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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.

Doubly or bisconjugated steroid metabolites have long been known as minor components of the steroid profile that have traditionally been studied by laborious and indirect fractionation, hydrolysis and gas chromatography-mass spectrometry (GC-MS) analysis. Recently, the synthesis and characterisation of steroid bis(sulfate) (aka disulfate or bis-sulfate) reference materials enabled the liquid chromatography-tandem mass spectrometry (LC-MS/MS) study of this metabolite class and the development of a constant ion loss (CIL) scan method for the direct and untargeted detection of steroid bis(sulfate) metabolites. Methods for the direct LC-MS/MS detection of other bisconjugated steroids, such as steroid bisglucuronide and mixed steroid sulfate glucuronide metabolites, have great potential to reveal a more complete picture of the steroid profile. However, access to steroid bisglucuronide or sulfate glucuronide reference materials necessary for LC-MS/MS method development, metabolite identification or quantification is severely limited. In this work, ten steroid bisglucuronide and ten steroid sulfate glucuronide reference materials were synthesised through an ordered combination of chemical sulfation and/or enzymatic glucuronylation reactions. All compounds were purified and characterised using NMR and MS methods. Chemistry for the preparation of stable isotope labelled steroid {13C6}-glucuronide internal standards has also been developed and applied to the preparation of two selectively mono-labelled steroid bisglucuronide reference materials used to characterise more completely MS fragmentation pathways. The electrospray ionisation and fragmentation of the bisconjugated steroid reference materials has been studied. Preliminary targeted ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis of the reference materials prepared revealed the presence of three steroid sulfate glucuronides as endogenous human urinary metabolites.

The steroid disulfates (akabis-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.

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.

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.

Hydroxytyrosol and tyrosol are dietary phenolic compounds present in virgin olive oil and wine. Both compounds are also endogenously synthesized in our body as byproducts of dopamine and tyramine metabolisms, respectively. Over the last decades, research into hydroxytyrosol and tyrosol has experienced an increasing interest due to the role that these compounds may play in the prevention of certain pathologies (e.g. cardiovascular, metabolic, neurodegenerative diseases and cancer). The translation of promising in vitro and in vivo biological effects from preclinical studies to the context of human disease prevention initially depends on whether the dose ingested becomes available at the site of action. In this regard, information regarding the bioavailability and metabolic disposition of hydroxytyrosol and tyrosol is of most importance to evaluate the impact they may have on human health. In this review, we discuss and summarize the state of the art of the scientific evidence regarding the processes of absorption, distribution, metabolism and excretion of both hydroxytyrosol and tyrosol. We also examine the impact of these compounds and their metabolites on biological activity in terms of beneficial health effects. Finally, we evaluate the different analytical approaches that have been developed to measure the plasma and urinary levels of hydroxytyrosol, tyrosol and their metabolites. This work was supported by grants from Instituto de Salud Carlos III FEDER, (PI14/00072) and from DIUE de la Generalitat de Catalunya (2014SGR 680). JRM was supported by a FI-DGR2012 predoctoral fellowship from the Generalitat de Catalunya and CPM was supported by a Juan Rodes fellowship (ISCIII, JR, 15/00005). CIBER de Fisiopatologıa de la Obesidad y Nutricion (CIBEROBN) is an initiative of the Instituto de Salud Carlos III, Madrid, Spain.

Background Recently, the European Food Safety Authority approved a claim concerning the benefits of olive oil polyphenols for the protection of LDL from oxidation. Polyphenols could exert health benefits not only by scavenging free radicals but also by modulating gene expression. Objective We assessed whether olive oil polyphenols could modulate the human in vivo expressions of atherosclerosis-related genes in which LDL oxidation is involved. Design In a randomized, crossover, controlled trial, 18 healthy European volunteers daily received 25 mL olive oil with a low polyphenol content (LPC: 2.7 mg/kg) or a high polyphenol content (HPC: 366 mg/kg) in intervention periods of 3 wk separated by 2-wk washout periods. Results Systemic LDL oxidation and monocyte chemoattractant protein 1 and the expression of proatherogenic genes in peripheral blood mononuclear cells [ie, CD40 ligand (CD40L), IL-23α subunit p19 (IL23A), adrenergic β-2 receptor (ADRB2), oxidized LDL (lectin-like) receptor 1 (OLR1), and IL-8 receptor-α (IL8RA)] decreased after the HPC intervention compared with after the LPC intervention. Random-effects linear regression analyses showed 1) a significant decrease in CD40, ADRB2, and IL8RA gene expression with the decrease of LDL oxidation and 2) a significant decrease in intercellular adhesion molecule 1 and OLR1 gene expression with increasing concentrations of tyrosol and hydroxytyrosol in urine. Conclusions In addition to reducing LDL oxidation, the intake of polyphenol-rich olive oil reduces CD40L gene expression, its downstream products, and related genes involved in atherogenic and inflammatory processes in vivo in humans. These findings provide evidence that polyphenol-rich olive oil can act through molecular mechanisms to provide cardiovascular health benefits. This trial was registered at www.controlled-trials.com as ISRCTN09220811.

