Universitat Rovira i Virgili


El grup de Nutrigenòmica publica anualment en revistes indexades del primer quartil els resultats de la seva recerca. A més a més, periòdicament assisteix a congressos internacionals i publica capítols en llibres especialitzats de l'àmbit d'investigació. També participa en diverses activitats de divulgació.

Articles Cientifics


Photoperiodic Remodeling of Adiposity and Energy Metabolism in Non‐Human Mammals

Èlia Navarro-Masip 1, Alexandre Caron 2, Miquel Mulero 1, Lluís Arola 1, Gerard Aragonès 1

Energy homeostasis and metabolism in mammals are strongly influenced by seasonal changes. Variations in photoperiod patterns drive adaptations in body weight and adiposity, reflecting changes in the regulation of food intake and energy expenditure. Humans also show distinct patterns of energy balance depending on the season, being more susceptible to gaining weight during a specific time of the year. Changes in body weight are mainly reflected by the adipose tissue, which is a key metabolic tissue and is highly affected by circannual rhythms. Mostly, in summer-like (long-active) photoperiod, adipocytes adopt a rather anabolic profile, more predisposed to store energy, while food intake increases and energy expenditure is reduced. These metabolic adaptations involve molecular modifications, some of which have been studied during the last years and are summarized in this review. In addition, there is a bidirectional relation between obesity and the seasonal responses, with obesity disrupting some of the seasonal responses observed in healthy mammals, and altered seasonality being highly associated with increased risk of developing obesity. This suggests that changes in photoperiod produce important metabolic alterations in healthy organisms. Biological rhythms impact the regulation of metabolism to different extents, some of which are already known, but further research is needed to fully understand the relationship between energy balance and seasonality.


In-Season Consumption of Locally Produced Tomatoes Decreases Cardiovascular Risk Indices

Ma Josefina Ruiz de Azua 1, Álvaro Cruz-Carrión 1, Begoña Muguerza 1, Gerard Aragonès 1, Anna Arola-Arnal 1, María Paz Romero 2, Francisca Isabel Bravo 1, Manuel Suarez 1

Tomatoes are widely consumed worldwide at any time of the year. However, depending on the variety, they have a characteristic season. We evaluated the consequences metabolic of consumption of Ekstasis tomatoes from different geographical origin and in different seasons in Fischer 344 rats. The hepatic gene expression of key enzymes in lipid metabolism was also evaluated. Animals were classified in three photoperiods (L6, L12, and L18) and in three treatments (vehicle: VH; local tomato: LT; and non-local tomato: nLT). We measured serum metabolic parameters and the gene expression of liver enzymes related to lipid metabolism (Acc1, Cpt1a, Had, Fas1, Srebp-1c, Fatp5, Cd36). LT consumption in season decreased cardiovascular risk 1 and coefficient atherogenic by 1.81 (p = 0.031) and in L6 decreased TAG and glucose (p = 0.046; p = 0.024). The L18-LT animals had decreased total cholesterol (p = 0.029) and gene expression of Srebp1-c (p = 0.022) but increased expression of Acc1 (p = 0.032). The treatments significantly affected the expression of Acc1 and Fas1 in the liver and the levels of serum TAG and glucose. A significant effect of photoperiod on serum concentration of glucose, insulin, HOMA index, and on the hepatic expression of Srep1-c, Fas1, and Acc1 was observed.


Organic vs. Non-Organic Plant-Based Foods-A Comparative Study on Phenolic Content and Antioxidant Capacity

Álvaro Cruz-Carrión 1, Ma Josefina Ruiz de Azua 1, Begoña Muguerza 1, Miquel Mulero 1, Francisca Isabel Bravo 1, Anna Arola-Arnal 1, Manuel Suarez 1

A plant's stress response involves the production of phytochemicals, including phenolic compounds. Their synthesis can be modulated by organic (ORG) or non-organic (NORG) farming systems in which they are grown. To examine this issue, thirteen plant-based foods cultivated in ORG and NORG systems were compared in terms of antioxidant capacity, total content of phenolics, anthocyanins, flavan-3-ols and flavonols. The results showed that NORG fruits tended to have higher phenolic compounds content, whereas ORG fruits had more antioxidant capacity. NORG legume stood out for having higher values from all the parameters analyzed in comparison to its ORG equivalent. ORG nuts showed more flavan-3-ols and flavonols than their NORG counterparts, nonetheless, tended to be less antioxidant. ORG vegetables displayed higher phenolics and anthocyanins, which reflected in higher antioxidant capacity than NORG ones. These findings suggest that farming systems differentially modulate phenolic compound composition and antioxidant capacity based on the plant species studied.


Fatty acid metabolism in liver and muscle is strongly modulated by photoperiod in Fischer 344 rats

Ma Josefina Ruiz de Azua 1, Francesca Manocchio 2, Álvaro Cruz-Carrión 2, Anna Arola-Arnal 2, Cristina Torres-Fuentes 2, Claudio Adrían Bernal 3, Juliana Saín 3, Manuel Suarez 4

Circadian and seasonal variations produce variations in physiological processes throughout the day and the year, respectively. In this sense, both the light and the moment of feeding are strong modulators of the central and peripheral clocks. However, little is known about its influence on certain metabolic parameters and on the composition of liver and muscle fatty acids (FA). In the present study, 24 Fischer 344 rats were exposed for 11 weeks to different photoperiods, L6, L12 and L18, with 6, 12 and 18 h of light/day, respectively. They were fed a standard diet. Serum metabolic parameters, gene expression of liver enzymes and gastrocnemius muscle involved in the synthesis, elongation, desaturation and β-oxidation of FA were analyzed. We have found that exposure to different hours of light has a clear effect on FA composition and gene expression in the liver. Mainly, the biosynthesis of unsaturated FA was altered in the L18 animals with respect to those exposed to L12, while the L6 did not show significant changes. At the muscle level, differences were observed in the concentration of mono and polyunsaturated FA. A multivariate analysis confirmed the differences between L12 and L18 in a significant way. We conclude that exposure to long days produces changes in the composition of liver and muscle FA, as well as changes in the gene expression of oxidative enzymes compared to exposure to L12, which could be a consequence of different seasonal eating patterns.


Brain N-Glycosylation and Lipidomic Profile Changes Induced by a High-Fat Diet in Dyslipidemic Hamsters

Beatrix Paton 1, Elisabet Foguet-Romero 1, Manuel Suarez 2, Jordi Mayneris-Perxachs 3 4, Noemí Boqué 5, Antoni Caimari 6, Núria Canela 1, Pol Herrero 1

The consumption of diets rich in saturated fats is known to be associated with higher mortality. The adoption of healthy habits, for instance adhering to a Mediterranean diet, has proved to exert a preventive effect towards cardiovascular diseases and dyslipidemia. Little is known about how a suboptimal diet can affect brain function, structure, and the mechanisms involved. The aims of this study were to examine how a high-fat diet can alter the brain N-glycan and lipid profile in male Golden Syrian hamsters and to evaluate the potential of a Mediterranean-like diet to reverse this situation. During twelve weeks, hamsters were fed a normal fat diet (CTRL group), a high-fat diet (HFD group), and a high-fat diet followed by a Mediterranean-like diet (MED group). Out of seventy-two identified N-glycans, fourteen were significant (p < 0.05) between HFD and CTRL groups, nine between MED and CTRL groups, and one between MED and HFD groups. Moreover, forty-nine lipids were altered between HFD and CTRL groups, seven between MED and CTRL groups, and five between MED and HFD groups. Our results suggest that brain N-glycan composition in high-fat diet-fed hamsters can produce events comparable to those found in some neurodegenerative diseases, and may promote brain ageing.


Grape-Seed Proanthocyanidins Modulate Adipose Tissue Adaptations to Obesity in a Photoperiod-Dependent Manner in Fischer 344 Rats

Èlia Navarro-MasipMarina Colom-PellicerFrancesca ManocchioAnna Arola-ArnalFrancisca Isabel BravoBegoña MuguerzaGerard Aragonès

Seasonal rhythms drive metabolic adaptations that influence body weight and adiposity. Adipose tissue is a key regulator of energy homeostasis in the organism, and its healthiness is needed to prevent the major consequences of overweight and obesity. In this context, supplementation with proanthocyanidins has been postulated as a potential strategy to prevent the alterations caused by obesity. Moreover, the effects of these (poly)phenols on metabolism are photoperiod dependent. In order to describe the impact of grape-seed proanthocyanidins extract (GSPE) on important markers of adipose tissue functionality under an obesogenic environment, we exposed Fischer 344 rats to three different photoperiods and fed them a cafeteria diet for five weeks. Afterwards, we supplemented them with 25 mg GSPE/kg/day for four weeks. Our results revealed that GSPE supplementation prevented excessive body weight gain under a long photoperiod, which could be explained by increased lipolysis in the adipose tissue. Moreover, cholesterol and non-esterified fatty acids (NEFAs) serum concentrations were restored by GSPE under standard photoperiod. GSPE consumption slightly helped combat the obesity-induced hypertrophy in adipocytes, and adiponectin mRNA levels were upregulated under all photoperiods. Overall, the administration of GSPE helped reduce the impact of obesity in the adipose tissue, depending on the photoperiod at which GSPE was consumed and on the type of adipose depots.



