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

William Kim and Nicole Kiess

Figure 1: Muscle Mass Associated with Aging

Need

Polyphenols

http://www.interdiscipline.org/Biochemistry/PNG_files/Chlorogenic_acid2.png

Chlorogenic Acid (CA)

Anthocyanins (ATC) Proanthocyanidins (PAC)http://harvardforest.fas.harvard.edu/research/leaves/images/anthocyanin_molecule.jpg

http://www.worldofmolecules.com/antioxidants/antho.gif

Caenorhabditis elegans

http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif

http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg

C. Elegans Life Cycle

http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif

Pharynx of C. elegans

http://www.wormbook.org/chapters/www_organformation/organformation.html

Fig. 2 Blueberry polyphenols extend lifespan and slow aging in C. elegans

Literature Review

Wilson et. al. 2006

Literature Review

Figure 3: Mean lifespan of different genotypes of C. elegans

Literature Review

Figure 4: Structural and functional declines in C. elegans pharynx from days 2-10.

Chow et. al. 2006

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

PurposeTherefore the purpose of this experiment is to test the ability of fruit polyphenols in the prolonging of lifespan in C. elegans.

HypothesisNull 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.

Methodology

Caenorhabditis elegansShort lifespan of about 2-3 weeks, rapid generation time, and experiemental flexibilty

Control MediumSample Size = 5

Medium + Fed crude blueberry extract (Vaccinium angustifolium)Sample Size = 5

MeasurementAverage pumping rate of pharynx contraction is recorded with the use of Motic Plus Camera attached to a stereomicroscope.

Statistical Analysis

Medium + Fed crude blackberry extract Sample Size = 5

Medium + Fed crude strawberry extractSample Size = 5

Medium + Fed crude Plum extractSample Size = 5

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

BudgetVendorVendor Catalogue Catalogue

##PagePage ItemItem Item #Item # QuantiQuanti

tytyUnit $$Unit $$ Total $Total $

$$

Carolina Carolina Science 2009Science 2009

7878 248248 C. C. eleganselegans N2 (wild N2 (wild

type)type)

NP-17-NP-17-35003500

11 $10.95 (per $10.95 (per culture)culture)

$10.95$10.95

Caroline Caroline Science 2009Science 2009

7878 248248 NematodNematode Growth e Growth

AgarAgar

NP-17-NP-17-35203520

55 $6.25 (per $6.25 (per 135 ml 135 ml bottle)bottle)

$31.25$31.25

https://https://www.waters.cwww.waters.c

om/waters/om/waters/partDetail.htmpartDetail.htm

??partNumber=partNumber=WAT066224WAT066224

n/an/a n/an/a SymmetrSymmetry C18 y C18

Column, Column, 3.5 µm, 3.5 µm, 4.6 x 75 4.6 x 75

mmmm

WAT06WAT0662246224

11 $543.00$543.00 $543.0$543.000

Carolina Science Website

n/an/a n/an/a MethanolMethanol 874950 11 $5.75 per $5.75 per

500 mL500 mL$5.75$5.75

Total Price: $590.95

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