In the present study we report on a UPLC-MRM validated method for the simultaneous direct analysis of main glucuronidated metabolites of olive oil phenols: tyrosol, hydroxytyrosol and its O-methyl metabolite homovanillyl alcohol in human urine after dietary olive oil ingestion. The developed method was linear within the concentration range 20–2000 ng/mL with adequate recovery of analytes (>86%). Intra- and inter-day precision and accuracy were according to standard requirements for method validation criteria. Using the developed method, urinary concentrations and excretion rates of glucuronides of olive oil phenols were successfully estimated in an intervention study with 11 healthy volunteers supplemented with a dietary dose of virgin olive oil (VOO) (50 mL). Therefore, about 13% of the consumed olive oil polyphenols were recovered in 24-h urine, where 75% of them were in the form of glucuronides (3′- and 4′-O-hydroxytyrosol glucuronides, 4′-O-glucuronides of tyrosol and homovanillyl alcohol) and 25% as free compounds., We would like to thank the volunteers who participated in this study and Esther Menoyo, R.N., for her valuable assistance in dealing with them. The authors thank the Fundación Patrimonio Comunal Olivarero and Hojiblanca SA, for donating the olive oil. This work was supported by CIBER de Fisiopatología de la Obesidad y Nutrición (CB06/03) an initiative of the Carlos III Institute, the Fondo de Investigaciones Sanitarias (FIS/ISCIII PI081913, FEDER-ERDF) and AGAUR 2009 SGR 718, the MICIN-AGL2009-13517-C03-01.

Hydroxytyrosol (HOTYR) and tyrosol (TYR), main phenolic compounds of olive oil, have been reported to contribute to the prevention of cardiovascular diseases due to their antioxidant activities, e.g., protection of low-density lipoprotein (LDL) oxidation. Their bioavailability in humans is poor, and they are found in biological fluids mainly as conjugated metabolites. Low concentrations of free phenols are unlikely to explain the biological activities seen in humans after olive oil intake. In this context, antioxidant activities of conjugated metabolites, in a range of concentrations compatible with their dietary consumption, were evaluated. Concentrations of metabolites and their core compounds were estimated in an intervention study of 11 healthy volunteers whose diet was supplemented with 50 ml of virgin olive oil, using high-performance liquid chromatography coupled to mass spectrometry for the simultaneous analysis of 3'-O- and 4'-O-HOTYR-glucuronides and 4'-O-glucuronides of TYR and homovanillyl alcohol in human urine. Glucuronides and core compounds were tested for their chemical (hydrogen donation by 1,1-diphenyl-2-picrylhydrazyl free radical test) and in vitro biological (inhibition of Cu-mediated LDL oxidation) antioxidant activities at the concentration ranges observed in human biological fluids (range, 0.01-10 muM) after dietary olive oil consumption. None of the glucuronides displayed significant antioxidant activities at the concentrations tested.

Virgin olive oil (VOO) is considered to be one of the main components responsible for the health benefits of the Mediterranean diet, particularly against atherosclerosis where peripheral blood mononuclear cells (PBMNCs) play a crucial role in atherosclerosis development and progression. The objective of this article was to identify the PBMNC genes that respond to VOO consumption in order to ascertain the molecular mechanisms underlying the beneficial action of VOO in the prevention of atherosclerosis. Gene expression profiles of PBMNCs from healthy individuals were examined in pooled RNA samples by microarrays after 3 weeks of moderate and regular consumption of VOO, as the main fat source in a diet controlled for antioxidant content. Gene expression was verified by qPCR. The response to VOO consumption was confirmed for individual samples (n = 10) by qPCR for 10 upregulated genes (ADAM17, ALDH1A1, BIRC1, ERCC5, LIAS, OGT, PPARBP, TNFSF10, USP48, and XRCC5). Their putative role in the molecular mechanisms involved in atherosclerosis development and progression is discussed, focusing on a possible relation with VOO consumption. Our data support the hypothesis that 3 weeks of nutritional intervention with VOO supplementation, at doses common in the Mediterranean diet, can alter the expression of genes related to atherosclerosis development and progression. The CNS Contract CP06/00100 (Montserrat Fitó) is acknowledged. This work has been supported by CICYT (SAF 2004-08173-C03-00) and partially supported by the Generalitat of Catalunya (2005 SGR 0057).

