Study of the Nutraceutical Properties and Health Benefits of Traditional Components of the Mediterranean Diet

Study Overview
=================
Brief Summary
-----------------
The present protocol is designed to conduct nutrigenomic and nutrigenetic studies on foods conforming to the Mediterranean diet, in order to determine the effect functional foods have on blood parameters (cholesterol metabolism, glucose metabolism, hepatic function, inflammation, nutritional status) and body composition in the context of four different diets (standard, high fat, high protein, low carbohydrate). The study focuses on the effect of these nutraceutical foods in relation to different diets. Diets were chosen to reflect the standard reference diets used by the general population, so as the outcomes of the addition of each interventional food element may be interpreted in the context of a variety of dietary patterns.

Detailed Description
-----------------
This study envisages the enrollment of subjects, to be recruited consecutively from May 2013 to May 2015 from the staff of the laboratory of the Evaluation of Nutritional State, Nutrigenetics and nutrigenomics of the Human Nutrition and Diet section of the University of Rome Tor Vergata. The study will be conducted in accordance with the Declaration of Helsinki, and approved by the Committee of the Ministero delle Politiche Alimentari e Forestali (MIPAAF) , Italy, approved by DM 06/12/2011 n° prot. 32064. All participants will be required to provide written informed consent for the study. At the time of recruitment, relevant subject data will be collected including clinical history, dietary habits, pharmacological therapy, smoking habit, alcohol consumption and level of physical activity. Exclusion criteria include history of coronary vascular disease, peripheral or cerebro- vascular disease, hepatic disease, diagnosis of diabetes mellitus, autoimmune disease, immunodeficiency syndrome or malignant neoplasia. The subjects enrolled will undergo: Anthropometric measures (body weight, height, waist, hip, abdominal, neck, arm and forearm circumferences). Skin fold measurements (bicipital, tricipital, subscapular, pectoral, supra iliac, abdominal, anterior thigh, internal thigh, supra patellar and popliteal); Blood Pressure; Body composition evaluation (bone, fat and lean mass), evaluated by DEXA (iDXA; GE Medical Systems, Milwaukee, WI) (Dual energy X-ray Absorptiometry; DPX Lunar Radiation Corp., Madison, Wisconsin, USA); Distribution of body fluids, evaluated by bioimpedensometry. Resistance, reactance, phase angle at 50 kHz will be measured using the Bioelectrical Impedance Analysis method (BIA 101S, by Akern/RIL System-Florence Analysis of genetic expression Participants will be randomized into groups. Each group will cycle through different interventions, according to a cross over study. The interventions are represented by different dietary plans based on different meals, consisting of: Standard Diet (SD) alone, and SD with functional food (40g of hazelnuts or 150 ml of biological red wine, or 100g of chestnuts, or 200g of wild mixed greens, or 100 g of olive oil, or 100 g of chocolate); High Fat diet (HF) alone, and HF with functional food (40g of hazelnuts or 150 ml of biological red wine, or 100g of chestnuts, or 200g of wild mixed greens, or 100 g of olive oil, or 100 g of chocolate). Low Carbohydrate diet (LC) alone, and LC with functional food (40g of hazelnuts or 150 ml of biological red wine, or 100g of chestnuts, or 200g of wild mixed greens, or 100 g of olive oil, or 100 g of chocolate). High Protein (HP) alone, and HP with functional food (40g of hazelnuts or 150 ml of biological red wine, or 100g of chestnuts, or 200g of wild mixed greens, or 100 g of olive oil, or 100 g of chocolate). The duration of the experimental period is 4 weeks for each diet, with a wash-out period of 3 weeks at the beginning of the study between one diet and the next.

Official Title
-----------------
Study of the Nutraceutical Properties and Health Benefits of Traditional Components of the Mediterranean Diet - A Cross Over Interventional Study

Conditions
-----------------
Obesity

Intervention / Treatment
-----------------
* Dietary Supplement: Hazelnuts
* Dietary Supplement: Chocolate
* Dietary Supplement: Red wine
* Dietary Supplement: Olive Oil
* Dietary Supplement: Wild mixed greens
* Dietary Supplement: Chestnut
* Other: No Dietary Supplement


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria:Age: 18 to 75 years inclusive BMI >19 kg/m2 - Exclusion Criteria: Past history of ischaemic coronary artery disease Peripheral or cerebral vasculopathy Hepatic disease Diabetes Mellitus Autoimmune disease HIV/AIDS Neoplastic disease Use of the following medications: lipid-lowering medications, oral anti-diabetic medication or insulin, nitroglycerin, corticosteroids.

