Polyphenols, precisely resveratrol, with red wine as the most substantial source, was associated with improvements in cognitive function. Also, the loss of muscle mass and strength in elderly, that significantly increases dependency of these people, it could be attributed to alterations in gut microbiota through the "gut-muscle axis" and this underline the urgent need to efficiently find out any intervention or preventive approach via modulation of gut microbiota to improve muscle function in elderly. In this context, red wine polyphenols exert their effects through interaction with gut microbiota following the well-known two-way interaction between polyphenols and gut microbiota by promoting the proliferation of beneficial bacteria and increasing their abundance. Similarly, aging cognitive decline can be modulate by microbiota, notably through "gut-brain axis". Additionally, dietary polyphenols can delay inflammation or/and oxidation on the onset of age-related cognitive decline or muscular oxidation or cardiovascular factors risk factors, all of them relevant factors for the onset of physical frailty and dependence in elderly. Moreover, wine is a singular alcoholic beverage with a high content of phenolic compounds of a very diverse nature on which numerous protective effects on health have been described. In fact, wine, in addition to alcohol, contains a complex mixture of polyphenols, including anthocyanins and non-coloured phenols as proanthocyanidins, flavonols, hydroxycinnamic and hydroxybenzoic acids, stilbenes and lignans. Thus, the bioprotective effects of wine polyphenols could be the consequence of the synergistic effect of this complex mixture of polyphenols from the grape and the winemaking process. That is why it is essential to clarify whether consumption of polyphenols from red wine provided by a nonalcohol red wine within a healthy diet can produce beneficial effects on health, differentiating this pattern from a general consumption of alcohol generally associated with negative effects. Based on the ethical limitations to carry out diet intervention studies with wine in humans, this project proposes the use of a nonalcoholic wine as vehicle of the complex mixture of red wine polyphenols. The hypothesis of our research is that regular consumption of red wine polyphenols, 150 mg/day, delivered through a nonalcoholic red wine, in the context of a Mediterranean diet (MD pattern), could promote protective mechanisms for a healthy aging, especially through its beneficial effects on cognitive and locomotor abilities and mediated by the modulation of the intestinal microbiota (composition, function and associated metabolome). The main objective of the WinAging project is to add knowledge concerning the diet modulation of molecular mechanisms of the aging process through multi-omic approaches based on the potential health effects of a dietary strategy by a sustained MD supplementation with nonalcoholic red wine rich in polyphenols to tackle cognitive and locomotor abilities in early elderly home-dwelling subjects. The specific objectives: * Objective 1. To develop a nonalcoholic red wine with high phenolic content and sensorial acceptability. * Objective 2. To evaluate the chronic effects of the intake of wine polyphenols (average dose 150 mg/day) delivered through a nonalcoholic red wine in the context of a MD in early elderly home-dwelling subjects, and applying participatory research to increase adherence of subjects in the clinical intervention study. * Objective 2.1. To identify selective biological phenolic metabolites in human urine samples to be used as biomarkers of nonalcoholic red wine intake. * Objective 2.2. To assess the effects of the diet supplementation with red wine polyphenols on the improvement of cognitive ability. * Objective 2.3. To assess the effects of the diet supplementation with red wine polyphenols on the improvement of locomotor ability * Objective 2.4. To evaluate the effect of the diet supplementation with red wine polyphenols on the improvement of cardiovascular disease (CVD) risk factors. * Objective 3. To unravel the underlying mechanisms involved in the potential beneficial effects of red wine polyphenols on aging. * Objective 3.1. To evaluate the influence on microbiota composition, function and microbial catabolites. * Objective 3.2. To evaluate the impact on inflammation and gut health. * Objective 3.3. To evaluate the impact on metabolic pathways related with aging * Objective 3.4. To analyze the impact on age-related epigenetic modifications * Objective 3.5. To deeply characterize the underlying muscle signalling pathways affected using an animal model of aging. * Objective 4. To apply integrative computational analyses for the identification of variables (clinical or gut-related) more determinant for a successful prevention of locomotor and cognitive abilities associated with wine polyphenols.