Metabolomics is one of the emerging “-omics” technologies that has great advantages for studying metabolic interaction between diet and human health. Over the last decade it has been successfully used in various wine nutritional studies, applying both hypothesis-driven and exploratory strategies; as a result, various aspects of the wine-human health interaction have been addressed. The exploration-driven findings provided original information on the complex metabolic relationship between wine and the human organism, reflected in changes in homeostatic metabolism and/or in immediate metabolic events, as well as in metabolic changes related to the efficiency and activity of gut microbiota. Targeted along with untargeted profiling studies provided new aspects on the bioavailability of wine-derived compounds and suggested the application of certain metabolic patterns as markers of wine dietary exposure. Finally, the outcomes of these studies showed the advantage and great potential of the application of a metabolomic approach in wine nutrition research with regard to understanding and revealing novel mechanisms underlying the health benefits of wine consumption and in searching for targets of wine benefits in human health.

Hydroxytyrosol (HT) from olive oil, a potent bioactive molecule with health benefits, has a poor bioavailability, its free form (free HT) being undetectable so far. This fact leads to the controversy whether attained HT concentrations after olive oil polyphenol ingestion are too low to explain the observed biological activities. Due to this, an analytical methodology to determine free HT in plasma is crucial for understanding HT biological activity. Plasma HT instability and low concentrations have been major limitations for its quantification in clinical studies. Here, we describe a method to detect and quantify free HT in human plasma by using liquid chromatography coupled to tandem mass spectrometry. The method encompasses different steps of sample preparation including plasma stabilization, protein precipitation, selective derivatization with benzylamine, and purification by solid-phase extraction. A high sensitivity (LOD, 0.3ng/mL), specificity and stability of HT is achieved following these procedures. The method was validated and its applicability was demonstrated by analyzing human plasma samples after olive oil intake. A pharmacokinetic comparison was performed measuring free HT plasma concentrations following the intake of 25mL of ordinary olive oil (nearly undetectable concentrations) versus an extra-virgin olive oil (Cmax=4.40ng/mL). To our knowledge, this is the first time that an analytical procedure for quantifying free HT in plasma after olive oil dietary doses has been reported. The present methodology opens the door to a better understanding of the relationship between HT plasma concentrations and its beneficial health effects. This work was supported by grants from Instituto de Salud Carlos III FEDER, (PI14/00072), the CICYT-FEDER AGL2009-13517- C03-01 and AGL2012-40144-C03-01), grants from DIUE of the Generalitat de Catalunya (2014 SGR 680). CIBER de Fisiopatología de la Obesidad y Nutrición (CIBEROBN) is an initiative of the Instituto de Salud Carlos III, Madrid, Spain. J.R-M. was supported by a FI-DGR2012 predoctoral fellowship from the Generalitat de Catalunya. C.P-M. was supported by a Juan Rodés contract (JR15/00005).

Diet plays an important role in the prevention and development of chronic diseases. Interventional and observational studies have provided evidence of the health benefits of dietary bioactive compo...

The biocatalytic synthesis and purification of O-β-D-monoglucuronide conjugates of hydroxytyrosol, tyrosol, homovanillic alcohol, and 3-(4′-hydroxyphenyl)propanol, using porcine liver microsomes, are described here. The glucuronides were synthesized, analyzed and separated by HPLC-UV, identified by HPLC-MS, and their structures unequivocally established by NMR techniques. The outcome of the glucuronidation reaction depends on the structure of the phenolic compounds. Thus, the glucuronidation of hydroxytyrosol, biocatalyzed with porcine liver microsomes, proceeded exclusively on the phenolic hydroxy groups. The regioselectivity was similar to that observed for human and rat liver microsomes, the 4′-hydroxy position being more favorable than the 3′-hydroxy one. In the case of tyrosol, homovanillic alcohol, and hydroxyphenylpropanol, two products were formed during microsomal glucuronidation: a major one, the phenolic O-β-D-glucuronidated derivative and, a minor one, the O-β-D-glucuronidated aliphatic alcohol.

