ASEA Research Summary Presentation - http://bit.ly/1j4wXnn
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Metabolomics: Goal is to measure the influence of ASEA on small molecules (metabolites) that shift in response to supplementation. The shift in metabolites, depending on the nutritional product, may represent effects on inflammation, oxidative stress, and physiologic stress.
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ASEA Research Summary Presentation - http://bit.ly/1j4wXnn
- 1. ASEA Metabolomics Results Nieman DC. Human Performance Laboratory, North Carolina Research Campus and Appalachian State University
- 2. Metabolomics Laboratory, North Carolina Research Campus, David H. Murdock Research Institute Slide 1 Appalachian state Human Performance Laboratory
- 3. BASELINE TESTING for VO2max, body composition (N=20 subjects) 1-week • 75-km cycling ASEA 1 week • 75-km Placebo cycling 1-h Blood/urine : 3-week washout Blood/urine : Pre-Ex Post- Ex Crossover Pre-Ex Post- Ex 1-h 1-week • 75-km cycling ASEA 1-week • 75-km Placebo cycling Metabolomics: Goal is to measure the influence of ASEA on small molecules (metabolites) that shift in response to supplementation. The shift in metabolites, depending on the nutritional product, may represent effects on inflammation, oxidative stress, and physiologic stress. Slide 2 Appalachian state Human Performance Laboratory
- 4. Working Summary • Seven days ingestion of ASEA (relative to placebo) caused an extensive mobilization of free fatty acids from adipose tissue in male cyclists. • Athletes on ASEA for 7-days started the 75-km cycling trial with high blood free fatty acids leading to increased fat oxidation and a sparing of amino acids (and potentially muscle glycogen). • ASEA intake was associated with a large increase in serum ascorbic acid levels (probably from the adrenal cortex). • Serum creatinine and urea also increased postexercise. Slide 3 Appalachian state Human Performance Laboratory
- 5. 2) Acute Effect: Increased fatty acid oxidation and mobilization during exercise (placebo condition was linked to a late mobilization). Triglyceride Mobilization: corresponding with the increase in free fatty acids, glycerol was higher at baseline (indicative of extensive adipose triglyceride hydrolysis). Least Square Mean Area 1) Chronic Effect: Higher fatty acid levels preexercise (several types of fatty acids --- see slides). Least Square Mean Area Finding 1: Ingestion of ASEA beverage for one week strongly increased serum fatty acids levels (most likely from adipose tissue). Myristic acid 14C Saturated Fatty Acid FDR=6.49E-32 2,000 1,500 1,000 500 - ASEA Placebo Myristic acid 14C Saturated Fatty Acid FDR=2.61E-20 2,000 1,500 1,000 500 - Pre ASEA Post 1H Post Placebo TG Hydrolysis FFAs Slide 4 + Glycerol with ASEA Appalachian state Human Performance Laboratory
- 6. Post 7-day Ingestion: Fatty Acids Higher in ASEA vs. Placebo 2,000 1,800 1,600 1,400 1,200 1,000 800 600 400 200 - Palmitic Acid 16C Saturated Fatty Acid FDR=1.86E-25 Least Square Mean Area Least Square Mean Area Myristic acid 14C Saturated Fatty Acid FDR=6.49E-32 ASEA Placebo Least Square Mean Area Oleic Acid 18C Monounsaturated n9 Fatty Acid FDR=5.21E-18 18000 16000 14000 12000 10000 8000 6000 4000 2000 0 Slide 5 25,000 20,000 15,000 10,000 5,000 - ASEA Placebo Stearic Acid 18C Saturated Fatty Acid FDR=3.22E-12 8000 7000 6000 5000 4000 3000 2000 1000 ASEA Placebo 0 ASEA Placebo Appalachian state Human Performance Laboratory
