The Effect of Fruit polyphenols on the lifespan of c. elegans

1. William Kim The Effect of Fruit polyphenols on the lifespan of c. elegans

2. Need Figure 1: Muscle Mass Associated with Aging

3. Polyphenols Anthocyanins (ATC) Proanthocyanidins (PAC) http://harvardforest.fas.harvard.edu/research/leaves/images/anthocyanin_molecule.jpg http://www.worldofmolecules.com/antioxidants/antho.gif http://www.interdiscipline.org/Biochemistry/PNG_files/Chlorogenic_acid2.png Chlorogenic Acid (CA)

4. Caenorhabditiselegans http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif

5. Pharynx of C. elegans http://www.wormbook.org/chapters/www_organformation/organformation.html

6. C. ElegansLife Cycle http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif

7. Literature Review Fig. 2Blueberry polyphenols extend lifespan and slow aging in C. elegans Wilson et. al. 2006

8. Literature Review Figure 3: Mean lifespan of different genotypes of C. elegans

9. Literature Review Figure 4: Structural and functional declines in C. eleganspharynx from days 2-10. Chow et. al. 2006

10. Literature Review • James Joseph (2007-2009) • Tested and still testing fruit polyphenols and their effects on aging and the brain of mice • Tested effects of blackberries, blueberries, strawberries, plum juice, and other berry fruits

11. Purpose Therefore the purpose of this experiment is to test the ability of fruit polyphenols in the prolonging of lifespanin C. elegans. Hypothesis Null Hypothesis: The fruit polyphenols will show no increase on the lifespan of C. elegans. Alternate Hypothesis: The fruit polyphenolswill successfully increase the lifespan of C. elegans.

12. Methodology Caenorhabditis elegans Short lifespan of about 2-3 weeks, rapid generation time, and experiemental flexibilty Control Medium Sample Size = 5 Medium + Fed crude blueberry extract (Vaccinium angustifolium) Sample Size = 5 Medium + Fed crude blackberry extract Sample Size = 5 Medium + Fed crude strawberry extract Sample Size = 5 Medium + Fed crude Plum extract Sample Size = 5 Measurement Average pumping rate of pharynx contraction is recorded with the use of Motic Plus Camera attached to a stereomicroscope. Statistical Analysis

13. Protocols for Measurement • Commercially prepared single strength wild blueberry juice (Vaccinium angustifolium) is applied to a preconditioned C18 Column • C18 column is washed with water to remove fructose, glucose, and organic acids of blueberries, then with 100% methanol to obtain the total polyphenolic fraction http://www.waters.com/waters/partDetail.htm?locale=101&partNumber=WAT066224

14. Protocols for Measurement • Fruit polyphenol extracts is added to NGM agar plates of C. elegans on the first day of the lifespan assay • Lifespan scoring is initiated after hermaphrodites complete their final larval molt, on the first day of adulthood • Pharynx pumping rates are measured on days 4, 6, 8, and 10 of C. elegans adulthood

15. Protocols for Measurement • Pharynx contraction rate recorded with Motic Plus camera attached to a stereomicroscope • Pumps per minute measured http://www.technicalvideo.com/photos/ElegansPharynx.jpg

16. Doability • Motic Plus with Stereomicroscope available in lab for pharynx contraction observation • Fruit polyphenols are easily obtainable • C. elegans are easily obtainable and culturable

17. Budget Total Price: $590.95

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