The Synergistic Effect of Quince Fruit and Probiotics (Lactobacillus and Bifidobacterium) on Reducing Oxidative Stress and Inflammation at the Intestinal Level and Improving Athletic Performance during Endurance Exercise.

Endurance exercise promotes damage at the intestinal level and generates a variety of symptoms related to oxidative stress processes, inflammatory processes, microbiota dysbiosis, and intestinal barrier damage. This study evaluated the effects of quince (Cydonia oblonga Mill.) and probiotics of the genera Lactobacillus and Bifidobacterium on intestinal protection and exercise endurance in an animal swimming model. Phytochemical characterization of the quince fruit demonstrated a total dietary fiber concentration of 0.820 ± 0.70 g/100 g and a fiber-bound phenolic content of 30,218 ± 104 µg/g in the freeze-dried fruit. UPLC-PDA-ESI-QqQ analyses identified a high content of polyphenol, mainly flavanols, hydroxycinnamic acids, hydroxybenzoic acids, flavonols, and, to a lesser extent, dihydrochalcones. The animal model of swimming was performed using C57BL/6 mice. The histological results determined that the consumption of the synbiotic generated intestinal protection and increased antioxidant (catalase and glutathione peroxidase enzymes) and anti-inflammatory (TNF-α and IL-6 and increasing IL-10) activities. An immunohistochemical analysis indicated mitochondrial biogenesis (Tom2) at the muscular level related to the increased swimming performance. These effects correlated mainly with the polyphenol content of the fruit and the effect of the probiotics. Therefore, this combination of quince and probiotics could be an alternative for the generation of a synbiotic product that improves exercise endurance and reduces the effects generated by the practice of high performance sports.

Effects of protein-carbohydrate vs. carbohydrate alone supplementation on immune inflammation markers in endurance athletes: a randomized controlled trial.

PURPOSE: The impact of ingesting carbohydrates alone or combined with proteins to support exercise immune adaptation in endurance athletes is scarcely investigated. The present study compares the effect of ingesting a combined protein-carbohydrate supplement vs. a carbohydrate-only supplement post-workout on immune inflammation markers following a 10 week periodized endurance training program in well-trained athletes. METHODS: Twenty-five men completed the study after being randomly assigned to one of the following intervention groups: combined protein-carbohydrate (PRO-CHO n = 12, 31 ± 9 years, [Formula: see text]O2peak 61.0 ± 5.6 ml.kg-1.min-1) or non-protein isoenergetic carbohydrate (CHO, n = 13, 33 ± 8 years, [Formula: see text]O2peak 60.6 ± 6.9 ml.kg-1.min-1). Treatment consisted of ingesting 24 g of assigned supplement, mixed with 250 ml of orange juice, once a day for 10 weeks immediately post-workout (or before breakfast on non-training days). Measurements were conducted pre- and post-intervention on total leukocytes, leukocyte subsets (i.e., neutrophils, eosinophils, basophils, monocytes, and lymphocytes), and platelets. The inflammatory status was assessed by the neutrophil-to-lymphocyte ratio (NLR), the platelet-to-lymphocyte ratio (PLR), and the systemic-immune inflammation index (SII). RESULTS: Post-intervention, significant increases were observed for CHO group only for the three inflammatory markers: NLR (p = 0.050, d = 0.58), PLR (p = 0.041, d = 0.60), and SII (p = 0.004, d = 0.81) but not for PRO-CHO (p > 0.05). CONCLUSION: Ingesting a post-workout protein-carbohydrate combined beverage promoted a more favourable immune status than carbohydrate-only ingestion by attenuating cellular inflammation over a 10 week training period in endurance male athletes. TRIAL REGISTRATION: The study was registered in ClinicalTrials.gov with the following ID: NCT02954367. The study was registered by 3 November 2016.

[Impact of vitamin D on health. Difficulties and strategies to reach the recommended intakes]. / Impacto de la vitamina D en la salud. Dificultades y estrategias para alcanzar las ingestas recomendadas.

