The Effect of Anthocyanins in The Effect of Anthocyanins in the form of Whole Berries in the form of Whole Berries in

the Obesity levels of the Obesity levels of C. C. eleganselegans

Douglas ChoeDouglas Choe

Manhasset Science ResearchManhasset Science Research

NeedNeed

Picture 1 – Obesity statisticshttp://www.obesity.org/statistics/trends2001.gif

Knowledge BaseKnowledge Base

Chart 1 – Body Mass Index (BMI) Chart

http://www.bmc.org/medicine/medicine/nutrition/images/BMI-Chart.png

Knowledge BaseKnowledge Base

Picture 2 – C. eleganshttp://www.kiwicrossing.com/drrussell/images/worm.jpg

Picture 3 – C. elegans anatomyhttp://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif

Literature ReviewLiterature Review• Tsuda, et al. (2003)Tsuda, et al. (2003)• Anthocyanin – cyanidin 3-glucoside-rich purple corn color Anthocyanin – cyanidin 3-glucoside-rich purple corn color

(PCC) (PCC) • Four groups:Four groups:

– Control dietControl diet– Purple corn color anthocyanin with regular (control) diet (PCC)Purple corn color anthocyanin with regular (control) diet (PCC)– high fat (HF)high fat (HF)– HF + PCCHF + PCC

Graph 1 – Tsuda, et al. (2003) resultshttp://jn.nutrition.org/cgi/content/full/133/7/2125/F2

Literature ReviewLiterature Review

Graph 2 – Prior, et al. (2008) Resultshttp://pubs.acs.org/appl/literatum/publisher/achs/journals/production/jafcau/2008/

jafcau.2008.56.issue-3/jf071993o/images/large/jf-2007-01993o_0001.jpeg

•Prior, et al (2000)

Literature ReviewLiterature Review• Elle, et al Elle, et al

(2008)(2008)• To view lipid stores To view lipid stores

in in C. elegansC. elegans::– Fluorescent Fluorescent

microscopemicroscope– Nile Red or Sudan Nile Red or Sudan

Black BBlack B

Picture 4 – C. elegans stained with Nile Red dyehttp://wormsrus.50webs.com/Nematodes/c

%20elegans%20nile%20red.JPG

Literature ReviewLiterature Review

• Lila (2004)Lila (2004)

• Anthocyanins can help in many areasAnthocyanins can help in many areas– Visual acuityVisual acuity– Improved night visionImproved night vision– Regenerate rhodopsinRegenerate rhodopsin– ObesityObesity– DiabetesDiabetes

HypothesisHypothesis

PurposeTo determine if anthocyanins fed as a whole berry can increase obesity in test subjects or if it has the same effect of purified anthocyanins and reduces obesity

• Null hypothesis: Anthocyanins fed as the whole blueberry will have no effect on the obesity levels in Caenorhabditis elegans • Alternate hypothesis: Anthocyanins fed as a whole berry will increase obesity

MethodologyMethodology

Statistical analysis by T-test, p<.05, will be used to determine if the collected data is significant.

Protocols for measurementProtocols for measurement• Use of fluorescence microscopy and vital dyes Use of fluorescence microscopy and vital dyes

such as Nile red to view stained lipid droplets in such as Nile red to view stained lipid droplets in the intestinal cells in live animalsthe intestinal cells in live animals

• Use of Sudan Black B to stain lipid stores in Use of Sudan Black B to stain lipid stores in dead animalsdead animals

• The Nile Red dye causes the lipid droplets to The Nile Red dye causes the lipid droplets to become highly fluorescentbecome highly fluorescent

Picture 5 – C. elegans stained with Nile Red dyehttp://wormsrus.50webs.com/Nematodes/c

%20elegans%20nile%20red.JPG

Do-AbilityDo-Ability

• Fluorescent microscopes are Fluorescent microscopes are available in the labavailable in the lab

