Olive Pomace Extract Contains Low Molecular Weight Peptides and Possesses ACE Inhibitory Activity.

The aim of the present study was to determine the ACE inhibitory activity of aqueous extracts of olive pomace and to understand whether they represent a good source of bioactive LMW peptides for nutritional and pharmacological applications. We produced a water extract from olive pomace (var. Picual) and obtained its low molecular weight (LMW) fraction (<3 kDa). The calculated yield of extraction was 100.2 ± 7.9 mg of LMW peptides per 100 g of olive pomace. The olive pomace LMW fraction possessed strong ACE inhibitory activity (IC50 = 3.57 ± 0.22 µg prot/mL). The LMW fraction (<3 kDa) was analysed by nanoscale liquid chromatography-Orbitrap coupled with tandem mass spectrometry and de novo sequencing. Thirty new peptides, containing between 7-17 amino acids and molecular masses ranging 778-1354 Da, were identified by the Peaks database algorithm using the available Olea europaea (cv. Farga) genome database. Ten new peptides were also identified by Peaks de novo sequencing. The protein sources of twelve peptides detected in the database by Peaks DB were identified by BLAST search. The ACE inhibitory activity of the identified peptides was predicted by BIOPEP software. We conclude that olive pomace possesses ACE inhibitory activity and contains low molecular weight peptides with (predicted) biological activity. Olive pomace may represent a good source of peptides for nutritional and pharmaceutical applications. In our study, it has been shown that olive pomace possesses ACE inhibitory activity and contains low molecular weight peptides with (predicted) biological activity. Olive pomace may represent a good source of peptides for nutritional and pharmaceutical applications. More research is needed in order to identify the in vivo effects of olive pomace bioactive peptides.

Beneficial Effects of Limosilactobacillus fermentum CECT 5716 Administration to Infants Delivered by Cesarean Section.

Cesarean section (CS) disrupts the natural microbiota colonization process in infants, which might compromise immune system maturation, leading to a higher risk of infections. We evaluated the effect of the probiotic Limosilactobacillus (L.) fermentum CECT 5716 on the incidence of gastrointestinal and respiratory infections in the CS infant subgroups (n = 173) of three randomized clinical trials in which this probiotic strain was demonstrated to be safe and effective for preventing infections. Therefore, the data for the CS infants were extracted to obtain the incidence rate ratio (IRR) and 95% CI for gastrointestinal and respiratory infections for each study and were then combined to obtain a pooled IRR and 95% CI using the generic inverse variance method. There was a significant reduction of 73% in the incidence of gastrointestinal infections in CS infants receiving L. fermentum CECT 5716 compared with those receiving the control formula [n = 173, IRR: 0.27 (0.13, 0.53), p = 0.0002]. Regarding respiratory infections, although pooled results showed a reduction of 14% in the probiotic group, the difference was not statistically significant [n = 173, IRR (95% CI): 0.86 (0.67, 1.11), p = 0.25]. In conclusion, the administration of L. fermentum CECT 5716 to CS-born infants protects them from gastrointestinal infections by reducing the risk by up to 73% in this population.

Characterisation of Endogenous Peptides Present in Virgin Olive Oil.

The low molecular weight peptide composition of virgin olive oil (VOO) is mostly unknown. We aimed to investigate the composition of the endogenous peptides present in VOO, the protein sources from which those peptides originate and their biological activities. A water-soluble extract containing peptides was obtained from VOO. The peptides were separated by size-exclusion using fast protein liquid chromatography, and the low molecular weight fraction (1600-700 kDa) was analysed by nanoscale liquid chromatography Orbitrap coupled with tandem mass spectrometry and de novo sequencing. Nineteen new peptides were identified by Peaks database algorithm, using the available Olea europaea (cv. Farga) genome database. Eight new peptides were also identified by Peaks de novo sequencing. The protein sources of the peptides detected in the database by Peaks DB were identified by BLAST-P search. Seed storage proteins were among the most frequent sources of VOO peptides. BIOPEP software was used to predict the biological activities of peptides and to simulate (in silico) the proteolytic activity of digestive enzymes on the detected peptide sequences. A selection of synthetic peptides was obtained for investigation of their bioactivities. Peptides VCGEAFGKA, NALLCSNS, CPANGFY, CCYSVY and DCHYFL possessed strong ACE-inhibitory and antioxidant activities in vitro. Antioxidant peptides could play a role in VOO quality.

Olive oil varieties and ripening stages containing the antioxidants hydroxytyrosol and derivatives in compliance with EFSA health claim.

