[Microbiota-diet interactions: towards personalized nutrition]. / Interacciones microbiota-dieta: hacia la personalización de la nutrición.

Introduction: The gut microbiota has emerged as a key factor on one hand as a mediator of the effects of diet on health and, on the other hand, as a source of intervariability of response to a diet. Moreover, there is a strong bidirectional interaction between our health and the microbiota that inhabit us, with each determining the presence of the other. In this review are named some of the metabolic functions in which the microbiota participates and which have an impact on our health, with particular emphasis on its ability to ferment fiber and produce short-chain fatty acids (SCFA) that provide numerous benefits to our health, but which have also been linked to obesity. Finally, some examples of dietary intervention in which the microbiota has been shown to play a key role in the results obtained are mentioned.

Introducción: La microbiota intestinal se ha revelado como un factor clave, por un lado, como mediador de los efectos de la dieta en la salud y, por otro lado, como fuente de intervariabilidad de respuesta a una dieta. Además, existe una fuerte interacción bidireccional entre nuestra salud y la microbiota que nos habita, determinando cada uno la presencia del otro. En esta revisión se nombran algunas de las funciones metabólicas en las que participa la microbiota y que tienen un impacto en nuestra salud, con especial hincapié en su capacidad para fermentar la fibra y producir ácidos grasos de cadena corta (AGCC), que aportan numerosos beneficios a nuestra salud, pero que también se han relacionado con la obesidad. Por último, se nombran algunos ejemplos de intervención dietética en los que se ha demostrado que la microbiota cumple un papel fundamental en los resultados obtenidos.

Interacciones microbiota-dieta: hacia la personalización de la nutrición / Microbiota-diet interactions: towards personalized nutrition

La microbiota intestinal se ha revelado como un factor clave, por un lado, como mediador de los efectos de la dieta en la salud y, por otro lado, como fuente de intervariabilidad de respuesta a una dieta. Además, existe una fuerte interacción bidireccional entre nuestra salud y la microbiota que nos habita, determinando cada uno la presencia del otro. En esta revisión se nombran algunas de las funciones metabólicas en las que participa la microbiota y que tienen un impacto en nuestra salud, con especial hincapié en su capacidad para fermentar la fibra y producir ácidos grasos de cadena corta (AGCC), que aportan numerosos beneficios a nuestra salud, pero que también se han relacionado con la obesidad. Por último, se nombran algunos ejemplos de intervención dietética en los que se ha demostrado que la microbiota cumple un papel fundamental en los resultados obtenidos. (AU)

The gut microbiota has emerged as a key factor on one hand as a mediator of the effects of diet on health and, on the other hand, as a source of intervariability of response to a diet. Moreover, there is a strong bidirectional interaction between our health and the microbiota that inhabit us, with each determining the presence of the other. In this review are named some of the metabolic functions in which the microbiota participates and which have an impact on our health, with particular emphasis on its ability to ferment fiber and produce short-chain fatty acids (SCFA) that provide numerous benefits to our health, but which have also been linked to obesity. Finally, some examples of dietary intervention in which the microbiota has been shown to play a key role in the results obtained are mentioned. (AU)

Green/Roasted Coffee May Reduce Cardiovascular Risk in Hypercholesterolemic Subjects by Decreasing Body Weight, Abdominal Adiposity and Blood Pressure.