A total of 72 home-dwelling early elderly volunteers (men and women) 60-74 years old will be included in the intervention (36 in each arm of the intervention). During the study there will be 11 visits in total. Visits will be every 4 weeks form the V1. The study visits will be the following: * Screening visit (V0). * Basal visit (V1). * Visits during intervention (V2, V3, V4, V5, V6, V7, V8, V9). * 3-month study visit (V4) * 6-month study visit (V7) * Telephone visits (V2, V3, V5, V6, V8, V9) * Final study visit (V10). In visits V0, V1, V4, V7 and V10 volunteers must present themselves in fasting conditions of 8 hours to obtain fasting blood samples. * In addition, future candidates will be provided with the nonalcoholic wine, selected for study and allowed to taste it for one week to check its tolerability and sensory acceptance and thus ensure that candidates can commit to drinking the study wine daily for 9 months and reduce follow-up losses during the intervention. Basal visit (V1; week 0): * Nuclear Magnetic Resonance (NMR) and muscle biopsy (±5 days V1)\*. * Isokinetic assessment (±5 days V1)\* * Revision of study clinical history. * Vital signs (blood pressure/resting heart rate). * Checking the concomitant medication. * Anthropometry waist circumference; hip; body weight and composition; height). * Checking the physical activity and sedentary habits (IPAQ-E). * Checking the quality of life (SF-36 Health survey questionnaire) * Checking depression symptoms (Geriatric Depression Scale Questionnaire) * Checking MD adherence * Checking the cognitive and locomotor abilities * Spanish validated of the Mini Mental State Examination (MMSE), Alzheimer disease questionnaire (AD8), Memorey Alteration tets (TAM test), Fototest * Sarcopenic parameters (muscle strength based on grip dynamometry; skeletal muscle mass index (SMI) based on bioimpedance analysis (BIA), kg/m2 and appendicular skeletal muscle mass index (ASMI) based on bioimpedance analysis (BIA), low physical performance or physical function based on 4 m gait speed, m/s. * Blood sample extraction. * Ultrasound (muscle mass and abdominal fat). * Checking the 3-day dietary record and Food Frequency Questionnaire (FFQ). * Collection of feaces samples. * Collection of saliva samples. * Collection of urine samples. * MD guidelines explanation. * Schedule the next visit and instructions. * Administration of red nonalcoholic wine through opaque bottles to protect from light. * In a subsample of n= 15 subjects/group, 5 days before or after the visit V1, volunteers must have a: * NMR, in order to measure changes in hippocampal volume, and volume of white matter hyperintense lesions, * Muscle biopsys, in order to measure changes in signalling pathways in skeletal muscle, * Isokinetics analysis in order to measure changes in muscle function through variables such as torque, work and power. Telephone visits: (V2 and V3; week 4 and 8): * Checking the FFQ * Checking changes in concomitant medication * Product intake control * Record adverse effects * Schedule the next visit and instructions. 3-month study visit (V4; week 12): * Revision of study clinical history. * Vital signs (blood pressure/resting heart rate). * Checking the concomitant medication. * Anthropometry waist circumference; hip; body weight and composition; height). * Checking the physical activity and sedentary habits (IPAQ-E). * Checking the quality of life (SF-36 Health survey questionnaire) * Checking depression symptoms (Geriatric Depression Scale Questionnaire) * Checking MD adherence * Checking the cognitive and locomotor abilities * Spanish validated of the MMSE, AD8, TAM test, Fototest * Sarcopenic parameters (muscle strength based on grip dynamometry; skeletal muscle mass index (SMI) based on bioimpedance analysis (BIA), kg/m2 and appendicular skeletal muscle mass index (ASMI) based on bioimpedance analysis (BIA), kg/m2; low physical performance or physical function based on 4 m gait speed, m/s. * Blood sample extraction. * Ultrasound (quadricep muscle mass and abdominal fat). * Checking the Food Frequency Questionnaire, Mediterranean adherence (FFQ). * Collection of urine samples. * Product intake control * Record adverse effects * MD guidelines explanation. * Schedule the next visit and instructions. * Administration of red nonalcoholic wine through opaque bottles to protect from light. Telephone visits: (V5 and V6; week 16 and 20): * Checking the FFQ * Checking changes in concomitant medication * Product intake control * Record adverse effects * Schedule the next visit and instructions. 