Gut Seasons: Photoperiod Effects on Fecal Microbiota in Healthy and Cafeteria-Induced Obese Fisher 344 Rats

Verónica Arreaza-Gil 1, Iván Escobar-Martínez 1, Manuel Suárez 1, Francisca Isabel Bravo 1, Begoña Muguerza 1, Anna Arola-Arnal 1, Cristina Torres-Fuentes 1

Gut microbiota and biological rhythms are emerging as key factors in the modulation of several physiological and metabolic processes. However, little is known about their interaction and how this may affect host physiology and metabolism. Several studies have shown oscillations of gut microbiota that follows a circadian rhythmicity, but, in contrast, variations due to seasonal rhythms have not been sufficiently investigated yet. Thus, the goal of this study was to investigate the impact of different photoperiods, which mimic seasonal changes, on fecal microbiota composition and how this interaction affects diet-induced obesity development. To this aim, Fisher 344 male rats were housed under three photoperiods (L6, L12 and L18) and fed with standard chow diet or cafeteria diet (CAF) for 9 weeks. The 16S ribosomal sequencing of collected fecal samples was performed. The photoperiod exposure significantly altered the fecal microbiota composition under L18, especially in CAF-fed rats. Moreover, these alterations were associated with changes in body weight gain and different fat parameters. These findings suggest a clear impact of seasonal rhythms on gut microbiota, which ultimately translates into different susceptibilities to diet-induced obesity development. This is the first time to our knowledge that the photoperiod impact on gut microbiota composition has been described in an obesity context although further studies are needed in order to elucidate the mechanisms involved.


Identification of novel antihypertensive peptides from wine lees hydrolysate

Francisca Isabel Bravo 1, Anna Mas-Capdevila 2, Raúl López-Fernández-Sobrino 3, Cristina Torres-Fuentes 4, Miquel Mulero 5, Juan María Alcaide-Hidalgo 6, Begoña Muguerza 7

Enzymatic-assisted extraction using Flavourzyme® has been demonstrated to be a useful methodology to obtain wine lees (WL) enriched in phenolic compounds and with enhanced antihypertensive activity. Nevertheless, taking into account that Flavourzyme® possess proteolytic activity, the release of bioactive peptides should not be ruled out. In this study, we investigate the presence of antihypertensive peptides in the WL hydrolysate. Peptides were separated into fractions by ultrafiltration and RP-HPLC. Next, peptide identification by nano-HPLC-(Orbitrap)MS/MS was performed in the fractions showing the highest angiotensin-converting enzyme inhibitory (ACEi) activities. Six peptides were identified; three of them showing ACEi (IC50) values lower than 20 µM. The peptide antihypertensive effect was evaluated in spontaneously hypertensive rats at an oral dose of 10 mg/kg bw. Peptides FKTTDQQTRTTVA, NPKLVTIV, TVTNPARIA, LDSPSEGRAPG and LDSPSEGRAPGAD exhibited antihypertensive activity, confirming that they could contribute to the blood pressure-lowering effect of the WL hydrolysate. These peptides have a great potential as functional ingredients to manage hypertension.


Serum lysophospholipidome of dietary origin as a suitable susceptibility/risk biomarker of human hypercholesterolemia: A cross-sectional study

Lorena Calderón-Pérez 1, Susana Suárez-García 2, Anna Pedret 3, Manuel Suárez 4, Elisabet Llauradó 5, Laura Rubió 6, Josep M Del Bas 7, Antoni Caimari 7, Francesc Puiggrós 7, Lluís Arola 2, Rosa Solà 8, Rosa M Valls 1

Background & aims: Whether bioactive lysophospholipids (lyso-PLs) and trimethylamine-N-oxide (TMAO) serve as non-invasive biomarkers in early human hypercholesterolemia (HC) is unknown. This study aimed to assess whether serum lyso-PLs and plasma TMAO may be suitable susceptibility/risk biomarkers of HC in humans. Secondarily, we aimed to evaluate the relationships between targeted metabolites, diet composition and circulating liver transaminases, and verify these results in hamsters.

Methods: A targeted metabolomics and lipidomics approach determined plasma TMAO and serum lysophosphatidylcholines (lyso-PCs) and lysophosphatidylethanolamines (lyso-PEs) in low (L-LDL-c) and moderate to high (MH-LDL-c) LDL-cholesterol subjects. Additionally, the relationships between targeted metabolites, liver transaminases and diet, particularly fatty acid intake, were tested. In parallel, plasma and liver lyso-PL profiles were studied in 16 hamsters fed a moderate high-fat (HFD) or low-fat (LFD) diet for 30 days.

Results: Predictive models identified lyso-PC15:0 and lyso-PE18:2 as the most discriminant lyso-PLs among groups. In MH-LDL-c (n = 48), LDL-cholesterol and saturated FAs were positively associated with lyso-PC15:0, whereas in L-LDL-c (n = 70), LDL-cholesterol and polyunsaturated fatty acids (PUFAs) were negatively and positively related to lyso-PE18:2, respectively. Interestingly, in MH-LDL-c, the lower lyso-PE 18:2 concentrations were indicative of higher LDL-cholesterol levels. Intrahepatic accumulation of lyso-PLs-containing essential n-6 PUFAs, including lyso-PE18:2, were higher in HFD-fed hamsters than LFD-fed hamsters.

Conclusions: Overall, results revealed a possible hepatic adaptive mechanism to counteract diet-induced steatosis in animal and hypercholesterolemia progression in humans. In particular, low serum lyso-PE18:2 suggests a suitable susceptibility/risk biomarker of HC in humans.


Proanthocyanidins Restore the Metabolic Diurnal Rhythm of Subcutaneous White Adipose Tissue According to Time-Of-Day Consumption

Marina Colom-Pellicer 1, Romina M Rodríguez 1, Jorge R Soliz-Rueda 1, Leonardo Vinícius Monteiro de Assis 2, Èlia Navarro-Masip 1, Sergio Quesada-Vázquez 3, Xavier Escoté 3, Henrik Oster 2, Miquel Mulero 1, Gerard Aragonès 1

Consumption of grape seed proanthocyanidin extract (GSPE) has beneficial effects on the functionality of white adipose tissue (WAT). However, although WAT metabolism shows a clear diurnal rhythm, whether GSPE consumption could affect WAT rhythmicity in a time-dependent manner has not been studied. Ninety-six male Fischer rats were fed standard (STD, two groups) or cafeteria (CAF, four groups) diet for 9 weeks (n = 16 each group). From week 6 on, CAF diet animals were supplemented with vehicle or 25 mg GSPE/kg of body weight either at the beginning of the light/rest phase (ZT0) or at the beginning of the dark/active phase (ZT12). The two STD groups were also supplemented with vehicle at ZT0 or ZT12. In week 9, animals were sacrificed at 6 h intervals (n = 4) to analyze the diurnal rhythms of subcutaneous WAT metabolites by nuclear magnetic resonance spectrometry. A total of 45 metabolites were detected, 19 of which presented diurnal rhythms in the STD groups. Although most metabolites became arrhythmic under CAF diet, GSPE consumption at ZT12, but not at ZT0, restored the rhythmicity of 12 metabolites including compounds involved in alanine, aspartate, and glutamate metabolism. These results demonstrate that timed GSPE supplementation may restore, at least partially, the functional dynamics of WAT when it is consumed at the beginning of the active phase. This study opens an innovative strategy for time-dependent polyphenol treatment in obesity and metabolic diseases.