In previous studies, after regularly consuming a green/roasted coffee blend, body weight, body fat%, glucose, plasminogen activator inhibitor-1 (PAI-1), resistin, leptin, ghrelin, diastolic (DBP) and systolic blood pressure (SBP) significantly changed in healthy and hypercholesterolemic subjects. However, glucagon, total-cholesterol (T-C), triglycerides (TG), LDL-cholesterol (LDL-C) and Homeostasis Model Assessment index to estimate insulin resistance (HOMA-IR) only changed in the hypercholesterolemics. This work looks into the antiobesity effects of coffee blend and into the relationship of antiobesity with the aforementioned cardiometabolic modifications in hypercholesterolemics. (1) Methods: Tricipital and subscapular skinfolds, hip, thigh, arm and waist circumference (WC) were measured in normocholesterolemic and hypercholesterolemics. To understand the relationship between cardiometabolic and antiobesity results in hypercholesterolemics, factor analysis was carried out using baseline values of the variables that changed. (2) Results: WC, WC/hip and WC/height showed significant coffee×group interaction, and in hypercholesterolemics tended to decrease. After factor analysis, three factors emerged, accounting for 29.46, 13.13 and 11.79% of variance. Only factor 1 (main loadings: WC, DBP and SBP, body weight, WC/hip and WC/height ratios, TG and ghrelin, inversely) decreased after coffee intake. (3) Conclusion: Regularly consuming green/roasted coffee may help to control body weight, and in hypercholesterolemics, may reduce cardiovascular risk by reducing abdominal adiposity and blood pressure.

Cocoa colonic phenolic metabolites are related to HDL-cholesterol raising effects and methylxanthine metabolites and insoluble dietary fibre to anti-inflammatory and hypoglycemic effects in humans.

BACKGROUND: In many cocoa intervention studies, health outcomes are related to cocoa components without taking into account the bioavailability of the main bioactive components: phenolic compounds and methylxanthines. METHODS: The present work associates the results of bioavailability and randomised controlled crossover studies in humans carried out with similar cocoa products, so that the main phenol and methylxanthine metabolites observed in plasma and urine are associated to the health effects observed in the chronic studies. We outstand that doses of cocoa and consumption rate used are realistic. In the bioavailability study, a conventional (CC) and a methylxanthine-polyphenol rich (MPC) cocoa product were used, whereas in the chronic study a dietary fibre-rich (DFC) and a polyphenol-rich (PC) product were studied in healthy and cardiovascular risk subjects. RESULTS AND DISCUSSION: The main phenolic metabolites formed after CC and MPC intake, 5-(4'-hydroxyphenyl)-γ-valerolactone-3'-sulfate, 3'-methyl-epicatechin-5-sulfate, 4-hydroxy-5-(4'-hydroxyphenyl)valeric acid-sulfate, 5-phenyl-γ-valerolactone--sulfate and 5-(4'-hydroxyphenyl)-γ-valerolactone-3'-glucuronide, may contribute to the changes in cholesterol (and indirectly HDL-cholesterol) observed after the regular intake of both DFC and PC, in healthy and cardiovascular risk subjects, whereas 7-methylxanthine (the main cocoa methylxanthine metabolite) and theobromine, together with its content in insoluble dietary fibre, may be responsible for the decrease of IL-1ß and hypoglycemic effects observed with DFC. With both phenolic and methylxanthine metabolites a strong dose-response effect was observed. CONCLUSION: After the regular consumption of both DFC and PC, positive changes were observed in volunteer's lipid profile, which may be related to the long-lasting presence of colonic phenolic metabolites in blood. In contrast, the anti-inflammatory and hypoglycemic effects were only observed with DFC, and these may be related to methylxanthine metabolites, and it is likely that insoluble dietary fibre may have also played a role.

Microbiota Features Associated With a High-Fat/Low-Fiber Diet in Healthy Adults.

