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William Kim Science Research – Period 4 Mrs. Flanagan. Polyphenols and Aging. Need. Figure 1: Number of people age 65 and over, by age group, selected years 1900-2006 and projected 2010-2050. Need. Figure 2: Life expectancy at ages 65 and 85, by sex, selected years 1900-2004.
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William Kim Science Research – Period 4 Mrs. Flanagan Polyphenols and Aging
Need Figure 1: Number of people age 65 and over, by age group, selected years 1900-2006 and projected 2010-2050
Need Figure 2: Life expectancy at ages 65 and 85, by sex, selected years 1900-2004
Knowledge Base http://www.mayday-info.dk/715.0.html
Knowledge Base http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif
Literature Review Fig. 3 Blueberry polyphenols extend lifespan and slow aging in Caenorhabditis elegans. Wilson et. al. 2006
Literature Review • James Joseph (2007-2009) • Tested and still testing fruit polyphenols and their effects on aging and the brain • Tested effects of blackberries, blueberries, strawberries, plum juice, and other berry fruits
Purpose Therefore the purpose of this experiment is to test the ability of blueberries, blackberries, and other berries in the prolonging of lifespan in C. elegans. Hypothesis Null Hypothesis: The blueberries, etc. will have no effect in prolonging the lifespan of C. elegans. Alternate Hypothesis: The blueberries, etc. will have a significant effect in prolonging the lifespan of C. elegans.
Polyphenols and Aging Methodology Caenorhabditis elegans Short lifespan of about 2-3 weeks, rapid generation time, and experiemental flexibilty Control Group Sample Size = 25 Not fed C. elegans medium with blueberry and other polyphenols Experimental Group Sample Size = 25 Fed crude blueberry extract (Vaccinium angustifolium) or a C18 column fraction containing bulk polyphenols in their medium Measurement Average pumping rate of pharynx contraction is recorded at days 8 and 10 of C. elegans life with the use of Motic Plus Camera attached to a stereo microscope. Statistical Analysis Statistical analysis by T-test, p<.05, will be used to determine if the collected data is significant.
Protocols for Measurement • Commercially prepared single strength wild blueberry juice (Vaccinium angustifolium) applied to a preconditioned C18 Column • C18 column washed with water to remove fructose, glucose, and organic acids of blueberries, then with 100% methanol to obtain the total polyphenolic fraction http://www.abnativeplants.com/index.cfm/fuseaction/plants.plantdetail/plant_ID/17/index.htm http://www.waters.com/waters/partDetail.htm?locale=101&partNumber=WAT066224
Protocols for Measurement • Lifespan scoring was initiated after hermaphrodites completed their final larval molt, on the first day of adulthood • Blueberry extracts were added to NGM agar plates of C. elegans on the first day of the lifespan assay • Pharynx contraction rate recorded with Motic Plus camera attached to a stereomicroscope