Time-of-day dependent effect of proanthocyanidins on adipose tissue metabolism in rats with diet-induced obesity

Marina Colom-Pellicer 1, Romina M Rodríguez 1, Èlia Navarro-Masip 1, Francisca Isabel Bravo 1, Miquel Mulero 1, Lluís Arola 1, Gerard Aragonès 2

Background: Grape-seed proanthocyanidin extract (GSPE) improve white adipose tissue (WAT) expansion during diet-induced obesity. However, because adipose metabolism is synchronized by circadian rhythms, it is plausible to speculate that the bioactivity of dietary proanthocyanidins could be influenced by the time-of-day in which they are consumed. Therefore, the aim of the present study was to determine the interaction between zeitgeber time (ZT) and GSPE consumption on the functionality of WAT in rats with diet-induced obesity.

Methods: Male Wistar rats were fed a cafeteria diet for 9 weeks. After 5 weeks, the animals were supplemented with 25 mg GSPE/kg for 4 weeks at the beginning of the light/rest phase (ZT0) or of the dark/active phase (ZT12). Body fat content was determined by nuclear magnetic resonance and histological analyses were performed in the epididymal (EWAT) and inguinal (IWAT) fat depots to determine adipocyte size and number. In addition, the expression of genes related to adipose metabolism and circadian clock function were analyzed by qPCR.

Results: GSPE consumption at ZT0 was associated with a potential antidiabetic effect without affecting adiposity and energy intake and downregulating the gene expression of inflammatory markers in EWAT. In contrast, GSPE consumption at ZT12 improved adipose tissue expansion decreasing adipocyte size in IWAT. In accordance with this adipogenic activity, the expression of genes involved in fatty acid metabolism were downregulated at ZT12 in IWAT. In turn, GSPE consumption at ZT12, but not at ZT0, repressed the expression of the clock gene Cry1 in IWAT.

Conclusions: The interaction between ZT and GSPE consumption influenced the metabolic response of WAT in a tissue-specific manner. Understanding the impact of circadian clock on adipose metabolism and how this is regulated by polyphenols will provide new insights for the management of obesity.


Administration Time Significantly Affects Plasma Bioavailability of Grape Seed Proanthocyanidins Extract in Healthy and Obese Fischer 344 Rats

Iván Escobar-Martínez 1, Verónica Arreaza-Gil 1, Begoña Muguerza 1, Anna Arola-Arnal 1, Francisca Isabel Bravo 1, Cristina Torres-Fuentes 1, Manuel Suárez 1

Scope: Phenolic compounds are bioactive molecules that are associated with several health benefits. Metabolization and absorption are the main determinants of their bioavailability and bioactivity. Thus, the study of the factors that modulate these processes, such as sex or diet is essential. Recently, it has been shown that biological rhythms may also play a key role. Hence, the aim of this study is to evaluate if the bioavailability of a grape proanthocyanidin extract (GSPE) is affected by the administration time in an animal model of metabolic syndrome (MetS).

Methods and results: Female and male Fischer 344 rats are fed either a standard or a cafeteria diet (CAF) for 9 weeks, and an oral dose of GSPE (25 mg kg-1 ) is daily administered either at 8:00 am (zeitgeber time (ZT)-0) or at 8:00 pm (ZT-12) during the last 4 weeks. Plasma phenolic compounds are then quantified by liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Phase-II and gut microbiota-derived phenolic metabolites are affected by ZT in all conditions or only in obese rats, respectively. CAF feeding affected the bioavailability of phenolic acids and free flavan-3-ols. Differences due to sex are also observed.

Conclusion: These findings demonstrate that ZT, diet, and sex are key factors influencing phenolic compounds bioavailability.


In vitro fermentability of a broad range of natural ingredients by fecal microbiota from lean and obese individuals: potential health benefits

Andreu Gual-Grau 1, Maria Guirro 2, Anna Crescenti 3, Noemí Boqué 3, Lluís Arola 1

The prevalence of obesity and related complications is continuously increasing while the gut microbiota might have a significant role to address this challenge. In this context, the food industry generates large amounts of residues that could be likely revalorised as functional ingredients. Hence, we evaluated the fermentability of food skins, husks, shells, trimming residues, mosses and mushrooms, which were subjected to in vitro fermentation with faecal microbiota from lean and obese adults. We demonstrated for the first time that pumpkin skin is highly fermented by human faecal microbiota showing pH-lowering effects and promoting gas and SCFA production. Furthermore, brewers' spent grain generated an inulin-like SCFA profile after microbial fermentation, whereas Irish moss, plum skin, quinoa husk and mushrooms, including Armillaria mellea and Boletus edulis, showed high fermentation rates. Remarkably, although propionate production was significantly higher in obese individuals, the fermentability of the ingredients was similar between lean and obese conditions.


Imbalances in TCA, Short Fatty Acids and One-Carbon Metabolisms as Important Features of Homeostatic Disruption Evidenced by a Multi-Omics Integrative Approach of LPS-Induced Chronic Inflammation in Male Wistar Rats

Julia Hernandez-Baixauli 1, Nerea Abasolo 2, Hector Palacios-Jordan 2, Elisabet Foguet-Romero 2, David Suñol 3, Mar Galofré 3, Antoni Caimari 1, Laura Baselga-Escudero 1, Josep M Del Bas 1, Miquel Mulero 4 5

Chronic inflammation is an important risk factor in a broad variety of physical and mental disorders leading to highly prevalent non-communicable diseases (NCDs). However, there is a need for a deeper understanding of this condition and its progression to the disease state. For this reason, it is important to define metabolic pathways and complementary biomarkers associated with homeostatic disruption in chronic inflammation. To achieve that, male Wistar rats were subjected to intraperitoneal and intermittent injections with saline solution or increasing lipopolysaccharide (LPS) concentrations (0.5, 5 and 7.5 mg/kg) thrice a week for 31 days. Biochemical and inflammatory parameters were measured at the end of the study. To assess the omics profile, GC-qTOF and UHPLC-qTOF were performed to evaluate plasma metabolome; 1H-NMR was used to evaluate urine metabolome; additionally, shotgun metagenomics sequencing was carried out to characterize the cecum microbiome. The chronicity of inflammation in the study was evaluated by the monitoring of monocyte chemoattractant protein-1 (MCP-1) during the different weeks of the experimental process. At the end of the study, together with the increased levels of MCP-1, levels of interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-α) and prostaglandin E2 (PGE2) along with 8-isoprostanes (an indicative of oxidative stress) were significantly increased (p-value < 0.05). The leading features implicated in the current model were tricarboxylic acid (TCA) cycle intermediates (i.e., alpha-ketoglutarate, aconitic acid, malic acid, fumaric acid and succinic acid); lipids such as specific cholesterol esters (ChoEs), lysophospholipids (LPCs) and phosphatidylcholines (PCs); and glycine, as well as N, N-dimethylglycine, which are related to one-carbon (1C) metabolism. These metabolites point towards mitochondrial metabolism through TCA cycle, β-oxidation of fatty acids and 1C metabolism as interconnected pathways that could reveal the metabolic effects of chronic inflammation induced by LPS administration. These results provide deeper knowledge concerning the impact of chronic inflammation on the disruption of metabolic homeostasis.


Role of Chrononutrition in the Antihypertensive Effects of Natural Bioactive Compounds

Néstor Ibarz-Blanch 1, Diego Morales 1, Enrique Calvo 1, Laura Ros-Medina 1, Begoña Muguerza 1, Francisca Isabel Bravo 1, Manuel Suárez 1

Hypertension (HTN) is one of the main cardiovascular risk factors and is considered a major public health problem. Numerous approaches have been developed to lower blood pressure (BP) in hypertensive patients, most of them involving pharmacological treatments. Within this context, natural bioactive compounds have emerged as a promising alternative to drugs in HTN prevention. This work reviews not only the mechanisms of BP regulation by these antihypertensive compounds, but also their efficacy depending on consumption time. Although a plethora of studies has investigated food-derived compounds, such as phenolic compounds or peptides and their impact on BP, only a few addressed the relevance of time consumption. However, it is known that BP and its main regulatory mechanisms show a 24-h oscillation. Moreover, evidence shows that phenolic compounds can interact with clock genes, which regulate the biological rhythm followed by many physiological processes. Therefore, further research might be carried out to completely elucidate the interactions along the time-nutrition-hypertension axis within the framework of chrononutrition.


