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Caffeic Acid and C. elegans’ Protection Against Toxin Induced Neurodegeneration

Caffeic Acid and C. elegans’ Protection Against Toxin Induced Neurodegeneration. By: Michelle Chyn. Need. Parkinson’s Disease (PD)- 500,000 people in U.S. $6,000,000,000 spent annually for PD treatment. National Institute of Neurological Disorders and Stroke.

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Caffeic Acid and C. elegans’ Protection Against Toxin Induced Neurodegeneration

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  1. Caffeic Acid and C. elegans’ Protection Against Toxin Induced Neurodegeneration By: Michelle Chyn

  2. Need • Parkinson’s Disease (PD)- 500,000 people in U.S. • $6,000,000,000 spent annually for PD treatment National Institute of Neurological Disorders and Stroke Figure 1: Cost of PD treatment per person annually Dodel RC, Pepperl S, Köhne-Volland R, Szucs T, Werhahn KJ, Noachtar S, Oertel WH. “Costs of drug treatment of neurologic diseases: Parkinson disease, dystonia, epilepsy.” European Journal of Neurology. Vol. 7. Pp. 479-85 1991.

  3. Need Cont. • Coffee consumption in North America and Europe = 1/3 of the tap water intake http://www.e-importz.com/images/coffee_consumption.gif Figure 3: Coffee Consumption http://www.parkinsonsdecisionaid.eu.com/images/ac/2008/incidence.gif Figure 2: PD in European Countries

  4. Knowledge Base Figure 4: http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=681&loc=ec_rcs Figure 5: 3-hydroxytyramine (Dopamine) http://en.wikipedia.org/wiki/Basal_ganglia • C. elegans biosynthesize dopamine in the corpus stratum (basil ganglia) (Braugart, 2004)

  5. Knowledge Base Cont. • Copper is a neurotoxin when consumed in excess amounts (Gaggelli et al., 2006) Figure 6: Copper http://en.wikipedia.org/wiki/Copper(I)_oxide

  6. Literature Review • Antioxidants in coffee decreases risk of development of age-related diseases (Eskelinen et. al, 2009) • Caffeine intake doesn’t show health benefits (Lopez-Garcia, et. al, 2008) Figure 7: Caffeic acid http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=689043&loc=ec_rcs

  7. Literature Review Cont. • Caenorhabditis elegans MPP+ model used as PD model for testing anti-PD drugs (Braungart, et al. 2004) Figure 8: Dose-response effect of MPP+ treatment on the mobility of wild-type C. elegans animals. http://content.karger.com/ProdukteDB/produkte.asp?Aktion=ShowPDF&ArtikelNr=80983&Ausgabe=230458&ProduktNr=229093&filename=80983.pdf

  8. Literature Review Cont. • Green Florescent Protein (GFP) florescence shown in dopaminergic neurons of transgenic strains of C. elegans (Braungart, et al., 2004) http://content.karger.com/ProdukteDB/produkte.asp?Aktion=ShowPDF&ArtikelNr=80983&Ausgabe=230458&ProduktNr=229093&filename=80983.pdf Figure 9: GFP expression in dopaminergic neurons of untreated and MPP+ treated worms

  9. Literature Review Cont. • Direct addition of dopamine-2-c did not help increase noradrenaline presence in PD patients (Goodall and Alton, 1969) Figure 10: % noradrenaline recovery after dopamine-2-c infusion http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=297487&blobtype=pdf

  10. Purpose • Observing the effect of caffeic acid on neurotoxin-induced degeneration in C. elegans • H0- caffeic acid will not affect the degeneration rate in the C. elegans • HA- caffeic acid will decrease degeneration in the C. elegans. Hypothesis

  11. Methodology Compounds in Coffee and C. elegans Protection Against Parkinson’s Disease Caenorhabditis elegans egIs1[Pdat-1::GFP] strain from Caenorhabditis Genetics Center (CGC) N=90 Group 3 addition of copper oxide caffeic acid- higher concentration according to LD 50 N=30 Group 2 addition of copper oxide caffeic acid- lower concentration according to LD 50 N=30 Control addition of copper oxide, concentratin according to LD 50 N=30 C. elegans cultured at 20°C in NGM plates in normal oxygen environment. Caffeic acid and MMP+ concentrations inserted in food of E. coli for 3 days at L1 stage. Organisms then moved out into normal NGM plates. Data collected through stereo florescent microscope viewing of GFP florescence on a scale of normal, weak, and none, and area through Adobe Photoshop on the 3rd day. Statistical analysis using SPSS and ANOVA

  12. Protocols Figure 11: Culturing of C. elegans on a petri dish Picture drawn by Victoria Wei • Eggs laid overnight at L4 stage on fresh NGM plates (Mehta, et al, 2009)

  13. Protocols Cont. • N=30 per group, total N=90 • Group 1: Copper oxide • Group 2: Copper oxide and Caffeic acid • Group 3: Copper oxide and Caffeic acid http://www.scq.ubc.ca/wp-content/uploads/2006/07/wormcycle.gif Figure 12: C. elegans growth stages

  14. Protocols Cont. • Data collect on scaled brightness of GFP expression (Braungart, et al., 2004) and area of florescence on Photoshop • Data analysis using SPSS Figure 14: Fluorescent Microscope http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=33557&loc=ec_rcs http://www.niams.nih.gov/Research/Ongoing_Research/images/ost_images/leica_dmr_fluor_micro.jpg Figure 13: Sodium azide (NaN3)