Virgin olive oils (VOO) and extra virgin olive oils (EVOO) contain a specific fraction of polyphenols (hydroxytyrosol and its derivatives) that produce beneficial physiological effects. The European Food Safety Authority (EFSA) authorised a health claim for olive oil (OO) containing at least 250 mg/Kg of those polyphenols. The specific polyphenol content of twelve varieties of EVOO extracted at three different maturation stages was investigated. The total concentration of specific polyphenols changed depending on the olive oil variety. The varieties showing the highest specific polyphenol content (all above 250 mg/Kg) were Lechín Sevilla (429.5 ± 5), Manzanilla Sevilla (407.6 ± 6) and Cornezuelo (394.0 ± 6) in the green phase; Cornicabra (362.0 ± 8), Nevadillo Negro (326.5 ± 4) and Picual 296.0 ± 6) in the turning phase; and Lechin Granada (382.8 ± 4), Picual (317.7 ± 3), Lechin Sevilla (294.4 ± 5) and Manzanilla Sevilla (278.0 ± 2) in the mature phase. These results could have potential application for the industrial production of a category of healthy antioxidant OO.

Changes in Glutathione, Ascorbate, and Antioxidant Enzymes during Olive Fruit Ripening.

The content of glutathione, ascorbate (ASC), and the enzymatic antioxidants, superoxide dismutase and catalase, and components of the ascorbate-glutathione cycle were investigated in the olive fruit (cv. Picual) selected at the green, turning, and mature ripening stages. The changes observed in total and reduced glutathione (GSH), oxidized glutathione (GSSG), the ratio GSH/GSSG, ASC, and antioxidant enzymes (mainly superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase) indicate a shift to a moderate cellular oxidative status during ripening and suggest a role for antioxidants in the process. The antioxidant composition of olive oils obtained from the olive fruits of the study was investigated. A model is proposed for the recycling of antioxidant polyphenols mediated by endogenous molecular antioxidants in the olive fruit.

Antihypertensive Effects of Virgin Olive Oil (Unfiltered) Low Molecular Weight Peptides with ACE Inhibitory Activity in Spontaneously Hypertensive Rats.

The low molecular weight peptide composition of virgin olive oil (VOO) is mostly unknown. We hypothesised that unfiltered VOO could possess low molecular weight peptides with antihypertensive activity. We produced unfiltered VOO and obtained a water-soluble peptide extract from it. The peptides were separated by size-exclusion using fast protein liquid chromatography, and the low molecular weight fraction was analysed by nanoscale liquid chromatography-Orbitrap coupled with tandem mass spectrometry and de novo sequencing. We selected 23 peptide sequences containing between 6 and 9 amino acids and molecular masses ranging 698-1017 Da. Those peptides were chemically synthesised and their angiotensin-converting enzyme (ACE) inhibitory activity was studied in vitro. Seven peptides showed a strong activity, with half maximal inhibitory concentration (IC50) <10 µm. The antihypertensive effects of the four most active synthesised ACE inhibitor peptides were studied in spontaneously hypertensive rats (SHR). Acute oral administration of synthetic peptides RDGGYCC and CCGNAVPQ showed antihypertensive activity in SHR. We conclude that unfiltered VOO naturally contains low molecular weight peptides with specific ACE inhibitory activity and antihypertensive effects in SHR.

Virgin olive oil (unfiltered) extract contains peptides and possesses ACE inhibitory and antihypertensive activity.

BACKGROUND & AIMS: The peptide and protein composition of olive oil is mostly unknown and the few studies available have not focused on the study of its low molecular weight peptides. We hypothesised that olive oil could naturally contain low molecular weight peptides with antihypertensive effect. METHODS: We produced virgin olive oil (unfiltered, var. Picual) and obtained a water-soluble peptide extract. We fractionated the peptide extract by FPLC and studied its angiotensin converting enzyme (ACE) inhibitory activity. We studied the antihypertensive effect of olive oil peptides on the systolic blood pressure (SBP) and diastolic blood pressure (DBP) using an animal model of hypertension (spontaneously hypertensive rats, SHR). The animals were randomly distributed into 3 study groups (n = 8 per group) and received an oral dose of olive oil peptides (0.425 mg/kg of BW), or a dose of Captopril (50 mg/kg of BW) or water. SBP and DBP were registered in the rats before administration and a at 2, 4, 6, 8, 24 and 48 h post-administration of the corresponding dose. RESULTS: The peptide extract and FPLC purified fractions possessed angiotensin converting enzyme (ACE) inhibitory activity. Acute oral administration of olive oil water-soluble extract produced an average blood pressure reduction of 10 mmHg at 4 h (P < 0.01) and reached a maximum antihypertensive effect of 20 mmHg at 6 h, compared with baseline. CONCLUSION: Unfiltered virgin olive oil contains peptides and a water-soluble extract obtained from this oil possesses ACE inhibitory activity and in vivo antihypertensive effect.

