Week 5 Flashcards
2 general approaches in immunoassays?
measuring specific Ag in sample
look for Ab specific for Ag of interest as evidence that the ind has been exposed to specific Ag
antiserum?
serum collected from an animal that has been exposed to Ag and thus contains Ab against that Ag
produced by immunizing the animal with the Ag of interest and collecting the serum
anti-species Ab?
produced when Ab from one species is collected and injected into another
ELISA?
enzyme linked immunosorbent assay indirect elisa is the most common: - coat wells with Ag - wash away unbound Ag - add serum - Ig binds Ag - wash away unbound Ab - enzyme conjugate - wash way unbound AB - substrate/ chemogen solution for coloraturas change
sanwhich ELISA?
2 Ab go on either side of target Ag
both Ab bind to different epitopes of Ag
substrate specific to enzyme linked Ab is added so we get coloraturas and a positive test which forms in Ag in the sample
SAT?
serum agglutination test
agglutinating Ab present in sera which react with specific Ag to form visible clumps
hemagglutination?
can chemically couple any Ag to RBC to detect Abs to that Ab
if sufficient Ab present to agglutinate and form cross links with Ag then Ab:Ag complex forms a mat at the bottom of the well
if insufficient Ab present then the cells roll down sides of well and forms a pellet down the bottom
direct coombs test?
detects Ab and complement on the surface of RBC
addition of anti-human Ig Ab to washed fetal RBC - they will agglutinate if maternal Rh-Ab were already coated on fetal RBCs
indirect coombs test?
detects Ab in serum
materna sera incubated with Rh+ Abs present in the maternal serum; if they agglutinate then anti-human Ig Ab are present
what immunoassay is used for syphillis testing?
serum agglutination test
what is a common test for HIV? why?
western blotting
AB to various HIV proteins appear at different stages post infection
point of care tests?
rapid test for Ab and Ag detection; lower specificity
drop mixture on sample pad
mixture flows on the membrane
positive signal detected as a line
what are the different flow cytometry tests?
elispot assay: for quantifying cells and producing cytokines
lymphocyte proliferation assay: for memory function and transplant matching
target cell killing assay: for measuring cellular cytotoxicity
microarrays or DNA/RNA chips: for screening large number of genes or samples
what does flow cytometry tell us about a cell?
relative size
granularity or internal complexity
how does flow cytometry work?
take blood sample and mix it with Ab that recognizes the Ag in which we are interested in
tag it with fluorescent tag so when its passed through flow cut-meter all the molecules will be counted (eg, CD4)
screening rules for avg risk of BrCa?
mammogram every 2 years from age 50-74
higher risk screening rules of BrCa?
mammogram every 2 years from age 50-74 and breast awareness (self-examination)
screening rules for cervical cancer?
HPV tests every 5 years from the age of 25 or 2 years after first sexual intercourse
end between 70-74 y/o
avg risk screening rules for Bowel cancer?
FOBT every 2 years from 50-74
higher risk screening rules for bowel cancer?
FOBT every 2 years from age 35-44
colonoscopy from 45-74 every 5 years
screening rules for skin cancer?
avg risk= primary prevention
moderate risk= primary prevention and skin exam
higher risk= preventative advice, examination of skin and advice on self-exam
what are some factors that come into play for primary prevention?
tobacco alcohol diet excercise/ obesity radiation chronic infections occupational exposure
whats the problem with PSA testing?
increased PSA is not solely specific for cancer but their is no precise way to tell
so men that undergo tx may experience side effects but don’t necessarily have PrCa = over tx and over dx
what are some scenarios for increased PSA/
carcinoma benign prostatic hypertrophy prostitis bike riding recent ejaculation
what is incidence?
the number of new cases of disease in a population/ the risk of contracting the disease
what is prevalence?
the number of existing cases in a population/ how widespread the disease is
what is attributable risk?
the amount of risk that is attributable to exposure
what is absolute risk?
probability that a specified event will occur in a specified population
what is relative risk?
ratio of risk to those exposed to risk factor vs those not exposed
what is odds ratio?
measure of association of disease in those exposed and not exposed
what are the 2 viral oncoprotein expressed by HIV?
