Human absorption of a supplement containing purified hydroxytyrosol,a natural antioxidant from olive oil,and evidence for its transient association with low-density lipoproteins

There is growing interest in the health effects of olive oil polyphenols, particularly hydroxytyrosol (HT), for their potential application in the treatment of inflammatory conditions such as cardiovascular disease (CVD). As oxidative modification of low-density lipoproteins (LDL) plays a central role in the development of CVD, natural antioxidants are a main target for the nutraceutical industry. In this study we firstly investigated the absorption of pure hydroxytyrosol (99.5%) administered as a supplement in an aqueous solution (2.5 mg/kg BW) in the plasma and urine of healthy volunteers (n = 10).Plasma C_max for HT and homovanillic alcohol (HvOH) were detected at 13.0 ± 1.5 and 16.7 ± 2.4 min, respectively. The HT and HvOH levels were undetectable 2-h after the administration. HT, HvOH, homovanillic acid and 3,4-dihydroxyphenylacetic acid were found as free forms (44%) or as glucuronide (34.4%) or sulphate (21.2%) conjugates in the 24-h urine samples of the subjects.In a second phase of the study, the same amounts of HT were administered to the subjects and the presence of HT in purified plasma lipoproteins was investigated in LDL fractions freshly isolated. 10 min after the ingestion of the HT supplement, more than 50% of the total amount detected was present in the LDL-purified fractions and its concentration declined in accordance with its presence in plasma but no changes were found in total antioxidant capacity, malondialdehyde or LDL lag time. These results indicate that pure HT transiently associates with LDL lipoproteins in vivo.     

The administration of a multivitamin/mineral fortified dairy product improves folate status and reduces plasma homocysteine concentration in women of reproductive age

BACKGROUND AND OBJECTIVES: Folate deficiency during the periconceptional period is related to the occurrence and recurrence of neural tube defects. The aim of the study was to assess whether the administration of folic acid and other vitamins and minerals as a fortified dairy product (400 microg per day of folic acid) improves the folate status in women of reproductive age. DESIGN AND METHODS: Plasma and red blood cell folate, plasma vitamin E, B12, total plasma homocysteine, plasma lipid profile, and serum ferritin and transferrin levels were investigated in 31 healthy nonpregnant women receiving 500 mL/day of the fortified dairy product for eight weeks. RESULTS: The women showed a significant increase in plasma levels of folate and vitamin B12 concentrations after four and eight weeks of supplementation. Moreover, we observed an increase in red blood cell folate concentration during the period of the study. Simultaneously, total plasma homocysteine levels decreased significantly during the intervention period. CONCLUSIONS: The regular consumption of a folic acid and other vitamins (mainly vitamins B6 and B12) and minerals in a fortified dairy product improves folate status and reduces total plasma homocysteine concentration in healthy women of childbearing age.     

Absorption of calcium from milks enriched with fructo-oligosaccharides, caseinophosphopeptides, tricalcium phosphate, and milk solids

BACKGROUND: Adequate intakes of calcium are required for optimal bone health and protection against chronic disease. Dairy products are an excellent source of calcium. OBJECTIVE: The absorption of calcium from a range of fortified milks was measured in humans with the use of stable isotopes. DESIGN: Fifteen volunteers participated in a randomized, controlled, double-blind crossover study. Five types of semi-skimmed (1.9% fat) milk drinks were administered with a light breakfast: standard milk (control milk); milk enriched with calcium from milk solids and tricalcium phosphate [(TCP) MSS milk]; milk enriched with calcium from concentrated milk (CON milk); milk with added fructo-oligosaccharides [(FOSs) FOS milk]; and milk with added caseinophosphopeptides [(CPPs) CPP milk]. All the milks were labeled with 42Ca as CaCl2. The MSS milk was also labeled with 44Ca as TCP. The quantity of calcium in each drink was kept the same by varying the volume given. RESULTS: Calcium absorption did not differ significantly between the control milk and the calcium-fortified milks (MSS and CON milk) or the FOS and CPP milks. However, calcium absorption from the TCP added to the MSS milk was significantly higher than that from the control milk (27.5 +/- 7.6% and 24.5 +/- 7.3%, respectively; P = 0.003). CONCLUSIONS: Calcium-enriched milks are a valuable source of well-absorbed calcium. Absorption of added calcium as TCP was higher than that of calcium from the control milk, but the addition of FOSs or CPPs did not significantly increase calcium absorption. Further research is needed to ascertain the cost-effectiveness and public health benefits of consuming fortified milks.     

Daily Intake of Milk Enriched with n-3 Fatty Acids,Oleic Acid,and Calcium Improves Metabolic and Bone Biomarkers in Postmenopausal Women

Objective: Nutritional strategies can be effective for the prevention of menopause-related diseases, such as osteoporosis and cardiovascular disease. Our aim was to evaluate the effects of a dairy product enriched in polyunsaturated fatty acids, calcium, oleic acid, and vitamins on cardiovascular markers and bone metabolism in postmenopausal women with moderate cardiovascular risk.

