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William Kim. The Effect of Fruit polyphenols on the lifespan of c. elegans. Need. Figure 1: Age Related Changes in Motor Skills and Plasticity . Knowledge Base. http://www.mayday-info.dk/715.0.html. Knowledge Base. http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg.
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William Kim The Effect of Fruit polyphenols on the lifespan of c. elegans
Need Figure 1: Age Related Changes in Motor Skills and Plasticity
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
Knowledge Base http://www.wormbook.org/chapters/www_organformation/organformation.html
Knowledge Base http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif
Literature Review Fig. 2Blueberry polyphenols extend lifespan in strains of C. elegans. Wilson et. al. 2006
Literature Review Figure 3: Blueberry polyphenols slow decline of pharynx contraction in C. elegans. Wilson et. al. 2006
Literature Review Figure 4: Mean lifespan of different genotypes of C. elegans
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
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 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 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
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
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
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
Doability • Motic Plus with Stereomicroscope available in lab for pharynx contraction observation • Fruit polyphenols are easily obtainable • C. elegans are easily obtainable and culturable
Budget Total Price: $590.95
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