New insights into the plasma and urinary metabolomic signatures of spontaneously hypertensive rats Celia Rodríguez-Pérez, Alejandra Vázquez-Aguilar, Oscar Daniel Rangel Huerta, Estefanía Sánchez-Rodríguez, Ángel Gil, Caridad Díaz, Félix Vargas, María D. Mesa Plos One, 2026 Background Hypertension is a major risk factor associated with cardiovascular diseases and one of the leading causes of premature death. Metabolomics is a useful tool for studying in vivo metabolic profiles to better understand the pathogenesis of diseases such as hypertension. This work aimed to explore the plasma and urinary non-targeted metabolic profile of 16-week-old spontaneously hypertensive rats (SHR) to identify new metabolomic profiles associated with hypertensive phenotypical characteristics. Methods Plasma and 24-hours urine samples were collected from 10 SHR and 10 age-matched normotensive Wistar-Kyoto 16-week-old male rats. Plasma and urinary metabolic profiles were investigated using high-performance liquid chromatography quadrupole time of flight coupled to mass spectroscopy followed by multivariate statistical analysis. The mummichog pathway enrichment analysis was used to integrate metabolomics data into biological contexts. Results A total of 16 differential metabolites were found in plasma and 13 differential metabolites in urine from SHR compared to normotensive rats. Differences in some microbiota-derived metabolites suggest changes in the gut microbiota associated with hypertension in our experimental model. The mummichog algorithm has recognized that hypertensive metabolism is associated with the altered metabolism of steroid hormones, bile acid, and purines. Conclusions This work highlights the importance of metabolomics as a tool for the identification of biomarkers related to hypertension and its consequences in SHR. The findings suggest that alterations in the metabolism of steroid hormones, bile acids, and purines, as well as metabolites derived from the intestinal microbiota are associated with the presence of hypertension. More research is needed to further understand their role during hypertension.
Chemometrics-guided plasma lipidomics insights underlying the decrease of plasma total cholesterol and LDLc in children with hypercholesterolemia following habitual intake of an EVOO-based phytosterols-enriched spreadable cream Pascual García-Pérez, Alejandra Vázquez-Aguilar, Estefanía Sánchez-Rodríguez, José Manuel Jurado-Castro, Belén Pastor-Villaescusa, Mercedes Gil-Campos, Óscar Daniel Rangel-Huerta, Ángel Gil, Concepción M. Aguilera, María D. Mesa, Gabriele Rocchetti, Luigi Lucini Metabolomics, 2025 BACKGROUND: Higher LDL cholesterol levels are associated with increased risk of cardiovascular disease, and early intervention in children and regular follow-up are keystones for its prevention. Hypercholesterolemia can be partially delayed adopting healthy lifestyles, including healthy diets such as the Mediterranean diet and physical activity. Extra-virgin olive oil (EVOO), the main edible oil in the Mediterranean diet, has demonstrated cardiovascular benefits. OBJECTIVE: The present randomized double-blind crossover clinical study aimed to evaluate the efficacy of the daily consumption of a spreadable cream based on extra virgin olive oil enriched in phytosterols (0.04-0.06 g plant sterols/kg of body weight/day) for 8 weeks separated by a wash-out period of 4 weeks on children (6-18 years old) with hypercholesterolemia. METHODS: Fifty children were included although only 23 reported at least 70% of compliance and were used for final analyses. We used a dual approach combining the determination of anthropometric and biochemical plasma parameters and the lipidomics profile of plasma samples. RESULTS: Spreadable cream enriched in phytosterols consumption led to a decrease in total cholesterol and low-density lipoprotein cholesterol plasma levels. Multivariate analysis of the lipidomic plasma profile indicated that phospholipids and sphingolipids were mostly involved in modulating the observed effects. CONCLUSIONS: Overall, this study provides a multi-level integrated approach for the characterization of a novel functional food to counteract hypercholesterolemia from a nutritional perspective. CLINICAL TRIAL REGISTRY: Clinical Trial Registry number and website where it was obtained https://clinicaltrials.gov/ct2/show/NCT05460208.