Winery by-products as a valuable source for natural antihypertensive agents

Raúl López-Fernández-Sobrino 1, Cristina Torres-Fuentes 1, Francisca Isabel Bravo 1, Begoña Muguerza 1

Hypertension (HTN) is one of the leading causes of death in the world. Agri-food by-products are emerging as a novel source of natural antihypertensive agents allowing for their valorization and making food and agricultural industries more environmentally friendly. In this regard, wine making process generates large amounts of by-products rich in phenolic compounds that have shown potential to exert several beneficial effects including antihypertensive properties. The aim of this study was to review the blood pressure-lowering effects of winery by-products. In addition, molecular mechanisms involved in their bioactivity were also evaluated. Among the winery by-products, grape seed extracts have widely shown antihypertensive properties in both animal and human studies. Moreover, recent evidence suggests that grape stem, skin and pomace and wine lees may also have great potential to manage HTN, although more studies are needed in order to confirm their potential in humans. Improvement of endothelial dysfunction and reduction of oxidative stress associated with HTN are the main mechanisms involved in the blood pressure-lowering effects of these by-products.


Structured Long-Chain Omega-3 Fatty Acids for Improvement of Cognitive Function during Aging

Ignasi Mora 1, Lluís Arola 2, Antoni Caimari 3, Xavier Escoté 4, Francesc Puiggròs 3

Although the human lifespan has increased in the past century owing to advances in medicine and lifestyle, the human healthspan has not kept up the same pace, especially in brain aging. Consequently, the role of preventive health interventions has become a crucial strategy, in particular, the identification of nutritional compounds that could alleviate the deleterious effects of aging. Among nutrients to cope with aging in special cognitive decline, the long-chain omega-3 polyunsaturated fatty acids (ω-3 LCPUFAs) docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), have emerged as very promising ones. Due to their neuroinflammatory resolving effects, an increased status of DHA and EPA in the elderly has been linked to better cognitive function and a lower risk of dementia. However, the results from clinical studies do not show consistent evidence and intake recommendations for old adults are lacking. Recently, supplementation with structured forms of EPA and DHA, which can be derived natural forms or targeted structures, have proven enhanced bioavailability and powerful benefits. This review summarizes present and future perspectives of new structures of ω-3 LCPUFAs and the role of "omic" technologies combined with the use of high-throughput in vivo models to shed light on the relationships and underlying mechanisms between ω-3 LCPUFAs and healthy aging.


Mediterranean natural extracts improved cognitive behavior in zebrafish and healthy rats and ameliorated lps-induced cognitive impairment in a sex dependent manner

Matteo M Pusceddu 1, Julia Hernandez-Baixauli 2, Francesc Puiggrós 2, Lluis Arola 2 3, Antoni Caimari 4, Josep M Del Bas 5, Laura Baselga 2

Background: Several findings suggest neuroinflammation as a contributing factor for the onset of psychiatric disorders such as Alzheimer's disease, depression, and anxiety. There is increasing evidence pointing out that the Mediterranean diet influences brain and behavior. Mediterranean herbs and spices have been shown to be within those components of the Mediterranean diet involved in cognitive enhancement. Thus, we investigated the influence of Mediterranean natural extracts (MNE), Rosemary extract (RE) and Glycyrrhiza glabra root extract (GGRE), on cognitive behavior.

Results: Adult zebrafish were exposed to RE or GGRE (100 and 250 mg/L) treatments. Both MNE improved memory retention during the T-maze test, although no improvements were observed during the novel object preference. Similarly, chronic administration of RE (150 mg/Kg) and GGRE (150 mg/Kg) improved, respectively, spatial and retention memory, as assessed by the Morris Water Maze (MWM), and the Elevated Plus Maze (EPM) in healthy male rats. However, no improvements were observed during the novel object recognition. Finally, male, and female rats were chronically treated with lipopolysaccharide [(LPS) 300 ug/kg] and orally administered with RE. Interestingly, RE reversed LPS-induced cognitive deficit during the MWM and EPM in female rats.

Conclusions: We found that MNE improved cognition in both zebrafish and rats. Moreover, MNE rescued LPS-induced cognitive impairment in a gender-specific manner. Therefore, our study supports the view that zebrafish represent a valuable preclinical model for drug discovery in neuroscience. These findings contribute to an exciting and growing body of research suggesting that MNE may play an important role in the prevention of cognitive impairment.


Recent Advances in Research on Polyphenols: Effects on Microbiota, Metabolism, and Health

Hiba N Rajha 1, Armelle Paule 2, Gerard Aragonès 3, Mariana Barbosa 4, Carla Caddeo 5, Espérance Debs 6, Rada Dinkova 7, Gunter P Eckert 8, Ariel Fontana 9, Prisca Gebrayel 2, Richard G Maroun 1, Alessandra Napolitano 10, Lucia Panzella 10, Giulio Maria Pasinetti 11, Jan F Stevens 12, Andreas Schieber 13, Marvin Edeas 14

Polyphenols have attracted huge interest among researchers of various disciplines because of their numerous biological activities, such as antioxidative, antiinflammatory, antiapoptotic, cancer chemopreventive, anticarcinogenic, and antimicrobial properties, and their promising applications in many fields, mainly in the medical, cosmetics, dietary supplement and food industries. In this review, the latest scientific findings in the research on polyphenols interaction with the microbiome and mitochondria, their metabolism and health beneficial effects, their involvement in cognitive diseases and obesity development, as well as some innovations in their analysis, extraction methods, development of cosmetic formulations and functional food are summarized based on the papers presented at the 13th World Congress on Polyphenol Applications. Future implications of polyphenols in disease prevention and their strategic use as prophylactic measures are specifically addressed. Polyphenols may play a key role in our tomorrow´s food and nutrition to prevent many diseases.


Grape-Seed Procyanidin Extract (GSPE) Seasonal-Dependent Modulation of Glucose and Lipid Metabolism in the Liver of Healthy F344 Rats

Romina M Rodríguez 1, Marina Colom-Pellicer 1, Jordi Blanco 2, Enrique Calvo 1, Gerard Aragonès 1, Miquel Mulero 1

Seasonality is gaining attention in the modulation of some physiological and metabolic functions in mammals. Furthermore, the consumption of natural compounds, such as GSPE, is steadily increasing. Consequently, in order to study the interaction of seasonal variations in day length over natural compounds' molecular effects, we carried out an animal study using photo-sensitive rats which were chronically exposed for 9 weeks to three photoperiods (L6, L18, and L12) in order to mimic the day length of different seasons (winter/summer/and autumn-spring). In parallel, animals were also treated either with GSPE 25 (mg/kg) or vehicle (VH) for 4 weeks. Interestingly, a seasonal-dependent GSPE modulation on the hepatic glucose and lipid metabolism was observed. For example, some metabolic genes from the liver (SREBP-1c, Gk, Acacα) changed their expression due to seasonality. Furthermore, the metabolomic results also indicated a seasonal influence on the GSPE effects associated with glucose-6-phosphate, D-glucose, and D-ribose, among others. These differential effects, which were also reflected in some plasmatic parameters (i.e., glucose and triglycerides) and hormones (corticosterone and melatonin), were also associated with significant changes in the expression of several hepatic circadian clock genes (Bmal1, Cry1, and Nr1d1) and ER stress genes (Atf6, Grp78, and Chop). Our results point out the importance of circannual rhythms in regulating metabolic homeostasis and suggest that seasonal variations (long or short photoperiods) affect hepatic metabolism in rats. Furthermore, they suggest that procyanidin consumption could be useful for the modulation of the photoperiod-dependent changes on glucose and lipid metabolism, whose alterations could be related to metabolic diseases (e.g., diabetes, obesity, and cardiovascular disease). Furthermore, even though the GSPE effect is not restricted to a specific photoperiod, our results suggest a more significant effect in the L18 condition.