Introduction: Vitamin D is a key nutrient for health. Recent research has shown that it is not only necessary for the maintenance of bone health, but also for the prevention of cardiovascular diseases, insulin resistance, respiratory diseases, infections, and cancer, among others. Solar synthesis of vitamin D is usually insufficient, so it is necessary to provide enough vitamin D through the diet in order to maintain an adequate nutritional status. Studies carried out in Spain and other countries have shown that serum levels of the vitamin were usually insufficient and that average dietary intakes were well below those marked as recommended, while the main food sources of vitamin D were fatty fish, breakfast cereals, eggs, and dairy. Food sources of vitamin D are scarce, and it is naturally located in the fatty part of foods of animal origin, and in the liver and viscera. Fortifying foods that are not natural food sources of vitamin D but are regularly consumed by a significant percentage of the population, is a strategy that could help to increase vitamin D intake. In this regard, dairy products and breakfast cereals are two of the most widely used food matrices for vitamin D fortification, and their incorporation into the usual diet has proven to be an effective strategy to improve the nutritional situation of the population in vitamin D.

Introducción: La vitamina D es clave en la salud y sus funciones se relacionan con el mantenimiento de la salud ósea, cardiovascular, la resistencia a la insulina, enfermedades respiratorias, infecciones y cáncer, entre otras. La síntesis solar de la vitamina D es con frecuencia insuficiente, por lo que para mantener un estatus adecuado de la vitamina es necesario que se aporten cantidades suficientes con la dieta. Los estudios realizados en España y en otros países muestran que los niveles séricos de la vitamina son con frecuencia insuficientes y que las ingestas dietéticas medias se encuentran muy por debajo de las marcadas como recomendadas. Las principales fuentes alimentarias de la vitamina D son los pescados grasos, los cereales de desayuno, los huevos y los lácteos. Las fuentes alimentarias de vitamina D son escasas y se encuentra especialmente en la parte grasa de alimentos de origen animal, en hígados y en vísceras. El enriquecimiento de alimentos que de forma natural no tienen un elevado contenido en vitamina D, pero que son consumidos habitualmente por un porcentaje significativo de la población, es una estrategia que ayuda a aumentar su ingesta. En este sentido, los lácteos y los cereales de desayuno son dos de las matrices alimentarias más utilizadas y su incorporación en la dieta habitual ha demostrado ser una estrategia eficaz para mejorar la situación nutricional de la población respecto a la vitamina D.

Evaluation of a Zingiber officinale and Bixa orellana Supplement on the Gut Microbiota of Male Athletes: A Randomized Placebo-Controlled Trial.

The gut microbiota has emerged as a factor that influences exercise performance and recovery. The present study aimed to test the effect of a polyherbal supplement containing ginger and annatto called "ReWin(d)" on the gut microbiota of recreational athletes in a pilot, randomized, triple-blind, placebo-controlled trial. Thirty-four participants who practice physical activity at least three times weekly were randomly allocated to two groups, a ReWin(d) group or a maltodextrin (placebo) group. We evaluated the gut microbiota, the production of short-chain fatty acids, and the serum levels of interleukin-6 and lipopolysaccharide at baseline and after 4 weeks. Results showed that ReWin(d) supplementation slightly increased gut microbiota diversity. Pairwise analysis revealed an increase in the relative abundance of Lachnospira (ß-coefficient = 0.013; p = 0.001), Subdoligranulum (ß-coefficient = 0.016; p = 0.016), Roseburia (ß-coefficient = 0.019; p = 0.001), and Butyricicoccus (ß-coefficient = 0.005; p = 0.035) genera in the ReWin(d) group, and a decrease in Lachnoclostridium (ß-coefficient = - 0.008; p = 0.009) and the Christensenellaceae R7 group (ß-coefficient = - 0.010; p < 0.001). Moreover, the Christensenellaceae R-7 group correlated positively with serum interleukin-6 (ρ = 0.4122; p = 0.032), whereas the Lachnospira genus correlated negatively with interleukin-6 (ρ = - 0.399; p = 0.032). ReWin(d) supplementation had no effect on short-chain fatty acid production or on interleukin-6 or lipopolysaccharide levels.

Chronic Consumption of Cocoa Rich in Procyanidins Has a Marginal Impact on Gut Microbiota and on Serum and Fecal Metabolomes in Male Endurance Athletes.