A high intake of dietary saturated fatty acids (SFAs) is related to an increased risk of obesity, inflammation and cancer-related diseases, and this risk is attenuated only when SFAs are replaced by unsaturated fats and unrefined carbohydrates. The gut microbiota has recently emerged as a new environmental factor in the pathophysiology of these disorders, and is also one of the factors most influenced by diet. We sought to determine whether the gut microbiota of healthy individuals whose intake of SFAs exceeds World Health Organization (WHO) recommendations exhibits features similar to those reported in people with obesity, inflammation, cancer or metabolic disease. Healthy non-obese subjects were divided into two groups based on their SFAs intake. Body composition and gut microbiota composition were analyzed, and associations between bacterial taxa, diet and body fat composition were determined globally and separately by sex. Metagenome functional pathways were predicted by PICRUSt analysis. Subjects whose SFAs intake exceeded WHO recommendations also had a dietary pattern of low fiber intake. This high saturated fat/low fiber diet was associated with a greater sequence abundance of the Anaerotruncus genus, a butyrate producer associated with obesity. Analysis of data of high SFAs intake by sex showed that females presented with a greater abundance of Campylobacter, Blautia, Flavonifractor and Erysipelatoclostridium, whereas males showed higher levels of Anaerotruncus, Eisenbergiella, a genus from the order Clostridiales (FamilyXIIIUCG_001) and two genera from the Lachnospiraceae family. PICRUSt analysis confirmed these data, showing a correlation with a decrease in the abundance of sequences encoding for transporters of some metals such as iron, which is needed to maintain a healthy metabolism. Thus, the microbiota of healthy people on a high SFAs diet contain bacterial taxa (Anaerotruncus, Lachnospiraceae Flavonifractor, Campylobacter, Erysipelotrichacea and Eisenbergiella) that could be related to the development of some diseases, especially obesity and other pro-inflammatory diseases in women. In summary, the present study identifies bacterial taxa that could be considered as early predictors for the onset of different diseases in healthy subjects. Also, sex differences in gut microbiota suggest that women and men differentially benefit from following a specific diet.

Stevia rebaudiana Bertoni bioactive effects: From in vivo to clinical trials towards future therapeutic approaches.

Stevia rebaudiana Bertoni, a plant from South America and indigenous of Paraguay, has shown several biological effects and healthy properties, although it is especially used in South America and some Asiatic regions. In addition, it is a natural sweetener, almost 300 times sweeter than sucrose, being attributed to its phytoconstituents prominent antioxidant, antimicrobial, antidiabetic (antihyperglycemic, insulinotropic, and glucagonostatic), antiplatelet, anticariogenic, and antitumor effects. In this sense, this work aims to provide an extensive overview on the historical practices of stevia and its effects in human health based on its chemical composition and applications for both food and pharmaceutical industries.

Flavanol Bioavailability in Two Cocoa Products with Different Phenolic Content. A Comparative Study in Humans.

Cocoa has beneficial health effects partly due to its high flavanol content. This study was aimed at assessing the absorption and metabolism of polyphenols in two soluble cocoa products: a conventional (CC) and a flavanol-rich product (CC-PP). A crossover, randomized, blind study was performed in 13 healthy men and women. On two different days, after an overnight fast, volunteers consumed one serving of CC (15 g) or CC-PP (25 g) in 200 mL of semi-skimmed milk containing 19.80 mg and 68.25 mg of flavanols, respectively. Blood and urine samples were taken, before and after CC and CC-PP consumption, and analyzed by high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight mass spectrometry (HPLC-ESI-QToF-MS). Up to 10 and 30 metabolites were identified in plasma and urine, respectively. Phase II derivatives of epicatechin were identified with kinetics compatible with small intestine absorption, although the most abundant groups of metabolites were phase II derivatives of phenyl-γ-valerolactone and phenylvaleric acid, formed at colonic level. 5-(4'-Hydroxyphenyl)-γ-valerolactone-sulfate could be a sensitive biomarker of cocoa flavanol intake. CC and CC-PP flavanols showed a dose-dependent absorption with a recovery of 35%. In conclusion, cocoa flavanols are moderately bioavailable and extensively metabolized, mainly by the colonic microbiota.

Moderate consumption of a soluble green/roasted coffee rich in caffeoylquinic acids reduces cardiovascular risk markers: results from a randomized, cross-over, controlled trial in healthy and hypercholesterolemic subjects.

