Human hydroxytyrosol's absorption and excretion from a nutraceutical

Among the various (poly)phenols that are being sold as such or as part of a more complex mixture, hydroxytyrosol (HT) is the only one that bears a European Food Safety Authority health claim. Therefore, several HT-based products are being developed and sold and it becomes necessary to evaluate its accessibility following ingestion. Twenty-one volunteers were recruited for a randomized, crossover, placebo-controlled, and double-blind intervention study.We performed a Latin square design: after one-week washout, i.e. olive-free diet, subjects were randomly assigned to the placebo (maltodextrin), 5, or 25 mg/day HT group. Twenty-four hour urine samples were collected after the intervention week, and baseline urines were collected the week before the study and during periods of washout. The results show that HT given as the foremost component of a nutraceutical preparation is bioavailable and is recovered in the urine chiefly as sulphate-3′

Metabolic fingerprint after acute and under sustained consumption of a functional beverage based on grape skin extract in healthy human subjects

Grape-derived polyphenols are considered to be one of the most promising ingredients for functional foods due to their health-promoting activities. We applied a HPLC-MS-based untargeted metabolomic approach in order to evaluate the impact of a functional food based on grape skin polyphenols on the urinary metabolome of healthy subjects. Thirty-one volunteers participated in two dietary crossover randomized intervention studies: with a single-dose intake (187 mL) and with a 15-day sustained consumption (twice per day, 187 mL per day in total) of a functional beverage (FB). Postprandial (4-hour) and 24-hour urine samples collected after acute consumption and on the last day of sustained FB consumption, respectively, were analysed using an untargeted HPLC-qTOF-MS approach. Multivariate modelling with subsequent application of an S-plot revealed differential mass features related to acute and prolonged consumption of FB. More than half of the mass features were shared between the two types of samples, among which several phase II metabolites of grape-derived polyphenols were identified at confidence level II. Prolonged consumption of FB was specifically reflected in urine metabolome by the presence of first-stage microbial metabolites of flavanols: hydroxyvaleric acid and hydroxyvalerolactone derivatives. Overall, several epicatechin and phenolic acid metabolites both of tissular and microbiota origin were the most representative markers of FB consumption. To our knowledge, this is one of the first studies where an untargeted LC-MS metabolomic approach has been applied in nutrition research on a grape-derived FB

Cocoa polyphenols and inflammatory markers of cardiovascular disease

Epidemiological studies have demonstrated the beneficial effect of plant-derived food intake in reducing the risk of cardiovascular disease (CVD). The potential bioactivity of cocoa and its polyphenolic components in modulating cardiovascular health is now being studied worldwide and continues to grow at a rapid pace. In fact, the high polyphenol content of cocoa is of particular interest from the nutritional and pharmacological viewpoints. Cocoa polyphenols are shown to possess a range of cardiovascular-protective properties, and can play a meaningful role through modulating different inflammatory markers involved in atherosclerosis. Accumulated evidence on related anti-inflammatory effects of cocoa polyphenols is summarized in the present review

Alternate steroid sulfation pathways targeted by LC-MS/MS 1 analysis of disulfates. Application to prenatal diagnosis of steroid synthesis disorders

The steroid disulfates (aka bis-sulfates) are a significant but minor fraction of the urinary steroid metabolome that have not been widely studied because major components are not hydrolyzed by the commercial sulfatases commonly used in steroid metabolomics. In early studies, conjugate fractionation followed by hydrolysis using acidified solvent (solvolysis) was used for the indirect detection of this fraction by GC–MS. This paper describes the application of a specific LC–MS/MS method for the direct identification of disulfates in urine, and their use as markers for the prenatal diagnosis of disorders causing reduced estriol production: STSD (steroid sulfatase deficiency), SLOS (Smith-Lemli-Opitz syndrome) and PORD (P450 oxidoreductase deficiency). Disulfates were detected by monitoring a constant ion loss (CIL) from the molecular di-anion. While focused on disulfates, our methodology included an analysis of intact steroid glucuronides and monosulfates because steroidogenic disorder diagnosis usually requires an examination of the complete steroid profile. In the disorders studied, a few individual steroids (as disulfates) were found particularly informative: pregn-5-ene-3β,20S-diol, pregn-5-ene-3β,21-diol (STSD, neonatal PORD) and 5α-pregnane-3β,20S-diol (pregnancy PORD). Authentic steroid disulfates were synthesized for use in this study as aid to characterization. Tentative identification of 5ξ-pregn-7-ene-3ξ,20S-diol and 5ξ-pregn-7-ene-3ξ,17,20S-triol disulfates was also obtained in samples from SLOS affected pregnancies. Seven ratios between the detected metabolites were applied to distinguish the three selected disorders from control samples. Our results show the potential of the direct detection of steroid conjugates in the diagnosis of pathologies related with steroid biosynthesis.Spanish Health National System is acknowledged for OP contract (CPII16/00027). Strategic Plan for Research and Innovation in Health (PERIS) of the Catalan government is acknowledge for OK contract (SLT002/16/00007)