Cocoa is used in the sports world as a supplement, although there is no consensus on its use. We investigated the effect of cocoa intake on intestinal ischemia (intestinal fatty acid-binding protein (I-FABP)), serum lipopolysaccharide (LPS) levels, gastrointestinal symptoms, and gut microbiota in endurance athletes during their training period on an unrestricted diet. We also performed a metabolomics analysis of serum and feces after a bout of exercise before and after supplementation. Cocoa consumption had no effect on I-FABP, LPS, or gastrointestinal symptoms. Cocoa intake significantly increased the abundance of Blautia and Lachnospira genera and decreased the abundance of the Agathobacter genus, which was accompanied by elevated levels of polyphenol fecal metabolites 4-hydroxy-5-(phenyl)-valeric acid and O-methyl-epicatechin-O-glucuronide. Our untargeted approach revealed that cocoa had no significant effects on serum and fecal metabolites and that its consumption had little impact on the metabolome after a bout of physical exercise.

Can study of the ADRB3 gene help improve weight loss programs in obese individuals? / ¿Puede el estudio del gen ADRB3 ayudar a mejorar los programas de pérdida de peso en individuos obesos?

INTRODUCTION: Obesity is a chronic disease of multifactorial origin characterized by excess weight and excess fat accumulation, and whose etiology includes intrinsic (genetic, physiological, and metabolic) and extrinsic (social and cultural) factors. Fat accumulation is caused by a prolonged imbalance in the energy balance influenced, among other factors, by adaptive thermogenesis, which is triggered by cold environmental conditions, or by hypercaloric intake. Thermogenesis is regulated by the sympathetic nervous system and occurs in the muscle and brown adipose tissue. There are adrenergic receptors in brown adipose tissue, including the beta-3 adrenergic receptor (ADRB3), the main receptor for the regulation of thermogenesis. The presence in heterozygosis of an SNP-type polymorphism in the ADRB3 gene (Trp64Arg; rs4994) is associated with a lower lipolytic activity, a predisposition to obesity, and resistance to weight loss. The objective of this study was to analyze through a systematic review the weight loss program most appropriate for carriers. METHODS: A retrospective study of published papers on rs4994 polymorphism in the SNP and PubMed databases was conducted. RESULTS: Most published studies suggest the presence of obesity and resistance to weight loss in carriers, and report significant improvements in anthropometric parameters when patients receive fat-rich hypocaloric diets. CONCLUSIONS: Based on these conclusions, specific nutritional and physical exercise guidelines are proposed for individuals carrying the Trp64Arg allele.

Polyphenol-Rich Foods for Human Health and Disease.

Polyphenols are a class of well-known bioactive compounds widely distributed in the plant kingdom and abundant in plant foods and derived food products [...].

Effects of gastrointestinal digested polyphenolic enriched extracts of Chilean currants (Ribes magellanicum and Ribes punctatum) on in vitro fecal microbiota.

Chilean currants (Ribes magellanicum and Ribes punctatum) are wild polyphenol-rich berries with interesting bioactivities in several in vitro models. The aim of this study was to investigate the effects of the pre-digested PEE (polyphenol-enriched extract) in a simulated colon model. Fruits were extracted, submitted to simulated gastrointestinal digestion and further colonic fermentation with feces from healthy human donors. Samples were taken at 1, 4, 8 and 24 h of incubation, monitoring pH, ammonia, branched-chain fatty acids (BCFA), short-chain fatty acids (SCFA) and bacterial growth. FOS (fructooligosaccharides) and fecal slurry without treatments were positive and negative control, respectively. Both Ribes species reduced (p < 0.05) both BCFA and SCFA at 24 h. R. punctatum promoted the growth (p < 0.05) of beneficial bacteria such as Clostridium cluster XIVa, and Faecalibacterium prausnitzii; while a trend to increase Akkermansia muciniphila was observed. R. magellanicum increased (p < 0.05) Clostridium cluster XIVa population. Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. remained unaffected. Our results suggest that polyphenols from R. punctatum and R. magellanicum may modulate both bacterial metabolism and some selected gut beneficial bacteria under simulated conditions. Therefore, Chilean currants might be useful as supplements to maintain a healthy colon; however, further in vivo studies are needed to confirm their effect and their mechanisms.

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.