MELI-POP Study: MEditerranean LIfestyle in Pediatric Obesity Prevention. Study protocol for a randomized controlled trial Pilar De Miguel-Etayo, Katherine Flores-Rojas, Rocío Vázquez-Cobela, Jose Manuel Jurado-Castro, Alicia Larruy-García, Rosaura Picans-Leis, Cristina Castro-Collado, Belén Pastor-Villaescusa, Francisco Jesus Llorente-Cantarero, Concepción M. Aguilera, Mercedes Gil-Campos, Rosaura Leis, Luis A. Moreno, , V. Vučinić, M. L. Miguel Berges, I. Maneschy, A. Jimeno-Martinez, D. Córdoba, A. Ayala-Marín, M. Seral-Cortés, I. Benedicto, G. Bueno Lozano, M. J. Blasco Pérez-Aramendia, M. L. Bartolomé Lalanza, P. Lobera, M. López-Campos, N. Martinez, I. Lostal, A. I. Ramírez, A. I. Gastón Faci, E. Atance Melendo, E. Zapata Mallen, R. Pardos Romano, L. M. Alutiz Lapeña, C. Viñas, E. Olmo Pérez, A. Santaliestra-Pasías, Á Matute Llorente, A. Agüero González de Lafuente, J. Rico Díaz, E. M. García García, O. Fernández Seijas, I. Izquierdo López, A. Romero Agrelo, L. Rivas Rodríguez, S. Mosteiro Losada, MJ. De la Torre-Aguilar, MA. Alarcón-Roldán, M. J. Cortés Sillero, A. Carmona Priego, N. Babio, S. De las Heras-Delgado, M. Gil-Segura, G. Mena-Sánchez, A. Pedraza, J. Salas-Salvadó, O. Salvadó, I. Valverde-Aguilera, M. Bustos-Aibar, Á. Torres-Martos, M. C. Rico, E. Sánchez-Rodríguez, I. Rueda-De Torre BMC Public Health, 2025 Childhood obesity is a significant public health challenge, with Mediterranean countries showing high prevalence rates. While genetic factors play a role, diet and physical activity (PA) are critical modifiable influences. Emphasizing healthy dietary patterns, like the Mediterranean diet, and promoting regular PA can help mitigate obesity risk. MELI-POP is a randomized controlled multi-center clinical trial in a cohort of children aged 3 to 6 years at baseline and being at obesity risk. The main objective consists on assessing the efficacy of an intervention during early childhood, considering a healthy lifestyle based on the promotion of a Mediterranean dietary pattern and regular PA, compared with a control group, on decreasing obesity incidence 5 and 10 years after the beginning of the intervention. It is expected to include 310 children, aged 3 to 6 years, having at baseline a normal weight or overweight according to the International Obesity Task Force (IOTF) criteria, and at least one parent having a body mass index > 25 kg/m2. The clinical trial has two arms and is performed in Spanish Primary Health Care centers. The control group receive usual care by healthcare professionals. The intervention group receive education on Mediterranean diet and PA, combined with the provision of extra-virgin olive oil and fish, in order to be consumed at least 2 times per week. Free PA sessions with a physical education teacher are also offered for the children (3 sessions of 60 min of moderate-vigorous PA per week). The participants’ adherence to the intervention is periodically monitored. The study primarily focuses on adiposity as the main outcome, with secondary outcomes encompassing dietary intake and eating habits, physical activity and lifestyle behaviors (including extracurricular sports, screen time, and sleep duration), physical fitness, biochemical parameters (such as inflammation and cardiometabolic biomarkers, fatty acids, and oxidation), as well as microbiota, genetic, omic and metabolomic analyses. Beneficial results are expected by preventing obesity during childhood, and associated comorbidities. ClinicalTrials.gov, ID: NCT04597281.
Genetic markers associated with bone strength and density in Rhode Island Red laying hens Qiaoxian Yue, Martin Johnsson, Peter W Wilson, Björn Andersson, Matthias Schmutz, Cristina Benavides, Nazaret Dominguez-Gasca, Estefania Sanchez-Rodriguez, Alejandro B Rodriguez-Navarro, Ian C Dunn, Dirk-Jan de Koning Poultry Science, 2025 Damage to the keel bone in commercial laying hens represent one of the greatest welfare issues in laying hens. This study aims to identify the DNA markers and candidate genes for bone strength and density traits in a Rhode Island Red laying hen population. We conducted genome-wide association studies (GWAS) on bone quality traits using a sample of 925 Rhode Island Red laying hens genotyped with a genotyping array consisting of 60 000 DNA markers. With a univariate linear mixed model, we identified 52 suggestive genetic markers located within 28 candidate genes that are associated with the humerus, keel, and tibia strength and density. We also found overlaps between the GWAS results for medullary bone score and tibia strength and density with published quantitative trait loci (QTL) for eggshell effective layer thickness and abdominal fat weight, respectively. Heritability estimates for the humerus stiffness, tibia stiffness, medullary bone score and minor bone diameter ranged from 0.21 to 0.34. Annotation term enrichment analysis of genes within 2 Megabases of suggestive markers found that mTOR signalling pathway, tryptophan metabolism, TGF-β signalling pathway, and apoptosis were significantly enriched. These loci do not overlap previously published associations, and thus appear to be novel.