Ages Eligible for Study
-----------------
Minimum Age: 18 Years
Maximum Age: 75 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Design Details

Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: STANDARD DIET<br>each patient is administered a food plan for four weeks. Every patient is required to consume per day: Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fiber) Three weeks of washout to avoid additive effects on treatments to follow. | Other: No Dietary Supplement<br>* Mediterranean Reference Diet, High Fat Diet, Low Carbohydrate or High Protein Diet without an added dietary supplement (hazelnut, chocolate, red wine, wild mixed greens, chestnuts, olive oil)<br>|
| Experimental: STANDARD DIET WITH HAZELNUTS<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fibers) with 40g Italian hazelnuts from Piedmont with Protected Geographical Indication Certification Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Hazelnuts<br>* Italian hazelnuts from Piedmont with Protected Geographical Indication Certification<br>* Other names: Corylus avellana;|
| Experimental: STANDARD DIET WITH RED WINE<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day : Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fibers) with 200 ml of Italian organic Red Wine. Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Red wine<br>* Italian Organic Red Wine<br>* Other names: Vitis vinifera;|
| Experimental: STANDARD DIET WITH CHESTNUTS<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fibers) with 100 g of Italian organic chestnuts. Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chestnut<br>* Italian Organic Chestnut<br>* Other names: Castanea sativa;|
| Experimental: STANDARD DIET WITH CHOCOLATE<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day : Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fibers) with 100 g of Extra-dark Italian Chocolate (min. 70% of organic cocoa solids) Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chocolate<br>* Extra-dark Italian Chocolate (min. 70% of organic cocoa solids)<br>* Other names: Theobroma cacao;|
| Experimental: STANDARD DIET WITH WILD MIXED GREEN<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fibers) with 200 g of Italian organic wild mixed green Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Wild mixed greens<br>* Italian organic wild mixed greens<br>* Other names: Chicory, rocket, nettle;|
| Experimental: STANDARD DIET WITH OLIVE OIL<br>Intervention type: each patient is administered a food plan for four weeks. Every patient patient is required to consume per day : Standard diet (Mediterrean reference: carbohydrates 55%-60%; protein 15% - 20% of which 50% are of vegetable derivation; total fats < 30% e 30 g of fibers) with 100g of Italian organic Olive Oil Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Olive Oil<br>* Italian Organic Olive Oil<br>* Other names: Olea europea;|
| Placebo Comparator: HIGH FAT DIET<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%) Three weeks of washout to avoid additive effects on treatments to follow. | Other: No Dietary Supplement<br>* Mediterranean Reference Diet, High Fat Diet, Low Carbohydrate or High Protein Diet without an added dietary supplement (hazelnut, chocolate, red wine, wild mixed greens, chestnuts, olive oil)<br>|
| Experimental: HIGH FAT WITH HAZELNUTS<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%) with 40g of Italian hazelnuts from Piedmont with Protected Geographical Indication Certification. Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Hazelnuts<br>* Italian hazelnuts from Piedmont with Protected Geographical Indication Certification<br>* Other names: Corylus avellana;|
| Experimental: HIGH FAT WITH RED WINE<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%)with 200 ml of Italian organic Red Wine Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Red wine<br>* Italian Organic Red Wine<br>* Other names: Vitis vinifera;|
| Experimental: HIGH FAT WITH CHESTNUT<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%) with 100 g of Italian organic chestnuts. Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chestnut<br>* Italian Organic Chestnut<br>* Other names: Castanea sativa;|
| Experimental: HIGH FAT WITH CHOCOLATE<br>Intervention type: each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%) with 100 g of Extra-dark Italian Chocolate (min. 70% of organic cocoa solids) Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chocolate<br>* Extra-dark Italian Chocolate (min. 70% of organic cocoa solids)<br>* Other names: Theobroma cacao;|
| Experimental: HIGH FAT WITH WILD MIXED GREEN<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%) with 200 g of Italian organic wild mixed green Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Wild mixed greens<br>* Italian organic wild mixed greens<br>* Other names: Chicory, rocket, nettle;|
| Experimental: HIGH FAT WITH OLIVE OIL<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High fat diet (carbohydrates 35%; protein 15%; total fats 50%) with 100 of Italian organic Olive oil Three weeks of washout to avoid additive effects on treatment following. | Dietary Supplement: Olive Oil<br>* Italian Organic Olive Oil<br>* Other names: Olea europea;|
| Placebo Comparator: LOW CARBOHYDRATE DIET<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) Three weeks of washout to avoid additive effects on treatments to follow. | Other: No Dietary Supplement<br>* Mediterranean Reference Diet, High Fat Diet, Low Carbohydrate or High Protein Diet without an added dietary supplement (hazelnut, chocolate, red wine, wild mixed greens, chestnuts, olive oil)<br>|
| Experimental: LOW CARBOHYDRATE DIET WITH HAZELNUT<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) with 40g Italian hazelnuts from Piedmont with Protected Geographical Indication Certification Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Hazelnuts<br>* Italian hazelnuts from Piedmont with Protected Geographical Indication Certification<br>* Other names: Corylus avellana;|
