1. High Incidence of Lipid Deposition in the Liver of Rats Fed a Diet Supplemented with Branched- Chain Amino Acids under Vitamin B 6 Deficiency Tae Kaimoto, Mayumi Shibuya, Kazutaka Nishikawa and Hideo Maeda Department of Living Science Shikoku University, Tokushima, Tokushima 779-1192 Japan Elizabeth Katzman 2.27.14

2. What are Branched Chain Amino Acids (BCAA)? http://agron-www.agron.iastate.edu/Courses/Agron317/AA_inhibitors.htm

3. How Does this Fit with B 6

4. Roles of BCAA  Suppresses fatigue in CNS.  Prevention of Hepatic Encephalopathy.  Recovery of burn victims.  Required for protein synthesis.  Contribute to energy metabolism.

5. Problems with Supplementation  Lack of information  High protein diets with B 6 deficiency  Biochemical parameters  Fatty liver

6. Previous Study Accumulation of triacylglycerol in the liver B6 Deficiency BCAAs 20% Casein

7. Hypothesis “It is possible that a disorder of amino acid metabolism in the presence of vitamin B 6 deficiency might influence the assembly of lipids and apolipoproteins and/or the secretion of lipoproteins.”

8. What are Apolipoproteins? https://www.mabtech.com/knowledge-center/applied-research/apolipoproteins/research- studies-apolipoproteins

9. Diets and Animals  20% vitamin-free casein diet  21 days  Sacrificed

10. Isolation of Lipoproteins and Purification of Apo E Pooled serum of male Wistar Rats VLDL Preparative Ultracentrifugation APO E Column Chromatography

11. Purification of Apo E  Column Chromatography http://www.waters.com/waters/en_US/HPLC---High-Performance-Liquid- Chromatography/nav.htm?cid=10048919&locale=en_US

12. Spectrophotometer

13. Serum and Hepatic Apolipoproteins http://virology-online.com/general/Test1.htm  Enzyme-Linked Immunosorbent Assay (ELISA)

14. Enzyme-Linked Immunosorbent Assay (ELISA) http://medical-dictionary.thefreedictionary.com/enzyme- linked+immunosorbent+assay

15. Lipoprotein Analysis  High Performance Liquid Chromatography (HPLC)

16. Growth Parameters

17. Serum and Hepatic Lipids

18. Serum and Hepatic Apolipoprotein Levels

19. HPLC Analysis of Serum

20. Conclusion “Therefore, it can be inferred from our data that the secretion of synthesized lipoproteins from the liver to the bloodstream might be impaired.”

21. What does it all mean?  Triacylglycerol and APO E are getting stuck in the liver  Fatty Liver Disease  Supplementation can do more harm than good.