Methods: One hundred seventeen healthy postmenopausal women (aged 45 ± 7.7 years) were allocated to 2 groups: the intervention group (IG; n = 63), who consumed 0.5 L/day of a low-lactose skimmed milk enriched with 40 mg/100 mL of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA), 0.54 g/100 mL oleic acid, and vitamins, and the control group (CG; n = 54), who consumed 0.5 L/day of semiskimmed milk 0.5 L/day enriched with vitamins A and D.

Results: After 12 months, in the IG there was an improvement in lipid profile: a ?5.78% decrease in total cholesterol (p = 0.010), ?9.79% (p = 0.004) in low-density lipoprotein (LDL) cholesterol, ?9.56% (p < 0.001) in total cholesterol (TC)/high-density lipoprotein (HDL) ratio, and ?3.38% in LDL/HDL ratio (p < 0.001). No changes were observed in the CG. In the IG we observed a decrease of ?28.20% in high-sensitivity C-reactive protein (hs-CRP; p = 0.012). There was no effect on bone turnover markers or serum osteoprotegerin (OPG) in either of the study groups. In the IG, receptor activator of nuclear factor κB ligand (RANKL) was reduced ?17.64% (p = 0.003), with no effect in the CG.

Conclusion: In postmenopausal women with moderate cardiovascular risk, dietary supplementation with a dairy drink enriched with fatty acids (EPA+DHA), oleic acid, minerals, and vitamins induces a positive effect on cardiovascular risk and parameters of bone metabolism. Its regular consumption may be a useful nutritional support for postmenopausal women.     

Improvement of bone formation biomarkers after 1-year consumption with milk fortified with eicosapentaenoic acid,docosahexaenoic acid,oleic acid,and selected vitamins

The hypothesis of this study was that the replacement of regular milk with fortified milk in hyperlipidemic adults for 1 year would improve bone biomarkers. The fortified milk contained eicosapentaenoic acid and docosahexaenoic acid from fish oils, oleic acid, vitamins A, B₆, and E, as well as folic acid. We believe that the fortified milk will improve the blood fatty acid profile and vitamin status in subjects to benefit bone health biomarkers. From the 84 patients who accepted to participate, 11 of these were excluded for the presence of metabolic diseases and 1 was excluded for noncompliance with the protocol. Seventy-two hyperlipidemic patients (35-65 years) were randomly divided between 2 study groups. The supplement group (E; n = 39) consumed 0.5 L/d of fortified milk that contained fish oil, oleic acid, and vitamins. The control group (C; n = 33) consumed 0.5 L/d of semiskimmed milk containing the same amount of total fat. Blood samples were taken at T₀, T₃, T₆, and T₁₂ months to determine plasma fatty acids, vitamins B₆, E, and 25-hydroxyvitamin D and serum folate, calcium, soluble osteoprotegerin (OPG), soluble receptor activator of NF-κB ligand (RANKL), osteocalcin, parathormone, type I collagen carboxy-terminal telopeptide, and malondialdehyde. After 1 year, the E group showed a significant increase in plasma eicosapentaenoic acid (42%), docosahexaenoic acid (60%), vitamin B6 (38%), OPG (18%), RANKL (7%), OPG/RANKL (10%), red blood cell folate (21%), serum folate (53%), calcium (4%), vitamin D (11%), and osteocalcin (22%). Dietary supplementation with the fortified milk drink improved nutritional status and bone formation markers in adult hyperlipidemic patients.     

Milk enriched with “healthy fatty acids” improves cardiovascular risk markers and nutritional status in human volunteers

ObjectiveThe main goal of the present study was to evaluate the effect of a commercially available milk containing small amounts of eicosapentaenoic acid and docosahexaenoic acid, oleic acid, and vitamins A, B6, D, E, and folic acid compared with semi-skimmed and skimmed milk in volunteers with moderate cardiovascular risk.MethodsTwo hundred ninety-seven subjects 25 to 65 y of age with moderate cardiovascular risk were randomly allocated into three groups. In addition to their diets, one group consumed 500 mL/d of the enriched milk, another group consumed 500 mL/d of skimmed milk, and a control group consumed 500 mL/d of semi-skimmed milk. All groups consumed the dairy drinks for 1 y and blood samples were taken at 0 and 12 mo.ResultsConsumption of enriched milk for 1 y produced significant (P < 0.05) increases in serum folate (58%) and high-density lipoprotein cholesterol (4%). Plasma triacylglycerols (10%), total cholesterol (4%), and low-density lipoprotein cholesterol (6%) were reduced significantly only in the supplemented group. Serum glucose, homocysteine, and C-reactive protein remained unchanged. In the skimmed milk and semi-skimmed milk groups, the only significant decreases were in serum folate (17% and 11%, respectively).ConclusionDaily intake of a milk enriched with fish oil, oleic acid, and vitamins improved the nutritional status and cardiovascular risk markers of volunteers, whereas skimmed milk and semi-skimmed milk did not.     

