HPLC‐QTOF method for quantifying 11‐ketoetiocholanolone, a cortisol metabolite, in ruminants' feces: Optimization and validation

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2018 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd[EN]Studies of animal ecology can benefit from a quantified understanding of eco-physiological processes and, in particular, of the physiological responses in free-ranging animals to potential stressors. The determination of fecal cortisol metabolites as a noninvasive method for monitoring stress has proved to be a powerful tool. High-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/ MS) has emerged as the most accurate method for avoiding problems related to the nonspecificity of immunoassays. In this study, we optimize and validate a reliable method using HPLC-MS/ MS for quantifying 11-ketoetiocholanolone (11-k), a representative fecal cortisol metabolite in ruminants. An appropriate extraction and purification procedure was developed taking into account the complex nature of feces. The final extract obtained was then analyzed with HPLC-MS/ MS using a quadrupole-time- of- fly (QTOF) tandem mass spectrometer with an electrospray ionization interface operating in positive mode, which allowed an unequivocal determination of the metabolite due to its accurate mass capabilities. After rigorous optimization of both sample extraction and the HPLC-QTOF parameters, making use of feces from free-ranging Iberian ibex, ideal conditions were established. Matrix-matched standards were used to calibrate the method. The limit of detection and quantification was 13-and 40-ng/ g, respectively. The validation of the method was performed with recoveries in the range of 85–110%, a figure much higher than the 60% obtained with the previous extraction methods used in our laboratory, and with relative standard deviations (RSDs) no higher than 15% for the complete analytical procedure, including extraction and analysis. The time required for the fecal 11-k analysis was greatly reduced in comparison with the previous work carried out in our laboratory. This is the first time that QTOF mass detection coupled with HPLC has been validated for 11-k quantification in feces from free-ranging ruminants such as Iberian ibex. Given the high selectivity and sensitivity attained, our method could become a useful tool for noninvasive stress quantification in ruminants.SIThis study was partly funded by the Fédération Nationale des Chasseurs (France), project FNC-PSN-PR4-2013. The technical and human support provided by the CICT of Jaén University (UJA, MINECO, Junta de Andalucía, FEDER) is gratefully acknowledged. The research activities of the authors were partially supported by the PAIDI, Junta de Andalucía (RNM-118 group and RNM-175 groups), as well as by the grant P07-RNM-03087 and the European Fund for Regional Development (FEDER). This study complied with all Andalusian, Spanish and European legal requirements and guidelines regarding experimentation and animal welfare. It was approved by the Committee on Ethics of Animal Experimentation of the University of Jaén and authorized by the General Direction of Agriculture and Livestock of the Ministry of Agriculture, Fisheries and Environment of the Junta de Andalucía

Light signals generated by vegetation shade facilitate acclimation to low light in shade-avoider plants

