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The Effect of Centrophenoxine on Parkinson’s Disease

The Effect of Centrophenoxine on Parkinson’s Disease . Victoria Wei. Need. Taken from Rajput AH, Offord KP, Beard CM, Kurland LT. Epidemiology of parkinsonism: incidence, classification, and mortality. Ann Neurol. 1984;16:278-282. .

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The Effect of Centrophenoxine on Parkinson’s Disease

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  1. The Effect of Centrophenoxine on Parkinson’s Disease Victoria Wei

  2. Need Taken from Rajput AH, Offord KP, Beard CM, Kurland LT. Epidemiology of parkinsonism: incidence, classification, and mortality. Ann Neurol. 1984;16:278-282. Figure 1 The amount of Parkinson’s disease cases per 100,000 people in the United States as age increases

  3. Knowledge Base • Parkinson’s disease is a brain disorder involving the nerves. Figure 2 The effects of Parkinson’s disease http://www.spinstudios.co.uk/sa/pa3.jpg

  4. Knowledge Base http://www.wormatlas.org/handbook/fig.s/IntroFIG6.jpg Figure 3 The life cycle of C. elegans

  5. Knowledge Base http://www.innovitaresearch.org/news/res/06042501_01.jpg Figure 4 Lipofuscin in neurons of the human brain.

  6. Knowledge Base http://resources.metapress.com/pdf-preview.axd?code=w35465l87q324635&size=largest Figure 5 structure of centrophenoxide and the products of the hydrolysis- which are dimethylethanolamine (DMAE) and p-chlorophenoxyacetic acid (PCPA)

  7. Literature Review • Caldwin, et. al. (2008) Figure 6 Figure 7 Both images taken from Caldwin, Guy A.; K.A. Caldwell. “Traversing a wormhole to Combat Parkinson’s disease.” Disease Models and Mechanisms. Volume 1. pp.000-000. 2008.

  8. Literature Review • Gerstbrein, et. al. (2008) • Lipofuscin is a marker of lifespan in C. elegans Figure 9 Auto fluorescent lipofuscin pigments present in Day 7 and Day 12 C. elegans. Gerstbrein, Beate; G. Stamatas; N. Kollias; M. Driscoll. “In vivspectrofluorimetry reveals endogenous biomarkers that report healthspan and dietary restriction in Caenorhabditis elegans.

  9. Literature Review • Nass, et. al., (2002) • GFP is a usable method of observing the dopamine neurons in C. elegans. Dopamine neurons are unaffected Dopamine neurons are progressively dying Figure 10 Degeneration of dopamine neurons http://docs.google.com/gview?a=v&q=cache%3AtdtxA_lhng0J%3Aweb.mac.com%2Frdblakely%2FBlakelyLab%2FBlakely_Lab_News_files%2FWorm%2520research%2520yields%2520powerful%2520model%2520for%2520Parkinson%27s%2520disease%25203.1.02.pdf+c+elegans+parkinson%27s+filetype%3Apdf&hl=en&gl=us

  10. Literature Review • Application of centrophenoxine to the C. elegans decreased the rate of lipofuscin accumulation by an average of 41.3%. (Shulkin, et. al., 1978) Figure 10 Table displaying effect of centrophenoxine on lipofuscin in C. elegans Shulkin, D.J.; B.M. Zuckerman. “Spectrofluorometric analysis of the effect of centrophenoxine on lipofuscin accumulation in the nematode C. elegans.” Age. Volume 5. Pp. 50-53. 1982.

  11. Purpose • The purpose of the experiment is to observe the effects of centrophenoxine on Parkinson’s disease in C. elegans. Hypothesis • Null- the symptoms of Parkinson’s disease will remain the same with or without the application of centrophenoxine. • Alternate- the symptoms of Parkinson’s disease will lessen with the application of centrophenoxine.