Life style and risk of atypical eating disorders in university students: Reality versus perception. / Estilo de vida y riesgo de trastorno alimentario atípico en estudiantes universitarios: realidad versus percepción.

OBJECTIVE: The objectives of this paper were to determine weight status, eating, and alcohol drinking and smoking habits of university students, to determine the association between these variables with negative self-perception of their eating habits and to assess the risk of developing eating disorders. METHOD: A cross-sectional study was carried out on 422 university students. The parameters analyzed were: nutritional status, eating habits, alcohol/ tobacco consumption, and risk of eating disorder. Logistic regression was applied to identify factors associated with a negative perception of eating habits. RESULTS: Out of the whole population that was analyzed, 5% were underweight, 16% overweight and 4% obese. Fifty-five percent of the sample analyzed did not consume five meals a day. The recommended foods for daily consumption were consumed below recommendations, while sausages/fatty meats, industrial pastries, lean meats, and fish were over-consumed. Overall, the population perceived their eating habits as good/very good (63%). Alcohol and tobacco consumption predominated at weekends. The girls were more image-conscious (80.6% vs. 66%) and fearful of gaining weight (52.5% vs. 23.9%). Almost 30% had a distorted perception of body image. There was a 12.8% risk of atypical anorexia nervosa and 4.7% of atypical bulimia nervosa. CONCLUSIONS: College students led unhealthy lifestyles, mainly due to eating habits that do not conform to the establish recommendations. More than 17% are at risk of developing an atypical eating disorder. This information may be of interest in developing preventive actions.

A rosemary extract enriched in carnosic acid improves circulating adipocytokines and modulates key metabolic sensors in lean Zucker rats: Critical and contrasting differences in the obese genotype.

SCOPE: Carnosic acid (CA) and rosemary extracts (REs) have antiobesity effects but the mechanisms are not understood. We investigated some of the potential mechanisms contributing to the metabolic effects of an RE enriched in CA. METHODS AND RESULTS: An RE (∼40% CA) was administered to lean (Le, fa/+) and obese (Ob, fa/fa) female Zucker rats for 64 days. Several adipocytokines, brain-derived neurotrophic factor, phosphorylated AMP-activated protein kinase, and hepatic gene expression changes were investigated. The RE significantly decreased circulating tumor necrosis factor alpha (RE/CT = 0.36, p < 0.0003), IL-1ß (0.48, p < 0.032), and leptin (0.48, p < 0.002), and upregulated adiponectin (1.47, p < 0.045) in the Le rats. The RE also induced phase I and phase II gene expression and the peroxisome proliferator-activated receptor gamma coactivator 1-alpha. Notably, the RE decreased adipose phosphorylated AMP-activated protein kinase and did not affect hepatic peroxisome proliferator-activated receptor gamma coactivator 1-alpha in the Ob rats. CONCLUSION: Our results show that an RE rich in CA exerts anti-inflammatory effects and affects hepatic metabolism in normal Le rats. We report significant differences in the expression and regulation of key metabolic sensors between Le and Ob rats that may contribute to explain the different ability of the two genotypes to respond to the RE.

Phase-II metabolism limits the antiproliferative activity of urolithins in human colon cancer cells.

PURPOSE: Urolithins, gut microbiota metabolites derived from ellagic acid and ellagitannins, reach micromolar concentrations in the colon lumen where can have anti-inflammatory and anticancer effects. The antiproliferative activity of urolithins (Uro-A, Uro-B, Uro-C and Uro-D) and their most relevant in vivo glucuronides were evaluated in three human colon cancer cell lines (Caco-2, SW480 and HT-29). METHODS: Cell proliferation was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide and Trypan blue exclusion assays. Cell cycle was evaluated by flow cytometry and urolithins metabolism by HPLC­MS/MS. RESULTS: Urolithins inhibited cell proliferation and cell cycle progression in a time- and dose-dependent manner and arrested the cells at S and G2/M phases, depending on the urolithin. Uro-A exerted the highest antiproliferative activity, followed by Uro-C, Uro-D and Uro-B. Unlike Caco-2 and SW480 cells, HT-29 cells partially overcame the effects after 48 h, which was related to the complete glucuronidation of urolithins. Uro-A or Uro-B glucuronides did not affect cell cycle and showed lower antiproliferative activity than their aglycone counterparts. Uro-A or Uro-B plus inhibitors of drug efflux ABC transporters partially prevented the glucuronidation of urolithins in HT-29 cells which became more sensitive. CONCLUSIONS: Uro-A, Uro-B, Uro-C and Uro-D exerted different antiproliferative effects depending on the colon cancer cell line. We also report here, for the first time, the role of ABC transporters and Phase-II metabolism in HT-29 cells as a mechanism of cancer resistance against urolithins due to their conversion to glucuronide conjugates that exerted lower antiproliferative activity.