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., This work has been supported by Spanish public grants AGL2006-14228-C03-02 and AGL2009-13906-C02-01 and the CONSOLIDER-INGENIO 2010 Programme, FUN-C-FOOD (CSD2007-063) from the Spanish Ministry of Economy and Competitiveness (MINECO), the PI13/01172 Project (Plan N de I+D+i 2013–2016) by ISCII-Subdirección General de Evaluación y Fomento de la Investigación as co-funded by FEDER (Fondo Europeo de Desarrollo Regional); the National Strategic Consortia for Technical Research Programme, in collaboration with the Bosch i Gimpera Foundation, FBG305132 and FBG305273; and Miguel Torres S.A. (Proyecto CENIT HIGEA CEN-20072003). We also thank the award of 2014SGR1566 from the Generalitat de Catalunya’s Agency AGAUR. The Juan de la Cierva postdoctoral fellowship contract from the Spanish government awarded by the Spanish Ministry to Olha Khymenets is acknowledged. Rosa Vazquez-Fresno, Rafael Llorach and Mireia Urpi-Sarda would like to thank, respectively, the FPI 2010 fellowship and the Ramon y Cajal programmes from MICINN and the European Social Funds.

This is an open access article distributed under the Creative Commons Attribution License.-- This article belongs to the Special Issue Chocolate and Cocoa in Human Health., 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. © 2014 by the authors; licensee MDPI, Basel, Switzerland., The authors are grateful for the support granted by the following Spanish government programmes: CICYT AGL2009-13906-C02-01, AGL2010-10084-E and FUN-C Food CSD2007-063 from the Ingenio-CONSOLIDER programme of the Spanish Government. The postdoctoral contract fellowship, awarded by the Ministry of Innovation and Science to Olha Khymenets and Sara Tulipani (Juan de la Cierva Program), and the predoctoral fellowships awarded by the Generalitat de Catalunya (FI-DRG) to Ximena Mora-Cubillos and Mar Garcia-Aloy are acknowledged. Mireia Urpí-Sardà and Rafael Llorach would like to thank Ramon y Cajal Program of the Spanish Ministry (MINECO) and the Fondo Social Europeo., We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).

Over the last decade, a growing body of interventional and epidemiological studies has furnished new evidence on the beneficial properties of dietary polyphenols in the reduction of the risk of cardiovascular disease (CVD). However, further research is required on the discovery and validation of new polyphenol intake biomarkers and their effect on the promotion of cardiovascular health. To successfully face the challenge of biomarker discovery, the application of emerging “omics” technologies and hypothesis-generating approaches are very encouraging. Metabolomics is a particularly promising strategy for obtaining a comprehensive overview of the low-molecular-weight metabolites present in a biological system at a given moment, and to explore unknown metabolic pathways impacted on diet. Despite being the youngest of the “omics” technologies, it already counts for a series of successful applications in the identification of potential biomarkers related to the prediction, early diagnosis and prognosis of CVD. This chapter will review the state of the art in the application of the metabolomics approach to polyphenol and CVD biomarker discovery, and their implications in cardiovascular risk assessment.

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., This study was supported by grants from the National Institutes of Health (NIH) grant no. 1 R01 DA017987, Grant 2005SGR00032, Fondo de Investigaciones Sanitarias (FIS-00/00777), Plan Nacional Sobre Drogas (INT/2012/2002) Spain, Plan Nacional Sobre Drogas: PNSD 2006/101(2007–2009) Spain.