Еffects of fortified milk on cognitive abilities in school-aged children: results from a randomized-controlled trial.

BACKGROUND: Micronutrients such as vitamins and minerals and long-chain polyunsaturated omega-3 fatty acids (PUFAs) are essential for children's brain development and cognitive functions. The current study investigated whether milk fortified with micronutrients and PUFA can result in improved cognitive function in mainstream school children. METHODS: One-hundred-and-nineteen children (age 8-14, 58 boys) were randomly allocated to a fortified milk group or a regular full milk control group. Participants consumed 0.6L/day of the milk for 5 months. We recorded relevant biochemical, anthropometric, and cognitive measures (working memory and processing speed) at the start of the study and at follow-up after 5 months. RESULTS: The fortified milk significantly increased docosahexaenoic acid (DHA) (change from baseline of 28% [95% CI 17-39%] vs. -6% [95% CI - 13 to 0%] in the control group) and serum 25OH-vitamin D concentrations (41% [95% CI 30-52%] vs. 21% [95% CI 11-30%] in the control group). The fortified milk improved working memory on one of two tests (32% [95% CI 17-47%] vs. 13% [95% CI 6-19%] in the control group). The fortified milk also indirectly increased processing speed on one of two tests; this effect was small and completely mediated by increases in 25OH-vitamin D concentrations. CONCLUSIONS: These results suggest that fortifying milk with micronutrients and PUFA could be an effective and practical way to aid children's cognitive development.

Hydroxytyrosol supplementation increases vitamin C levels in vivo. A human volunteer trial.

Hydroxytyrosol (HT) is a main phenolic component of olive oil. In this study, we investigated the safety and effects produced by HT purified (99.5%) from olive mill waste. HT was administered at a daily dosage of 45mg for 8 weeks to volunteers with mild hyperlipidemia (n=14). We measured markers of cardiovascular disease risk, enzyme markers of several clinical conditions, hematology, antioxidant parameters, vitamins and minerals at baseline (T0), 4 weeks (T4) and 8 weeks (T8). The values obtained at T4 and T8 were compared with baseline. We found that the HT dose administered was safe and mostly did not influence markers of cardiovascular disease, blood lipids, inflammatory markers, liver or kidney functions and the electrolyte balance. Serum iron levels remained constant but a significant (P<0.05) decrease in ferritin at T4 and T8 was found. Serum folate and red blood cell folate levels were also reduced at T4 and T8. Finally, vitamin C increased by two-fold at T4 and T8 compared with levels at baseline. These results indicate a physiologically relevant antioxidant function for HT through increasing endogenous vitamin C levels.

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.

ROS Generation in Peroxisomes and its Role in Cell Signaling.

In plant cells, as in most eukaryotic organisms, peroxisomes are probably the major sites of intracellular H2O2 production, as a result of their essentially oxidative type of metabolism. In recent years, it has become increasingly clear that peroxisomes carry out essential functions in eukaryotic cells. The generation of the important messenger molecule hydrogen peroxide (H2O2) by animal and plant peroxisomes and the presence of catalase in these organelles has been known for many years, but the generation of superoxide radicals (O2·- ) and the occurrence of the metalloenzyme superoxide dismutase was reported for the first time in peroxisomes from plant origin. Further research showed the presence in plant peroxisomes of a complex battery of antioxidant systems apart from catalase. The evidence available of reactive oxygen species (ROS) production in peroxisomes is presented, and the different antioxidant systems characterized in these organelles and their possible functions are described. Peroxisomes appear to have a ROS-mediated role in abiotic stress situations induced by the heavy metal cadmium (Cd) and the xenobiotic 2,4-D, and also in the oxidative reactions of leaf senescence. The toxicity of Cd and 2,4-D has an effect on the ROS metabolism and speed of movement (dynamics) of peroxisomes. The regulation of ROS production in peroxisomes can take place by post-translational modifications of those proteins involved in their production and/or scavenging. In recent years, different studies have been carried out on the proteome of ROS metabolism in peroxisomes. Diverse evidence obtained indicates that peroxisomes are an important cellular source of different signaling molecules, including ROS, involved in distinct processes of high physiological importance, and might play an important role in the maintenance of cellular redox homeostasis.

