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William Kim and Nicole Kiess

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1 William Kim and Nicole Kiess
The Effect of Fruit Polyphenols on the Longevity of Caenorhabditis elegans William Kim and Nicole Kiess

2 Abstract The Effect of Fruit Polyphenols on the Longevity of C. elegans Sarcopenia is defined as the loss of muscle mass, strength, and function associated with aging. As aging takes place, there is a sharp decrease in muscle mass and strength seen in both physically active individuals and physically inactive individuals. Sarcopenia is a major feature of aging in both humans and C. elegans, a model organism used for studying aging and longevity. Blueberry polyphenols have been found to increase the lifespan of C. elegans, seen in the pharynx contraction rate in adult C. elegans. C. elegans pumping rate of the pharynx is an indication of the age for the pumping rate of the pharynx decreases with age. Using various concentrations of different fruit polyphenols the lifespan of C. elegans will be measured by observing the pharynx contraction rate. Seventy-five worms will be fed two different concentrations of plum polyphenols, fifty C. elegans will be fed three different concentrations of strawberry polyphenols, and there will be one-hundred fifty worms in the control group. They all will be fed an equal amount of each fruit polyphenol. The average pumping rate of the pharynx per minute will be measured with the use of a Motic camera attached to a Zeiss Axiovert 40 CFL microscope.

3 Need Figure 1: Muscle Mass Associated with Aging

4 Caenorhabditis elegans
Image 1: C. elegans external appearance Image 2: C. elegans internal appearance

5 C. Elegans Life Cycle Image 3: C. elegans Life Cycle

6 Pharynx of C. elegans Image 4: Pharyngeal Structure of Caenorhabditis elegans

7 Literature Review Wilson et. al. (2006) found that blueberry polyphenols were successful in increasing the lifespan and thermotolerance in C. elegans James Joseph Fig. 2 Blueberry polyphenols extend lifespan and slow aging in C. elegans Wilson et. al. 2006

8 Literature Review Figure 3: Blueberry polyphenols extend lifespan and slow aging in C. elegans

9 Literature Review Figure 4: Structural and functional declines in C. elegans pharynx from days 2-10. Chow et. al. 2006

10 Literature Review Figure 5: Visible Structural Decline in C. elegans as a result of Sarcopenia Herndon et. al. (2002)

11 Purpose Therefore the purpose of this experiment is to test the ability of different concentrations of fruit polyphenols on the prolonging of lifespan in C. elegans. Hypothesis H(0): The pelargonidin chloride will show no effect on the prolonging of lifespan in C. elegans. H(0): The cyanidin chloride will show no effect on the prolonging of lifespan in C. elegans H(a): The pelargonidin chloride will show a positive increase in the lifespan of C. elegans. H(a): The cyanidin chloride will show a positive increase in the lifespan of C. elegans.

12 Pelgargonidin Choride
Methodology Caenorhabditis elegans Control n = 150 Pelargonidin Chloride 200 ug/ml n = 50 Measurement Aging of C. elegans measurement possible with the use of FUDR. Statistical Analysis ANOVA run by SPSS followed by a Scheffe Post Hoc Test Pelargonidin Chloride 100ug/ml Pelargonidin Chloride 50ug/ml Cyanidin Chloride 200ug/mg n = 75 Cyanidin Chloride 50ug/ml N= 75 Pelgargonidin Choride N = 150 Cyanidin Choride N = 150

13 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

14 Protocols for Measurement
Pharynx contraction rate recorded with Motic Plus camera attached to a stereomicroscope Pumps per minute measured

15 Doability Motic Plus with Stereomicroscope available in lab for pharynx contraction observation Fruit polyphenols are easily obtainable C. elegans are easily obtainable and culturable

16 Budget Vendor Catalogue # Page Item Item # Quantity Unit $$ Total $$ Carolina Science 2009 78 248 C. elegans N2 (wild type) NP 1 $10.95 (per culture) $10.95 Caroline Science 2009 Nematode Growth Agar NP 5 $6.25 (per 135 ml bottle) $31.25 https://www.waters.com/waters/partDetail.htm?partNumber=WAT066224 n/a Symmetry C18 Column, 3.5 µm, 4.6 x 75 mm WAT066224 $543.00 Carolina Science Website Methanol 874950 $5.75 per 500 mL $5.75 Total Price: $590.95

17 Bibliography “Aging Statistics.” Administration on Aging. (2009) 24 March <http://www.aoa.gov/AoARoot/Aging_Statistics/index.aspx> “Antioxidants.” McGraw-Hill Yearbook of Science & Technology. McGraw-Hill, “Antioxidants.” MedlinePlus. (2009) 9 Feb <http://www.nlm.nih.gov/medlineplus/antioxidants.html#cat1> Carlson, Rick E., Kirby, Brett S., Voyles, Wyatt F., and Dinenno, Frank A. “Evidence for impaired skeletal muscle contraction-induced rapid vasodilation in aging humans.” American Journal of Physiology – Heart and Circulatory Physiology 294 (2008): H1963-H1970. 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, Larsson, L., Grimby, G., and Karlsson, J.. “Muscle Strength and Speed of Movement in Relation to Age and Muscle Morphology.” Journal of Applied Physiology 46 (1979): "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): 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 < USDA/Agricultural Research Service. "Adding Walnuts To Good Diet May Help Older People Improve Motor And Behavioral Skills." ScienceDaily 25 April May 2009 <http://www.sciencedaily.com­ /releases/2009/04/ htm>. “What Is an Antioxidant?.” American Dietetic Association. (2006). 14 Sept <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):


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