12 páginas, 3 figuras, 5 tablas.-- et al., The aim of the study was to assess whether benefits associated with the traditional Mediterranean diet (TMD) and virgin olive oil (VOO) consumption could be mediated through changes in the expression of atherosclerosis-related genes. A randomized, parallel, controlled clinical trial in healthy volunteers (n=90) aged 20 to 50 yr was performed. Three-month intervention groups were as follows: 1) TMD with VOO (TMD+VOO), 2) TMD with washed virgin olive oil (TMD+WOO), and 3) control with participants’ habitual diet. WOO was similar to VOO, but with a lower polyphenol content (55 vs. 328 mg/kg, respectively). TMD consumption decreased plasma oxidative and inflammatory status and the gene expression related with both inflammation [INF-γ (INFγ), Rho GTPase-activating protein15 (ARHGAP15), and interleukin-7 receptor (IL7R)] and oxidative stress [adrenergic β2-receptor (ADRB2) and polymerase (DNA-directed) κ (POLK)] in peripheral blood mononuclear cells. All effects, with the exception of the decrease in POLK expression, were particularly observed when VOO, rich in polyphenols, was present in the TMD dietary pattern. Our results indicate a significant role of olive oil polyphenols in the down-regulation of proatherogenic genes in the context of a TMD. In addition, the benefits associated with a TMD and olive oil polyphenol consumption on cardiovascular risk can be mediated through nutrigenomic effects.—Konstantinidou, V., Covas, M.-I., Muñοz-Aguayo, D., Khymenets, O., de la Torre, R., Saez, G., del Carmen Tormos, M., Toledo, E., Marti, A., Ruiz-Gutiérrez, V., Ruiz Mendez, M. V., Fito, M. In vivo nutrigenomic effects of virgin olive oil polyphenols within the frame of the Mediterranean diet: a randomized controlled trial., This work was supported by Fóndo de Investigación Sanitaria–Fóndo Europeo de Desarrollo Regional (FIS-FEDER; PI041308), by Sistema National de Salud (SNS) contract Miguel Servet (CP06/ 00100) Instituto de Salud Carlos III, and by the Greek State Scholarship Foundation (Athens, Greece), and partially supported by the Generalitat of Catalunya (2005 SGR 00577).

Aims: Differences in adverse drug reactions can be explained by genetic variations, especially if they determine the expression of certain protein effectors and/or drug-metabolizing enzymes. Over the last decade, several tests screening for the most frequent polymorphisms in drug-metabolizing enzymes have been marketed for research and diagnostic purposes. The aim of this study was to assess the suitability of PHARMAchip™ for the genotyping of polymorphisms of genes associated with drug metabolism and response as an alternative to Jurilab Ltd’s DrugMEt® Test. Materials & methods: In this observational study, performed using 100 previously genotyped DNA samples, we report on common genes included in the two different tests examined: the former DrugMEt test and the recently introduced PHARMAchip test. Results & conclusion: Although these tests are based on different methodological approaches, we have found a high concordance of results between both methods. Some of the discrepancies between tests were related to allelic variants not monitored in a particular microarray and the quality of the genomic DNA used.

Publisher Summary The beneficial effects of olive oil on cardiovascular risk factors are now recognized and often only attributed to its high levels of monounsaturated fatty acids (MUFA). Olive oil is, however, more than a MUFA fat. Olive oil is a functional food, which besides having a high level of MUFA contains other minor components with biological properties. The content of the minor components of an olive oil varies, depending on several conditions such as the cultivar, climate, ripeness of the olives at harvesting, and the processing system employed to produce the olive oil. Three types of olive oil are currently present on the market: virgin, ordinary, or pomace. Virgin olive oils are produced by direct pressing or centrifugation of the olives, among them those with an acidity greater than or equal to 3.3 degrees (2 degrees in the European Union) are submitted to a refining process in which some components, mainly phenolic compounds, and to a lesser degree squalene, are lost. By mixing virgin and refined olive oil, an ordinary olive oil (olive oil, UE 1991) is produced and marketed. After virgin olive oil production, the rest of the olive drupe and seed is processed and submitted to a refining process, resulting in pomace olive oil, to which a certain quantity of virgin olive oil is added before marketing. The minor components of virgin olive oil are classified into two types: the unsaponificable fraction, defined as the fraction extracted with solvents after the saponification of the oil, and the soluble fraction, which includes the phenolic compounds. The major phenolic compounds in olive oil are (1) simple phenols, (2) secoiridoids, and (3) polyphenols.

The pharmacokinetics of resveratrol in 11 healthy male volunteers has been assessed in a randomized, crossover, controlled clinical trial after the administration of three grape products: red wine (250 mL), grape juice (1/L), or tablets (red wine extracts enriched with trans-resveratrol). Doses of trans-resveratrol independently of the product administered, were about 14 μg/kg. Biological samples were collected and analysed by capillary gas chromatography–mass spectrometry. cis-Resveratrol, trans-resveratrol and dihydroresveratrol were determined in plasma and urine. Plasma concentration of trans-resveratrol after hydrolysis increased as a response to all grape products and that of cis-resveratrol after wine and grape juice. Free forms of these phenolic compounds were deemed undetectable in plasma. Despite similar trans-resveratrol doses being administered, its bioavailability from wine and grape juice was 6-fold higher, than that from tablets. Resveratrol was better absorbed from natural grape products than from tablets, pointing out the importance of the matrix in its bioavailability.

