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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 • Lipofuscin is an auto-fluorescent age pigment which is found in people with neurodegenerative diseases. (Gray, et. al., 2005) http://www.innovitaresearch.org/news/res/06042501_01.jpg Figure 4 Lipofuscin in neurons of the human brain.

  6. Knowledge Base • Centrophenoxine is an anti-aging medicine which slows the accumulation of lipofuscin. • Schneider, et. al. (1977) http://commons.wikimedia.org/wiki/File:Centrophenoxine.svg Figure 5 Molecular structure of centrophenoxine

  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 • Sutphin, et. al. (2009) Figure 8 Auto fluorescent pigments present in Day 4 and Day 8 C. elegans Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009.

  9. Literature Review • Gerstbrein, et. al. (2008) Figure 9 Auto fluorescent pigments present in Day 4 and Day 8 C. elegans. Gerstbrein, Beate; G. Stamatas; N. Kollias; M. Driscoll. “In vivspectrofluorimetry reveals endogenous biomarkers that report healthspan and dietary restriction in Caenorhabditis elegans.

  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 Centrophenoxine on the development of Parkinson’s disease in C. elegans Methodology Wild type C. elegans and ham-1 (ot339) C. elegans obtained from the Caenorhabditis Genetics Center N=80 Wild type C. elegans: N=40 ham-1(ot339) C. elegans: N=40 Given 6.8 mMcentrophenoxine N=20 Control N=20 Given 6.8 mMcentrophenoxine N=20 Control N=20 6.8 mMcentrophenoxine will be applied to the Nematode Growth Media for 21 days. Use of 4',6-diamidino-2-phenylindole (DAPI) to observe the amount of auto fluorescent pigment- lipofuscin- in C. elegans. GFP filters may also be used to observe whether the C. elegans DA neurons will be affected by the application of centrophenoxine Statistical analysis

  13. Protocol • C. elegans are grown in petri dishes containing Nematode Growth Media (NGM) from Carolina Biological and fed U.V. killed Escherichia coli. Picture by author Figure 11 Culturing the C. elegans in Petri dishes

  14. Protocol • Both Ampicillin and 5-Fluoro-2′-deoxyuridin will be used with NGM in the petri dishes with C. elegans http://upload.wikimedia.org/wikipedia/commons/b/b6/Ampicillin_structure.svg Figure 13 FUDR Figure 12 Ampicillin http://www.sigmaaldrich.com/structureimages/30/mfcd00006530.gif E.coli + NGM with centrophenoxine+ Ampicillin + FUDR  using the DAPI and GFP filter to observe amount of lipofuscin and fluorescence present in both C. elegans groups

  15. Protocol Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009. Figure 14 Age synchronization of C. elegans

  16. Protocol http://upload.wikimedia.org/wikipedia/commons/7/7a/DAPI.png Figure 15 4',6-diamidino-2-phenylindole (DAPI) Figure 16 Fluorescence microscope http://www.bcgsc.ca/people/msleumer/htdocs/fact_sheet/image004.jpg Figure 17 GFP expressed in C. elegans

  17. Budget

  18. Do-ability Available for Purchase: • ham-1(ot339) and wild type C. elegans strains from CGC • DAPI and Sodium Azide from Sigma • NGM and OP50 E.coli from Carolina Biological • Centrophenoxine purchasable from Science Lab.com Equipment already Acquired: • DAPI filter, GFP filter, fluorescent microscope

  19. 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. • 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.