6-month study visit (V7; week 24): * Revision of study clinical history. * Vital signs (blood pressure/resting heart rate). * Checking the concomitant medication. * Anthropometry waist circumference; hip; body weight and composition; height). * Checking the physical activity and sedentary habits (IPAQ-E). * Checking the quality of life (SF-36 Health survey questionnaire) * Checking depression symptoms (Geriatric Depression Scale Questionnaire) * Checking MD adherence * Checking the cognitive and locomotor abilities * Spanish validated of the MMSE, AD8, TAM test, Fototest * Sarcopenic parameters (muscle strength based on grip dynamometry; skeletal muscle mass index (SMI) based on bioimpedance analysis (BIA), kg/m2 and appendicular skeletal muscle mass index (ASMI) based on bioimpedance analysis (BIA), kg/m2; low physical performance or physical function based on 4 m gait speed, m/s. * Blood sample extraction. * Ultrasound (quadricep muscle mass and abdominal fat). * Checking the 3-day dietary record and Food Frequency Questionnaire (FFQ). * Collection of feaces samples. * Collection of saliva samples. * Collection of urine samples. * MD guidelines explanation. * Schedule the next visit and instructions. * Administration of red nonalcoholic wine through opaque bottles to protect from light. Telephone visits: (V8 and V9; week 28, and 32): * Checking the FFQ * Checking changes in concomitant medication * Product intake control * Record adverse effects * Schedule the next visit and instructions. Final visit (V10; week 36): * NMR and muscle biopsy (±5 days V1)\*. * Isokinetic assessment (±5 days V1)\* * Revision of study clinical history. * Vital signs (blood pressure/resting heart rate). * Checking the concomitant medication. * Anthropometry waist circumference; hip; body weight and composition; height). * Checking the physical activity and sedentary habits (IPAQ-E). * Checking the quality of life (SF-36 Health survey questionnaire) * Checking depression symptoms (Geriatric Depression Scale Questionnaire) * Checking MD adherence * Checking the cognitive and locomotor abilities * Spanish validated of the MMSE, AD8, TAM test, Fototest * Sarcopenic parameters (muscle strength based on grip dynamometry; skeletal muscle mass index (SMI) based on bioimpedance analysis (BIA), kg/m2 and appendicular skeletal muscle mass index (ASMI) based on bioimpedance analysis (BIA), kg/m2; low physical performance or physical function based on 4 m gait speed, m/s. * Blood sample extraction. * Ultrasound (quadricep muscle mass and abdominal fat). * Checking the 3-day dietary record and Food Frequency Questionnaire (FFQ). * Collection of feaces samples. * Collection of saliva samples. * Collection of urine samples. \*In a subsample of n= 15 subjects/group, 5 days before or after the visit V10. * Nuclear Magnetic Resonance (NMR), in order to measure changes in hippocampal volume, and volume of white matter hyperintense lesions, * Muscle biopsys, in order to measure changes in signalling pathways in skeletal muscle, * Isokinetics analysis in order to measure changes in muscle function through variables such as torque, work and power.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
PREVENTION
Masking
SINGLE
Enrollment
72
Mediteranean diet + nonalcoholic red wine (300 mL wine/day, equivalent to a daily dose of 150 mg red wine polyphenols/day, during meals)
Mediterranean diet + drinking water (300 mL/day, during meals)
Instituto de Ciencias de la Vid y del Vino-ICVV (Consejo Superior de Investigaciones Científicas-CSIC, Universidad de La Rioja, Gobierno de La Rioja)
Logroño, Spain, Spain
ACTIVE_NOT_RECRUITINGLyfestyle, Microbiota and Health Unit, Health Sciences Faculty, University of La Rioja
Logroño, Spain, Spain
ACTIVE_NOT_RECRUITINGUniversitat Rovira i Virgili, Facultat de Medicina i Ciències de la Salut, Functional Nutrition, Oxidation, and Cardiovascular Diseases Group (NFOC-Salut)
Reus, Spain, Spain
RECRUITINGCognitive function
The MMSE test for cognitive function. The total test score ranges from 0 (impaired) to 30 (normal) (Beaman et al., 2004).