PURPOSE: Coffee is rich in bioactive compounds with health beneficial properties, with green coffee presenting higher phenol content than roasted. We evaluated the effects of regularly consuming realistic amounts of a green/roasted coffee blend on cardiovascular health-related biomarkers. METHODS: A randomized, cross-over, controlled study was carried out in 25 normocholesterolemic [total cholesterol (TC) < 200 mg/dL] and 27 hypercholesterolemic (TC 200-240 mg/dL) subjects. During 8 weeks, volunteers consumed 6 g/day of soluble green/roasted (35:65) coffee or a control beverage (water or an isotonic drink). Blood pressure, heart rate and body weight were monitored at the end of each intervention, and serum lipids [TC, HDL-C, LDL-C, VLDL-C, triglycerides and phospholipids], cytokines and chemokines (IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12, IL-13, IL-17, G-CSF, GM-CSF, MCP-1, MIP-1ß, TNF-α, INF-γ), adhesion molecules (ICAM-1, VCAM-1), and C-reactive protein were measured. Plasma antioxidant capacity (FRAP, ORAC and ABTS methods), and lipid (malondialdehyde, MDA) and protein (carbonyl groups, CG) oxidation were also determined. RESULTS: Attending to the general lineal model of variance for repeated measures, after the green/roasted coffee intervention significant reductions in TC, LDL-C, VLDL-C and triglycerides levels (p = 0.006, 0.001, 0.003 and 0.017, respectively), and a significant group effect were observed (0.001, < 0.001, 0.019 and 0.027, respectively). Only within the hypercholesterolemic group, attending to the Bonferroni test, the aforementioned lipid parameters were significantly lower after regular green/roasted coffee intake compared to baseline values. Moreover, after the coffee stage, plasma antioxidant capacity improved, according to the increase in ORAC and FRAP values (p < 0.001 and p < 0.001, respectively) and decrease of MDA (p = 0.015) and CG (p < 0.001) levels, without differences between groups. Systolic (p = 0.001) and diastolic (p < 0.001) blood pressure, heart rate (p = 0.035), and body weight (p = 0.017) were reduced in both normo- and hypercholesterolemic groups. CONCLUSION: Regular consumption of moderate amounts of a soluble green/roasted (35:65) coffee blend may contribute to improve cardiovascular health in moderately hypercholesterolemic people, as reducing serum lipids, blood pressure and body weight effects, as well as increasing plasma antioxidant capacity, have been observed. Moreover, positive influences on blood pressure, body weight, and plasma antioxidant capacity were obtained in the healthy group. Therefore, incorporation of green coffee beans into the coffee brew can be recommended as part of a dietary strategy to protect from cardiovascular disease.

Absorption and metabolism of yerba mate phenolic compounds in humans.

Bioavailability of yerba mate phenolic compounds was assessed in healthy humans. More than 34 metabolites were identified in biological fluids, mainly sulfated conjugates of caffeic and ferulic/isoferulic acids, in addition to non-metabolized caffeoyl-, feruloyl- and p-coumaroilquinic acids, with rapid appearance and clearance in plasma indicative of small intestinal absorption. These compounds amounted to 13.1% of the urinary metabolites. Delayed absorption of dihydrocaffeic, dihydroferulic and dihydrocoumaric acids and their phase II metabolites, in addition to feruloylglycine, pointed to their microbial origin and colonic absorption, accounting for 81.0% of excreted metabolites. Phase II flavonol metabolites (0.2%) derived mainly from rutin after colonic transformation and absorption were also detected. Additionally, dihydroferuloyl-, dihydrocaffeoyl- and dihydrocoumaroylquinic acids (5.7%) were identified, showing the most delayed kinetics. Total phenolic excretion (147.6µmol) corresponded to 13.2% of ingested phenols. In conclusion, yerba mate polyphenols are partially bioavailable and extensively metabolized, mainly by the colonic microbiota.

Regularly consuming a green/roasted coffee blend reduces the risk of metabolic syndrome.

