the effect of fruit polyphenols on the lifespan of c. elegans
DESCRIPTION
William Kim and Nicole Kiess. The Effect of Fruit polyphenols on the lifespan of c. elegans. Need. Figure 1: Muscle Mass Associated with Aging. Polyphenols. Anthocyanins (ATC). Proanthocyanidins (PAC). http://harvardforest.fas.harvard.edu/research/leaves/images/anthocyanin_molecule.jpg. - PowerPoint PPT PresentationTRANSCRIPT
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William Kim and Nicole Kiess
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Figure 1: Muscle Mass Associated with Aging
Need
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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
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Caenorhabditis elegans
http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif
http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg
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C. Elegans Life Cycle
http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif
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Pharynx of C. elegans
http://www.wormbook.org/chapters/www_organformation/organformation.html
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Fig. 2 Blueberry polyphenols extend lifespan and slow aging in C. elegans
Literature Review
Wilson et. al. 2006
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Literature Review
Figure 3: Mean lifespan of different genotypes of C. elegans
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Literature Review
Figure 4: Structural and functional declines in C. elegans pharynx from days 2-10.
Chow et. al. 2006
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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
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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.
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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
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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
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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
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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
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Doability
Motic Plus with Stereomicroscope available in lab for pharynx contraction observation
Fruit polyphenols are easily obtainable C. elegans are easily obtainable and
culturable
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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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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.
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"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.