Resveratrol in primary and secondary prevention of cardiovascular disease: a dietary and clinical perspective.

Primary prevention of cardiovascular disease (CVD) aims to avoid a first event in subjects that are at risk but have not yet been diagnosed with heart disease. Secondary prevention of CVD aims to avoid new events in patients with established heart disease. Both approaches involve clinical intervention and implementation of healthy lifestyles. The grape and wine polyphenol resveratrol (3,5,4'-trihydroxy-trans-stilbene) has shown cardioprotective benefits in humans. Most of these approaches deal with rather high doses and short follow-ups, and do not address the issue of long-term resveratrol consumption safety, especially in medicated individuals. Here, we review the trials conducted with resveratrol in patients at risk for or with established CVD, focusing on the two longest human clinical trials reported so far (1-year follow-up). We also discuss the expectations for resveratrol from a dietary and clinical perspective in relation to CVD. However, statistically significant changes in CVD-risk markers do not necessarily equal clinical significance in the daily care of patients.

Biological significance of urolithins, the gut microbial ellagic Acid-derived metabolites: the evidence so far.

The health benefits attributed to pomegranate have been associated with its high content in polyphenols, particularly ellagitannins. This is also the case for other ellagitannin-containing fruits and nuts including strawberry, raspberry, blackberry, walnuts, and muscadine grapes. The bioavailability of ellagitannins and ellagic acid is however very low. These molecules suffer extensive metabolism by the gut microbiota to produce urolithins that are much better absorbed. Urolithins circulate in plasma as glucuronide and sulfate conjugates at concentrations in the range of 0.2-20 µ M. It is therefore conceivable that the health effects of ellagitannin-containing products can be associated with these gut-produced urolithins, and thus the evaluation of the biological effects of these metabolites is essential. Recent research, mostly based on in vitro testing, has shown preliminary evidence of the anti-inflammatory, anticarcinogenic, antiglycative, antioxidant, and antimicrobial effects of urolithins, supporting their potential contribution to the health effects attributed to pomegranate and ellagitannin-rich foods. The number of in vivo studies is still limited, but they show preventive effects of urolithins on gut and systemic inflammation that encourage further research. Both in vivo and mechanistic studies are necessary to clarify the health effects of these metabolites. Attention should be paid when designing these mechanistic studies in order to use the physiologically relevant metabolites (urolithins in gut models and their conjugated derivatives in systemic models) at concentrations that can be reached in vivo.

Nutraceuticals for older people: facts, fictions and gaps in knowledge.

In the last decades nutraceuticals have entered the health market as an easy and attractive means of preventing diseases. These products are of interest for an increasingly health-concerned society and may be especially relevant for preventing or delaying a number of age-related diseases, i.e. arthritis, cancer, metabolic and cardiovascular diseases, osteoporosis, cataracts, brain disorders, etc. Nutraceuticals are marketed in a variety of forms, composition and potential applications which have made their definition ambiguous and their use uncontrolled and poorly funded. Although epidemiological, animal and in vitro studies have given evidence of the potential benefits of some of these nutraceuticals or of their components, definitive proof of their effects in appropriate human clinical trials is still lacking in most cases, more critically among people above 65 years of age. We cover the well-established nutraceuticals (polyvitamins, omega-3 fatty acids, etc.) and will focus on many other 'novel' commercial nutraceuticals where the scientific evidence is more limited (food extracts, polyphenols, carotenoids, etc.). Solid scientific evidence has been reported only for a few nutraceuticals, which have some health claims approved by the European Food Safety Authority (EFSA). Further well-designed trials are needed to improve the current knowledge on the health benefits of nutraceuticals in the elderly. Overall, there are some facts, a lot of fiction and many gaps in the knowledge of nutraceutical benefits.

