Deciphering the Effect of Moderate Wine Consumption on Healthy Aging Through Postprandial Extracellular Vesicles.

Concentration of plasma-derived extracellular vesicles measured after acute intake of red wine, white wine, or water. Extracellular vesicles will be isolated by immunoaffinity methods and quantified using nanoparticle-based analysis. Results will be expressed as particles per milliliter (particles/mL).

Mean diameter of plasma-derived extracellular vesicles measured after acute intake of red wine, white wine, or water. Extracellular vesicle size will be assessed using dynamic light scattering. Results will be expressed in nanometers (nm).

Postprandial changes in the protein composition of plasma-derived extracellular vesicles following acute intake of red wine, white wine, or water. EV-associated proteins will be analyzed using quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results will be expressed as normalized protein abundance.

Postprandial changes in the lipid composition of plasma-derived extracellular vesicles following acute intake of red wine, white wine, or water. EV-associated lipid species will be analyzed using quantitative lipidomics by liquid chromatography-mass spectrometry (LC-MS). Results will be expressed as normalized lipid abundance.

Venous blood plasma glucose concentration measured using a validated clinical chemistry assay (enzymatic method). Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: mg/dL

Venous blood plasma insulin concentration measured using a validated immunoassay. Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: µIU/mL

Venous blood plasma triglyceride concentration measured using a validated clinical chemistry assay (enzymatic method). Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: mg/dL

Venous blood plasma total cholesterol concentration measured using a validated clinical chemistry assay (enzymatic method). Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: mg/dL

Venous blood plasma HDL cholesterol concentration measured using a validated clinical chemistry assay (enzymatic method). Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: mg/dL

Venous blood plasma LDL cholesterol concentration measured using a validated clinical chemistry assay (enzymatic method). Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: mg/dL

Venous blood plasma IL-6 concentration measured using a validated immunoassay. Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: pg/mL

Venous blood plasma TNF-α concentration measured using a validated immunoassay. Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: pg/mL

Venous blood plasma IL-1β concentration measured using a validated immunoassay. Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: pg/mL

Venous blood plasma MCP-1/CCL2 concentration measured using a validated immunoassay. Primary metric is change from baseline at each postprandial time point (time point value minus baseline value).

Unit of Measure: pg/mL

lood-brain barrier (BBB) integrity assessed by transendothelial electrical resistance (TEER) in a human in vitro BBB model after exposure to extracellular vesicles (EVs) isolated from plasma at baseline (0 hours) and postprandial peak (2 hours).

Unit of Measure: Ω·cm²

Tight junction protein expression assessed in the BBB model by Claudin-5 (CLDN5) protein abundance after exposure to EVs isolated at baseline (0 hours) and 2 hours post intake. Protein expression quantified using immunoblotting or immunofluorescence and reported relative to the control condition.

Unit of Measure: Fold change (dimensionless)

Tight junction protein expression assessed in the BBB model by occludin (OCLN) protein abundance after exposure to EVs isolated at baseline (0 hours) and 2 hours post intake. Protein expression quantified using immunoblotting or immunofluorescence and reported relative to the control condition.

Unit of Measure: Fold change (dimensionless)

Microglial activation assessed by IL-6 concentration in culture supernatant after exposure to EVs isolated at baseline (0 hours) and 2 hours post intake. Cytokine concentration quantified using a validated immunoassay.

Unit of Measure: pg/mL

Microglial activation assessed by TNF-α concentration in culture supernatant after exposure to EVs isolated at baseline (0 hours) and 2 hours post intake. Cytokine concentration quantified using a validated immunoassay.

Unit of Measure: pg/mL

Microglial activation assessed by IL1B mRNA expression after exposure to EVs isolated at baseline (0 hours) and 2 hours post intake. Gene expression quantified by RT-qPCR and reported as relative expression versus control condition.

Unit of Measure: Fold change (dimensionless)

Endothelial activation assessed by VCAM-1 protein expression in cultured human umbilical vein endothelial cells after exposure to extracellular vesicles isolated from plasma at baseline (0 hours) and postprandial peak (2 hours). Protein expression quantified by immunoblotting or immunofluorescence and reported relative to control condition.

Unit of Measure: Fold change (dimensionless)

Identification and quantification of polyphenolic compounds encapsulated within plasma-derived extracellular vesicles following acute intake of red wine or white wine. Polyphenols will be analyzed using ultra-high-performance liquid chromatography coupled to quadrupole-Orbitrap mass spectrometry. Results will be expressed as normalized peak areas and concentrations relative to internal standards.

Association assessed using Spearman rank correlation (ρ) between EV cargo score (PC1 derived from normalized EV proteomic and lipidomic features measured by mass spectrometry) and the systemic response measure defined as change from baseline in plasma glucose concentration at postprandial time points. The outcome measure is Spearman's ρ.

Unit of Measure: Spearman's ρ (dimensionless)

Endothelial activation assessed by ICAM-1 protein expression in cultured human umbilical vein endothelial cells after exposure to extracellular vesicles isolated from plasma at baseline (0 hours) and postprandial peak (2 hours). Protein expression quantified by immunoblotting or immunofluorescence and reported relative to control condition.

Unit of Measure: Fold change (dimensionless)

Endothelial response assessed by nitric oxide production after exposure to extracellular vesicles isolated from plasma at baseline (0 hours) and postprandial peak (2 hours). NO production quantified in culture supernatant using a validated colorimetric assay (e.g., nitrite/nitrate).

Unit of Measure: µM