Genetic markers associated with bone composition in Rhode Island Red laying hens Moh Sallam, Peter W. Wilson, Björn Andersson, Matthias Schmutz, Cristina Benavides, Nazaret Dominguez‑Gasca, Estefania Sanchez‑Rodriguez, Alejandro B. Rodriguez‑Navarro, Ian C. Dunn, Dirk‑Jan De Koning, Martin Johnsson Genetics Selection Evolution, 2023 Background Bone damage has welfare and economic impacts on modern commercial poultry and is known as one of the major challenges in the poultry industry. Bone damage is particularly common in laying hens and is probably due to the physiological link between bone and the egg laying process. Previous studies identified and validated quantitative trait loci (QTL) for bone strength in White Leghorn laying hens based on several measurements, including bone composition measurements on the cortex and medulla of the tibia bone. In a previous pedigree-based analysis, bone composition measurements showed heritabilities ranging from 0.18 to 0.41 and moderate to strong genetic correlations with tibia strength and density. Bone composition was measured using infrared spectroscopy and thermogravimetry. The aim of this study was to combine these bone composition measurements with genotyping data via a genome-wide association study (GWAS) to investigate genetic markers that contribute to genetic variance in bone composition in Rhode Island Red laying hens. In addition, we investigated the genetic correlations between bone composition and bone strength. Results We found novel genetic markers that are significantly associated with cortical lipid, cortical mineral scattering, medullary organic matter, and medullary mineralization. Composition of the bone organic matter showed more significant associations than bone mineral composition. We also found interesting overlaps between the GWAS results for tibia composition traits, particularly for cortical lipid and tibia strength. Bone composition measurements by infrared spectroscopy showed more significant associations than thermogravimetry measurements. Based on the results of infrared spectroscopy, cortical lipid showed the highest genetic correlations with tibia density, which was negative (− 0.20 ± 0.04), followed by cortical CO3/PO4 (0.18 ± 0.04). Based on the results of thermogravimetry, medullary organic matter% and mineral% showed the highest genetic correlations with tibia density (− 0.25 ± 0.04 and 0.25 ± 0.04, respectively). Conclusions This study detected novel genetic associations for bone composition traits, particularly those involving organic matter, that could be used as a basis for further molecular genetic investigations. Tibia cortical lipids displayed the strongest genetic associations of all the composition measurements, including a significantly high genetic correlation with tibia density and strength. Our results also highlighted that cortical lipid may be a key measurement for further avian bone studies.
Metabolomic-Based Studies of the Intake of Virgin Olive Oil: A Comprehensive Review Alejandra Vazquez-Aguilar, Estefania Sanchez-Rodriguez, Celia Rodriguez-Perez, Oscar Daniel Rangel-Huerta, Maria D. Mesa Metabolites, 2023 Virgin olive oil (VOO) is a high-value product from the Mediterranean diet. Some health and nutritional benefits have been associated with its consumption, not only because of its monounsaturated-rich triacylglycerols but also due to its minor bioactive components. The search for specific metabolites related to VOO consumption may provide valuable information to identify the specific bioactive components and to understand possible molecular and metabolic mechanisms implicated in those health effects. In this regard, metabolomics, considered a key analytical tool in nutritional studies, offers a better understanding of the regulatory functions of food components on human nutrition, well-being, and health. For that reason, the aim of the present review is to summarize the available scientific evidence related to the metabolic effects of VOO or its minor bioactive compounds in human, animal, and in vitro studies using metabolomics approaches.