Resveratrol and clinical trials: the crossroad from in vitro studies to human evidence.

Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a non-flavonoid polyphenol that may be present in a limited number of foodstuffs such as grapes and red wine. Resveratrol has been reported to exert a plethora of health benefits through many different mechanisms of action. This versatility and presence in the human diet have drawn the worldwide attention of many research groups over the past twenty years, which has resulted in a huge output of in vitro and animal (preclinical) studies. In line with this expectation, many resveratrol- based nutraceuticals are consumed all over the world with questionable clinical/scientific support. In fact, the confirmation of these benefits in humans through randomized clinical trials is still very limited. The vast majority of preclinical studies have been performed using assay conditions with a questionable extrapolation to humans, i.e. too high concentrations with potential safety concerns (adverse effects and drug interactions), short-term exposures, in vitro tests carried out with non-physiological metabolites and/or concentrations, etc. Unfortunately, all these hypothesis-generating studies have contributed to increased the number of 'potential' benefits and mechanisms of resveratrol but confirmation in humans is very limited. Therefore, there are many issues that should be addressed to avoid an apparent endless loop in resveratrol research. The so-called 'Resveratrol Paradox', i.e., low bioavailability but high bioactivity, is a conundrum not yet solved in which the final responsible actor (if any) for the exerted effects has not yet been unequivocally identified. It is becoming evident that resveratrol exerts cardioprotective benefits through the improvement of inflammatory markers, atherogenic profile, glucose metabolism and endothelial function. However, safety concerns remain unsolved regarding chronic consumption of high RES doses, specially in medicated people. This review will focus on the currently available evidence regarding resveratrol's effects on humans obtained from randomized clinical trials. In addition, we will provide a critical outlook for further research on this molecule that is evolving from a minor dietary compound to a possible multi-target therapeutic drug.

Grape resveratrol increases serum adiponectin and downregulates inflammatory genes in peripheral blood mononuclear cells: a triple-blind, placebo-controlled, one-year clinical trial in patients with stable coronary artery disease.

PURPOSE: The grape and wine polyphenol resveratrol exerts cardiovascular benefits but evidence from randomized human clinical trials is very limited. We investigated dose-depending effects of a resveratrol-containing grape supplement on stable patients with coronary artery disease (CAD) treated according to currently accepted guidelines for secondary prevention of cardiovascular disease. METHODS: In a triple-blind, randomized, placebo-controlled, one-year follow-up, 3-arm pilot clinical trial, 75 stable-CAD patients received 350 mg/day of placebo, resveratrol-containing grape extract (grape phenolics plus 8 mg resveratrol) or conventional grape extract lacking resveratrol during 6 months, and a double dose for the following 6 months. Changes in circulating inflammatory and fibrinolytic biomarkers were analyzed. Moreover, the transcriptional profiling of inflammatory genes in peripheral blood mononuclear cells (PBMCs) was explored using microarrays and functional gene expression analysis. RESULTS: After 1 year, in contrast to the placebo and conventional grape extract groups, the resveratrol-containing grape extract group showed an increase of the anti-inflammatory serum adiponectin (9.6 %, p = 0.01) and a decrease of the thrombogenic plasminogen activator inhibitor type 1 (PAI-1) (-18.6 %, p = 0.05). In addition, 6 key inflammation-related transcription factors were predicted to be significantly activated or inhibited, with 27 extracellular-space acting genes involved in inflammation, cell migration and T-cell interaction signals presenting downregulation (p < 0.05) in PBMCs. No adverse effects were detected in relation to the study products. CONCLUSIONS: Chronic daily consumption of a resveratrol-containing grape nutraceutical could exert cardiovascular benefits in stable-CAD patients treated according to current evidence-based standards, by increasing serum adiponectin, preventing PAI-1 increase and inhibiting atherothrombotic signals in PBMCs.

Alternative method for gas chromatography-mass spectrometry analysis of short-chain fatty acids in faecal samples.

