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Slide 1 - Antioxidants & Obesity in C. elegans Manhasset Science Research Madelyn Sayed
Slide 2 - Need: Obesity (American Heart Association, 2004) Figure 1: Prevalence of Obesity in adults ages 20-74 by sex. (1960-2004)
Slide 3 - Need: Obesity (2) Figure 2: Obese and overweight men and women in India, China, Mexico, & USA.
Slide 4 - Need: Diabetes Statistics Figure 2.5: Diabetes in patients over 18 by race and sex. (Non-Hispanic Whites, Non-Hispanic Blacks, and Mexican Americans).
Slide 5 - Knowledge Base Hyperlipidemia Phytochemicals Antioxidant Free Radical Beta-carotene. Thiol Hypodermis Figure 3: Layers of the skin in the human body are shown. The hypodermis is the lowest layer in this diagram.
Slide 6 - Knowledge Base (2) Figure 4: The lifecycle of C. elegans in hours at a constant temperature of 25°C.
Slide 7 - Knowledge Base (3) Figure 5: C. elegans at various stages in its lifecycle in hours at a constant temperature of 25°C.
Slide 8 - Seymour et al., 1997 Rats fed low and high fat diets with and without blueberries Rats that received the blueberry-enriched powder ‘healthier’ Also true with grapes and cherries in other experiments
Slide 9 - Jones et al., 2009 crucial regulators of lipid homeostasis in mammals serve similar functions in C. elegans. fat metabolism, & feeding/food-related behaviors intestinal and hypodermal cells in C. elegans (red) Systems transport fats between cells and tissues (green). Figure 6: C. elegans fat transport and storage areas.
Slide 10 - Jones et al., 2009 (2) Nile Red Sudan Black B LipidTOX neutral lipid staining Nile Red and fatty acid lipid conjugated BODIPY staining fatty acid lipid conjugated BODIPY staining Both Nile red and BODIPY florescence Figure 7: C. elegans fat deposits shown-stained with Nile Red, Sudan Black B, and other stains.
Slide 11 - Hildebrandt, 2004 Effect of thiol antioxidant on body fat and insulin reactivity 2 studies: obese & hyperlipidemic/normolipidemic treated with antioxidants such as x-lipoic acid and NAC Their studies showed that the effect of NAC was statistically significant, but the effect of creatine was not. Figure 8
Slide 12 - Ashrafi et al., 2003 & 2007 (1) Figure 9: C. elegans stained with Nile Red in C, D, and E, with Sudan Black B in F and G. Fat appears fluorescent red (N.R.) and blue-black (S.B.)
Slide 13 - Ashrafi et al., 2003 & 2007 (2) 10
Slide 14 - Halliwell et al., 2009 Pine bark extract on metabolism and lifespan of C. elegans Figure 11: Antioxidant balance in body, affect on metabolism
Slide 15 - Purpose / Hypothesis Purpose: to test the affect of antioxidants on fat deposition in C. elegans. Null Hypothesis: the blueberry polyphenol compounds will not reduce fat deposition in C. elegans. Alternate Hypothesis: the blueberry polyphenol compounds will reduce fat deposition.
Slide 16 - Methodology Cultured axenically Grown under monoxenic conditions Pine Bark Extract E. coli strain OP50 Stain: Sudan Black B
Slide 17 - Flow Chart Caenorhabditis elegans Lifespan: (2-3 weeks) Rapid generation time Experimental flexibility Control Group Sample Size N = ? Not fed Pine Bark Extract Experimental Group Sample Size N = ? Fed Pine Bark Extract Measurement: Sudan Black B fat staining Motic Plus camera with Stereomicroscope Amount of fat (using Adobe Photoshop) deposited in the worm (cm3)
Slide 18 - Do Ability Fluorescent Dissection microscope is available in the lab Pycnogenol® easily obtainable C. elegans easily cultured Sudan Black B reliable fat stain Motic plus camera with stereomicroscope for photography Adobe Photoshop available for fat measurement
Slide 19 - Protocols for Measurement C. elegans will be fed E.coli OP50 Pycnogenol® 50 mg capsules Sudan Black: 0.7 gm Propylene Glycol: 100.0 ml Figure: E. Coli bacteria
Slide 20 - Budget
Slide 21 - Bibliography "Antioxidant Definition." WebContent. 07 May 2009 . Ashrafi, Kaveh. "Genome-wide RNAi analysis of Caenorhabditis elegans fat regulatory genes." Nature (2003): 268-72. "Blueberries Health". US Highbush Blueberry Council. 07 May 2009 . Childers, Norman F. "Blueberry." Encyclopedia Americana. 2009. Grolier Online. 4 May 2009 . Clarkson, Priscilla M. "Antioxidants: what role do they play in physical activity and health?" American Journal of Clinical Nutrition 72 (2000). The American Journal of Clinical Nutrition. 15 May 2009 . Diabetes Information. American Diabetes Association. . Eisenmann, D. M., Wnt signaling (June 25, 2005), WormBook, ed. The C. elegans Research Community, WormBook, doi/10.1895/wormbook.1.7.1, Halliwell, Barry. Free Radicals in Biology and Medicine. 3rd ed. New York: Oxford University, 1999. Hope, Ian A. C. elegans A Practical Approach. New York: Oxford UP, USA, 1999. Knutson, Mitchell D. "Resveratrol and novel potent activators of SIRT1: effects on aging and age-related diseases." Nutrition Reviews 66 (2008): 591-96. Wiley InterScience.. "Maintenance of C. elegans." WormBook. 06 May 2009 McKay, R.M., McKay, J.P., Avery, L., and Graff, J.M. (2003). C. elegans: a model for exploring the genetics of fat storage. Dev. Cell 4, 131–142. Neto, Catherine C. "Cranberry and blueberry: Evidence for protective effects against cancer and vascular diseases." Molecular Nutrition & Food Research 51: 652-64. Wiley InerScience. "Obesity and Overweight: Defining Overweight and Obesity | DNPAO | CDC." Centers for Disease Control and Prevention. 13 May 2009 . “Obesity”. Companies, Mcgraw-Hill. McGraw-Hill Multimedia Encyclopedia of Science and Technology: Version 2.0. New York: Mcgraw-hill Companies, 1997. (278-280) "Obesity Statistics: Number of Obese Americans Increasing." American Sports Data - Sports Participation Marketing Research. 03 May 2009 . "Phytochemicals and Cardiovascular Disease." American Heart Association. 03 May 2009 . Riddle, Donald L. Introduction to C.elegans. 1997. Rizza, Robert A. "Diabetes Mellitus." Encyclopedia Americana. 2009. Grolier Online. 4 May 2009 . The Obesity Society. 13 May 2009 . University of Michigan. "Blueberries May Help Reduce Belly Fat, Diabetes Risk." ScienceDaily 20 April 2009. 3 May 2009 . "Welcome to the Institute for Diabetes, Obesity and Metabolism." University of Pennsylvania || School of Medicine. 13 May 2009 .