Intake of fish oil, oleic acid, folic acid, and vitamins B-6 and E for 1 year decreases plasma C-reactive protein and reduces coronary heart disease risk factors in male patients in a cardiac rehabilitation program

Certain nutrients have been shown to be effective in preventing coronary heart disease. We hypothesized that a daily intake of low amounts of a number of these nutrients would exert beneficial effects on risk factors and clinical variables in patients that suffered from myocardial infarction (MI) and were following a cardiac rehabilitation program. Forty male MI patients were randomly allocated into 2 groups. The supplemented group consumed 500 mL/d of a fortified dairy product containing eicosapentaenoic acid, docosahexaenoic acid, oleic acid, folic acid, and vitamins A, B-6, D, and E. The control group consumed 500 mL/d of semi-skimmed milk with added vitamins A and D. The patients received supervised exercise training, lifestyle and dietary recommendations, and they were instructed to consume the products in addition to their regular diet. Blood extractions and clinical examinations were performed after 0, 3, 6, 9, and 12 mo. Plasma concentrations of eicosapentaenoic acid, docosahexaenoic acid, oleic acid, folic acid, vitamin B-6, and vitamin E increased after supplementation (P<0.05). Plasma total and LDL-cholesterol, apolipoprotein B, and high-sensitivity C-reactive protein concentrations decreased in the supplemented group (P<0.05), and plasma total homocysteine decreased in both groups. There were no changes in heart rate, blood pressure, or cardiac electrocardiographic parameters in either group. Therapeutic lifestyle changes, effected through a CR program comprising regular exercise and the intake of a combination of dietary nutrients, reduced a variety of risk factors in MI patients, which supports the rationale for nutritional programs in the secondary prevention of coronary heart disease.     

Oral intake of Lactobacillus fermentum CECT5716 enhances the effects of influenza vaccination

OBJECTIVE: We studied the coadjuvant capability of oral consumption of the breast-milk-isolated strain Lactobacillus fermentum (CECT5716) for an anti-influenza vaccine. METHODS: A randomized, double-blinded, placebo-controlled human clinical trial including 50 volunteers (31 male and 19 female) was performed to address the immunologic effects of an intramuscular anti-influenza vaccine in adults (33.0 +/- 7.7 y old). Fifty percent of volunteers received an oral daily dose of methylcellulose (placebo) or probiotic bacteria (1 x 10(10) colony-forming units/d) 2 wk before vaccination and 2 wk after vaccination. RESULTS: Two weeks after vaccination there was an increase in the proportion of natural killer cells in the probiotic group but not in the placebo group. The vaccination induced an increase in T-helper type 1 cytokine concentrations and in T-helper and T-cytotoxic proportions in both groups; however, the probiotic group showed a significant higher induction in some of these parameters. Regarding the humoral effects, induction of antibody response in the placebo group could not be detected. In the case of the probiotic group, a significant increase in antigen specific immunoglobulin A was detected. Although an increase in total immunoglobulin M was observed, changes in anti-influenza antigen specific immunoglobulin M were not observed. The incidence of an influenza-like illness during 5 mo after vaccination (October to February) was lower in the group consuming the probiotic bacteria. CONCLUSION: Oral administration of the strain L. fermentum CECT5716 potentates the immunologic response of an anti-influenza vaccine and may provide enhanced systemic protection from infection by increasing the T-helper type 1 response and virus-neutralizing antibodies.     

Cardiovascular effects of milk enriched with omega-3 polyunsaturated fatty acids, oleic acid, folic acid, and vitamins E and B6 in volunteers with mild hyperlipidemia

OBJECTIVE: Results from epidemiologic studies and clinical trials have indicated that consumption of omega-3 fatty acids, oleic acid, and folic acid have beneficial effects on health, including decreased risk of cardiovascular disease. We evaluated the combined effects of these nutrients through the consumption of milk enriched with omega-3 polyunsaturated fatty acids, oleic acid, vitamins E and B6, and folic acid on risk factors for cardiovascular disease in volunteers with mild hyperlipidemia. METHODS: Thirty subjects ages 45 to 65 y (51.3 +/- 5.3 y) were given 500 mL/d of semi-skimmed milk for 4 wk and then 500 mL/d of the enriched milk for 8 wk. Plasma and low-density lipoproteins were obtained at the beginning of the study and at 4, 8, and 12 wk. RESULTS: Consumption of enriched milk for 8 wk increased plasma concentrations of docosahexaenoic acid and eicosapentaenoic acid and significantly (P < 0.05) decreased plasma concentrations of triacylglycerol (24%), total cholesterol (9%), and low-density lipoprotein cholesterol (13%). Plasma and low-density lipoprotein oxidation and vitamin E concentration remained unchanged throughout the study. Significant decreases in plasma concentrations of vascular cell adhesion molecule-1 (9%) and homocysteine (17%) were found, accompanied by a 98% increase in plasma concentration of folic acid. CONCLUSIONS: Dairy supplementation strategies with omega-3 polyunsaturated fatty acids, oleic acid, and vitamins may be useful for decreasing risk factors for cardiovascular disease.