  12. The Effects of Centrophenoxineon Parkinson’s disease in Caenorhabditiselegans Methodology Caenorhabditis elegansham-1(ot339) obtained from the Caenorhabditis Genetics Center (CGC) N=100 Given 6.8 mMcentrophenoxine: N=50 No centrophenoxine is applied: N=50 C. elegans will be cultured in the Nematode Growth Media at 20 °C , and fed Escherichia coli(op50). 6.8 mMcentrophenoxine will be applied to the NGM where the C. elegans are cultured and observed throughout lifespan. Qualitative data will be collected on how strong both the GFP and lipofuscin fluorescence is-using a scale that ranges the strength of fluorescence. Statistical analysis with SPSS

  13. Protocol • C. elegans are grown in petri dishes at 20°C with NGM and fed U.V. killed E. coli. • Both Ampicillin and 5-Fluoro-2′-deoxyuridin (FUDR) are added to the plates Picture by author Figure 11 Culturing the C. elegans in a petri dish

  14. Protocol E.coli + NGM with centrophenoxine + Amp + FUDR using the DAPI and GFP filter to observe amount of lipofuscin and GFP fluorescence present in both C. elegans ham-1(ot339)groups

  15. Protocol Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009. Figure 12 Age synchronization of C. elegans

  16. Budget

  17. Do-ability Available for Purchase: • ham-1(ot339) C. elegans strain from CGC • DAPI and Sodium Azide from Sigma • NGM and OP50 E.coli from Carolina Biological • Centrophenoxinefrom Science Lab.com Equipment already Acquired: • DAPI filter, GFP filter, fluorescent microscope, UV lights

  18. Bibliography •  "About Parkinson Disease." National Parkinson Foundation. <”http://www.parkinson.org/Page.aspx?pid=225”>. 1996-2007. • Braungart, Evelyn; Gerlach, Manfred; Riederer; Peter, Baumeister, Ralf; and Hoener, Marius C. “Caenorhabditis elegans MPP+ Model of Parkinson’s Disease for High-throughout Drug Screening.” Neurodegenerative Disease. 2004. Volume 1: pgs 175-183. • Caldwin, Guy A.; K.A. Caldwell. “Traversing a wormhole to Combat Parkinson’s disease.” Disease Models and Mechanisms. Volume 1. pp.000-000. 2008. • Colleta, Susan. Introduction to C. elegans. Waksman Student Scholars. <http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html>. 2009 • Gerstbrein, Beate; G. Stamatas; N. Kollias; M. Driscoll. “In vivspectrofluorimetry reveals endogenous biomarkers that report healthspan and dietary restriction in Caenorhabditis elegans. • Hall, D. H.; Z. F. Altun. “C. elegans Atlas.” Genetics Research,90 , pp 375-376. 2008. • Hunt, Sara S. The Aging Process. Washington D.C. April 2004. • Kenyon, Cynthia. “Environmental Factors and Gene Activities That Influence Life Span” C. elegans II. Cold Spring Harbor Press. 1997. • Kisiel, Marion J.; B. Zuckerman. “Effects of Centrophenoxine on the Nematode CaenorhabditisBriggsae” Age. Volume 1. Pp.17-20. January 1978. • Mc Naught, KS; P. Jenner. “Proteasomal function is impaired in substantianigra in Parkinson's disease “ Neuroscience Letters. Volume 297. pp. 191-194. 2001. • Nass, Richard; R. Blakely; D. Miller. “Worm yields powerful research model for Parkinson’s disease.” The Reporter. 2002. • O'Riordan ; A.M. Burnell. Intermediary metabolism in the dauer larva. II. The glyoxylate cycle and fatty acid oxidation. Comp. Biochem. Physiol. Volume 95. pp. 125-130. 1990. • Rajput AH, Offord KP, Beard CM, Kurland LT. Epidemiology of parkinsonism: incidence, classification, and mortality. Ann Neurol. 1984;16:278-282. • Schneider, Howard F.; C. Nandy. “Effects of Centrophenoxine on Lipofuscin Formation in Neuroblastoma Cells in Culture” Journal of Gerontology. Volume 32. Pp. 132-139. 1997. • Shulkin, D.J.; B.M. Zuckerman. “Spectrofluorometric analysis of the effect of centrophenoxine on lipofuscin accumulation in the nematode C. elegans.” Age. Volume 5. Pp. 50-53. 1982. • Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009. • “What is Parkinson’s?” American Parkinson Disease Association West Coast Office. <“http://www.apdawest.org/WhatIsParkinsons.html#2”>. 2009.

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