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Antioxidants, Diabetes

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Title: Antioxidants, Diabetes


1
Antioxidants, Diabetes Obesity in C. elegans
  • Manhasset Science Research
  • Madelyn Sayed

2
Need Obesity
Figure 1
http//www.americanheart.org/downloadable/heart/11
03834132259FS16OVR5.pdf
(American Heart Association, 2004)
3
Need Obesity (2)
Figure 2
http//www.oxha.org/knowledge/backgrounders/overwe
ight_and_obesity.gif/view
4
Diabetes Statistics
Figure 3
http//www.oxha.org/knowledge/backgrounders/overwe
ight_and_obesity.gif/view
5
Knowledge Base
  • Hyperlipidemia
  • Phytochemicals
  • Antioxidant
  • Beta-carotene. (Clarkson et. al., 2000)
  • Diabetes mellitus
  • Thiol
  • (also called Mercaptan)
  • (ex Methyl mercaptan, Ethyl mercaptan, Coenzyme
    A, Lipoamide, Glutathione, Cysteine)
  • "thiol." Encyclopædia Britannica. 2009.
    Encyclopædia Britannica Online. 17 May. 2009
    lthttp//www.britannica.com/EBchecked/topic/592252/
    thiolgt.

6
Seymour et al., 1997
  • Rats fed low and high fat diets with and without
    blueberries
  • Rats that received the blueberry-enriched powder
    healthier
  • Also found to be true with grapes and cherries in
    other experiments of Seymour et. al., 1997.

7
Jones et al., 2009
  • crucial regulators of lipid homeostasis in
    mammals serve similar functions in C. elegans.
  • fat metabolism, feeding and food-related
    behaviors
  • intestinal and hypodermal cells in C. elegans
    (red)
  • Systems transport fats between cells and tissues
    (green).

8
Jones et al., 2009 (2)
  1. Nile Red
  2. Sudan Black B
  3. LipidTOX neutral lipid staining
  4. Nile Red and fatty acid lipid conjugated BODIPY
    staining
  5. fatty acid lipid conjugated BODIPY staining
  6. Both Nile red and BODIPY florescence

9
Hildebrandt, 2004
  • Effect of thiol antioxidant on body fat and
    insulin reactivity
  • diabetic patients with hypoglycemia can be
    treated with antioxidants such as x-lipoic acid
    and NAC

Figure 5
Figure 4
10
Ashrafi et al., 2003 2007 (1)
  • Nile Red staining of wild type C. elegans

11
Ashrafi et al., 2003 2007 (2)
12
Ashrafi et al., 2003 2007 (3)
13
Purpose/Hypothesis
  • Purpose to test the affect of antioxidants and
    polyphenols 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.

14
Methodology
  • Cultured axenically
  • Usually grown under monoxenic conditions in labs.
  • Fed E. coli strain OP50
  • Stains Nile Red, Sudan Black B, BODIPY-labeled
    fatty acids

15
Flow Chart
Caenorhabditis elegans Lifespan (2-3 weeks)
Rapid generation time Experimental flexibility
Control Group Sample Size N ? Not fed
blueberry compounds
Experimental Group Sample Size N ? Fed crushed
frozen blueberry powder Fed crude blueberry
extract (Vaccinium angustifolium) or a C18 column
fraction containing bulk polyphenols
Measurement Amount of fat (using Adobe
Photoshop) deposited in the worm (cm3)
16
Do Ability
  • Flourescent microscope is available in the lab
  • Dissection microscope is available in the lab
  • Camera connected to microscope is available for
    photography
  • Adobe Photoshop is available to use for fat
    measurements

17
Budget
Item Vendor Catalogue Page Unit Price Qty Total Price
C. elegans N2 (wild type) Carolina Science 2009 NP-17-3500 248 10.95 (per culture) 1
NGM Carolina Science 2009 NP-17-3520 248 6.25 (per 135 mL bottle) 2
Sudan Black III (Fatstain Herxheimer Lab Grade) Carolina Science 2009 NP-89-2963 821 6.65 (120 mL bottle) 2
Nile Red Marker Gene Technologies Inc. 7385-67-3 (CAS ) __ 55.50 (10 mg) 1
79.35
18
Bibliography
  • "Antioxidant Definition." WebContent. 07 May 2009
    lthttp//www.webcontent.com/articles/113/1/Antioxid
    ant-Definition/Page1.htmlgt.
  • 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 lthttp//www.blueberry.org/hea
    lth.htmgt.
  • Childers, Norman F. "Blueberry." Encyclopedia
    Americana. 2009. Grolier Online. 4 May 2009
    lthttp//ea.grolier.com/cgi-bin/article?assetid005
    1620-00gt.
  • 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
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  • Diabetes Information. American Diabetes
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    es.jspgt.
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    WormBook, ed. The C. elegans Research Community,
    WormBook, doi/10.1895/wormbook.1.7.1,
    http//www.wormbook.org.
  • 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.
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    potent activators of SIRT1 effects on aging and
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  • Neto, Catherine C. "Cranberry and blueberry
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    Food Research 51 652-64. Wiley InerScience.
    lthttp//www3.interscience.wiley.com/journal/114274
    034/abstract?CRETRY1SRETRY0.gt
  • "Obesity and Overweight Defining Overweight and
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  • Obesity. Companies, Mcgraw-Hill. McGraw-Hill
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  • "Phytochemicals and Cardiovascular Disease."
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19
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