Time-of-Day Circadian Modulation of Grape-Seed Procyanidin Extract (GSPE) in Hepatic Mitochondrial Dynamics in Cafeteria-Diet-Induced Obese Rats

Romina M Rodríguez 1, Antonio J Cortés-Espinar 1, Jorge R Soliz-Rueda 1, Christine Feillet-Coudray 2, François Casas 2, Marina Colom-Pellicer 1, Gerard Aragonès 1, Javier Avila-Román 3, Begoña Muguerza 1, Miquel Mulero 1, Maria Josepa Salvadó 1

Major susceptibility to alterations in liver function (e.g., hepatic steatosis) in a prone environment due to circadian misalignments represents a common consequence of recent sociobiological behavior (i.e., food excess and sleep deprivation). Natural compounds and, more concisely, polyphenols have been shown as an interesting tool for fighting against metabolic syndrome and related consequences. Furthermore, mitochondria have been identified as an important target for mediation of the health effects of these compounds. Additionally, mitochondrial function and dynamics are strongly regulated in a circadian way. Thus, we wondered whether some of the beneficial effects of grape-seed procyanidin extract (GSPE) on metabolic syndrome could be mediated by a circadian modulation of mitochondrial homeostasis. For this purpose, rats were subjected to "standard", "cafeteria" and "cafeteria diet + GSPE" treatments (n = 4/group) for 9 weeks (the last 4 weeks, GSPE/vehicle) of treatment, administering the extract/vehicle at diurnal or nocturnal times (ZT0 or ZT12). For circadian assessment, one hour after turning the light on (ZT1), animals were sacrificed every 6 h (ZT1, ZT7, ZT13 and ZT19). Interestingly, GSPE was able to restore the rhythm on clock hepatic genes (Bmal1, Per2, Cry1, Rorα), as this correction was more evident in nocturnal treatment. Additionally, during nocturnal treatment, an increase in hepatic fusion genes and a decrease in fission genes were observed. Regarding mitochondrial complex activity, there was a strong effect of cafeteria diet at nearly all ZTs, and GSPE was able to restore activity at discrete ZTs, mainly in the diurnal treatment (ZT0). Furthermore, a differential behavior was observed in tricarboxylic acid (TCA) metabolites between GSPE diurnal and nocturnal administration times. Therefore, GSPE may serve as a nutritional preventive strategy in the recovery of hepatic-related metabolic disease by modulating mitochondrial dynamics, which is concomitant to the restoration of the hepatic circadian machinery.


Systematic analysis of nutrigenomic effects of polyphenols related to cardiometabolic health in humans - Evidence from untargeted mRNA and miRNA studies

Tatjana Ruskovska 1, Irena Budić-Leto 2, Karla Fabiola Corral-Jara 3, Vladimir Ajdžanović 4, Anna Arola-Arnal 5, Francisca Isabel Bravo 5, Georgia-Eirini Deligiannidou 6, Jaroslav Havlik 7, Milkica Janeva 1, Elena Kistanova 8, Christos Kontogiorgis 6, Irena Krga 9, Marika Massaro 10, Marko Miler 4, Hicham Harnafi 11, Verica Milosevic 4, Christine Morand 3, Egeria Scoditti 10, Manuel Suárez 5, David Vauzour 12, Dragan Milenkovic 13

Cardiovascular and metabolic disorders present major causes of mortality in the ageing population. Polyphenols present in human diets possess cardiometabolic protective properties, however their underlying molecular mechanisms in humans are still not well identified. Even though preclinical and in vitro studies advocate that these bioactives can modulate gene expression, most studies were performed using targeted approaches. With the objective to decipher the molecular mechanisms underlying polyphenols cardiometabolic preventive properties in humans, we performed integrative multi-omic bioinformatic analyses of published studies which reported improvements of cardiometabolic risk factors following polyphenol intake, together with genomic analyses performed using untargeted approach. We identified 5 studies within our criteria and nearly 5000 differentially expressed genes, both mRNAs and miRNAs, in peripheral blood cells. Integrative bioinformatic analyses (e.g. pathway and gene network analyses, identification of transcription factors, correlation of gene expression profiles with those associated with diseases and drug intake) revealed that these genes are involved in the processes such as cell adhesion and mobility, immune system, metabolism, or cell signaling. We also identified 27 miRNAs known to regulate processes such as cell cytoskeleton, chemotaxis, cell signaling, or cell metabolism. Gene expression profiles negatively correlated with expression profiles of cardiovascular disease patients, while a positive correlation was observed with gene expression profiles following intake of drugs against cardiometabolic disorders. These analyses further advocate for health protective effects of these bioactives against age-associated diseases. In conclusion, polyphenols can exert multi-genomic modifications in humans and use of untargeted methods coupled with bioinformatic analyses represent the best approach to decipher molecular mechanisms underlying healthy-ageing effects of these bioactives.


Grape Seed Proanthocyanidins Mitigate the Disturbances Caused by an Abrupt Photoperiod Change in Healthy and Obese Rats

Jorge R Soliz-Rueda 1, Raúl López-Fernández-Sobrino 1, Francisca Isabel Bravo 1, Gerard Aragonès 1, Manuel Suarez 1, Begoña Muguerza 1

Variations in the light/dark cycle and obesogenic diets trigger physiological and behavioral disorders. Proanthocyanidins, in addition to their healthy properties, have recently demonstrated a modulating effect on biological rhythms. Therefore, the aim of this study was to evaluate the administration of a grape seed proanthocyanidin-rich extract (GSPE) to mitigate the disruption caused by a sudden photoperiod change in healthy and cafeteria (CAF)-diet obese rats. For this, 48 photoperiod-sensitive Fischer 344 rats were fed standard or CAF diets for 6 weeks under a standard (12 h light/day, L12) conditions. Then, rats were switched to a long (18 h light/day, L18) or short (6 h light/day, L6) photoperiod and administered vehicle or GSPE (25 mg/kg) for 1 week. Body weight (BW) and food intake (FI) were recorded weekly. Animal activity and serum hormone concentrations were studied before and after the photoperiod change. Hormone levels were measured both at 3 h (ZT3) and 15 h (ZT15) after the onset of light. Results showed the impact of the CAF diet and photoperiod on the BW, FI, activity, and hormonal status of the animals. GSPE administration resulted in an attenuation of the changes produced by the photoperiod disruption. Specifically, GSPE in L6 CAF-fed rats reduced serum corticosterone concentration, restoring its circadian rhythm, increased the T3-to-T4 ratio, and increased light phase activity, while under L18, it decreased BW and testosterone concentration and increased the animal activity. These results suggest that GSPE may contribute to the adaptation to the new photoperiods. However, further studies are needed to elucidate the metabolic pathways and processes involved in these events.


Cardioprotective Properties of Phenolic Compounds: A Role for Biological Rhythms

Cristina Torres-Fuentes 1, Manuel Suárez 1, Gerard Aragonès 1, Miquel Mulero 1, Javier Ávila-Román 1, Anna Arola-Arnal 1, Maria Josepa Salvadó 1, Lluís Arola 1, Francisca Isabel Bravo 1, Begoña Muguerza 1

Cardiovascular diseases (CVD) are the leading cause of deaths worldwide and their prevalence is continuously increasing. Available treatments may present several side effects and therefore the development of new safer therapeutics is of interest. Phenolic compounds have shown several cardioprotective properties helpful in reducing different CVD risk factors such as inflammation, elevated blood pressure, hyperlipidemia, or endothelial dysfunction. These factors are significantly influenced by biological rhythms which are in fact emerging as key modulators of important metabolic and physiological processes. Thus, increased events of CVD have been observed under circadian rhythm disruption or in winter versus other seasons. These rhythms can also affect the functionality of phenolic compounds. Indeed, different effects have been observed depending on the administration time or under different photoperiods. Therefore, in this review the focus will be on the potential of phenolic compounds as therapeutics to prevent CVD via biological rhythm modulation.


Effects of an Optimized Aged Garlic Extract on Cardiovascular Disease Risk Factors in Moderate Hypercholesterolemic Subjects: A Randomized, Crossover, Double-Blind, Sustainedand Controlled Study

Rosa M Valls 1, Judit Companys 2, Lorena Calderón-Pérez 2, Patricia Salamanca 1, Laura Pla-Pagà 2, Berner Andrée Sandoval-Ramírez 1, Antonio Bueno 3, Jose Puzo 3, Anna Crescenti 2, Josep M Del Bas 2, Antoni Caimari 4, Aurora Salamanca 5, Alberto E Espinel 5, Anna Pedret 1, Lluís Arola 2 6, Rosa Solà 1 7

The consumption of aged black garlic (ABG) has been related to improvements in several cardiovascular disease (CVD) risk factors. However, the extent of the beneficial effects depends on the garlic aging process and the amount and type of chemical compounds accumulated. The main objective of this study was to assess the effect of daily intake of a well-characterized ABG extract with a standardized S-allyl-L-cysteine (SAC) yield in combination with dietary recommendations regarding CVD risk factors in individuals with moderate hypercholesterolemia. Sixty-seven hypercholesterolemic individuals with low-density lipoprotein cholesterol levels ≥115 mg/dL were randomized in a crossover, double-blind, sustained, and controlled intervention study. The participants consumed 250 mg (1.25 mg SAC)/tablet/day ABG or a placebo for 6 weeks, with 3 weeks of washout. Blood and pulse pressure and other CVD risk biomarkers were determined at the beginning and end of each intervention. At 6 weeks, ABG extract reduced diastolic blood pressure (DBP) (mean (95% CI) -5.85 (-10.5; -1.3) mm Hg) compared to the placebo, particularly in men with a DBP > 75 mm Hg. The consumption of an improved ABG extract with 1.25 mg of SAC decreased DBP, particularly in men with moderate hypercholesterolemia. The potential beneficial effects of ABG may contribute to obtaining an optimal DBP.