Short-chain fatty acids are the major end products of bacterial metabolism in the large bowel. They derive mostly from the bacterial breakdown of carbohydrates and are known to have positive health benefits. Due to the biological relevance of these compounds it is important to develop efficient, cheap, fast, and sensitive analytical methods that enable the identification and quantification of the short-chain fatty acids in a large number of biological samples. In this study, a gas chromatography-mass spectrometry method was developed and validated for the analysis of short-chain fatty acids in faecal samples. These volatile compounds were extracted with ethyl acetate and 4-methyl valeric acid was used as an internal standard. No further cleanup, concentration, and derivatization steps were needed and the extract was directly injected onto the column. Recoveries ranged between 65 and 105%, and no matrix effects were observed. The proposed method has wide linear ranges, good inter- and intraday variability values (below 2.6 and 5.6%, respectively) and limits of detection between 0.49 µM (0.29 µg/g) and 4.31 µM (3.8 µg/g). The applicability of this analytical method was successfully tested in faecal samples from rats and humans.

Consumption of a grape extract supplement containing resveratrol decreases oxidized LDL and ApoB in patients undergoing primary prevention of cardiovascular disease: a triple-blind, 6-month follow-up, placebo-controlled, randomized trial.

SCOPE: The cardioprotective role of resveratrol as part of the human diet is not yet clear. Our aim was to investigate the effect of a grape supplement containing 8 mg resveratrol in oxidized LDL (LDLox), apolipoprotein-B (ApoB), and serum lipids on statin-treated patients in primary cardiovascular disease prevention (PCP). METHODS AND RESULTS: A triple-blind, randomized, placebo-controlled trial was conducted. Seventy-five patients (three parallel arms) consumed one capsule (350 mg) daily for 6 months containing resveratrol-enriched grape extract (GE-RES, Stilvid®), grape extract (GE, similar polyphenolic content but no resveratrol), or placebo (maltodextrin). After 6 months, no changes were observed in the placebo group and only LDL cholesterol (LDLc) decreased by 2.9% (p = 0.013) in the GE group. In contrast, LDLc (-4.5%, p = 0.04), ApoB (-9.8%, p = 0.014), LDLox (-20%, p = 0.001), and LDLox/ApoB (-12.5%, p = 0.000) decreased in the Stilvid® group, whereas the ratio non-HDLc (total atherogenic cholesterol load)/ApoB increased (8.5%, p = 0.046). No changes were observed in hepatic, thyroid, and renal function. No adverse effects were observed in any of the patients. CONCLUSION: This GE-RES reduced atherogenic markers and might exert additional cardioprotection beyond the gold-standard medication in patients from PCP. The presence of resveratrol in the GE was necessary to achieve these effects.

Ellagitannin metabolites, urolithin A glucuronide and its aglycone urolithin A, ameliorate TNF-α-induced inflammation and associated molecular markers in human aortic endothelial cells.

SCOPE: Numerous in vitro and in vivo studies indicate that ellagitannins exhibit anti-inflammatory, anti-atherosclerotic and anti-angiogenic activity which support their potential preventive effect against cardiovascular diseases. Ellagitannins exhibit low bioavailability and are transformed in the gut to ellagic acid and its microbiota metabolites urolithin A (Uro-A) and urolithin B (Uro-B). Urolithins are found in plasma mostly as glucuronides at low µM concentrations. We investigated whether urolithin glucuronides and their aglycones exhibit vascular protective effects. METHODS AND RESULTS: Human aortic endothelial cells were exposed to tumor necrosis factor alpha and to Uro-A glucuronide, Uro-B glucuronide or their corresponding aglycones at low µM concentrations to determine their effects on monocytes adhesion and endothelial cell migration. The levels of related adhesion cytokines and growth molecular markers were also measured. Uro-A glucuronide (∼5-15 µM) inhibited monocyte adhesion and endothelial cell migration in a significant manner. These effects were associated with a moderate but significant down-regulation of the levels of chemokine (C-C motif) ligand 2 (CCL2) and plasminogen activator inhibitor-1 (PAI-1). Uro-A inhibited endothelial cell migration and was able to decrease the expression of CCL2 and interleukin-8 (IL-8). CONCLUSION: Our results suggest that these metabolites might be involved, at least in part, in the beneficial effects against cardiovascular diseases attributed to the consumption of ellagitannin-containing foods.