Our aim was to examine whether an acute fat load could induce changes in the expression of insulin sensitivity-related genes in human peripheral blood mononuclear cells. Selection of candidate genes was based on previous studies with sustained virgin olive oil (VOO) consumption and biological plausibility in relation to insulin sensitivity. Eleven healthy volunteers ingested raw VOO (50 mL). Blood samples were collected at 0, 1 and 6 h. Plasma glucose, insulin and hydroxytyrosol increased at 1 h and decreased at 6 h. Lipid oxidative damage increased at 6 h (p0.05). Gene expression changes were characterized based on quantification of the samples relative to a reference sample [i.e., relative quantification (RQ) method]. A 1 h downregulation was observed in O-linked-N-acetylglucosamine transferase (OGT, RQ: 0.62 +/- 0.32) and arachidonate-5-lipoxygenase-activating protein (ALOX5AP, RQ: 0.64 +/- 0.31) genes (p0.005). OGT was upregulated at 6 h (RQ: 1.88 +/- 0.28, p0.05). CD36 (thrombospondin receptor) was upregulated at 1 h (RQ: 1.6 +/- 0.8, p0.05) returning to the basal values at 6 h. Lipoic acid synthetase (LIAS), peroxisome proliferator-activated receptor binding protein (PPARBP), a disintegrin and metallopeptidase domain 17 (ADAM17), and adrenergic beta-2-receptor (ADRB2) genes were upregulated at 6 h (range for the mean RQ: 1.33-1.56) following an increasing linear trend (p0.05) from baseline to 6 h. ALOX5AP and OGT genes inversely correlated with insulin and glucose levels at 1 h. ADAM17 and ADRB2 inversely correlated with oxLDL at 6 h (p0.05). Taken together, these observations may inform the future clinical nutrigenomics study designs and indicate that a single dose of VOO can elicit quantifiable and rapid changes in gene expression in targets that are mechanistically relevant for insulin sensitivity and the metabolic syndrome.

Objectives: This study investigates how some of human physiological parameters, in particular sex and daytime, can contribute to the normal variation in blood gene expression. Design and methods: The expressions of two genes: SOD1 (Cu/Zn) and SOD2 (Mn) in white blood cells of 19 healthy individuals (10 men and 9 women) were examined by qPCR at 09.00, 12.00, and 15.00 h over three consecutive weeks at 7-day intervals. SOD1 and SOD2 expressions differ between sexes, higher expression being exhibited in males (Pb0.005). In the male group both genes had lower expression (Pb0.005) with smaller variation at 09.00 h. In females, both inter- and intra-subject, variability in SOD1 expression was increased at ovulation. Intra-individually, the expressions of genes positively correlated in males, but less in females. Conclusions: SOD1 and SOD2 expressions demonstrate how sex and daytime, merit being controlled when human gene expression analyses are evaluated, particularly within the framework of clinical trials or cohort studies.

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. Financial support was received from the TV3 Marató (01/810), Fondo de Investigaciones Sanitarias de la Seguridad Social, FIS (PI040632, PI040619), and the Instituto Carlos III (RTA 06/0001/1009). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Deficits in verbal fluency associated with ecstasy use have been well established; however, the mechanisms underlying this impairment have yet to be elucidated. In this study we investigated for the first time whether there was a disproportionate impairment in two cognitive subcomponents of verbal fluency: clustering (ability to generate words within the same subcategory) and switching (ability to change the subcategory). We also investigated a possible association between ecstasy use and verbal fluency in subjects genotyped for 5-HTT (5-HTTLPR and 5-HTTVNTR) and COMT (val108/158met, rs165599 and rs2097603) polymorphisms, in order to find a potential implication of genetic factors. Ecstasy polydrug users (n = 30) and non-ecstasy users (n = 41) were evaluated in both semantic and phonemic fluency. Results showed that ecstasy users had poorer semantic (but not phonemic) fluency performance than controls. Detailed analysis of clustering and switching performance revealed that this impairment was associated with poorer clustering mechanisms. Clustering was also modulated by the COMT rs165599 polymorphism independently of the group. A specific effect of the 5-HTTLPR polymorphism on switching performance was also found, with ss carriers performing significantly worse than ls and ll carriers, suggesting a serotonin modulation of frontal-executive flexibility. Based on the impaired clustering and switching strategies observed in ecstasy users, it might be proposed that both semantic knowledge and retrieval are impaired in this population. The verbal fluency deficit in ecstasy users may be attributable to a disruption of frontal-striatal circuits directly related with the serotonin function as well as a depletion of lexical-semantic stores mediated by temporal structures.

Journal Article; Research Support, Non-U.S. Gov't; 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. Yes