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

1. William Kim and Nicole Kiess 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. Caenorhabditis elegans http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif

5. C. Elegans Life Cycle http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif

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

7. Literature Review Fig. 2 Blueberry 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. elegans pharynx 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 lifespan in C. elegans. Hypothesis Null Hypothesis: The fruit polyphenols will show no increase on the lifespan of C. elegans. Alternate Hypothesis: The fruit polyphenols will 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

18. Bibliography “Aging Statistics.” Administration on Aging. (2009) 24 March 2009. <http://www.aoa.gov/AoARoot/Aging_Statistics/index.aspx> “Antioxidants.” McGraw-Hill Yearbook of Science & Technology. McGraw-Hill, 2001. “Antioxidants.” MedlinePlus. (2009) 9 Feb. 2009. <http://www.nlm.nih.gov/medlineplus/antioxidants.html#cat1> Carlson, Rick E., Kirby, Brett S., Voyles, Wyatt F., and Dinenno, Frank A. “Evidence for impaired skeletal muscle contraction-induced rapid vasodilation in aging humans.” American Journal of Physiology – Heart and Circulatory Physiology294 (2008): H1963-H1970. Joseph, James. "ARS Project: The effects of walnuts on age related changes in neuroprotection, inflammatory signaling and behavior (414597)." <http://www.ars.usda.gov/research/projects/projects.htm?ACCN_NO=414597>. Kenney, Richard A. Physiology of Aging: A Synopsis. Chicago, London. Year Book Medical Publishers, INC, 1982. Larsson, L., Grimby, G., and Karlsson, J.. “Muscle Strength and Speed of Movement in Relation to Age and Muscle Morphology.” Journal of Applied Physiology46 (1979): 451-456. "NCLD - Dyspraxia." NCLD - Home. 04 May 2009 <http://www.ncld.org/index.php?option=content&task=view&id=466>. Oliver, Rush H., Albury, Acchia N.J., and Mousseau, Timothy A.. “Programmed Cell Death in Flight Muscle Histolysis of the House Cricket.” Journal of Insect Physiology 53 (2007): 30-39. Scalbert, Augustin, Johnson, Ian T., and Saltmarsh, Mike. “Polyphenols: antioxidants and beyond.” The American Journal of Clinical Nutrition 81 (2005): 215S-7S. “Types of contractions.” Muscle Physiology. (2006). University of California. 31 May 2006. < http://muscle.ucsd.edu/musintro/contractions.shtml> USDA/Agricultural Research Service. "Adding Walnuts To Good Diet May Help Older People Improve Motor And Behavioral Skills." ScienceDaily 25 April 2009. 5 May 2009 <http://www.sciencedaily.com­ /releases/2009/04/090419201207.htm>. “What Is an Antioxidant?.” American Dietetic Association. (2006). 14 Sept. 2006. <http:/www.eatright.org/cps/rde/xchg/ada/hs.xsl/home_9660_ENU_HTML.htm>. Wilson, Mark A., Shukitt-Hale, Barbara, Kalt, Wilhemina, Ingram, Donald K., Joseph, James A., and Wolkow, Catherine A. “Blueberry polyphenols increase lifespan and thermotolerance in Caenorhabditis elegans.” Aging Cell. 5 (2006): 59-68.