Dietary Epicatechin Is Available to Breastfed Infants through Human Breast Milk in the Form of Host and Microbial Metabolites

Polyphenols play an important role in human health. To address their accessibility to a breastfed infant, we planned to evaluate whether breast milk (BM) (colostrum, transitional, and mature) epicatechin metabolites could be related to the dietary habits of mothers. The polyphenol consumption of breastfeeding mothers was estimated using a food frequency questionnaire and 24 h recalls. Solid-phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS) was applied for direct epicatechin metabolite analysis. Their bioavailability in BM as a result of dietary ingestion was confirmed in a preliminary experiment with a single dose of dark chocolate. Several host and microbial phase II metabolites of epicatechin were detected in BM among free-living lactating mothers. Interestingly, a modest correlation between dihydroxyvalerolactone sulfate and the intake of cocoa products was observed. Although a very low percentage of dietary polyphenols is excreted in BM, they are definitely in the diet of breastfed infants. Therefore, evaluation of their role in infant health could be further promoted

An NMR-based metabolomics approach reveals a combined-biomarkers model in a wine interventional trial with validation in free-living individuals of the PREDIMED study

The development of robust biomarkers of consumption would improve the classification of participants with regard to their dietary exposure. In addition, validation of them in free-living individuals remains an important challenge. The aim of this study is to assess wine intake biomarkers using an NMR metabolomic approach to measure the utility of these biomarkers in a wine interventional study (WIS, n = 56) and also to evaluate them in a free-living individuals (PREDIMED study, n = 91). Nine metabolites showed a significantly higher presence in urinary excretion in WIS after wine intake: five food metabolome metabolites (tartrate, ethyl glucuronide [EtG], 2,3-butanediol, mannitol, and ethanol); one related to the endogenous response to wine exposure (3-methyl-2-oxovalerate) and three unidentified compounds. Receiver operating characteristic (ROC) curve for each single metabolite were evaluated and exhibited areas under the curves (AUC) between 67.4 and 86.3 % when they were evaluated individually. Then, a logistic regression model was fitted to generate a combined-biomarkers model using these metabolites. The model generated which included tartrate-EtG, showed an AUC of 90.7 % in WIS. Similarly, the AUC in the PREDIMED study was 92.4 %. Results showed that a model combining tartrate-EtG is more useful for evaluating exposure to wine than single biomarkers, both in interventional studies and epidemiological data. To our knowledge, this is the first time that a combined-biomarker model using an NMR platform in wine biomarkers' research has been generated and reproduced in a free-living population

The Influence of Genetic and Environmental Factors among MDMA Users in Cognitive Performance

This study is aimed to clarify the association between MDMA cumulative use and cognitive dysfunction, and the potential role of candidate genetic polymorphisms in explaining individual differences in the cognitive effects of MDMA. Gene polymorphisms related to reduced serotonin function, poor competency of executive control and memory consolidation systems, and high enzymatic activity linked to bioactivation of MDMA to neurotoxic metabolites may contribute to explain variations in the cognitive impact of MDMA across regular users of this drug. Sixty ecstasy polydrug users, 110 cannabis users and 93 non-drug users were assessed using cognitive measures of Verbal Memory (California Verbal Learning Test, CVLT), Visual Memory (Rey-Osterrieth Complex Figure Test, ROCFT), Semantic Fluency, and Perceptual Attention (Symbol Digit Modalities Test, SDMT). Participants were also genotyped for polymorphisms within the 5HTT, 5HTR2A, COMT, CYP2D6, BDNF, and GRIN2B genes using polymerase chain reaction and TaqMan polymerase assays. Lifetime cumulative MDMA use was significantly associated with poorer performance on visuospatial memory and perceptual attention. Heavy MDMA users (>100 tablets lifetime use) interacted with candidate gene polymorphisms in explaining individual differences in cognitive performance between MDMA users and controls. MDMA users carrying COMT val/val and SERT s/s had poorer performance than paired controls on visuospatial attention and memory, and MDMA users with CYP2D6 ultra-rapid metabolizers performed worse than controls on semantic fluency. Both MDMA lifetime use and gene-related individual differences influence cognitive dysfunction in ecstasy users