| Experimental: LOW CARBOHYDRATE DIET WITH RED WINE<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) with 200 ml of Italian organic Red Wine Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Red wine<br>* Italian Organic Red Wine<br>* Other names: Vitis vinifera;|
| Experimental: LOW CARBOHYDRATE DIET WITH CHESTNUT<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) with 100 g of Italian organic chestnuts Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chestnut<br>* Italian Organic Chestnut<br>* Other names: Castanea sativa;|
| Experimental: LOW CARBOHYDRATE DIET CHOCOLATE<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) with 100 g of Extra-dark Italian Chocolate (min. 70% of organic cocoa solids) Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chocolate<br>* Extra-dark Italian Chocolate (min. 70% of organic cocoa solids)<br>* Other names: Theobroma cacao;|
| Experimental: LOW CARBOHYDRATE DIET WITH WILD MIXED GREEN<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) with 200 g of Italian organic wild mixed green Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Wild mixed greens<br>* Italian organic wild mixed greens<br>* Other names: Chicory, rocket, nettle;|
| Experimental: LOW CARBOHYDRATE DIET WITH OLIVE OIL<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: Low carbohydrate diet (carbohydrates 35%; protein 30%; total fats 35%) with 100 of Italian organic Olive Oil Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Olive Oil<br>* Italian Organic Olive Oil<br>* Other names: Olea europea;|
| Placebo Comparator: HIGH PROTEIN DIET<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) Three weeks of washout to avoid additive effects on treatments to follow. | Other: No Dietary Supplement<br>* Mediterranean Reference Diet, High Fat Diet, Low Carbohydrate or High Protein Diet without an added dietary supplement (hazelnut, chocolate, red wine, wild mixed greens, chestnuts, olive oil)<br>|
| Experimental: HIGH PROTEIN DIET WITH HAZELNUT<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) with 40g Italian hazelnuts from Piedmont with Protected Geographical Indication Certification | Dietary Supplement: Hazelnuts<br>* Italian hazelnuts from Piedmont with Protected Geographical Indication Certification<br>* Other names: Corylus avellana;|
| Experimental: HIGH PROTEIN DIET WITH RED WINE<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) with 200 ml of Italian organic Red Wine Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Red wine<br>* Italian Organic Red Wine<br>* Other names: Vitis vinifera;|
| Experimental: HIGH PROTEIN DIET WITH CHESTNUT<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) with 100 g of Italian organic chestnuts Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chestnut<br>* Italian Organic Chestnut<br>* Other names: Castanea sativa;|
| Experimental: HIGH PROTEIN DIET WITH CHOCOLATE<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) with 100 g of Extra-dark Italian Chocolate (min. 70% of organic cocoa solids) Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Chocolate<br>* Extra-dark Italian Chocolate (min. 70% of organic cocoa solids)<br>* Other names: Theobroma cacao;|
| Experimental: HIGH PROTEIN DIET WITH ITALIAN ORGANIC WILD MIXED GREEN<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) with 200 g of Italian organic wild mixed green Three weeks of washout to avoid additive effects on treatments to follow. | Dietary Supplement: Wild mixed greens<br>* Italian organic wild mixed greens<br>* Other names: Chicory, rocket, nettle;|
| Experimental: HIGH PROTEIN DIET WITH OLIVE OIL<br>Intervention type: Each patient is administered a food plan for four weeks. Every patient is required to consume per day: High protein diet (carbohydrates 30%; protein 40%; total fats 30%) with 100 of Italian organic Olive Oil This is followed by a 3 week wash out period, to neutralize any additive effects. | Dietary Supplement: Olive Oil<br>* Italian Organic Olive Oil<br>* Other names: Olea europea;|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sarcopenic index | To calculate the sarcopenic index three calculations will be made: Body Cell Mass Index from bioelectrical impedance analysis (body cell mass in kilograms divided by height in metres squared),and the Appendicular Skeletal Muscle Index (sum of appendicular muscle mass divided by height in metres squared)and Relative Skeletal Muscle Index (total body lean mass divided by height in metres squared)both calculated from dual-energy X-ray asorptiometry. | Variation in sarcopenic index from baseline to 4 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Cardiovascular Risk Index | Cardiovascular Risk Index will be calculated from 3 formulas: Total Cholesterol (mg/dl) divided by High Density Lipoprotein (mg/dl), Low Density Lipoprotein (mg/dl) divided by High Density Lipoprotein (mg/dl), Log10 Triglycerides (mg/dl) divided by High Density Lipoprotein (mg/dl) | Variation in cardiovascular risk index from baseline to 4 weeks | 
| Homeostatic Model Assessment (HOMA-IR) | HOMA will be calculated as follows: Blood glucose (mmol/L) x Blood Insulin (mU/L) divided by 22.5 | Variation in HOMA from baseline to 4 weeks | 
| Prognostic Inflammatory and Nutritional Index (PINI) | PINI will be calculated as follows: [C-reactive Protein (mg/L) multiplied by alpha-1 acid glycoprotein (g/L)] divided by [albumin (g/L) multiplied by prealbumin (g/L)] | Variation in PINI from baseline to 4 weeks | 
| Nuclear factor kappa-light-chain-enhancer of activated B cells (NFkappaB) | Measurement of gene up-regulation or down-regulation in order of magnitude set at ±2FC (fundamental-costant-based system of units) | Variation in expression of NFkappaB from baseline to 4 weeks | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
mediterranean diet, nutrigenomics, body composition, inflammation, oxidative stress