the effect of fruit polyphenols on the lifespan of c. elegans
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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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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.
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