Time frame: Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Muscle strength
The muscle strength based on handgrip dynamometry (Jamar dynamometer; Sammons Preston Rolyan, Bolingbrook, IL)
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Anthropometric measures
WC: waist circumference (cm) measured by steel measuring tape (at the umbilicus)
Time frame: Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Anthropometric measures
Height (cm) measured by wall-mounted stadiometer (Tanita Leicester Portable; Tanita Corp., Barcelona, Spain)
Time frame: Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Anthropometric measures
Weight (kg) measured by calibrated scale (TANITA MC-780MA; Tanita Corp., Tokyo, Japan).
Time frame: Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Anthropometric measures
BMI (kg/m2) calculated by weight (kg) divided to height (m2).
Time frame: Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Anthropometric measures
Body composition (Fat mass (% and kg)) measured by calibrated scale (TANITA MC-780MA; Tanita Corp., Tokyo, Japan).
Time frame: Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Physical activity compliance
International physical activity questionnaire for elderly: IPAQ for elderly IPAQ-E Spanish version (Rubio-Castañeda et al., 2017). Higher scores mean a better outcome.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Sedentary habits
Questionnaire developed by Marshall et al 2010. Higher amount of hours mean bad outcome.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Sleep quality
Pittsburgh questionnaire (Buysse et al., 1989). The maximum score is 21 points. More than 5 points are considered bad outcomes and sleep problems, and less than 5 points are considered better outcomes and no sleep problems.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Life quality
SF-36 Health survey questionnaire (Ware \& Sherbourne, 1992). Higher scores mean a better outcome.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Depressive symptoms
Geriatric Depression Scale Questionnaire (Yesavage et al., 1983). The maximum puntuation is 15. The score is 0-4 no depression, 5-8 mild depression, 9-11 moderate depression and 12-15 severe depression.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Nutritional intake
3-day dietary record
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Nutritional intake
FFQ MD adherence to assess food groups intake (Martinez-Gonzalez et al., 2012) Higher scores mean a better outcome.
Time frame: Visit 1 (week 0), visit 2 (week 4), visit 3 (week 8), visit 4 (week 12), visit 5 (week 16), visit 6 (week 20), visit 7 (week 24), visit 8 (week 28), visit 9 (week 32), visit 10 (week 36))
Nutritional status
Mini Nutrition Assessment (MNA)
Time frame: Visit 0 (week -1)
Nutritional status
Total cholesterol (mmol/L) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Nutritional status
Albumin (g/dl) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Nutritional status
Haemoglobin (g/dl) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Nutritional status
Vitamin D (ng/ml) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Nutritional status
Vitamin A (ug/dl) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Nutritional status
Vitamin E (mg/L) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Nutritional status
Carotenoids (ug/dl) by standardized methods in an automated analyzer (Beckman Coulter-Synchron (Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Cognitive function
Spanish validated Mini Mental State Examination test (MMSE). The MMSE test various cognitive. The total test score ranges from 0 (impaired) to 30 (normal) (Beaman et al., 2004).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Cognitive function
AD8. The AD8 (Alzheimer disease questionnaire) aids in identifying cognitive impairment. This test in combination with the Fototest, significantly increases the diagnosis accuracy of both tests (Carnero Pardo et al., 2012). The optimal cut-off point was 3/4.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Cognitive function
TAM test. TAM test (Memory alteration test) provides efficient and valid screening for amnestic Mild Cognitive Impairment and early stage Alzheimer's disease, and discriminates between them. The cut-off score is of 28 points (Rami et al., 2007).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Cognitive function
Fototest. The Fototest is useful to identify cognitive impairment and dementia in routine clinical practice. (Carnero Pardo et al., 2007). The cut off points are 25/26 for dementia and 28/29 for cognitive impairment.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Brain volume
Nuclear Magnetic Resonance (NMR) Total brain volume, hippocampal volume, and volume of white matter hyperintense lesions will be evaluated in a subsample of participants. NMR have allowed important advances in the understanding of age-related brain changes. NMR is a non-invasive instrument that allows the study of the normal aging of individuals at different times of their lives (Barnes et al. 2023).
