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what are three types of endocytosis
- receptor mediated cytosis
heparan sulfate proteoglycans
glycoproteins with 2-3 heparin sulfate chains attached to cell surface and extracellular matrix (tau fibrils can bind, triggering intracellular fibril aggregation)
how do heparin bind?
- heparin binds tau and prevents its interaction with heparin moieties of HSPGs
- create more oligomeric forms
- sound waves to agitate particles
- human embryonic kidney cells, easy to grow and transfect
- expression of recombinant proteins (recomb tau)
what are luciferase
oxidative enzymes used to create bioluminescence
- two luciferase fragments are nonfunctional individually but when connected can create bioluminescence.
- if fragments are on separate particles, this can be used to detect when particles are interacting with each other.
why was tau concentration and fluorescence showed together
as protein concentration increased, so did tau
Fluorescence Correlation Spectroscopy (FCS)
- measures the fluctuations of fluorescent signal intensity
- confirm SEC was accurate in identifying tau oligomer sizes
- det size and concentration
laser based, depending on light scattering (analyzed with computer) to obtain cell sorting, cell counting, biomarker detection
seeds=tau assemblies with ability to cause aggregation inside cells
neurodegenerative diseases related to aggregation of tau protein.
how are tau aggregates brought into cell?
diff between macro and micropinocytosis
- macropinocytosis: nonselective uptake of molecules, forms macropinosomes
- micropinocytosis: mediated by clathrin coated vesicles and other vesicles
tau would dissociate into monomers when undergoing ionization, but insolubility of PFA keeps selected subjects of oligomers bound to each other during mass spec.
having more than one site of attachment.
- measures protein content
- protein concentration increase, absorbance increases
how to isolate BD tau?
papain and DNAseI
- papine protease used for cell dissociation to study neurons (leaves cells viable)
- DNAseI (digest DNA)
bacteria expression vector, used to introduce foreign genetic material into host DNA
what word does Alice forgets in lecture?
what was the topic Alice forgets in lecture?
dan's wife, red dress, Alice forgets
how did Alice test her own memory
choosing words from dictionary to remember
- boosts cholinergic functioning
- acetylcholinesterase inhibitor
what are definitive diagnosis for AD
histology of brain tissue (autopsy or biopsy)
- acts on glutamatergic system by blocking NMDA receptors
- overactivity initially leads to excitotoxicity
important in learning and memory
what did Alice's father die of
cirrhosis (liver failure)
what mutation was alice pos for
name of mom, sister and dad
louise, anne, peter daly
what kind of medications was alice taking?
statin (cholesterol lowering), aricpet, namenda, antioxidant vitamins, aspirin
AD more quickly felt by which group and which group lives longer?
early onset, but early onset lives longer
bind soluble amyloid beta and prevent its aggregation
intravenous immunoglobin (IVIg therapy)
- derived from donated plasma blood
- obtain antibodies that neutralize pathogens
- some antibody in patients will be effective against bad antigens in AD patients
where does Alice find Moby Dick?
what does her knowing how to swirl wine say?
retains automatic memory, perceptual part is intact
long adventure that teaches the hero lessons