DiAgRaMs- RAAK PRO. Shirley Kartaram

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1 DiAgRaMs- RAAK PRO Standardization of the bicycle ergometer test as challenge model to assess nutritional effects on intestinal health and the immune system Shirley Kartaram Department Innovative Testing in Life Sciences and Chemistry Institute for Life Sciences and Chemistry DAS_Conference_

2 Content Introduction intestinal health, epithelial integrity and permeability Translational Strategy In vitro experiments DiAgRaMs: aim, study design, relevance Protégé study Wageningen Acknowledgements DAS_Conference_

3 Intestinal health and diseases Epithelial integrity vs permeability Passage macromolecules Antigens Gut-immune system Inflammation Gut-nervous system Nervus vagus Microbiota Inflammatory bowel diseases(ibd) Arthritis Food allergy Obesity Diabetes Alzheimer Cancer DAS_Conference_

4 Epithelial integrity - Tight Junctions DAS_Conference_

5 Translational Research Strategy In vitro Caco2 cells In vivo Human study DiAgRaMs Stressor model Biomarker epithelial integrity (TEER) Distribution TJ proteins Gene expression and pathways Effects food compounds and probiotics Stressor model Biomarkers intestinal permeability Biomarkers inflammation Effects food compounds and probiotics DAS_Conference_

6 In vitro experiments TEER measurement Caco2 cells EVOM - Epithelial Voltohmmeter DAS_Conference_

7 Day 14: In vitro experiments TEER measurement : t=0h 2 h preincubation with/without probiotic strain Stressor model: + cytomix (IFN-γ, TNF-α, IL-1β) Day 15: TEER measurement: t=16h Collection supernatants, Cell lysates for WB Membrane filters for IF staining DAS_Conference_

8 TEER development In vitro experiments experiment TEER (Ω/cm2) strain Days after seeding DAS_Conference_

9 DiAgRaMs Standardization of the bicycle ergometer test as challenge model to assess nutritional effects on intestinal and immune system DAS_Conference_

10 Main aim: Standardize exercise as challenge model to test health products Primary Objective: To determine the effect of exercise on a selection of relevant biomarkers of intestinal permeability and inflammation in healthy young men Secondary Objective: To standardize the exercise model with the selected relevant biomarkers of intestinal permeability and inflammation (within three laboratories) Tertiary Objective: To study the effects of natural food compounds on biomarkers of intestinal permeability and inflammation DAS_Conference_

11 Health Health Human study Bicycle ergometer Intestinal barrier & Immune system Health- Inflammation status Biomarkers Protocol Challenge model Standardization Nutritional effects Inflammation Inflammation DAS_Conference_

12 Biomarkers Intestinal permeability Sugartest lactulose/rhamnose Zonulin Inflammatory responses Cytokines TNF-α, IFN-γ, IL1β IL6, IL7, IL8, IL4, IL5, IL13, IL18, IL10, IL2, IL33 I-FABP BLOOD SALIVA FAECES Urine CRP Cellular responses (macrophages, neutrophils, NK cells) Microbial components (LPS) Calprotectin DAS_Conference_

13 Study design Phase 1 1a: Protégé - study, WUR 1b: DiAgRaMs - study, HU/WUR, set up challenge model Phase 2 2a: Standardization challenge model HU, WUR, HANZE 2b: Nutritional intervention probiotica mix cross over study design DAS_Conference_

14 Phase 1a Phase 1a WUR: Protégé-study protein digestion and uptake after intense physical exercise To determine interpersonal variation power calculation phase 1b/2 final selection parameters Final timetable samples DAS_Conference_

15 Phase 1b dag 1 dag 2 dag 3 tijdstip conditie bepalingstest 1 week 9.00 u u geen inspanning nacht met vasten u u inspanning nacht u bloed suiker drankje u u 9.30 u u urine elk uur tot 5 u na test van 5 tot 24 u na test elk uur tot 5 u na test van 5 tot 24 u na test faeces DAS_Conference_

16 Z Z Phase 2 WUR, HU and Hanze 2a: Standardization of the exercise challenge model by determining inter- and intralab variation 2b: Nutritional intervention cross over study design darm permeabiliteits test uithoudings test W darm permeabiliteits test uithoudings test C darm permeabiliteits test uithoudings test W C darm permeabiliteits test uithoudings test 14 weken voedingsinterventie (N) of placebo (C) W wash out periode van 4 weken, zonder inname probiotica mix DAS_Conference_

17 Partners Partners DAS_Conference_

18 Relevance human study A standardized test allows to: Support health claims of food products and natural compounds (e.g. EFSA) Evaluate health in specific patient groups (e.g. obese, alzheimer) DAS_Conference_

19 Acknowledgements Ir. Laura M Rabet Lonneke Janssen Duijghuijsen Prof. Johan Garssen DAS_Conference_

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