PURPOSE: Preventive health effects of coffee could have a widespread impact on public health. Green coffee has more phenols than roasted, and thus is healthier, although with less acceptable organoleptic properties. Therefore, the effects of regularly consuming a green/roasted coffee blend (35/65) on the main components of MetS in humans were evaluated. METHODS: A crossover, randomized, controlled study was performed in 25 normocholesterolaemic and 27 hypercholesterolaemic men and women aged 18-45 years with BMI 18-25 kg/m2. Three servings/day of the blend, providing 510.6 mg hydroxycinnamic acids and 121.2 mg caffeine/day, were consumed versus a control drink, during 8 weeks each. Polyphenol and methylxanthine-rich foods were restricted along the study. At the beginning (baseline) and end of the control and coffee interventions, blood samples were collected and glucose, HDL-cholesterol, triglycerides, insulin, leptin, plasminogen activator inhibitor-1 (PAI-1), resistin and visfatin were analysed; waist circumference, %body fat, and blood pressure were measured and dietary records and physical activity questionnaires completed. RESULTS: Systolic and diastolic blood pressure decreased (p = 0.001 and p < 0.001, respectively) in both groups as well as %body fat (p = 0.001) which may be related to the lower leptin (p = 0.001), PAI-1 (p < 0.001) and resistin (p = 0.034) levels in the two groups after coffee consumption. Glucose concentration (p = 0.030) and insulin resistance (p = 0.011; HOMA-IR) also decreased, as well as triglyceride levels (p = 0.017), so that the reduction was much greater in the hypercholesterolaemics (group effect, p = 0.027). CONCLUSION: Regular consumption of the green/roasted coffee blend may be recommended to healthy and hypercholesterolaemic subjects to prevent MetS, as it produces positive effects on blood pressure, glucose and triglyceride levels.

Bioavailability of hydroxycinnamates in an instant green/roasted coffee blend in humans. Identification of novel colonic metabolites.

Roasting greatly reduces the phenolic content in green coffee beans. Considering the beneficial effects of coffee polyphenols, blends containing green coffee beans are being consumed as a healthier alternative to roasted coffee. This study was aimed at assessing the absorption and metabolism of hydroxycinnamates in an instant green/roasted (35/65) coffee blend in healthy humans. Twelve fasting men and women consumed a cup of coffee containing 269.5 mg (760.6 µmol) of chlorogenic acids. Blood and urine samples were taken before and after coffee consumption at different times and analyzed by LC-MS-QToF. Up to 25 and 42 metabolites were identified in plasma and urine, respectively, mainly in the form of sulfate and methyl derivatives, and to a lower extent as glucuronides. Un-metabolized hydroxycinnamate esters (caffeoyl-, feruloyl-, and coumaroylquinic acids), hydroxycinnamic acids (caffeic, ferulic and coumaric acids) and their phase II metabolites, in addition to phase II derivatives of lactones, represented a minor group of metabolites (16.3% of the metabolites excreted in urine) with kinetics compatible with small intestine absorption. Dihydrohydroxycinnamic acids and their phase II derivatives, in addition to feruloylglycine, showed delayed kinetics due to their colonic origin and represented the most abundant group of metabolites (75.7% of total urinary metabolites). Dihydrohydroxycinnamate esters (dihydroferuloyl-, dihydrocaffeoyl- and dihydrocoumaroylquinic acids) have been identified for the first time in both plasma and urine, with microbial origin (excreted 8-12 h after coffee intake) amounting to 8% of total urinary metabolites. In conclusion, coffee polyphenols are partially bioavailable and extensively metabolized, mainly by the colonic microbiota.

Hydroxytyrosol in functional hydroxytyrosol-enriched biscuits is highly bioavailable and decreases oxidised low density lipoprotein levels in humans.