Metabolic Signatures Associated with Severity in Hospitalized COVID-19 Patients

The clinical evolution of COVID-19 pneumonia is poorly understood. Identifying the metabolic pathways that are altered early with viral infection and their association with disease severity is crucial to understand COVID-19 pathophysiology, and guide clinical decisions. This study aimed at assessing the critical metabolic pathways altered with disease severity in hospitalized COVID-19 patients. Forty-nine hospitalized patients with COVID-19 pneumonia were enrolled in a prospective, observational, single-center study in Barcelona, Spain. Demographic, clinical, and analytical data at admission were registered. Plasma samples were collected within the first 48 h following hospitalization. Patients were stratified based on the severity of their evolution as moderate (N = 13), severe (N = 10), or critical (N = 26). A panel of 221 biomarkers was measured by targeted metabolomics in order to evaluate metabolic changes associated with subsequent disease severity. Our results show that obesity, respiratory rate, blood pressure, and oxygen saturation, as well as some analytical parameters and radiological findings, were all associated with disease severity. Additionally, ceramide metabolism, tryptophan degradation, and reductions in several metabolic reactions involving nicotinamide adenine nucleotide (NAD) at inclusion were significantly associated with respiratory severity and correlated with inflammation. In summary, assessment of the metabolomic profile of COVID-19 patients could assist in disease severity stratification and even in guiding clinical decisions

Contribution of Cytochrome P450 and ABCB1 Genetic Variability on Methadone Pharmacokinetics, Dose Requirements, and Response

Although the efficacy of methadone maintenance treatment (MMT) in opioid dependence disorder has been well established, the influence of methadone pharmacokinetics in dose requirement and clinical outcome remains controversial. The aim of this study is to analyze methadone dosage in responder and nonresponder patients considering pharmacogenetic and pharmacokinetic factors that may contribute to dosage adequacy. Opioid dependence patients (meeting Diagnostic and Statistical Manual of Mental Disorders, [4th Edition] criteria) from a MMT community program were recruited. Patients were clinically assessed and blood samples were obtained to determine plasma concentrations of (R,S)-, (R) and (S)- methadone and to study allelic variants of genes encoding CYP3A5, CYP2D6, CYP2B6, CYP2C9, CYP2C19, and P-glycoprotein. Responders and nonresponders were defined by illicit opioid consumption detected in random urinalysis. The final sample consisted in 105 opioid dependent patients of Caucasian origin. Responder patients received higher doses of methadone and have been included into treatment for a longer period. No differences were found in terms of genotype frequencies between groups. Only CYP2D6 metabolizing phenotype differences were found in outcome status, methadone dose requirements, and plasma concentrations, being higher in the ultrarapid metabolizers. No other differences were found between phenotype and responder status, methadone dose requirements, neither in methadone plasma concentrations. Pharmacokinetic factors could explain some but not all differences in MMT outcome and methadone dose requirements

Health benefits of olive oil: contribution of phenolic compunds and transcriptomic response in humans

The evaluation of olive oil antioxidants, hydroxytyrosol and tyrosol, in vivo biological activities is challenged due to scarce data on their metabolic disposition and activities of their glucuronides, main metabolites found in humans in different biological matrices after olive oil consumption. In addition, the in vivo gene expression activity of virgin olive oil (VOO) as a dietary component has been never investigated in humans.Therefore, this Thesis was focused on three main aspects: (i) analysis of bioavailability of hydroxytyrosol and tyrosol glucuronides in humans; (ii) evaluation of the impact of glucuronidation on antioxidant activities of olive oil phenolics; and (iii) identification mechanisms underlying beneficial action of VOO analyzing induced in vivo transcriptome response in humans. To complete with the objectives, the glucuroconjugated standards, required for bioavailability and antioxidant activities studies were synthesized, and the preparative methodological studies for VOO-transcriptomic experiment were carried out. As a result of experimental work performed within this Dissertation, the glucuronidation was shown to account for 75% of recovered in urine olive oil phenols, and to have negative impact on their antioxidants activities, diminishing their free radical and inhibitory against LDL oxidation activities. The transcriptome studies revealed 10 genes as potential targets of VOO action against atherosclerosis.La avaluació in vivo de les activitats biològiques dels polifenols del oli d'oliva (OVV) hidroxitirosol i tirosol es un repte degut a les dades molt limitades que tenim de la seva depuració metabòlica i de les activitats biològiques dels seus principals metabòlits en matrius biològiques: els seus glucuronoconjugats. A més a més s'ha avaluat l'expressió gènica induïda en humans per la ingesta de OOV. Així la present tesi doctoral s'ha focalitzat en els següents aspectes: l'avaluació de la biodisponibilitat del hidroxitirosol i tirosol en humans; l'impacte de la glucuronoconjugació sobre les activitats antioxidants dels polifenols del OOV; i la identificació dels mecanismes subjacents a las accions benèfiques per la salut humana, analitzant la resposta transcriptòmica in vivo resultant de la ingesta OOV. Per complir amb els objectius de la tesi, ha estat necessari, sintetitzar patrons dels glucurònids i realitzar diversos estudis metodològics per tal d'estandarditzar l'avaluació de l'expressió gènica. S'ha demostrat que la glucuronoconjugació es un 75% dels polifenols recuperats en orina i que aquesta comporta la pèrdua de la capacitat bescanviadora de radicals i de la seva capacitat antioxidant (test ex-vivo d'oxidació de la LDL i DPPH). Els estudis transcriptòmics han detectat 10 gens rellevants pels efectes antiateroscleròtics induïts per OVV