- 7. 7-days Ingestion of ASEA Fatty Acids and Glycerol Backbone: Higher in ASEA vs. Placebo Least Square Mean Area Palmitelaidic Acid 16C Trans Fatty Acid FDR=1.13E-08 Capric Acid 10C Saturated Fatty Acid FDR=5.39E-07 1,200 160 1,000 140 120 800 100 600 80 400 60 40 200 - 20 ASEA Placebo 0 ASEA Placebo Glycerol Backbone of Triglycerides FDR=9.49E-07 25000 20000 15000 10000 5000 0 Slide 6 ASEA Placebo Appalachian state Human Performance Laboratory
- 8. Least Square Mean Area Myristic acid 14C Saturated Fatty Acid FDR=2.61E-20 2,000 1,500 1,000 500 - Pre Post Least Square Mean Area ASEA 20,000 15,000 10,000 5,000 - Pre Placebo Post ASEA 1,200 1H Post Placebo Oleic Acid 18C Monunsaturated n9 Fatty Acid FDR = 7.96E-10 20,000 1,000 15,000 800 600 10,000 400 200 Pre Post ASEA Slide 7 25,000 1H Post Palmitelaidic Acid 16C Trans Fatty Acid FDR=1.66E-16 - Palmitic Acid 16C Saturated Fatty Acid FDR=1.56E-20 Least Square Mean Area Least Square Mean Area Serum Fatty Acids During Exercise Placebo 1H Post 5,000 - Pre Post ASEA 1H Post Placebo Appalachian state Human Performance Laboratory
- 9. Capric Acid 10C Saturated Fatty Acids FDR=0.0059 12,000 10,000 8,000 6,000 4,000 2,000 - Pre Post Least Square Mean Area ASEA 1H Post Least Square Mean Area Stearic Acid 18C Saturated Fatty Acids FDR=1.33E-06 1,000 800 600 400 200 Post ASEA Slide 8 100 50 - Pre Placebo Post 1H Post Placebo Glyercol Monosterate FDR = 0.0060 1,200 Pre 150 ASEA Lauric Acid 12C Saturated Fatty Acids FDR=0.0281 - 200 Placebo 1H Post Least Square Mean Area Least Square Mean Area Serum Fatty Acids During Exercise 900 800 700 600 500 400 300 200 100 - Pre Post ASEA 1H Post Placebo Appalachian state Human Performance Laboratory
- 10. Serum Serine FDR=0.0273 Serum Aspartate FDR=0.0075 Serum Glycine FDR=0.0162 Pyruvate Pre Post Pre 1H Post In urea cycle Post Oxaloacetate Pre Malate Post Succinate 1H Post 1H Post Serum Threonine FDR=0.0108 Pre Post 1H Post Via Beta Oxidation alphaKetoglutarate Fumarate Serum Leucine FDR=0.0004 Succinyl CoA Pre Via Odd Chain Beta Oxidation Serum Valine FDR=0.0089 Pre Slide 9 Post Placebo 1H Post Finding 2: High levels of blood free fatty acids were linked to a sparing of amino acid catabolism, and increased Krebs Cycle intermediates, post-exercise Post Isocitrate Graph Key ASEA Pre Pre Citrate 1H Post Serum Malate FDR= 0.0004 1H Post Serum Citrate FDR = 0.0037 Acetyl Co-A Serum Fumarate FDR=1.06E-06 Pre Post Post 1H Post Post 1H Post Via Glutamate Serum Proline FDR=0.0066 Pre Post 1H Post Appalachian state Human Performance Laboratory
- 11. Serum Amino Acids at Pre, Post, and 1H Post-Exercise “Sparing” of Amino Acids with ASEA Proline Krebs Entry: alpha Ketoglutarate FDR = 0.0066 30,000 25,000 20,000 15,000 10,000 5,000 - Pre Post ASEA 1H Post Lesat Square Mean Area Least Square Mean Area Leucine Krebs Entry: alpha Ketoglutarate FDR =0.0004 50,000 40,000 30,000 20,000 10,000 - Placebo 300 200 100 ASEA Slide 10 Placebo 1H Post Lesat Square Mean Area Lesat Square Mean Area 400 Post 1H Post Placebo Valine Krebs Entry: Succinyl CoA FDR = 0.0089 500 Pre Post ASEA Aspartate Krebs Entry: Oxaloacetate FDR = 0.0074 - Pre 50,000 40,000 30,000 20,000 10,000 - Pre Post ASEA 1H Post Placebo Appalachian state Human Performance Laboratory
- 12. Serum Amino Acids at Pre, Post, and 1H Post-Exericse “Sparing” of Amino Acids with ASEA Serine Krebs Entry: Pyruvate FDR= 0.0273 7,000 Lesat Square Mean Area Lesat Square Mean Area Threonine Krebs Entry: Pyruvate FDR= 0.0108 6,000 5,000 4,000 3,000 2,000 1,000 - Pre Post 1H Post 12,000 10,000 8,000 6,000 4,000 2,000 - Pre Post 1H Post Lesat Square Mean Area Glycine Krebs Entry: Pyruvate FDR= 0.0162 5 5 4 4 3 3 2 2 1 1 - Slide 11 Graph Key ASEA Pre Post 1H Post Placebo Appalachian state Human Performance Laboratory