Genetics of tibia bone properties of crossbred commercial laying hens in different housing systems Martin Johnsson, Helena Wall, Fernando A Lopes Pinto, Robert H Fleming, Heather A McCormack, Cristina Benavides-Reyes, Nazaret Dominguez-Gasca, Estefania Sanchez-Rodriguez, Ian C Dunn, Alejandro B Rodriguez-Navarro, Andreas Kindmark, Dirk-Jan de Koning G3 Genes Genomes Genetics, 2023 Osteoporosis and bone fractures are a severe problem for the welfare of laying hens, with genetics and environment, such as housing system, each making substantial contributions to bone strength. In this work, we performed genetic analyses of bone strength, bone mineral density, and bone composition, as well as body weight, in 860 commercial crossbred laying hens from 2 different companies, kept in either furnished cages or floor pens. We compared bone traits between housing systems and crossbreds and performed a genome-wide association study of bone properties and body weight. As expected, the 2 housing systems produced a large difference in bone strength, with layers housed in floor pens having stronger bones. These differences were accompanied by differences in bone geometry, mineralization, and chemical composition. Genome scans either combining or independently analyzing the 2 housing systems revealed no genome-wide significant loci for bone breaking strength. We detected 3 loci for body weight that were shared between the housing systems on chromosomes 4, 6, and 27 (either genome-wide significant or suggestive) and these coincide with associations for bone length. In summary, we found substantial differences in bone strength, content, and composition between hens kept in floor pens and furnished cages that could be attributed to greater physical activity in pen housing. We found little evidence for large-effect loci for bone strength in commercial crossbred hens, consistent with a highly polygenic architecture for bone strength in the production environment. The lack of consistent genetic associations between housing systems in combination with the differences in bone phenotypes could be due to gene-by-environment interactions with housing system or a lack of power to detect shared associations for bone strength.
Bone quality and composition are influenced by egg production, layer line, and oestradiol-17ß in laying hens Beryl Katharina Eusemann, Reiner Ulrich, Estefania Sanchez-Rodriguez, Cristina Benavides-Reyes, Nazaret Dominguez-Gasca, Alejandro B. Rodriguez-Navarro, Stefanie Petow Avian Pathology, 2022 Keel bone fractures are a serious animal welfare problem in laying hens. The aim of the current study was to assess the influence of egg production, estradiol-17ß, and selection for high laying performance on bone quality. Hens of two layer lines differing in laying performance (WLA: 320 eggs per year, G11: 200 eggs per year) were allocated to four treatment groups. Group S received a deslorelin acetate implant that suppressed egg production. Group E received an implant with the sexual steroid estradiol-17ß. Group SE received both implants and group C did not receive any implant. In the 63rd week of age, composition and characteristics of the tibiotarsi were assessed using histological analysis, three-point bending test, thermogravimetric analysis, infrared spectroscopy, and two-dimensional X-ray diffraction, respectively. Non-egg laying hens showed a higher total bone area and a higher relative amount of cortical bone compared to egg laying hens. Hens of layer line G11 showed a higher relative amount of medullary bone and a higher degree of mineralization of the cortical bone compared to hens of layer line WLA. These differences in bone composition may explain different susceptibility for keel bone fractures in non-egg laying compared to egg laying hens as well as in hens of layer lines differing in laying performance. The effect of exogenous estradiol-17ß on bone parameters varied between the layer lines indicating a genetic influence on bone physiology and the way it can be modulated by hormone substitution.
No evidence that selection for egg production persistency causes loss of bone quality in laying hens Ian C. Dunn, Dirk-Jan De Koning, Heather A. McCormack, Robert H. Fleming, Peter W. Wilson, Björn Andersson, Matthias Schmutz, Cristina Benavides, Nazaret Dominguez-Gasca, Estefania Sanchez-Rodriguez, Alejandro B. Rodriguez-Navarro Genetics Selection Evolution, 2021 Background The physiological adaptations that have evolved for egg laying make hens susceptible to bone fractures and keel bone damage. In modern laying hen breeds, longer periods of egg laying could result in a greater risk of poor bone quality, and selection for increased egg production has frequently been stated to be a cause. However, the existing literature does not support this hypothesis. To test the hypothesis that egg production is associated with quality, breaking strength and density of bone, genetic correlations between these traits were estimated in White Leghorn and Rhode Island Red breeds. Genetic correlations of cortical and medullary bone material chemical properties with bone quality were also estimated, in order to identify methods to improve bone quality with appropriately targeted measurement of key traits. Results Estimates of heritability for bone quality traits were moderate (0.19–0.59) for both White Leghorn and Rhode Island Red breeds, except for the keel bone trait, which had a heritability estimate equal to zero. There was no evidence for genetic or phenotypic relationships between post-peak egg production and bone quality. In the White Leghorn breed, the estimate of the genetic correlation between pre-peak production/age at first egg and bone quality was significant and negative (− 0.7 to − 0.4). Estimates of heritability of thermogravimetric measurements of tibial medullary bone mineralisation were significant (0.18–0.41), as were estimates of their genetic correlations with tibia breaking strength and density (0.6–0.9). Conclusions The low genetic correlation of post-peak egg production with bone quality suggests that selection for increased persistency of egg production may not adversely affect bone quality. Onset of puberty and mineralisation of the medullary bone, which is a specialised adaptation for egg laying, were identified as important factors associated with the quality of the skeleton later during egg production. These are traits for which genetic, as well as environmental and management factors can positively impact the overall quality of the skeleton of laying hens.