Hydroxytyrosol (HT) and its derivatives in olive oil protect low-density lipoproteins (LDL) against oxidation. Biscuits could be a convenient alternative to broaden consumers' choice of HT-rich foods, although the biscuit matrix could affect HT bioavailability. We performed a crossover, randomized, double-blind study to evaluate HT bioavailability in HT-enriched biscuits (HT-B) versus non-enriched biscuits (C-B), and the effects on oxidative postprandial status. On two separate days, 13 subjects consumed 30 g of C-B or HT-B (5.25mg HT) after overnight-fasting. Blood and urine were collected at different intervals and analysed by LC-MS-QToF. After HT-B consumption, plasma metabolites peaked between 0.5 and 1h and were extensively excreted in urine. HT-sulphate and 3,4-dihydroxyphenylacetic acid (DOPAC)-sulphate were the main metabolites, followed by DOPAC and homovanillic acid (HVA). HT-glucuronide, DOPAC-glucuronide, HVA-glucuronide and HVA-sulphate were also detected. Postprandial oxidised-LDL concentrations decreased with HT-B. HT is a promising functional ingredient and, in biscuits, it is highly bioavailable and lowers postprandial oxidised-LDL levels.

Effects of bioactive constituents in functional cocoa products on cardiovascular health in humans.

Cocoa manufacturers are producing novel products increasing polyphenols, methylxanthines or dietary fibre to improve purported health benefits. We attempt to explain the contribution of cocoa bioactive compounds to cardiovascular effects observed in previous studies, placing particular emphasis on methylxanthines. We focused on a soluble cocoa product rich in dietary fibre (DFCP) and a product rich in polyphenols (PPCP). Effects of regularly consuming DFCP (providing daily 10.17 g, 43.8 mg and 168.6 mg of total-dietary-fibre, flavanols and methylxanthines, respectively) as well as PPCP (providing daily 3.74 g, 45.3 mg and 109.8 mg of total-dietary-fibre, flavanols and methylxanthines, respectively) on cardiovascular health were assessed in two controlled, cross-over studies in free-living normocholesterolemic and moderately hypercholesterolemic subjects. Both products increased HDL-cholesterol concentrations, whereas only DFCP decreased glucose and IL-1ß levels in all subjects. Flavanols appeared to be responsible for the increase in HDL-cholesterol, whereas insoluble-dietary-fibre and theobromine in DFCP were associated with the hypoglycemic and anti-inflammatory effects observed.

Comparative evaluation of the metabolic effects of hydroxytyrosol and its lipophilic derivatives (hydroxytyrosyl acetate and ethyl hydroxytyrosyl ether) in hypercholesterolemic rats.

Hydroxytyrosol (HT), a virgin olive oil phenolic phytochemical with proven health benefits, has been used to generate new lipophilic antioxidants to preserve fats and oils against autoxidation. The aim of this work is to comparatively evaluate the physiological effects of HT and its lipophilic derivatives, hydroxytyrosyl acetate (HT-Ac) and ethyl hydroxytyrosyl ether (HT-Et), in high-cholesterol fed animals. Male Wistar rats (n = 8) were fed a standard diet (C group), a cholesterol-rich diet (Chol group) or a cholesterol-rich diet supplemented with phenolic compounds (HT group, HT-Ac group and HT-Et group) for 8 weeks. Body and tissue weights, the lipid profile, redox status, and biochemical, hormonal, and inflammatory biomarkers were evaluated. Plasma levels of total cholesterol, LDL cholesterol, glucose, insulin and leptin, as well as malondialdehyde in serum increased in Chol compared to C (p < 0.05). Rats fed the test diets had improved glucose, insulin, leptin and MDA levels and antioxidant capacity status, with HT-Ac being the most effective compound. The studied phenolic compounds also modulated TNF-α and IL-1ß plasma levels compared to Chol. HT-Ac and HT-Et improved adipose tissue distribution and adipokine production, decreasing MCP-1 and IL-1ß levels. Our results confirm the metabolic effects of HT, which are maintained and even improved by hydrophobic derivatives, particularly HT-Ac.

Realistic intake of a flavanol-rich soluble cocoa product increases HDL-cholesterol without inducing anthropometric changes in healthy and moderately hypercholesterolemic subjects.