- 13. Serum Krebs Intermediate at Pre, Post, and 1H Post-Exercise Higher Levels with ASEA Fumarate FDR = 1.06E-06 Malate FDR= 0.0004 200 Lesat Square Mean Area Lesat Square Mean Area 600 500 400 300 200 100 - Pre Post 1H Post 180 160 140 120 100 80 60 40 20 - Pre Post 1H Post Citrate FDR = 0.0037 Least Squre Mean Area 60.00 50.00 40.00 30.00 Graph Key 20.00 10.00 - Slide 12 ASEA Pre Post Placebo 1H Post Appalachian state Human Performance Laboratory
- 14. 3) Ascorbic Acid Metabolism: ASEA supplementation appears to be affecting ascorbic acid both acutely and chronically. Fructose FDR=1.12E-05 Least Square Mean Area Least Square Mean Area Chronic Differences: ASEA group has lower baseline levels of fructose and lower levels of threonic acid. Fructose is broken down into ascorbic acid which is further metabolized into threonic acid. This could be suggestive of higher ascorbic acid production but no differences in groups were detected at baseline. 1500 1000 500 0 ASEA Placebo Threonic Acid FDR=1.46E-98 8 6 4 2 0 ASEA Placebo Acute Differences: ASEA group has higher levels of ascorbic acid, an antioxidant, and lower levels of both fructose and threonic acid. 1,000 500 Pre Post 1H Post 1,500 1,000 500 - Pre Graph Key Slide 13 ASEA Placebo Post 1H Post Least Square Mean Area 1,500 - Threonic Acid FDR=0.0031 Fructose FDR = 0.0002 Least Square Mean Area Least Square Mean Area Ascorbic Acid FDR = 5.6E-06 10 8 6 4 2 - Pre Post 1H Post Appalachian state Human Performance Laboratory
- 15. 4) Other Changes. Aminomalonic Acid FDR = 1.13E-05 400 Serum Creatinine FDR=2.55 E-06 Least Square Mean Area Least Squre Mean Area Some other changes were found both acutely and chronically that require further investigation into implications. 350 300 250 200 150 100 50 900 800 700 600 500 400 300 200 100 - - Pre Post Pre 1H Post Plays role in binding calcium to protein Post 1H Post Breakdown product of creatine Graph Key ASEA Slide 14 Placebo Least Square Mean Area Urea FDR = 0.0108 120 100 80 60 40 20 - Pre Post 1H Post Formed in liver; Removal of nitrogen and ammonia Appalachian state Human Performance Laboratory
- 16. Blood Urinary Nitrogen (BUN) Normal Range: 8-20 mg/dl 25 mg/dl 20 15 Placebo 10 ASEA 5 0 Pre Post 1H BUN levels did not differ between treatment (treatment x time p-value=0.9743) Slide 15 Appalachian state Human Performance Laboratory
- 17. Creatinine Normal Range: 0.7-1.2 mg/dl 1.4 1.2 mg/dl 1 0.8 Placebo 0.6 ASEA 0.4 0.2 0 Pre Post 1H Creatinine levels did not differ between treatment (treatment x time p-value=0.7717) Slide 16 Appalachian state Human Performance Laboratory
- 18. Bilirubin Normal Range: 0.3-1.2 mg/dl 1 mg/dl 0.8 0.6 Placebo 0.4 ASEA 0.2 0 Pre Post 1H Bilirubin levels did not differ between treatment (treatment x time p-value=0.9971) Slide 17 Appalachian state Human Performance Laboratory
- 19. Alkaline Phosphatase Normal Range: 39-117 IU/L 68 66 IU/L 64 62 Placebo 60 ASEA 58 56 Pre Post 1H Alkaline Phosphatase levels did not differ between treatment (treatment x time p-value=0.8819) Slide 18 Appalachian state Human Performance Laboratory
- 20. IU/L AST Normal Range: 15-41 IU/L 40 35 30 25 20 15 10 5 0 Placebo ASEA Pre Post 1H AST levels did not differ between treatment (treatment x time p-value=0.9546) Slide 19 Appalachian state Human Performance Laboratory
- 21. IU/L ALT Normal Range: 17-63 IU/L 40 35 30 25 20 15 10 5 0 Placebo ASEA Pre Post 1H ALT levels did not differ between treatment (treatment x time p-value=0.9739) Slide 20 Appalachian state Human Performance Laboratory