Time frame: Visit 1 (week 0), visit 10 (week 36)
Cognitive ability assessment
BDNF Brain-derived Neutrophic Factor (pg/ml) will be analyzed by Enzyme-linked immunosorbent assay ELISA kit in blood samples
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Skeletal muscle strength
Isokinetic. Isokinetic test: the evaluation was conducted with an isokinetic dynamometer using a gold standard method (Biodex System 4; Biodex Medical Systems, NewYork, USA) by five repetitions sat two angular velocities (180°s-1, and240°s-1) (Solà et al., 2015). This allows a quantitative evaluation of muscle function through variables such as torque, work and power.
Time frame: Visit 1 (week 0), visit 10 (week 36)
Muscle mass, wasting and turnover
Muscle mass (skeletal muscle mass (kg)) assessed by Bioimpedance TANITA (MC-780MA; Tanita Corp., Tokyo, Japan).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Muscle mass (appendicular skeletal muscle mass (kg)) assessed by Bioimpedance TANITA (MC-780MA; Tanita Corp., Tokyo, Japan).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Muscle mass (skeletal muscle mass index (kg/m2)) assessed by Bioimpedance TANITA (MC-780MA; Tanita Corp., Tokyo, Japan).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Muscle mass (appedicular skeletal muscle mass index (kg/m2)) assessed by Bioimpedance TANITA (MC-780MA; Tanita Corp., Tokyo, Japan).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Muscle mass assessed by ultrasound (VINNO 5 (Vinno (Suzhou) Co., Ltd., China)
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Creatine kinase (U/L) assessed by enzymatic assays in a Cobas Mira Plus autoanalyzer (Roche Diagnostics Systems, Madrid, Spain).
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Cystatin-C (mg/L) assessed by enzymatic assays in a Cobas Mira Plus autoanalyzer (Roche Diagnostics Systems, Madrid, Spain).
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Serum Myostatin (ng/ml) assessed by commercial ELISA kits (Muscari et al., 2023)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Muscle mass, wasting and turnover
Serum Follistatin (ng/ml) assessed by commercial ELISA kits (Muscari et al., 2023)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Physical performance
Gait speed (m/s) assessed by the length of the walking course divided by the time (Ladang et al., 2023).
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Signalling pathways in skeletal muscle
Muscle gene expression. A muscle biopsy (200-225 mg) will be obtained from a subgroup of volunteers (n=15/group) by trained physicians (Harper et al., 2021). After collection, these samples will be immediately snap frozen in liquid nitrogen and stored at -80°C prior to analysis. To confirm RNA sequencing results obtained in the animal model of age-associated sarcopenia (transcriptomic approach that will be carried out in gastrocnemius skeletal muscle as described in task 5.2), real-time gene expression analysis (qPCR) will be performed, as previously described (Pérez-Matute et al., 2016).
Time frame: Visit 1 (week 0), visit 10 (week 36)
Biological age marker (Aging biomarker)
Serum N-terminal pro-brain natriuretic peptide (NT-proBNP) (pg/ml) in blood samples (Muscari et al., 2023) is used to diagnose and establish prognosis for heart failure and is a promising biomarker of biological age as it correlates with chronological age and predict life span better than chronological age.
