1. The Effect of Anthocyanins in the form of Whole Berries in the Obesity levels of C. elegans Douglas Choe Manhasset Science Research

2. Need Picture 1 – Obesity statistics http://www.obesity.org/statistics/trends2001.gif

3. Knowledge Base Chart 1 – Body Mass Index (BMI) Chart http://www.bmc.org/medicine/medicine/nutrition/images/BMI-Chart.png

4. Knowledge Base Picture 2 – C. elegans http://www.kiwicrossing.com/drrussell/images/worm.jpg Picture 3 – C. elegans anatomy http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif

5. Literature Review Tsuda, et al. (2003) Tsuda, et al. (2003) Anthocyanin – cyanidin 3-glucoside-rich purple corn color (PCC) Anthocyanin – cyanidin 3-glucoside-rich purple corn color (PCC) Four groups: Four groups: –Control diet –Purple corn color anthocyanin with regular (control) diet (PCC) –high fat (HF) –HF + PCC Graph 1 – Tsuda, et al. (2003) results http://jn.nutrition.org/cgi/content/full/133/7/2125/F2

6. Literature Review Graph 2 – Prior, et al. (2008) Results http://pubs.acs.org/appl/literatum/publisher/achs/journals/production/jafcau/2008/jafcau.200 8.56.issue-3/jf071993o/images/large/jf-2007-01993o_0001.jpeg Prior, et al (2000)

7. Literature Review Elle, et al (2008) Elle, et al (2008) To view lipid stores in C. elegans: To view lipid stores in C. elegans: –Fluorescent microscope –Nile Red or Sudan Black B Picture 4 – C. elegans stained with Nile Red dye http://wormsrus.50webs.com/Nematodes/c%20el egans%20nile%20red.JPG

8. Literature Review Lila (2004) Lila (2004) Anthocyanins can help in many areas Anthocyanins can help in many areas –Visual acuity –Improved night vision –Regenerate rhodopsin –Obesity –Diabetes

9. Hypothesis Purpose To 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

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

11. Protocols for measurement Use of fluorescence microscopy and vital dyes such as Nile red to view stained lipid droplets in the intestinal cells in live animals Use of fluorescence microscopy and vital dyes such as Nile red to view stained lipid droplets in the intestinal cells in live animals Use of Sudan Black B to stain lipid stores in dead animals Use of Sudan Black B to stain lipid stores in dead animals The Nile Red dye causes the lipid droplets to become highly fluorescent The Nile Red dye causes the lipid droplets to become highly fluorescent Picture 5 – C. elegans stained with Nile Red dye http://wormsrus.50webs.com/Nematodes/c%20el egans%20nile%20red.JPG

12. Do-Ability Fluorescent microscopes are available in the lab Fluorescent microscopes are available in the lab The Nile Red and Sudan III dyes are easily obtainable by ordering online and through the Carolina Science 2009 catalogue The Nile Red and Sudan III dyes are easily obtainable by ordering online and through the Carolina Science 2009 catalogue The C. elegans can be bought from the Carolina Science 2009 catalogue The C. elegans can be bought from the Carolina Science 2009 catalogue

13. Budget Vendor Catalogue # PageItem Item # Quantity Unit $$ Total $$ Carolina Science 2009 78248 C. elegans H2 (wild type) NP-17- 3500 1 $10.95 (per culture) $10.95 Caroline Science 2009 78248 Nematode Growth Agar NP-17- 3520 5 $6.25 (per 135 ml bottle) Carolina Science 2009 78821 Sudan III NP-89- 2950 2$26.85 http://www.mark ergene.com/Prod uctDetails.php/M 1441 http://www.mark ergene.com/Prod uctDetails.php/M 1441 Nile Red 1 $55.50 (for 10 mg) $55.50

14. Bibliography 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. 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. 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. 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/