1. Wendy Russell Natural Bioactives Inverness, Scotland Metabolic Profiling Approach to Identifying Natural Bioactives

2. Primary Metabolome (host genome) Co-Metabolome (microbiome) Metabolic Phenotype (Metabotype) Diet Environment Stress

3. Strongest prospect for predictive nutrition O OH OH O Identify bioactive molecules Assess contribution of diet …the good & the bad Why determine the human metabotype?

4. Metabolomic Technology Summation of the individual metabolomes metabolites (< 1 kDa) in body fluids Measured by LC/GC-MS and NMR Multi/mega-variate analysis

5. Bioavailability Bioactivity (at in vivo concentration) Redox Activity (Dr Charles Bestwick) Genomic Stability (Dr Susan Duthie) Inflammation Gut Microbiology Microbial Transformation (Dr Sylvia Duncan) Effect of Diet on the Microbiota (Prof Harry Flint’s Group)

6. Human Interventions 1. Protein Concentration 2. Protein Source 3. Carbohydrate Source Protein Carbohydrate

7. Protein Concentration pre-study health screen Maintenance Diet (3 days) High Protein Low Carbohydrate (4 weeks) High Protein Medium Carbohydrate (4 weeks) High Protein Low Carbohydrate (4 weeks) High Protein Medium Carbohydrate (4 weeks) Volunteers: n = 8; male BMI > 27 Study Design: Diet (g/d): Protein Carb. HPLC Diet 138 23 HPMC Diet 138 182 Faecal, blood and urine samples collected at end of each dietary period

8. Protein Concentration High Protein; Low Carbohydrate anti-inflammatory molecules protein metabolites High Protein; Medium Carbohydrate anti-inflammatory molecules protein metabolites

9. Protein Source pre-study health screen Maintenance Diet (3 days) Meat Diet (2 weeks) Meat Diet (2 weeks) Soya Diet (2 weeks) Soya Diet (2 weeks) Volunteers: n = 20 male BMI > 30 Study Design: Diet (g/d): Protein Carb. Fat Soya Diet 70 218 153 Meat Diet 71 220 155 Faecal, blood and urine samples collected at end of each dietary period

10. Protein Source Meat Diet anti-inflammatory molecules protein metabolites Soya Diet heterocyclic amines polyamines anti-inflammatory molecules protein metabolites heterocyclic amines polyamines isoflavones

11. Carbohydrate Source pre-study health screen Maintenance Diet (3 days) Resistant Starch (3 weeks) Non-Starch Polysaccharide (3 weeks) Resistant Starch (3 weeks) Non-Starch Polysaccharide (3 weeks) Volunteers: n = 14; male BMI > 27 Diet (g/d): NSP RS NSP Diet 42 13 RS Diet 2 26 Study Design: Faecal, blood and urine samples collected at end of each dietary period

12. Carbohydrate Source Faecal Resistant Starch Diet anti-inflammatory molecules protein metabolites Non-starch Polysaccharide heterocyclic amines cancer-promoting bile acids anti-inflammatory molecules chemoprotective bile acids indoles

13. Carbohydrate Source Urine Plasma Non-starch Polysaccharide Diet anti-inflammatory molecules Non-starch Polysaccharide Diet anti-inflammatory molecules

14. Fruit & Vegetable Study pre-study health screen Normal Diet (2 weeks) Normal Diet (12 weeks) Fruit & Veg Diet (12 weeks) Volunteers: n = 60, BMI = 18-30 low habitual fruit & vegetable consumption (< 3 per day) Study Design: Biomarkers: Vascular Function, Oxidative Stress, Inflammation, HDL, LDL, cholesterol, Genomic Stability Dr Charles Bestwick, Dr Susan Duthie, Dr Janet Kyle & Prof Garry Duthie (RINH) Normal Diet (6 weeks) Normal Diet (6 weeks)

15. Blaeberry Study pre-study health screen Normal Diet (2 weeks) Placebo (3 weeks) Blaeberry Supplement (3 weeks) Volunteers: n = 60; BMI > 30 Type 2 Diabetes (diet controlled) Study Design: Biomarkers: Insulin Sensitivity Oxidative Stress Inflammation Dr Nigel Hoggard (RINH)

16. Human Gut Model GenomeLab System custom multiplex assays EXPLANT PREPEXPLANT CULTURE multiplex inflammatory cytokine analysis metabolic/gut function analyses RNA extraction SUPERNATENTEXPLANT 1EXPLANT 2EXPLANT 3 Dr Janice Drew (RINH)

17. Rodent Models Dr Janice Drew and Dr Andreas Kolb (RINH) Diet-Induced Obesity Chemical-Induced Colorectal Cancer Programming of Body Weight and Ageing Custom PCR Array Strategy: Leptin/adiponectin/ insulin targets Colon mitochondrial proteomics Proteomics/Metabolomics Gene expression profiling In situ hybridisation proximal transverse distal Regional Colon Analysis precancerous lesion

18. Development of Therapeutics Dr Charles Bestwick (RINH) Prof Paul Kong (RGU) Uptake of potential therapeutics into cancer cells 6 hours 12 hours Compound 1Compound 2Compound 3

19. Summary Quantitatively identifying dietary metabolites Key active metabolites Assess bioactivity at in vivo concentrations Inform our therapeutic development work

20. Acknowledgements Laboratory Lorraine Scobbie Hyun Sang Shin Analytical Gary Duncan Louise Cantlay David Brown Human Studies Dr Alex Johnstone Dr Gerald Lobley Dr Sylvia Stephen Dr Vanessa Rungapamestry Dr Silvia Gratz David Bremner Claire Fife Ninewells; Dundee Prof Frank Carey Prof Robert Steele Tayside Tissue Bank