Nutrient And Non-Nutrient Intake, Relative Biomarkers and Their Health Effects In Apparently Healthy Participants

Two repeated 24h recalls performed at least 3 weeks apart will be used to assess energy, nutrient and non-nutrient intake.

Anthropometric measures including weight (in kilograms) and height (in meters) will be measured and combined to report body mass index (BMI) in kg/m^2.

Bio-Impendence Analysis (BIA) was used to assess fat free mass (FFM) in kg and fat mass (FM) in kg.

Bio-Impendence Analysis (BIA) was used to measure phase angle in degree.

The questionnaire consists of 9 items assessing the daily consumption of fruit, vegetables, cereals, meat and meat products, dairy products, alcohol, and olive oil, and the weekly consumption of legumes and fish. Each item provides a score ranging from 0 to 2 points. The final score ranges from 0 (low adherence) to 18 (high adherence to the Mediterranean diet).

The questionnaire consists of 14 items evaluating the consumption of olive oil for cooking and dressing; daily intake of vegetables, fruits, butter and red meat; weekly intake of wine, legumes, fish, nuts, sweets, pastries or sugar-sweetened beverages; intake of white meat instead of red or processed red meat and the use of "sofrito" sauce. Each item provides a score ranging from 0 to 1 point. The final score ranges from 0 (low adherence) to 14 (high adherence to the Mediterranean diet).

The short form of the International Physical Activity Questionnaire (IPAQ) consists of 7 items, exploring the intensity of physical activity based on metabolic equivalent of task (MET) consumption and sedentary behavior.

The NFFQ (NOVA food frequency questionnaire) consists of 94-item grouped in 9 sections headings: (1) fruit and nuts, (2) vegetables and legumes, (3) cereals and tubers, (4) meat and fish, (5) milk, dairy products, and eggs, (6) oils, fats, and seasonings, (7) sweets and sweeteners, (8) beverages and (9) other. Data are reported as g/day and total daily intake for each NOVA category.

Serum proteins involved in the assessment of nutritional status including albumin in mg/L and transferrin in mg/L as well as serum iron in mg/L will be evaluated through standardized analytical methods.

Markers involved in glucose and lipid metabolisms such as fasting glucose in mg/dl, total cholesterol in mg/dl, LDL-cholesterol in mg/dl, HDL-cholesterol in mg/dl and triglycerides in mg/dl will be evaluated in serum through standardized analytical methods.

Markers involved in hepatic functions such as aspartate transaminase in U/L, alanine transaminase in U/L and gamma glutamyl transferase in U/L will be evaluated in serum through standardized analytical methods.

Markers involved in renal functions including uric acid, azotemia and creatinine in mg/dl will be evaluated in serum through standardized analytical methods.

Plasma vitamin A and vitamin E in µg/mL as well as bioactive compounds such as carotenoids in µg/mL will be evaluated through standardized analytical methods.

Plasma vitamin D in ng/mL will be evaluated through standardized analytical methods.

Plasma vitamin B12 in ng/mL and folate in ng/mL will be evaluated through standardized analytical methods.

C-reactive protein in mg/L will be measured in serum through standardized analytical methods.

Markers involved in the inflammatory status such as TNF-α in pg/mL, IL-6 in pg/mL will be measured in serum through ELISA kits.

Intestinal permeability will be assessed by analyzing serum zonulin in ng/mL and ZO-1 in ng/mL using ELISA kits.

Serum calprotectin in µg/mL and fecal calprotectin in µg/g will be analyzed.

Microbiota composition in fecal samples will be evaluated through 16S rRNA sequencing methods.

Proteomic will be performed on serum samples using Somascan.

Metabolomic will be performed on urine samples by mass spectrometry.

To assess damage at the cellular DNA level the Comet assay technique will be employed.