In-Season Consumption of Locally Produced Tomatoes Decreases Cardiovascular Risk Indices

Ma Josefina Ruiz de Azua 1, Álvaro Cruz-Carrión 1, Begoña Muguerza 1, Gerard Aragonès 1, Anna Arola-Arnal 1, María Paz Romero 2, Francisca Isabel Bravo 1, Manuel Suarez 1

Tomatoes are widely consumed worldwide at any time of the year. However, depending on the variety, they have a characteristic season. We evaluated the consequences metabolic of consumption of Ekstasis tomatoes from different geographical origin and in different seasons in Fischer 344 rats. The hepatic gene expression of key enzymes in lipid metabolism was also evaluated. Animals were classified in three photoperiods (L6, L12, and L18) and in three treatments (vehicle: VH; local tomato: LT; and non-local tomato: nLT). We measured serum metabolic parameters and the gene expression of liver enzymes related to lipid metabolism (Acc1, Cpt1a, Had, Fas1, Srebp-1c, Fatp5, Cd36). LT consumption in season decreased cardiovascular risk 1 and coefficient atherogenic by 1.81 (p = 0.031) and in L6 decreased TAG and glucose (p = 0.046; p = 0.024). The L18-LT animals had decreased total cholesterol (p = 0.029) and gene expression of Srebp1-c (p = 0.022) but increased expression of Acc1 (p = 0.032). The treatments significantly affected the expression of Acc1 and Fas1 in the liver and the levels of serum TAG and glucose. A significant effect of photoperiod on serum concentration of glucose, insulin, HOMA index, and on the hepatic expression of Srep1-c, Fas1, and Acc1 was observed.


Microbiota Dysbiosis and Gut Barrier Dysfunction Associated with Non-Alcoholic Fatty Liver Disease Are Modulated by a Specific Metabolic Cofactors' Combination

Sergio Quesada-Vázquez 1, Caitlin Bone 2, Shikha Saha 3, Iris Triguero 1, Marina Colom-Pellicer 4, Gerard Aragonès 4, Falk Hildebrand 2 5, Josep M Del Bas 6, Antoni Caimari 6, Naiara Beraza 2 3, Xavier Escoté 1

The gut is a selective barrier that not only allows the translocation of nutrients from food, but also microbe-derived metabolites to the systemic circulation that flows through the liver. Microbiota dysbiosis occurs when energy imbalances appear due to an unhealthy diet and a sedentary lifestyle. Dysbiosis has a critical impact on increasing intestinal permeability and epithelial barrier deterioration, contributing to bacterial and antigen translocation to the liver, triggering non-alcoholic fatty liver disease (NAFLD) progression. In this study, the potential therapeutic/beneficial effects of a combination of metabolic cofactors (a multi-ingredient; MI) (betaine, N-acetylcysteine, L-carnitine, and nicotinamide riboside) against NAFLD were evaluated. In addition, we investigated the effects of this metabolic cofactors' combination as a modulator of other players of the gut-liver axis during the disease, including gut barrier dysfunction and microbiota dysbiosis. Diet-induced NAFLD mice were distributed into two groups, treated with the vehicle (NAFLD group) or with a combination of metabolic cofactors (NAFLD-MI group), and small intestines were harvested from all animals for histological, molecular, and omics analysis. The MI treatment ameliorated gut morphological changes, decreased gut barrier permeability, and reduced gene expression of some proinflammatory cytokines. Moreover, epithelial cell proliferation and the number of goblet cells were increased after MI supplementation. In addition, supplementation with the MI combination promoted changes in the intestinal microbiota composition and diversity, as well as modulating short-chain fatty acids (SCFAs) concentrations in feces. Taken together, this specific combination of metabolic cofactors can reverse gut barrier disruption and microbiota dysbiosis contributing to the amelioration of NAFLD progression by modulating key players of the gut-liver axis.


Reduction of Obesity and Insulin Resistance through Dual Targeting of VAT and BAT by a Novel Combination of Metabolic Cofactors

Sergio Quesada-Vázquez 1, Anna Antolín 1, Marina Colom-Pellicer 2, Gerard Aragonès 2, Laura Herrero 3 4, Josep Maria Del Bas 5, Antoni Caimari 5, Xavier Escoté 1

Obesity is an epidemic disease worldwide, characterized by excessive fat accumulation associated with several metabolic perturbations, such as metabolic syndrome, insulin resistance, hypertension, and dyslipidemia. To improve this situation, a specific combination of metabolic cofactors (MC) (betaine, N-acetylcysteine, L-carnitine, and nicotinamide riboside) was assessed as a promising treatment in a high-fat diet (HFD) mouse model. Obese animals were distributed into two groups, orally treated with the vehicle (obese + vehicle) or with the combination of metabolic cofactors (obese + MC) for 4 weeks. Body and adipose depots weights; insulin and glucose tolerance tests; indirect calorimetry; and thermography assays were performed at the end of the intervention. Histological analysis of epidydimal white adipose tissue (EWAT) and brown adipose tissue (BAT) was carried out, and the expression of key genes involved in both fat depots was characterized by qPCR. We demonstrated that MC supplementation conferred a moderate reduction of obesity and adiposity, an improvement in serum glucose and lipid metabolic parameters, an important improvement in lipid oxidation, and a decrease in adipocyte hypertrophy. Moreover, MC-treated animals presented increased adipose gene expression in EWAT related to lipolysis and fatty acid oxidation. Furthermore, MC supplementation reduced glucose intolerance and insulin resistance, with an increased expression of the glucose transporter Glut4; and decreased fat accumulation in BAT, raising non-shivering thermogenesis. This treatment based on a specific combination of metabolic cofactors mitigates important pathophysiological characteristics of obesity, representing a promising clinical approach to this metabolic disease.


Do You Have Problems When Reproducing Bioactivities of Food or Food Components? The Importance of Biological Rhythms

Anna Arola-Arnal 1, Manuel Suárez 1

With the onset of omics sciences, in the 20th century, nutritional studies evolved to investigate the effects of diet at a molecular level, giving rise to nutritional genomics, which includes both nutrigenomics and nutrigenetics [...].


The effects of grape seed proanthocyanidins in cafeteria diet-induced obese Fischer 344 rats are influenced by faecal microbiota in a photoperiod dependent manner

Verónica Arreaza-Gil 1, Iván Escobar-Martínez 1, Begoña Muguerza 1, Gerard Aragonès 1, Manuel Suárez 1, Cristina Torres-Fuentes 1, Anna Arola-Arnal 1

Polyphenols are of high interest due to their beneficial health effects, including anti-obesity properties. The gut microbiota may play an important role in polyphenol-mediated effects as these bacteria are significantly involved in the metabolism of polyphenols. Moreover, seasonal rhythms have been demonstrated to influence both the gut microbiota composition and polyphenol bioavailability. Thus, the goal of this study was to evaluate the impact of photoperiods and microbiota on polyphenol functionality in an obesogenic context. Towards this aim, cafeteria diet-fed Fischer 344 rats were housed under three different photoperiod conditions (L6: 6 h of light, L12: 12 h of light and L18: 18 h of light) for 9 weeks. During the last 4 weeks of the experiment, rats were daily administered with an oral dose of a grape seed proanthocyanidin extract (GSPE) (25 mg per kg body weight). Additionally, rats treated with GSPE and an antibiotic cocktail (ABX) in their drinking water were included for a better understanding of the gut microbiota role in GSPE functionality. Vehicle and non-ABX treated rats were included as controls. GSPE decreased body weight gain and fat depots only under L18 conditions. Interestingly, the gut microbiota composition was strongly altered in this photoperiod. GSPE + ABX-treated rats gained significantly less body weight compared to the rats of the rest of the treatments under L18 conditions. These results suggest that GSPE functionality is modulated by the gut microbiota in a photoperiod dependent manner. These novel findings corroborate seasonal rhythms as key factors that must be taken into account when investigating the effects of polyphenols in the treatment or prevention of chronic diseases.