To assess whether antioxidant, anti-inflammatory and other cardio-protective effects attributed to cocoa are achieved when regularly consuming moderate amounts of a flavanol-rich soluble cocoa product, a non-randomized, controlled, crossover, free-living study was carried out in healthy (n = 24; 25.9 ± 5.6 years) and moderately hypercholesterolemic (200-240 mg dL(-1); n = 20; 30.0 ± 10.3 years) volunteers. Participants consumed two servings per day (7.5 g per serving) of a soluble cocoa product (providing 45.3 mg flavanols per day) in milk, which was compared with consuming only milk during a 4 week period. The effects on systolic and diastolic blood pressure and heart rate were determined, as well as on serum lipid and lipoprotein profiles, interleukins (IL)-1ß, IL-6, IL-8, IL-10, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), vascular (VCAM-1) and intercellular cell adhesion molecules (ICAM-1), serum malondialdehyde (MDA), carbonyl groups (CG), ferric reducing/antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and free radical scavenging capacity (ABTS). During the study, the volunteers' diets and physical activity were also evaluated, as well as any changes in weight, skin folds, circumferences and related anthropometric parameters. Cocoa and certain polyphenol-rich fruits and vegetables and their derivatives were restricted. After consuming the cocoa product positive effects were observed such as an increase in serum HDL-C (p < 0.001) and dietary fiber intake (p = 0.050), whereas IL-10 decreased (p = 0.022). Other cardiovascular-related biomarkers and anthropometric parameters were unaffected. We have therefore concluded that regular consumption of this cocoa product in a Spanish-Mediterranean diet may protect against cardiovascular disease in healthy and hypercholesterolemic subjects without producing any weight gain or other anthropometric changes.

Regular consumption of a cocoa product improves the cardiometabolic profile in healthy and moderately hypercholesterolaemic adults.

Cocoa products present great health potential due to their high content of polyphenols, mainly of flavanols. However, the antioxidant, anti-inflammatory and other health effects of regularly consuming cocoa products seem to depend on the intake and health status of the consumer, etc. and need to be further clarified. A randomised, controlled, cross-over, free-living study was carried out in healthy (n 24) and moderately hypercholesterolaemic (>2000 mg/l, n 20) subjects to assess the influence of regularly consuming (4 weeks) two servings (15 g each) of a cocoa product rich in fibre (containing 33·9 % of total dietary fibre (TDF) and 13·9 mg/g of soluble polyphenols) in milk v. consuming only milk (control) on (1) serum lipid and lipoprotein profile, (2) serum malondialdehyde levels, carbonyl groups, ferric reducing/antioxidant power, oxygen radical absorbance capacity and free radical-scavenging capacity, (3) IL-1ß, IL-6, TNF-α, IL-10, IL-8, monocyte chemoattractant protein-1, and vascular and intracellular cell adhesion molecule levels, and (4) systolic and diastolic blood pressure and heart rate. Throughout the study, the diet and physical activity of the volunteers, as well as any possible changes in weight or other anthropometric parameters, were also evaluated. The intake of TDF increased (P< 0·001) to the recommended levels. Serum HDL-cholesterol (HDL-C) levels were increased (P< 0·001), whereas glucose (P= 0·029), IL-1ß (P= 0·001) and IL-10 (P= 0·001) levels were decreased. The rest of the studied cardiovascular parameters, as well as the anthropometric ones, remained similar. In conclusion, regularly consuming a cocoa product with milk improves cardiovascular health by increasing HDL-C levels and inducing hypoglycaemic and anti-inflammatory effects in healthy and hypercholesterolaemic individuals without causing weight gain.

Pharmacokinetics of caffeine and its metabolites in plasma and urine after consuming a soluble green/roasted coffee blend by healthy subjects.

