Effect of filtered coffee intake in healthy subjects.

INTERVENTION: Twenty health volunteers, with age between 20 and 65 years old, will be selected and undergo 28 days of washout before the start the treatment. Then they will be divided in two groups: one will ingest from 3 to 4 cups daily (100 mL each one for people over 60 years old and 150 mL for people under 60 years old) of coffee brew with light roasting and another with dark roasting for 4 weeks. After, will be made a cross‐over and the volunteers will ingest the coffee brew for over 4 weeks. Coffee will be provided to participants in daily portion. There will not have a washout between the two coffee groups, because studies show that phenolic acids bioavailability is short (After 4 hours of coffee intake, phenolic acid are not present in plasma in detectable amount), therefore 4 weeks are sufficient to eliminate the effect of one rosting on the other). CONDITION: Antioxidant activity of the coffee phenolic acids atherogenesis prevention risk reduction of Diabetes mellitus risk reduction of cardiovascular disease PRIMARY OUTCOME: Blood pressure will be measured by "Arterial blood pressure ambulatorial Monitoring" Evaluation of the antioxidant enzymes (catalase, glutathione peroxidase, and superoxide dismutase) activity. Catalase activity will be determined in erithrocytes, using the method described by AEBI H (Catalase in vitro. Methods ; Enzymol. 1984; 105:121‐6). Glutathione peroxidase and superoxide dismutase activity will be determined by RANSEL and RANSOD kit (Randox Lab, San Diego, CA), respectively Improvement of insulin sensitivity and blood glucose stabilization. Fasting and postprandial blood glucose and insulin will be determined by commercial kit. Inhibition of lipid peroxidation, verified by conjugated diene formation and by malonaldehyde quantification in High performance liquid chromatography (HPLC). Oxidized Low density lipoprotein (LDL) in plasma. Quantification of ox‐LDL in plasma will be performed by sandwich enzyme‐linked immunosorbent assay (ELISA) (Mercodia AB, Uppsala, Sweden). Phenolic acids in plasma. To determine and quantify these acids, will be selected other 6 healthy volunteers, who will ingest 200mL of coffee beverage in a single dose. Reduction of inflammatory biomarkers (fibrinogen, C‐reactive protein, Interleukin 6 (IL‐6), Interleukin 1b (IL‐1b), soluble Intercellular adhesion molecule‐1 (sICAM‐1), soluble Vascular cell adhesion ; molecule‐1 (sVCAM‐1), soluble E‐selectin (sE‐selectin), Adiponectin, Monocyte chemotactic protein‐1 (MCP‐1), Tumor necrosis factor‐alpha (TNF‐a) and Plasminogen activator inhibitor ‐1 (PAI‐1). These biomarkers will be determined by panels for cardiovascular disease (LINCO Research, Inc.), using the Luminex xMAP technology. Total homocysteine in plasma, quantified in High performance liquid chromatography (HPLC). Total phenolic compounds in plasma. For this determination, plasma proteins will be precipitated and phenolic compounds quantified by Folin‐ciocalteau method. SECONDARY OUTCOME: Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) values will be determined by commercial kits in an automated machine Hematocrit will be determined in whole blood by an automated machine. Hemoglobin and glycated hemoglobin quantification. Hemoglobin will be quantified by commercial kit (Labtest) and this result will be used for enzymes antioxidant activity calculation. Glycated hemoglobin will be quantified to glycemic control monitoring in long term. This test provides an accurate indication of the average plasma glucose concentration within 2 to 3 months before testing, complementing the more traditional measures of glucose control, ; as a measure of glucose in plasma or urine. For this determination, turbidimetric assay using a commercial kit and automated machine will be used. Leukocytes (basophils, monocytes, white blood cell count and neutrophils) values will be determined in the hemogram (whole blood) by an automated electronic counter with two main sensors: a light detector and an electrical impedance. Nutritional status (weight, height, abdominal circunference and body fat percentage, measured by bioimpedance). Platelet aggregation test and coagulation time. The speed and extent to which platelets dispersed in a plasma sample form settlements after the addition of a material that stimulates the aggregation will be measured in a machine that measures the changes in turbidity "opacity" and print a graphic record of results. Total cholesterol, triacylglycerols, High density lipoprotein (HDL‐c) and Very low density lipoprotein (VLDL‐c) will be quantified in serum by enzymatic‐colorimetric automated method, using commercial kits. Low density lipoprotein (LDL‐c) will be calculated by Friedewald equation. Urea and creatinine are routine tests and they are important parameters to evaluate renal function. Biochemical testing of urea and creatinine will be performed in whole blood by using commercial kits (colorimetric method) in an automated machine. Variation in sodium, potassium, magnesium, phosphorus and folate concentration will be dosed in the serum by an ion‐selective electrode method. INCLUSION CRITERIA: All races, non‐smokers or former‐smokers for more than 2 years, non‐alcoholics, serum cholesterol below 240 mg/dL, fasting glucose below 100 mg/dL, normal liver enzymes (Aspartate aminotransferase ‐ AST and Alanine aminotransferase ‐ ALT) and normal plasma homocysteine, absence of continuous medicine intake. No history of cardivascular, kidney or liver disease, diabetes mellitus, hypertension or hyperthyroidism.