  15. Protocols

  16. Do-Ability • Equipment available: • GFP filter and florescent microscope • UV lamp • Petri dishes • Photoshop • Equipment needed: • C. elegans pdat-1::GFP strain from CGC • E. coli OP50 strain from CGC • Copper, sodium azide, and caffeic acid from Sigma • NGM from Carolina Biological

  17. Budget

  18. Bibliography • American College of Physicians. “Coffee Drinkers Have Slightly Lower Daeth Rates, Study Finds.” ScienceDaily 17 June 2008. 3 May 2009 <http://www.sciencedaily.com/releases/2008/06/080616170839.htm>. • Biology The Dynamics of Life. New York: McGraw-Hill Glencoe, 2004. • Braungart, Evelyn; Gerlach, Manfred; Riederer, Peter; Baumeister, Ralf; and Hoener, Marius C. “Caenorhabditis elegans MPP+ Model of Parkinson’s Disease for High-Throughput Drug Screening.” Neurodegenerative Diseases. 2004. Vol. 1: pgs 175-183. • Brown, Marishka K.; Evans, Joseph L.; Yuan, Luo. “Beneficial effects of natural antioxidants EGCG and α-lipoic acid on life span and age-dependent behavioral declines in Caenorhabditis elegans” Elsevier Science. Vol. 85. pp. 620-628. 2006 • Colleta, Susan. Introduction to C. elegans. Waksman Student Scholars. <http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html>. 2009. • “Flavonoids.” Phytochemicals. 2005. 4 May 2009. <http://www.phytochemicals.info/phytochemicals/flavonoids.php>. • Goodall, McC. and Alton, Harold. “Dopamine (3-Hydroxytyramine) Metabolism in Parkinsonism.” The Journal of Clinical Investigation. Volume 48. pp. 2300-2308. 1969. • Halliwell, Barry and Gutterridge, John M.C. “Free Radicals in Biology and Medicine.” Third Edition. Oxford Science Publications. Oxford Unitersity Press. 1999. • Hoch, Daniel B.; Zieve, David. “Parkinson’s Disease.” U.S. National Library of Medicine. January 21, 2009. May 6, 2009. <http://www.nlm.nih.gov/medlineplus/ency/article/000755.htm>. • Jessica C. Greene; Whitworth, Alexander J.; Kuo, Isabella; Andrews, Laurie A.; Feany, Mel B.; Pallanck, Leo J. “Mitochondrial pathology and apoptotic muscle degeneration inDrosophila parkinmutants.” Davidson College Review Papers. 2003. • Karolinska Institute. “Midlife Coffee And Tea Drinking May Protect Against Late-life Dementia”. ScienceDaily 15 January 2009. 3 May 2009. <http://www.sciencedaily.com/releases/2009/01/090114200005.htm>.

  19. Bibliography Cont. • Kiontke, Karin and Sudhaus, Walter. “Ecology of Caenorhabditis species.” WormBook The online review of C. elegans Biology. <http://www.wormbook.org/chapters/www_ecolCaenorhabditis/ecolCaenorhabditis.htm> • Lagasse, Paul. Columbia Encyclopedia. “Coffee”. Columbia University Press. May 2, 2009. • Lopez-Garcia, Esther; van Dam, Rob M.; Li, Tricia Y.; Rodriguez-Artalejo, Fernando; and Hu, Frank B. “The Relationship of Coffee Consumption with Mortality.” Annals of Internal Medicine, June 16, 2008 • Margreet R. Olthof, Peter C.H. Hollman, and Martijn B. Katan. “Chlorogenic Acid and Caffeic Acid are Absorbed in Humans.” Human Nutrition and Metabolism. 2000. <http://jn.nutrition.org/cgi/reprint/131/1/66>. • Nass R, Blakely RD. 2003. The Caenorhabditis Elegans Dopaminergic System: Opportunities for Insights into Dopamine Transport and Neurodegeneration. Annual Review of Pharmacology and Toxicology 43: 521‐544 • Natarajan, K.; Singh, Sanjaya; Burke, Terrence R. Jr.; Grunberger, Dezider. “Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF- κB”. Immunology. Vol 93, pp. 9090-9095. August 1996. • “Parkinson’s Disease: Glossary of Parkinson’s Disease Terms.” WebMD. Parkinson’s Disease Health. May 7, 2009. < http://www.webmd.com/parkinsons-disease/parkinsons-glossary?page=3>. • “Parkinson's Disease: Hope Through Research”. National Institute of Neurological Disorders and Stroke. April 24, 2009. May 5, 2009. <http://www.ninds.nih.gov/disorders/parkinsons_disease/detail_parkinsons_disease.htm> • Shanam, Shai. “Worming into the cell: Viral reproduction in Caenorhabditis elegans.” Proceedings of the National Academy of Sciences. Vol. 103. n. 11. pgs. 3955-3956. • Sutphin GL, Kaeberlein M (2009). Measuring Caenorhabditis elegans Life Span on Solid Media. JoVE. 27. <http://www.jove.com/index/details.stp?id=1152> • Tipton KF, Singer TP. “Advances in our understanding of the mechanisms of the neurotoxicity of MPTP and related compounds.” J Neurochem 1993;61:1191–1206.

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