Time frame: Visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36)
Glucose homeostasis
FBG (mmol/L) assessed by standardized methods in an automated analyzer (Beckman Coulter-Synchron, Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Glucose homeostasis
Insulin (IU/mL) assessed by standardized methods in an automated analyzer (Beckman Coulter-Synchron, Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Glucose homeostasis
HGBA1c (%) assessed by standardized methods in an automated analyzer (Beckman Coulter-Synchron, Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Glucose homeostasis
HOMA index calculated (Winell et al., 2010) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Glucose homeostasis
IGF-1 (ng/ml) levels were measured using Human IGF-1 Quantikine ELISA Kit (Pérez-Matute et al., 2022) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
Total cholesterol (mmol/L) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
HDLc (mmol/L) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
LDLc (mmol/L) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
VLDLc (mmol/L) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
Total TG (mmol/L) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
NEFA (mmol/L) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
ApoA1 (mg/dL) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
ApoB100 (mg/dL) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
Apo B100/Apo A1 ratio in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Lipid profile
Lp (a) (mg/dL) in blood samples assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Stress Response and Mitochondria
GDF15 (pg/mL) Growth differentiating factor 15 is recgonized in mitochondria dysfunction, and as biomarker of aging (Justice et al., 2018) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Oxidative markers
OxLDL (ug/mL) assessed by commercial ELISA in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Oxidative markers
MetSO/Met (ng/mL) assessed by commercial ELISA in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Oxidative markers
SOD (U/g Hb) 20uL (abcam) in plasma EDTA blood samples assessed by enzymatic assays (Hissin et al., 1976)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Oxidative markers
GSHPx (nmol/mL) 50uL (abcam) plasma EDTA blood samples enzymatic assays (Hissin et al., 1976)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Oxidative markers
CoQ10 (ug/mL) in blood samples assessed by HPLC
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Inflammation markers
IL-6 (pg/mL) assessed by immunoturbidimetry on an autoanalyzer (Roche Diagnostics Systems, Madrid, Spain) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Inflammation markers
hsCRP (mg/L) assessed by immunoturbidimetry on an autoanalyzer (Roche Diagnostics Systems, Madrid, Spain) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Inflammation markers
TNFRII (pg/mL) assessed by commercial ELISA in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Inflammation markers
Adiponectin (ug/mL) assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Inflammation markers
Uric acid (mg/dL) assessed by standardized enzymatic automated methods in an autoanalyzer (Beckman Coulter-Synchron, Galway, Ireland) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Vascular parameters
Systolic and diastolic blood pressure (SBP and DBP) (mm Hg) and pulse pressure (PP) (mm Hg) assessed by automatic sphygmomanometer (OMRON HEM-907; Peroxfarma, Barcelona, Spain).
Time frame: Visit 0, visit 1Visit 0 (week -1), visit 1 (week 0), visit 4 (week 12), visit 7 (week 24), visit 10 (week 36), visit 4, visit 7, visit 10
Biomarkers of endothelial dysfunction
E-selectin (CD62E) (ng/mL) assessed by Commercial ELISA and multi-analyte ELISArray kits as described in our previous work (Catalán et al., 2015) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Biomarkers of endothelial dysfunction
P-selectin (CD62P) (ng/mL) assessed by Commercial ELISA and multi-analyte ELISArray kits as described in our previous work (Catalán et al., 2015) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Biomarkers of endothelial dysfunction
ICAM-1 (CD54) (ng/mL) assessed by Commercial ELISA and multi-analyte ELISArray kits as described in our previous work (Catalán et al., 2015) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Biomarkers of endothelial dysfunction
VCAM-1 (CD106) (ng/mL) assessed by Commercial ELISA and multi-analyte ELISArray kits as described in our previous work (Catalán et al., 2015) in blood samples
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Faecal microbial composition
Metataxonomic profile.
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Faecal microbial function
Microbial functional profile
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Faecal metabolome
SCFA acetic, butyric, propionic, isobutyric, valeric, isovaleric and caproic acid and phenol colonic metabolites
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
DNA methylation
Epigeneitc age. Total genomic DNA will be extracted from saliva samples using the Norgen Saliva DNA Isolation Kit.
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Gut permeability determined on plasma
Zonulin (ng/mL). Human zonulin levels will be quantified using an ELISA kit (Cusabio® Technology Llc, Houston, TX, USA)
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
Biomarkers of wine intake compliance
Quantification of urine phenol metabolites in human elderly
Time frame: Visit 1 (week 0), visit 7 (week 24), visit 10 (week 36)
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