturing!
incubator, laminar hood flow, fridge & freezer, autoclave (sterilizer), inverted microscope, water purification unit, liquid handelling devices.
what is the purpose of an inverted microscope?
to see cell cultures at the bottom of conical flasks, light source and condenser above the sample with the objective below the sample to the lenses.
why do RBC's undergo haemolysis whilst in an isotonic solution?
because the solution is isotonic to a certain ION, all other ions are thus in an "unbalenced" state in relation to RBC's

Hence we use ISOIONIC solution (ringers) which is specific for RBC's
Where do we use Monoclonal Antibodies in medicine?
1 - IHC for cell staining or radiotherapy

2- Tumour marker diagnostics

3- Differential diagnostics of viral, bacterial, parasitic infections.
what is PCR?
polymerase chain reaction

it is a method to exponentially amplify DNA invitro using DNA polymerases and primers (variable temp)

Denaturation, Annealing, Extension & repeat....
what is flow cyctometry?
A technique used to count & examine microscopic particles in cells / chromosomes > suspended in fluid stream and detected by electronic device by light penetration.
what is filliopodia?
cytoplasmic projections of plasmamembrane in a migrating cell containing bundles of microfilaments.
what is an enhancer?
it is a region of DNA that can bind proteins to enchance trascription of genes
Agar diffusion principle is a test of what? How does it work?
cytotoxicity test on cell viability:

effect of substance on bacteria growth culture.
define passage
primary culture dissaggregated and reseeded into a fresh vessel
confocal lens characteristics:
laser / photomultiplier / pinhole filter
Epiflourescent lens characteristics:
xenon arc lamp / ocular detection / diaphragmatic filter.
phosphorescence
emission of light persists after the excitation by light
epi-flourescence
light ot the objective illuminates the specimen and emission light is reabsorbed by the objective to the lenses
dichromatic mirror
efficiently reflects shorter wavelengths of light and efficiently passes longer wavelengths of light (emission)
stokes law
difference in wavelength between excitation and emission spectra
what is quenching?
FRET - flourescence resonance energy transfer - the excited flurophore excites the other molecules around it = other cellular interactions
light sources for flourescence microscopy>
xenon arc lamp

mercury arc lamp

argon ion laser
what is CBS
chromatic beam splitter (dichomatic mirror)
light budget is what?
3.6x 10^4 photons are emitted before destruction of the flurophore (with molecular oxygen)
what is CCD?
charge coupled device- used to enchance quantum efficiency for detecting emission
what is TRIF?
total internal reflection flourescence - it decreases detector noise and enhances the optical background
give three examples of physiologically occuring apoptosis
1- development of the embryo- formation of all tubular structures

2- homeostasis: postitive and negative selection of B + T lymphocytes inthe thymus

3- deletion of damaged and dangerous cells (DNA damaged)
what is porphyria?
it is a hetergeneous group of inherited disorders of haem biosynthesis
what are the three most common presenting symptoms of porphyria?
cutaneous lesions, neuropsychiatric disorder, acute abdominal pain.
How would you diagnose porphyria?>
urine test for increased

1- aminolaevulinic acid
2- porphobilinogen
What is the treatment for porphyria?
Avoid risk factors - sunlight/ alcohol/ illicit drugs

haem arginate and glucose (attack)
chloroquinine (inc porphyrin excretion in urine)
What is the volume of water in a 70kg human body?
42L

65% intracellular
35% ECM
What does AQP mean?
aquaporin protein
what is GLP?
aquaglyceroporin
what is NDI?
nephrogenic diabetes insipidis
what does one osmole mean?
The ammount of substance which dissociates in solution to form one mole of osmotically active particles
when do you find and decrease in AQP2?
1 - diabetes inspidius
2 - renal failure
3 - water loading
when do you find an increase in AQP2?
1 - thirst
2 - pregnancy
3 - cirrhosis of the liver
Which type of aquaporin is present in the kidney collecting duct?
AQP2 regulated by ADH (antidiuretic hormone)
in what phase of the cell cycle do microtubule inhibitors most likely arrest the process?
G2/M phase
what is NLP?
natural language processing -

it converts information from computers into human language (speech / syntax)
what is text mining?
Automatic extraction of information about genes, proteins

bioNLP = natural language processing about biological data
what is PMID?
the unique identification number of a pubmed citation.