Research Note: Changes in eggshell quality and microstructure related to hen age during a production cycle Cristina Benavides-Reyes, Elisa Folegatti, Nazaret Dominguez-Gasca, Gilberto Litta, Estefania Sanchez-Rodriguez, Alejandro B. Rodriguez-Navarro, Murtala Umar Faruk Poultry Science, 2021 We have studied in detail the changes that occur in eggshell structure and composition during a production cycle in order to better understand the deterioration of eggshell quality with hen age (at 33, 45, and 67 wk). To study changes in eggshell ultrastructure and microstructure characteristics (mammillary density, palisade layer thickness, size, and orientation of calcite crystals) and the cuticle composition, we used complementary analytical techniques such optical and electron microscopy, X-ray diffraction, and infrared spectrometry. The marked decrease in eggshell breaking strength from 5.8 Kg at 33 wk to 4.4 Kg at 67 wk (25% reduction) could not be solely explained by the modest reduction in eggshell thickness (6-10% reduction) and seems to be associate to abrupt changes in eggshell ultra- and microstructure characteristics (i.e., decreased mammillary density; increased size of crystal units), occurring in older hens. Particularly, the decrease in mammillary density reduces the attachment points of the eggshell mineral to the membranes and therefore should negatively impact eggshell mechanical properties. Also, the observed increase in the calcite crystal size making the shell could also reduce the cohesion of crystals and eggshell resistance against impacts. Additionally, there was a decrease in the amount of cuticle and internal egg quality parameters (egg albumen height) with hen age that could have a negative impact in egg safety and quality.
May bioactive compounds from the olive fruit improve the postprandial insulin response in healthy adults? Estefania Sanchez-Rodriguez, Laura Alejandra Vazquez-Aguilar, Sara Biel-Glesson, Jose Ramon Fernandez-Navarro, Juan Antonio Espejo-Calvo, Jose Maria Olmo-Peinado, Rafael de la Torre, Monserrat Fito-Colomer, Maria Isabel Covas, Concepcion Romero, Manuel Brenes, Emilio Martinez de Victoria, Angel Gil, Maria Dolores Mesa Journal of Functional Foods, 2021
An eQTL in the cystathionine beta synthase gene is linked to osteoporosis in laying hens Dirk-Jan De Koning, Nazaret Dominguez-Gasca, Robert H. Fleming, Andrew Gill, Dominic Kurian, Andrew Law, Heather A. McCormack, David Morrice, Estefania Sanchez-Rodriguez, Alejandro B. Rodriguez-Navarro, Rudolf Preisinger, Matthias Schmutz, Veronica Šmídová, Frances Turner, Peter W. Wilson, Rongyan Zhou, Ian C. Dunn Genetics Selection Evolution, 2020
The NUTRAOLEOUM Study, a randomized controlled trial, for achieving nutritional added value for olive oils Sara Biel, Maria-Dolores Mesa, Rafael de la Torre, Juan-Antonio Espejo, Jose-Ramón Fernández-Navarro, Montserrat Fitó, Estefanía Sánchez-Rodriguez, Carmen Rosa, Rosa Marchal, Juan de Dios Alche, Manuela Expósito, Manuel Brenes, Beatriz Gandul, Miguel Angel Calleja, María-Isabel Covas BMC Complementary and Alternative Medicine, 2016
