1. Identification of 2nd
Site Suppressor Mutations in the HIV-1 Capsid
Precious de Verteuil1
, Ernest Yufenyuy2
, Christopher Aiken3
Abstract
Introduction
Methods
References
Acknowledgements
Kenneth Wong 1
Laura Mudd2
Barry University, Miami Shores, Florida 33161
The purpose of this experiment is to try to
stop and maybe even reduce the effects of
ethanol on neurons In this experiment we
performed immunocytochemistry to see the
effects folic acid had on neurons as they were
being exposed to ethanol. I let the cells
incubate for 48 hours then we “fixed” them. I
added florcence and performed microscopy to
see how the folate affected the cells.
Folate deficiency are found in most patients
with alcoholic liver disease.Alcoholis, are
characterized by degradation of executive
control over behavior and increased
compulsion drug seeking. Ethanol-induced
neuronal apoptosis causes brain shrinkage
and cognitive defects Ethanol exposure
severely impairs the development of the cell
bodies and projection of serotonin Ethanol
also disrupts the cognitive process in the
prefrontal cortex. Over-expression of
antiapoptotic BCL2 can protect the neonatal
cerebellum from ethanol-induced apoptosis
Folic acid stimulates neural stem cell
proliferation in Vitro and enhances
Hippocampal Neurogenesis in rats.
Liu H,Cao J Zhang, Qin S, Yu M Zhang X, Wang X, Gao Y Folic
acid stimulates proliferation of transplanted neural stem cells after
focal cerbral ischemia in rats. © 2013 july 11 Cited 9/26/2013
available from : www.ncbi.nlm.nih.gov/pubmed/23850087
Rundio, A. Understanding alcoholism. 2013, Sept. [cited 2013,
Sept. 5]. Available from:
http://www.ncbi.nlm.nih.gov/pubmed/23998765
Understanding alcoholism. [Nurs Clin North Am. 2013] - PubMed –
NCBI
Eriksen, JL. Gillespie, RA. Druse, MJ. Effects of in utero ethanol
exposure and maternal treatment with a 5-HT (1A) agonist on
S100B- containing glial cells. 2000, Jun. 30. [cited 2013, Sept. 5].
Available from: http://www.ncbi.nlm.nih.gov/pubmed/10876026
Effects of in utero ethanol exposure... [Brain Res Dev Brain Res.
2000] - PubMed – NCBI
Depoy, L. Daut, R. Brigman, JL. Macpherson, K. Crowley, N.
Gunduz-Cinar, O. Pickens, CL. Cinar, R. Saksida, LM. Kunos, G.
Lovinger, DM. Bussey, TJ. Camp, MC. Holmes, A. Chronic
alcohol produces neuroadaptations to prime dorsal striatal
learning. 2013, Sept. 3. [cited 2013, Sept. 5]. Available from:
http://www.ncbi.nlm.nih.gov/pubmed/23959891.
In this experiment we created solutions such phosphate buffer solution which saw made from
sodium phosphate and diluted with 100 mL of water. After we made the buffer solution we made a
33x folate solution out of folic acid and neural basal medium which we use to treat the cells. Then
made a tripson solution out of tripison and water and sterile filtered it. Then we treated cells with the
folate solution and ethanol twice for a two day intervial. Next immunocytochemistry was performed.
The cells were fixed and the cover slips and placed them 4% paraformaldehyde for 15 min at 0.5
degrees Celsius. Permeablize so antibodies can go trough cells for 20 min with 0.1% triton x100 in
HBS for 20 min to stop non-specfic binding. Then I incubated the cells using antibody BCL2 1:500
for 48hours at four degrees Celsius . The cells were then washed in 0.1% PBS/Tween-20 3 times.
Then I put Goat anti mouse #448(green florescence in the well on the cover slip and let incubate
and dry. Cover slips were then placed on slides and fluorescence microscopy was performed.