9-PAHPA long term intake in DIO and db/db mice ameliorates insulin sensitivity but has few effects on obesity and associated metabolic disorders

Béatrice Bonafos 1, Antonio J Cortés-Espinar 2, Laurence Balas 3, Laurence Pessemesse 1, Karen Lambert 4, Melha Benlebna 1, Sylvie Gaillet 1, François Pelletier 1, Pierre Delobel 1, Javier Ávila-Román 5, Miquel Mulero Abellán 2, Christelle Bertrand-Gaday 1, Thierry Durand 3, Charles Coudray 1, François Casas 6, Christine Feillet-Coudray 7

Branched fatty acid esters of hydroxy fatty acids are endogenous lipids reported to have antidiabetic and anti-inflammatory effects. Recently, we showed that 9-palmitic acid esters of hydroxypalmitic acid (9-PAHPA) and 9-oleic acid esters of hydroxypalmitic acid increased insulin sensitivity in mice when incorporated to a chow diet or to a high fat and high sucrose diet. However, preventive supplementation with 9-PAHPA and 9-oleic acid esters of hydroxypalmitic acid in high fat and high sucrose diet mice did not impair significant weight gain or the development of hyperglycemia. The aim of this work was therefore to study whether in two animal models of obesity, namely the classical diet-induced obesity (DIO) and the db/db mice, 9-PAHPA may have beneficial effects against obesity and liver and skeletal muscle metabolic dysfunction. In DIO mice, we observed that 9-PAHPA increased body weight and fat mass. In line with this observation, we found that 9-PAHPA supplementation decreased energy expenditure. In liver and in skeletal muscle, mitochondrial activities and oxidative stress parameters were not modified by 9-PAHPA supplementation. In db/db mice, 9-PAHPA had no effect on the dramatic weight gain and hyperglycemia. In addition, 9-PAHPA supplementation did not correct either the hepatomegaly and hepatic steatosis or the severe muscle atrophy recorded compared with db/+ animals. Likewise, supplementation with 9-PAHPA did not impact the different metabolic parameters analyzed, either in the liver or in the skeletal muscles. However, it decreased insulin resistance in DIO and db/db mice. In conclusion, our study indicated that a long-term intake of 9-PAHPA in DIO and db/db mice improved insulin sensitivity but had only few effects on obesity and associated metabolic disorders.


(Poly)phenolic composition of tomatoes from different growing locations and their absorption in rats: A comparative study

Álvaro Cruz-Carrión 1, Luca Calani 2, Ma Josefina Ruiz de Azua 1, Pedro Mena 3, Daniele Del Rio 1, Manuel Suárez 4, Anna Arola-Arnal 1

The aim of this work was to address whether the growing location of tomato could generate a different (poly)phenol profile able to affect both in vivo absorption and (poly)phenol metabolite pattern upon tomato consumption. uHPLC-MSn analyses allowed to obtain a detailed (poly)phenol profile of tomatoes from two locations in Spain, quantifying 57 (poly)phenolic compounds. However, local and non-local tomatoes showed a different concentration of their native (poly)phenols, which could be attributed to diverse cultivation origin. Rat serum was analysed after an acute tomato feeding. Seven phenolic metabolites were quantified through uHPLC-MSn. Pharmacokinetic parameters were further evaluated, revealing different serum concentrations of (poly)phenolic metabolites between tomatoes. The maximum peak serum concentrations, reached mainly after 2 h after ingestion, led to suppose that serum metabolites were mostly derived from absorption in the upper gastrointestinal tract. The growing location of tomatoes affected both the content of native (poly)phenols and their in vivo absorption.


Eat Fruits In-Season to Give Rhythm to Your Life

Álvaro Cruz-CarriónMa Josefina Ruiz de AzuaFrancisca Isabel BravoBegoña MuguerzaManuel SuáRezAnna Arola-Arnal

Do you think that eating fruit in summer has the same effect on your body as does eating it in winter? Scientific evidence says no. Fruits contain polyphenols, which are substances produced by plants in response to the growing conditions. When animals eat these fruits, polyphenols signal animals' bodies to adapt to the seasons. For example, bears eat berries in late summer because these fruits provide essential substances needed during hibernation. It has been observed that the effect of these fruit substances is affected by biological rhythms, which are chemical cycles that animals' bodies follow that vary throughout the year. Thus, eating fruit in- or out-of-season generates different effects in your body. Eating fruit in-season is associated with optimal health effects. Hence, we must eat fruits in-season so that the rhythms of our lives are synchronized with the seasons.


Impact of Seasonal Consumption of Local Tomatoes on the Metabolism and Absorption of (Poly)Phenols in Fischer Rats

Álvaro Cruz-Carrión 1 2 3, Luca Calani 4, Ma Josefina Ruiz de Azua 1, Pedro Mena 4, Daniele Del Rio 4, Anna Arola-Arnal 1, Manuel Suárez 1

Consuming (poly)phenol-rich fruits and vegetables, including tomato, is associated with health benefits. The health effects of tomato (poly)phenolic compounds have been attributed to their metabolites rather than parent compounds and their bioavailability can be modulated by several factors. This study aimed to evaluate the effect of seasonal consumption of local tomatoes on their (poly)phenol bioavailability. For this, (poly)phenol absorption and metabolism were evaluated by ultra-high-performance liquid chromatography coupled with mass spectrometry and linear ion trap mass spectrometric (uHPLC-MSn) after chronic tomato consumption in Fischer rats exposed to three photoperiods mimicking the seasonal daylight schedule. Tomatoes from two locations in Spain (LT, local tomatoes and NLT, non-local tomatoes) were used in this in vivo feeding study. The bioavailability of tomato (poly)phenols depended on the photoperiod to which the rats were exposed, the metabolite concentrations significantly varying between seasons. In-season tomato consumption allowed obtaining the highest concentration of total circulating metabolites. In addition, the origin of the tomato administered generated marked differences in the metabolic profiles, with higher serum concentrations reached upon NLT ingestion. We concluded that in-season tomato consumption led to an increase in (poly)phenol circulation, whereas LT consumption showed lower circulating metabolites than NLT ones. Thus, the origin of the tomato and the seasonal daylight schedule affect the bioavailability of tomato (poly)phenols, which could also affect their bioactivity.


Alterations in Metabolome and Microbiome Associated with an Early Stress Stage in Male Wistar Rats: A Multi-Omics Approach

Julia Hernandez-Baixauli 1, Pere Puigbò 1 2 3, Nerea Abasolo 4, Hector Palacios-Jordan 4, Elisabet Foguet-Romero 4, David Suñol 5, Mar Galofré 5, Antoni Caimari 1, Laura Baselga-Escudero 1, Josep M Del Bas 1, Miquel Mulero 2 6

Stress disorders have dramatically increased in recent decades becoming the most prevalent psychiatric disorder in the United States and Europe. However, the diagnosis of stress disorders is currently based on symptom checklist and psychological questionnaires, thus making the identification of candidate biomarkers necessary to gain better insights into this pathology and its related metabolic alterations. Regarding the identification of potential biomarkers, omic profiling and metabolic footprint arise as promising approaches to recognize early biochemical changes in such disease and provide opportunities for the development of integrative candidate biomarkers. Here, we studied plasma and urine metabolites together with metagenomics in a 3 days Chronic Unpredictable Mild Stress (3d CUMS) animal approach that aims to focus on the early stress period of a well-established depression model. The multi-omics integration showed a profile composed by a signature of eight plasma metabolites, six urine metabolites and five microbes. Specifically, threonic acid, malic acid, alpha-ketoglutarate, succinic acid and cholesterol were proposed as key metabolites that could serve as key potential biomarkers in plasma metabolome of early stages of stress. Such findings targeted the threonic acid metabolism and the tricarboxylic acid (TCA) cycle as important pathways in early stress. Additionally, an increase in opportunistic microbes as virus of the Herpesvirales was observed in the microbiota as an effect of the primary stress stages. Our results provide an experimental biochemical characterization of the early stage of CUMS accompanied by a subsequent omic profiling and a metabolic footprinting that provide potential candidate biomarkers.