• The Nile Red and Sudan III dyes are The Nile Red and Sudan III dyes are easily obtainable by ordering online easily obtainable by ordering online and through the Carolina Science and through the Carolina Science 2009 catalogue2009 catalogue

• The The C. elegansC. elegans can be bought from can be bought from the Carolina Science 2009 cataloguethe Carolina Science 2009 catalogue

BudgetBudget

VendorVendor Catalogue Catalogue ##

PagePage ItemItem Item #Item # QuantitQuantityy

Unit $$Unit $$ Total $Total $$$

Carolina Carolina Science 2009Science 2009

7878 248248 C. C. eleganselegans H2 (wild H2 (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)

Carolina Carolina Science 2009Science 2009

7878 821821 Sudan IIISudan III NP-89-NP-89-29502950

2 2 $26.85$26.85

http://http://www.markergewww.markerge

ne.com/ne.com/ProductDetails.ProductDetails.

php/M1441 php/M1441

Nile RedNile Red 11 $55.50 (for $55.50 (for 10 mg)10 mg)

$55.50$55.50

BibliographyBibliography•Biggs, Alton, Peter Rillero, and Dinah Zike. Biology Dynamics of Life. Columbus, OH: Mcgraw-Hill/Glencoe, 2005. Pg 928 •"Blueberries May Help Reduce Belly Fat, Diabetes Risk." Science Daily: News & Articles in Science, Health, Environment & Technology. 5 May 2009 <http://www.sciencedaily.com/ •Brito, E., Araujo, M., Alves, R., Clevidence, B.A., Novotny Dura, J. 2007. Anthocyanins present in some tropical fruits. Journal of Agricultural and Food Chemistry. 55:9389-9394. •Hope. C. elegans: A Practical Approach (Practical Approach Series). New York: Oxford University Press, USA, 2000. Pgs 2-5 •Cassada, R.C., Russell, and R.L. (1976). Dev. Biol. "Maintenance of C. elegans ." WormBook. 6 May 2009 <http://www.wormbook.org/chapters/www_strainmaintain/strainmaintain.html>. •Charron, C.S., Clevidence, B.A., Britz, S.J., Novotny Dura, J. 2007. The effect of dose size on bioavailability of acylated and nonacylated anthocyanins from red cabbage (Brassica oleracea L. var. capitata). Journal of Agricultural and Food Chemistry. 55(13):5354-5362.

•Elle, Ida, Louise Olsen, Mai-Britt Mosbeech, Steven Rodkaer, Dennis Pultz, Sanne Boelt, Julius Fredens, Pernille Sorensen, and Nils Faergeman. "C. elegans: A Model for Understanding Lipid Accumulation." Lipids Insights 2008.1 (2008): 13-21. •Lila, Mary. "Anthocyanins and Human Health: An In Vitro Investigative Approach." PubMed Central Homepage. 7 May 2009 <http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1082894>. •Lila, Mary. "Anthocyanins and Human Health: An In Vitro Investigative Approach.." Journal of biomedicine & biotechnology 2004.5 (2004): 306-313. •"Natural Purple Pigments In Fruits, Vegetables And Berries, Such As Blueberries, May Help Prevent Obesity." Science Daily: News & Articles in Science, Health, Environment & Technology. 5 May 2009 <http://www.sciencedaily.com/releases/2008/02/080211091354.htm>.

•Nestle, Marion. McGraw-Hill Encyclopedia of Science and Technology. New York: McGraw-Hill, Inc., 1993. •Prior, Ronald, Xianli Wu, Liwei Gu, Tiffany Hager, Aaron Hager, and Luke Howard. "Whole Berries versus Berry Anthocyanins: Interactions with Dietary Fat Levels in the C57BL/6J Mouse Model of Obesity." J. Agric. Food Chem. 56.3 (2008): 647-653. •"WHO | World Health Organization." 3 May 2009 <http://www.who.int/en/>.