Characterization of antioxidant enzymes and peroxisomes of olive (Olea europaea L.) fruits.

The presence of peroxisomes in olive (Olea europaea L.) fruits and different antioxidant enzymes occurring in this plant tissue is reported for the first time. Ultrastructural analysis showed that olive cells were characterized by the presence of large vacuoles and lipid drops. Plastids, mitochondria and peroxisomes were placed near the cell wall, showing some type of association with it. Olive fruit peroxisomes were purified by sucrose density-gradient centrifugation, and catalase, glutathione reductase and ascorbate peroxidase were found in peroxisomes. In olive fruit tissue the presence of a battery of antioxidant enzymes was demonstrated, including catalase, four superoxide dismutase isozymes (mainly an Fe-SOD plus 2 Cu,Zn-SOD and a Mn-SOD), all the enzymes of the ascorbate-glutathione cycle, reduced and oxidized glutathione, ascorbate, and four NADPH-recycling dehydrogenases. The knowledge of the full composition of antioxidants (enzymatic and non-enzymatic) in olive fruits is crucial to be able to understand the processes regulating the antioxidant composition of olive oil.

Pepper fruit as a model to study the metabolism of antioxidants, ROS and RNS.

Pepper (Capsicum annuum L.) is the second worldwide most consumed vegetable nowadays, with southern European countries being among the major producer areas. Pepper fruits contain high levels of vitamins A and C (ascorbate) and low calories what makes this produce greatly appropriate for human diet. In fact, fruits have enormous interest from the culinary and gastronomic points of view and can be used as raw, canned, condiment and food colorant, among others. Ripening of pepper fruits, an ethylene-independent process in this plant species, is somehow modulated by nitric oxide (NO), and the profile of several reactive nitrogen species (RNS) could be used as an index of this physiological stage. Regarding to the antioxidants' metabolism in fruits [1,2], it has been hypothesized that ascorbate plays an important role as a redox buffer during the ripening process. Furthermore, the interplay among enzymatic antioxidants protects pepper fruits against damage promoted by chilling conditions. The oxidative metabolism of peroxisomes seems to also participate in the regulation of the ripening process of fruits.

Characterization of Antioxidant Enzymes and Peroxisomes of Olive (Olea europaea L.) Fruits.

The presence of peroxisomes in olive (Olea europaea L.) fruits and different antioxidant enzymes occurring in this plant tissue is reported for the first time. Ultrastructural analysis showed that olive cells were characterized by the presence of large vacuoles and lipid drops. Plastids, mitochondria and peroxisomes were placed near the cell wall, showing some type of association with it. Olive fruit peroxisomes were purified by sucrose density-gradient centrifugation, and catalase, glutathione reductase and ascorbate peroxidase were found in peroxisomes. In olive fruit tissue the presence of a battery of antioxidant enzymes was demonstrated, including catalase, four superoxide dismutase (SOD) isozymes (mainly an Fe-SOD plus 2 Cu,Zn-SODs and a Mn-SOD), all the enzymes of the ascorbate-glutathione cycle, reduced and oxidized glutathione, ascorbate, and four NADPH-recycling dehydrogenases. Knowing the full composition of antioxidants (enzymatic and non-enzumatic) in olive fruits is crucial to be able to understand the processes regulating the antioxidant composition of olive oil.

The effect of EPA and DHA on metabolic syndrome patients: a systematic review of randomised controlled trials.

Metabolic syndrome (MS) is characterised by accumulation of CVD risk factors. The use of very long chain n-3 polyunsaturated fatty acids (VLC n3 PUFA) could potentially benefit MS by reducing risk factors. To better understand the possible VLC n3 PUFA benefits, the literature was systematically reviewed for randomised controlled trials (RCT) that published effects of VLC n3 PUFA on MS patients. 17 RCT fulfilled the inclusion criteria and were analysed for relevance to the research question. The available RCT convincingly show that the administration of VLC n3 PUFA doses > 1 g for at least 3 months produces a significant reduction of triglycerides ranging from 7 % to 25 %. These results confirm the hypotriglyceridemic effect of VLC n3 PUFA in MS patients. The triglyceride lowering may produce further benefits by reducing the % of pro-atherogenic small dense LDL particles (sdLDL) and also perhaps by ameliorating the inflammatory process associated with MS. High doses of VLC n3 PUFA ( ≥ 3 g/day) may produce further TAG reductions but could raise other risk factors such as LDL-C. No clear effects were found on other MS markers. The combination of VLC n3 PUFA plus a statin may be useful to prevent the occurrence of coronary events. More studies are needed using different amounts of VLC n3 PUFA, time lengths, dietary backgrounds and different profiles of MS patients before clear recommendations can be made.