E6 - binds p53
E7 binds Rb gene
leads to active cell proliferation and gene transcription
what is stroma made up of and functions?
non-neoplastic cells, CT and BV
provides growth and support for parenchyma
what are the 3 ways in which malignant neoplasms spread?
blood
lymphatics
seeding of body cavities
name for benign mesenchymal tissue?
-oma
name for benign epithelial tissue
adenoma
papilloma
malignant mesenchymal tissue?
sarcoma
malignant epithelial tissue?
carcinoma
seminoma?
carcinoma of testicualr germ cells
teratoma?
malignant neoplasm comprising a variety of parenchymal cell types
common functions of photo-oncogenes?
GF and GF-R
signal transduction, cell cycle and apoptotic proteins
nuclear transcription factors
common functions of TSG?
DNA damage repair protein degradation cell cycle regulation apoptosis cell adhesion cell signalling
which point in the cell cycle does the Rb protein act?
transition between G1-S
where are the alterations in the Rb1 gene in retinoblastoma?
Q arm of Chromosome 13
how do some viruses stimulate proliferation of hose cell:
insertion of strong promoter in the vicinity of a host ell proto-oncogene
expression of proteins that neutralize host cell TSGs
expression of proteins that prevent or delay apoptosis
what is a histogram in flow cytometer?
single parameter histogram
distribution of cells expressing a single measured parameter (size, granularity, fluorescent intensity)
what is a 2D dot plot in flow cytometry?
provides more info than the histogram
2 parameters can be displayed simultaneously in a plot (eg, IgM ad IgG)
what is pleomorphism?
present in malignant tumours - variability in size and group: large cells hyperchromic nuclei and multinucleated cells growing in disorganized pattern frequent and abnormal mitosis increased nuclear-cytoplasmic ratio
function of elispot assay?
quantifying cells producing cytokines
function of lymphocyte proliferation assay?
for memory function and transplant matching
function of target cell killing assays?
for measuring cellular cytotoxicity (eg level of function in phagocytes)
function of microarrays for DNA/ RNA chips?
for screening large numbers of genes or samples
benign stratified squamous cell?
squamous cell papilloma
malignant stratified squamous cell?
squamous cell carcinoma
benign lung?
bronchial adenoma
malignant lung?
bronchogenic carcinoma
benign kidney?
renal tubular adenoma
malignant kidney?
renal cell carcinoma
malignant liver?
hepatocellular carcinoma
benign liver?
liver cell adenoma
malignant bladdER?
urothelial carcinoma
benign bladder?
urothelial papilloma
benign and malignant bone?
osteoma
osteogenic sarcoma
benign blood vessel?
hemangioma
malignant blood vessel?
angiosarcoma
benign lymph vessel?
lymphangioma
malignant lymph vessel?
lymphangiosarcoma
malignant hematopoietic cells?
leukemia
malignant lymphoid tissue?
lymphoma
benign and malignant smooth muscle?
leiomyoma
leiomyosarcoma
benign and malignant striated muscle?
rhabdomyoma
rhabdomyosarcoma
histology of retinoblastoma?
small round blue tumour (basophilic)
more viable cells = closer to BV core
pink fuzzy cells = necrotic, they rapidly outgrow blood supply
functions of cyclin dependant kinases?
drive progression through phases of the cell cycle by phosphorylating and activating downstream kinases
function of cyclins?
activating subunits what work with CDKs
specific cyclins for different parts of the cell cycle
example of carcinoma in situ?
chronic smokers show dysplasia in airways
ductal carcinoma in situ
What types of mutational events are commonly associated with proto-oncogenes?
overexpression
overamplification
point mutation
translocation