Coffee is widely consumed worldwide; therefore, the methylxanthines contained in coffee, mainly caffeine (CF), are among the most abundant bioactive compounds in our diet. In the present work, the bioavailability and metabolism of methylxanthines in a commercial soluble green/roasted coffee blend was studied. After a 3-day restriction of methylxanthine-containing foods, fasting healthy subjects (12 men and women) consumed the coffee product containing 70.69mg CF and 0.119mg theobromine (TB). Plasma samples were taken before (t=0h) and after coffee consumption at different time points (0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, 9, 10 and 12h). Urine was collected at baseline (-2-0h) and at different intervals (0-2, 2-5, 5-8, 8-12 and 12-24h). Samples were analyzed by HPLC-DAD and LC-MS-QToF, and pharmacokinetic parameters were calculated. CF was the main methylxanthine found in plasma (Cmax=10.50µM, Tmax=1.2h). In addition, seven methylxanthines and methyluric acids were detected between 0.5 and 12h after coffee intake, paraxanthine (PX) being the major metabolite (Cmax=3.36µM), followed by 1-methyluric acid (1-MU; Cmax=1.44µM) and 1-methylxanthine (1-MX; Cmax=1.27µM), identified in plasma samples for the first time. In 24h urine, eleven methylxanthines and methyluric acids were detected, 1-MU being the major metabolite (Cmax=150.52µM, Tmax=12h) amounting to 67.7% of the total urinary metabolites. In conclusion, a rapid absorption, metabolization and excretion of caffeine and its derived methylxanthines and methyluric acids have been observed after consumption of a green/roasted coffee product.

Effects of regularly consuming dietary fibre rich soluble cocoa products on bowel habits in healthy subjects: a free-living, two-stage, randomized, crossover, single-blind intervention.

BACKGROUND: Dietary fibre is both preventive and therapeutic for bowel functional diseases. Soluble cocoa products are good sources of dietary fibre that may be supplemented with this dietary component. This study assessed the effects of regularly consuming two soluble cocoa products (A and B) with different non-starch polysaccharides levels (NSP, 15.1 and 22.0% w/w, respectively) on bowel habits using subjective intestinal function and symptom questionnaires, a daily diary and a faecal marker in healthy individuals. METHODS: A free-living, two-stage, randomized, crossover, single-blind intervention was carried out in 44 healthy men and women, between 18-55 y old, who had not taken dietary supplements,laxatives, or antibiotics six months before the start of the study. In the four-week-long intervention stages, separated by a three-week-wash-out stage, two servings of A and B, that provided 2.26 vs. 6.60 g/day of NSP respectively, were taken. In each stage, volunteers' diet was recorded using a 72-h food intake report. RESULTS: Regularly consuming cocoa A and B increased fibre intake, although only cocoa B significantly increased fibre intake (p < 0.001) with respect to the non-cocoa stage. No changes in body weight were observed in either of the 4 week interventions. With cocoa product B, the number of daily bowel movements increased (p = 0.002), the frequency of having a bowel movement once a day increased (p = 0.009), the time to have a bowel movement was lower (p = 0.016) as well as the feeling of constipation (p = 0.046) without inducing adverse gastrointestinal symptoms, only flatulence increased (p = 0.019). CONCLUSIONS: Regular consumption of the cocoa products increases dietary fibre intake to recommended levels and product B improves bowel habits. The use of both objective and subjective assessments to evaluate the effects of food on bowel habits is recommended.

Anion exchange liquid chromatography for the determination of nucleotides in baby and/or functional foods.

A sensitive, selective and solvent-free procedure is proposed for the rapid determination of monophosphate nucleotides (cytidine 5'-monophosphate, uridine 5'-monophosphate, adenosine 5'-monophosphate, and guanosine 5'-monophosphate) in baby foods. The method is based on the deproteinization of foods and direct analysis by anion exchange liquid chromatography (LC). Nucleotides were separated on an SAX anion exchange column with isocratic elution using 0.01 M dihydrogenphosphate buffer (pH 3.5) as mobile phase at a flow-rate of 1 mL min(-1) and detected by diode-array detection (DAD). The LC method rapidly separated the nucleotides (9 min) and was validated for linearity, detection and quantitation limits, selectivity, accuracy, and precision. The recoveries obtained for spiked samples were satisfactory for all the analytes. The proposed procedure allowed the only authorized nucleotides in infant formulas to be determined and was successfully applied to the analysis of different baby and/or functional food samples, including infant formulas, fermented milk, cereals, and purees.