Lactobacillus fermentum CECT 5716 is safe and well tolerated in infants of 1-6 months of age: a randomized controlled trial.

The objective of the study was to evaluate the safety and tolerance of an infant formula supplemented with Lactobacillus fermentum CECT5716, a probiotic strain isolated from breast milk, in infants of 1-6 months of age. A randomized double blinded controlled study including healthy infants was conducted. One month aged infants received a prebiotic infant formula supplemented with L. fermentum (experimental group) or the same formula without the probiotic strain (control group) for 5 months. The primary outcome of the study was average daily weight gain between baseline and 4 months of age. Secondary outcomes were other anthropometric data (length and head circumference), formula consumption, and tolerance. Incidence of infections was also recorded by pediatricians. No significant differences in weight gain were observed between both groups, neither at 4 months of age (29.0±7.8 vs 28.9±5.7g/day) nor at 6 months (25.1±6.1 vs 24.7±5.2g/day). There were no statistically significant differences in the consumption of the formulae or symptoms related to the tolerance of the formula. The incidence rate of gastrointestinal infections in infants of the control group was 3 times higher than in the probiotic group (p=0.018). Therefore, consumption of a prebiotic infant formula enriched with the human milk probiotic strain L. fermentum CECT5716 from 1 to 6 months of life is well tolerated and safe. Furthermore, the consumption of this formula may improve the health of the infants by reducing the incidence of gastrointestinal infections.

Human milk probiotic Lactobacillus fermentum CECT5716 reduces the incidence of gastrointestinal and upper respiratory tract infections in infants.

OBJECTIVES: The aim of the study was to examine the effects of a follow-on formula containing Lactobacillus fermentum CECT5716 (L. fermentum) on the incidence of infections in infants between the ages of 6 and 12 months. PATIENTS AND METHODS: A randomized double-blinded controlled study including infants at the age of 6 months was conducted. Infants were assigned randomly to either follow-on formula supplemented with L. fermentum plus galactooligosaccharide (experimental group, EG), or the same formula supplemented with only galactooligosaccharide (control group, CG). The main outcome was the incidence of infections for the 6-month duration of the study. RESULTS: The EG showed a significant 46% reduction in the incidence rate (IR) of gastrointestinal infections (EG: 0.196 ±â€Š0.51, CG: 0.363 ±â€Š0.53, IR ratio 0.54, 95% confidence interval [CI] 0.307-0.950, P = 0.032), 27% reduction in the incidence of upper respiratory tract infections (EG: 0.969 ±â€Š0.96, CG: 1.330 ±â€Š1.23, IR ratio 0.729, 95% CI 0.46-1.38, P = 0.026), and 30% reduction in the total number of infections (EG: 1.464 ±â€Š1.15, CG: 2.077 ±â€Š1.59, IR ratio 0.70, 95% CI 0.46-1.38, P = 0.003), at the end of the study period compared with CG. CONCLUSIONS: Administration of a follow-on formula with L. fermentum CECT5716 may be useful for the prevention of community-acquired gastrointestinal and upper respiratory infections.

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, B(6), 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 T(0), T(3), T(6), and T(12) months to determine plasma fatty acids, vitamins B(6), E, and 25-hydroxyvitamin D and serum folate, calcium, soluble osteoprotegerin (OPG), soluble receptor activator of NF-kappaB 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.

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 (2.5mg/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+/-2.4min, 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. 10min 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.

Health effects of oleic acid and long chain omega-3 fatty acids (EPA and DHA) enriched milks. A review of intervention studies.

Substitution of dietary saturated fat by oleic acid and/or polyunsaturated fatty acids (PUFA) has been described to reduce the cardiovascular risk by reducing blood lipids, mainly cholesterol. Additional benefits have been described for long chain omega-3 PUFA (eicosapentaenoic acid-EPA and docosahexaenoic acid-DHA) from fish oils. In recent years, food technology has been used to produce dairy drinks with a reduced content of saturated fat in favour of those fatty acids, most of them claiming cardiovascular benefits. This review summarises all the scientific evidence regarding the effects of milks enriched with long chain omega-3 PUFA (EPA+DHA) and/or oleic acid on cardiovascular health. Nine controlled intervention studies with enriched milks have reported effects on healthy volunteers, subjects with increased risk factors and cardiovascular patients. The main effects observed were reductions of blood lipids, mainly cholesterol, LDL-cholesterol and triglycerides.