Bioavailability of N-acylethanolamines: an Ileostomy Study (NAE Study)

Study Overview
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Brief Summary
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The endocannabinoids (ECs) and N-acylethanolamines (NAEs) are a group of endogenous lipid mediators which have a pleiotropic activity in the body modulating several biological pathways such as: appetite cues, food intake, blood pressure, inflammation, glycaemia, cognition and immunity. The ECs consist of N-arachidonoylethanolamide (AEA) and 2-arachidonoylglycerol (2-AG). They may have agonist activity on cannabinoid receptors CB1 and CB2 which are located in the central nervous system (CNS) and in peripheral tissues such as in the enteric nervous system (ENS), in the liver and in the adipose tissue. NAEs are known as endocannabinoid-like molecules and include oleoylethanolamine (OEA), linoleylethanolamine (LEA), and palmitoyletahanolamine (PEA). Evidence indicates that diet composition may affect fasting and post-prandial plasma ECs, N-acylphosphatidylethanolamines (NAPEs) and NAEs profile due to the content of their precursors, fatty acids and amines. It is hypothesized that the concentration of NAPEs, NAEs and ECs in a meal could influence the intestinal concentrations of these lipid mediators that could bind the receptors located on the intestinal mucosa and in turn, differently modulate appetite and energy metabolism. The study is an acute randomized crossover feeding study in ileostmists (n=14), having a breakfast meal low or high in NAPEs, NAEs and ECs. The meals are designed on a database published by our collaborators (University of Naples) and detailed in the research proposal. Concentrations of NAEs and ECs in urine, plasma and ileal fluid, beside the blood glucose, hormonal response, appetite feelings and food intake will be monitored over the experimental days.

Official Title
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Bioavailability of N-acylethanolamines: an Ileostomy Study.

Conditions
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Healthy Nutrition

Intervention / Treatment
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* Dietary Supplement: High-N-acylethanolamine meal
* Dietary Supplement: Low-N-acylethanolamine meal


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Participant must have previously undergone an ileostomy and be more than 1.5-years post-operative Male or female Aged 18-70 years at recruitment Exclusion Criteria: Participants not undergone an ileostomy and/or is less 1.5-years post-operative Adults <18 or >70 years at recruitment Pregnant/lactating female Current smokers Lactose intolerant Allergic to nuts

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: High-N-acylethanolamines meal<br>Milk (150 mL), white bread (46 g), jam (10 g), cocoa powder (15 g), whole-grain cereals (30 g). | Dietary Supplement: High-N-acylethanolamine meal<br>* Milk (150 mL), white bread (46 g), jam (10 g), cocoa powder (15 g), whole-grain cereals (30 g).<br>|
| Active Comparator: Low-N-acylethanolamines meal<br>Milk (150 mL), whole-grain bread (80 g), jam (10 g), butter (5 g), instant coffee (2 g), dried apples (30 g). | Dietary Supplement: Low-N-acylethanolamine meal<br>* Milk (150 mL), whole-grain bread (80 g), jam (10 g), butter (5 g), instant coffee (2 g), dried apples (30 g).<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| N-acylphosphatidylethanolamines (NAPEs) levels in biofluids | Significant changes from baseline in plasma, urines and, Ileal fluids levels of NAPEs by HPLC-MS analysis. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| N-acylethanolamines (NAEs) levels in biofluids | Significant changes from baseline in plasma, urines and, Ileal fluids levels of NAEs by HPLC-MS analysis. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Endocannabinoids levels in biofluids | Significant changes from baseline in plasma, urines and, Ileal fluids levels of ECs by HPLC-MS analysis. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Glycaemia | Measure of glycaemia by using a bedside glucometer. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Appetite sensations | Significant changes from baseline in hunger, satiety, fullness and prospective of consumption. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Glucagon-like peptide 1 (GLP-1) plasmatic levels | Measure of GLP-1 by using of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Glucose-dependent insulinotropic peptide (GIP) plasmatic levels | Measure of GIP by mean of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Insulin plasmatic levels | Measure of insulin by mean of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Glucagon plasmatic levels. | Measure of glucagon by mean of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| C-peptide plasmatic levels. | Measure of c-peptide by mean of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Ghrelin plasmatic levels. | Measure of ghrelin by mean of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Leptin plasmatic levels | Measure of leptin by mean of Luminex kits in plasma samples pre-treated with protease inhibitor cocktail. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 
| Energy intake during a buffet meal test | Kilojoules | 0 hours | 
| Gut microbiota composition | Microbiota composition will be determined by high throughput sequencing of the 16S ribosomal ribonucleic acid (rRNA) gene. The massive number of sequences obtained will be analyzed by using state of the art bioinformatics tools and the presence and relative abundance of the microbial species occurring in each sample will be determined. | Change from baseline at 2, 4, 6 and, 8 hours after breakfast intake | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Lipid mediators, Nutrient sensing,, Bioavailability,, N-acylethanolamines,, Satiety.