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

Jan 14, 2016

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Page 1: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

The Effect of Fruit Polyphenols on the Longevity of

Caenorhabditis elegans

William Kim and Nicole Kiess

Page 2: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Abstract

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.

The Effect of Fruit Polyphenols on the Longevity of C. elegans

Page 3: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Figure 1: Muscle Mass Associated with Aging

Need

Page 4: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Caenorhabditis elegans

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

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

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

Page 5: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

C. Elegans Life Cycle

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

Image 3: C. elegans Life Cycle

Page 6: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Pharynx of C. elegans

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

Image 4: Pharyngeal Structure of Caenorhabditis elegans

Page 7: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

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

Literature Review

Wilson et. al. 2006

Page 8: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Literature Review

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

Page 9: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Literature Review

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

Chow et. al. 2006

Page 10: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Literature Review

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

Page 11: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

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

HypothesisH(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.

Page 12: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Methodology

Caenorhabditis elegans

Control n = 150

Pelargonidin Chloride 200 ug/mln = 50

MeasurementAging of C. elegans measurement possible with the use of FUDR.

Statistical AnalysisANOVA run by SPSS followed by a Scheffe Post Hoc Test

Pelargonidin Chloride 100ug/ml n = 50

Pelargonidin Chloride 50ug/mln = 50

Cyanidin Chloride 200ug/mgn = 75

Cyanidin Chloride

50ug/ml

N= 75

Pelgargonidin ChorideN = 150

Cyanidin ChorideN = 150

Page 13: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

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

Page 14: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

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

Page 15: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

Doability

Motic Plus with Stereomicroscope available in lab for pharynx contraction observation

Fruit polyphenols are easily obtainable C. elegans are easily obtainable and

culturable

Page 16: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

BudgetVendor Catalogue

#Page Item Item # Quanti

tyUnit $$ Total $

$

Carolina Science 2009

78 248 C. elegans N2 (wild

type)

NP-17-3500

1 $10.95 (per culture)

$10.95

Caroline Science 2009

78 248 Nematode Growth

Agar

NP-17-3520

5 $6.25 (per 135 ml bottle)

$31.25

https://www.waters.c

om/waters/partDetail.htm

?partNumber=WAT066224

n/a n/a Symmetry C18

Column, 3.5 µm, 4.6 x 75

mm

WAT066224

1 $543.00 $543.00

Carolina Science Website

n/a n/a Methanol 874950 1 $5.75 per 500 mL

$5.75

Total Price: $590.95

Page 17: The Effect of Fruit  Polyphenols  on the  Longevity  of Caenorhabditis elegans

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“Antioxidants.” McGraw-Hill Yearbook of Science & Technology. McGraw-Hill, 2001.

 

“Antioxidants.” MedlinePlus. (2009) 9 Feb. 2009. <http://www.nlm.nih.gov/medlineplus/antioxidants.html#cat1>

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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.

 

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): 451-456.

 

"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.