Serum lysophospholipidome of dietary origin as a suitable susceptibility/risk biomarker of human hypercholesterolemia: Letter to the editor

Lorena Calderón-Pérez 1, Susana Suárez-García 2, Anna Pedret 3, Manuel Suárez 4, Elisabet Llauradó 5, Laura Rubió 6, Josep M Del Bas 7, Antoni Caimari 7, Francesc Puiggrós 7, Lluís Arola 2, Rosa Solà 8, Rosa M Valls 1


Hesperidin Bioavailability Is Increased by the Presence of 2S-Diastereoisomer and Micronization-A Randomized, Crossover and Double-Blind Clinical Trial

Anna Crescenti 1, Antoni Caimari 2, Juan María Alcaide-Hidalgo 1, Roger Mariné-Casadó 1, Rosa M Valls 3, Judit Companys 1, Patricia Salamanca 3, Lorena Calderón-Pérez 1, Laura Pla-Pagà 1, Anna Pedret 3, Antoni Delpino-Rius 4, Pol Herrero 4, Iris Samarra 4, Lluís Arola 1 5, Rosa Solà 3 6, Josep M Del Bas 1

Hesperidin is a flavanone abundantly found in citrus fruits for which health beneficial effects have been reported. However, hesperidin shows a low bioavailability among individuals. The aim of this study was to evaluate the effects of the micronization process and 2R- and 2S-hesperidin diastereoisomers ratio on hesperidin bioavailability. In a first phase, thirty healthy individuals consumed 500 mL of orange juice with 345 mg of hesperidin, and the levels of hesperidin metabolites excreted in urine were determined. In the second phase, fifteen individuals with intermediate hesperidin metabolite levels excreted in urine were randomized in a crossover, postprandial and double-blind intervention study. Participants consumed 500 mg of the hesperidin-supplemented Hesperidin epimeric mixture (HEM), the micronized Hesperidin epimeric mixture (MHEM) and micronized 2S-Hesperidin (M2SH) in each study visit with 1 week of washout. Hesperidin metabolites and catabolites were determined in blood and urine obtained at different timepoints over a 24 h period. The bioavailability-relative urinary hesperidin excretion (% of hesperidin ingested)-of M2SH (70 ± 14%) formed mainly by 2S-diastereoisomer was significantly higher than the bioavailability of the MHEM (55 ± 15%) and HEM (43 ± 8.0%), which consisted of a mixture of both hesperidin diastereoisomers. Relative urinary excretion of hesperidin metabolites for MHEM (9.2 ± 1.6%) was significantly higher compared to the HEM (5.2 ± 0.81%) and M2SH (3.6 ± 1.0%). In conclusion, the bioavailability of 2S-hesperidin extract was higher compared to the standard mixture of 2S-/2R-hesperidin extract due to a greater formation of hesperidin catabolites. Furthermore, the micronization process increased hesperidin bioavailability.


"Som la Pera"; A School-Based, Peer-Led Social Marketing Intervention to Engage Spanish Adolescents in a Healthy Lifestyle: A Parallel-Cluster Randomized Controlled Study

Magaly Aceves-Martins 1, Elisabet Llauradó 1, Lucia Tarro 1, Ignasi Papell-Garcia 2, Jordi Prades-Tena 3, Helle Kettner-Høeberg 3, Francesc Puiggròs 2, Lluís Arola 2 4, Amy Davies 5, Montse Giralt 1, Rosa Solà 1 2 6

Background: Encouraging adolescents to adopt healthy lifestyles can be challenging. The aim of the "Som la Pera" study was to engage adolescents by applying new strategies to increase both their fruit and vegetable consumption and their physical activity (PA), while reducing their sedentary behavior (screen time per day). Methods: In disadvantaged neighborhoods of Reus (Spain), two high schools were randomly assigned to the intervention (n = 169 adolescents, 13- to 16-year old) and two were assigned to the control group (n = 223 adolescents, 13- to 16-year old). The intervention, which lasted 12 months and spanned two academic years (2013-2015), used social marketing (SM) to improve healthy choices. The peer-led strategy involved 5 adolescents, who designed and implemented 10 activities as challenges for their 169 school-aged peers. The control group received no intervention. To assess self-reported lifestyles in both groups, the Health Behavior in School-aged Children survey was used at baseline and end of study. Results: After 12 months, the weekly PA improved in the intervention group vs. the control one (p = 0.047). When male and female groups were examined separately, the improvement in PA remained with a borderline significance only in the male group (p = 0.050). Conclusions: A school-based, peer-led SM intervention designed and implemented by adolescents attending high schools in low-income neighborhoods promoted an increase in PA, particularly in male adolescents 13 to 16 years of age. No effect on fruit or vegetable consumption or screen time was observed.


Maternal Supplementation with a Cocoa Extract during Lactation Deeply Modulates Dams' Metabolism, Increases Adiponectin Circulating Levels and Improves the Inflammatory Profile in Obese Rat Offspring

Roger Mariné-Casadó 1, Cristina Domenech-Coca 1, Anna Crescenti 1, Miguel Ángel Rodríguez Gómez 2, Josep Maria Del Bas 3, Lluís Arola 4, Noemí Boqué 1, Antoni Caimari 3

High-flavonoid cocoa consumption has been associated with beneficial properties. However, there are scarce data concerning the effects of maternal cocoa intake on dams and in their progeny. Here, we evaluated in rats whether maternal supplementation with a high-flavan-3-ol cocoa extract (CCX) during lactation (200 mg.kg-1.day-1) produced beneficial effects on dams and in their normoweight (STD-CCX group) and cafeteria-fed obese (CAF-CCX group) adult male offspring. Maternal intake of CCX significantly increased the circulating levels of adiponectin and decreased the mammary gland lipid content of dams. These effects were accompanied by increased energy expenditure and circulating free fatty acids, as well as by a higher expression of lipogenic and adiponectin-related genes in their mammary glands, which could be related to a compensatory mechanism to ensure enough lipid supply to the pups. CCX consumption programmed both offspring groups towards increased plasma total adiponectin levels, and decreased liver weight and lean/fat ratio. Furthermore, CAF-CCX progeny showed an improvement of the inflammatory profile, evidenced by the significant decrease of the monocyte chemoattractant protein-1 (MCP-1) circulating levels and the mRNA levels of the gene encoding the major histocompatibility complex, class II invariant chain (Cd74), a marker of M1 macrophage phenotype, in the epididymal white adipose tissue. Although further studies are needed, these findings can pave the way for using CCX as a nutraceutical supplement during lactation.


Effects of enriched seafood sticks (heat-inactivated B. animalis subsp. lactis CECT 8145, inulin, omega-3) on cardiometabolic risk factors and gut microbiota in abdominally obese subjects: randomized controlled trial

Judit Companys 1 2, Lorena Calderón-Pérez 1 2, Laura Pla-Pagà 1 2, Elisabet Llauradó 2, Berner Andrée Sandoval-Ramirez 2, Maria José Gosalbes 3 4, Ainara Arregui 5, Maddi Barandiaran 5, Antoni Caimari 6, Josep Maria Del Bas 7, Lluís Arola 8, Rosa M Valls 9 10, Rosa Solà 11 12 13, Anna Pedret 1 2

Purpose: To assess the effects of enriched seafood sticks with postbiotic and bioactive compounds on CMD risk factors and the gut microbiota in abdominally obese individuals.

Methods: Randomized, double-blind, parallel, placebo-controlled trial with abdominally obese individuals. Participants (n = 120) consumed 50 g/day of enriched seafood sticks containing SIAP: (1010 colony forming units (CFUs) of heat-inactivated B. animalis subsp. lactis CECT8145, 370 mg/day omega 3 and 1.7 g/day inulin), or 50 g/day of placebo seafood sticks for 12 weeks. At 12 weeks, an acute single-dose study of 4 h was performed.

Results: Sustained SIAP2 consumption significantly decreased the insulin by - 5.25 mg/dL and HOMA-IR (homeostatic Model Assessment of Insulin Resistance) by - 1.33. In women, SIAP2 consumption significantly decreased the pulse pressure (PP) by - 4.69 mmHg. Gut microbiota analysis showed a negative association between glycemic parameter reduction and Alistipes finegoldii and Ruminococcaceae, and between PP reduction and Prevotella 9-ASV0283 and Christensenellaceae. In the acute single dose-study 4-h, SIAP2 consumption produced a lower increase in the postprandial circulating triglyceride levels [23.9 (7.03) mg/dL (mean [standard error])] than the observed with placebo [49.0 (9.52)] mg/dL.

Conclusion: In abdominally obese individuals, enriched seafood sticks induce a potential protection against type 2 diabetes development by the reduction in the insulin and HOMA-IR; and in cardiovascular disease, in women, by the PP reduction. These effects are accompanied by partial changes in the gut microbiota composition. The enriched seafood sticks reduce the atherogenic triglyceride postprandial concentrations. Our results support the use of enriched seafood sticks as a complementary strategy in the management of CMD risk factors.