[EN] When growing in search for light, plants can experience continuous or occasional shading by other plants. Plant proximity causes a decrease in the ratio of R to far-red light (low R:FR) due to the preferential absorbance of R light and reflection of FR light by photosynthetic tissues of neighboring plants. This signal is often perceived before actual shading causes a reduction in photo-synthetically active radiation (low PAR). Here, we investigated how several Brassicaceae species from different habitats respond to low R:FR and low PAR in terms of elongation, photosynthesis, and photoacclimation. Shade-tolerant plants such as hairy bittercress (Cardamine hirsuta) displayed a good adaptation to low PAR but a poor or null response to low R:FR exposure. In contrast, shade-avoider species, such as Arabidopsis (Arabidopsis thaliana), showed a weak photosynthetic performance under low PAR but they strongly elongated when exposed to low R:FR. These responses could be genetically uncoupled. Most interestingly, exposure to low R:FR of shade-avoider (but not shade-tolerant) plants improved their photoacclimation to low PAR by triggering changes in photosynthesis-related gene expression, pigment accumulation, and chloroplast ultrastructure. These results indicate that low R:FR signaling unleashes molecular, metabolic, and developmental responses that allow shade-avoider plants (including most crops) to adjust their photosynthetic capacity in anticipation of eventual shading by nearby plants.L.M. received a predoctoral fellowships from La Caixa Foundation (INPhINIT fellowship LCF/BQ/IN18/11660004). W.Q. is a recipient of a predoctoral Chinese Scholarship Council (CSC) fellowship. A.I.-S. is supported by a predoctoral fellowship from MICINN (PRE2018-083610). I.F.-S. has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement no. 753301. Our research is supported by grants from MICINN-FEDER (BIO2017-85316-R, and BIO2017-84041-P) and AGAUR (2017-SGR1211, 2017-SGR710 and Xarba) to J.F.M.-G. and M.R.-C. We also acknowledge the support of the MINECO for the "Centro de Excelencia Severo Ochoa 2016-2019" award SEV-2015-0533 and by the CERCA Programme/Generalitat de Catalunya.Morelli, L.; Paulisic, S.; Qin, W.; Iglesias-Sanchez; Roig-Villanova, I.; Florez-Sarasa, I.; Rodriguez Concepción, M.... (2021). Light signals generated by vegetation shade facilitate acclimation to low light in shade-avoider plants. Plant Physiology (Online). 186(4):2137-2151. https://doi.org/10.1093/plphys/kiab206S21372151186

The Lack of Mitochondrial Thioredoxin TRXo1 Affects In Vivo Alternative Oxidase Activity and Carbon Metabolism under Different Light Conditions

International audienceThe alternative oxidase (AOX) constitutes a nonphosphorylating pathway of electron transport in the mitochondrial respiratory chain that provides flexibility to energy and carbon primary metabolism. Its activity is regulated in vitro by the mitochondrial thioredoxin (TRX) system which reduces conserved cysteines residues of AOX. However, in vivo evidence for redox regulation of the AOX activity is still scarce. In the present study, the redox state, protein levels and in vivo activity of the AOX in parallel to photosynthetic parameters were determined in Arabidopsis knockout mutants lacking mitochondrial trxo1 under moderate (ML) and high light (HL) conditions, known to induce in vivo AOX activity. In addition, 13C- and 14C-labeling experiments together with metabolite profiling were performed to better understand the metabolic coordination between energy and carbon metabolism in the trxo1 mutants. Our results show that the in vivo AOX activity is higher in the trxo1 mutants at ML while the AOX redox state is apparently unaltered. These results suggest that mitochondrial thiol redox systems are responsible for maintaining AOX in its reduced form rather than regulating its activity in vivo. Moreover, the negative regulation of the tricarboxylic acid cycle by the TRX system is coordinated with the increased input of electrons into the AOX pathway. Under HL conditions, while AOX and photosynthesis displayed similar patterns in the mutants, photorespiration is restricted at the level of glycine decarboxylation most likely as a consequence of redox imbalance

Prescripción de ejercicio físico en la depresión por parte de médicos de familia : factores involucrados

Introducción: El problema del sedentarismo es acuciante en sociedades desarrolladas y sus consecuencias para la salud de la población constituyen uno de los principales problemas de salud pública actuales. La depresión es un motivo de consulta muy habitual en las consultas de atención primaria de países occidentales. Se ha planteado la prescripción de ejercicio físico como una estrategia antidepresiva útil en depresiones leves-moderadas, con frecuencia de forma complementaria a los tratamientos de primera elección: psicoterapia y/o farmacoterapia. Sin embargo, los médicos de atención primaria no siempre lo prescriben y se desconoce de qué depende que lo hagan. Contribuir a aclarar esta cuestión es el objetivo del presente trabajo. Nuestra hipótesis era que el grado de prescripción depende de sus opiniones y experiencia sobre el tratamiento de la depresión.Método: Estudio descriptivo transversal en el que participaron 13 centros de salud. Utilizamos el cuestionario Depression Questionaire Attitude para valorar la actitud de los médicos hacia la depresión, y recogimos otras variables relacionadas con la importancia que otorgaban al ejercicio físico y a otros aspectos del estilo de vida en la salud física y mental.RESULTADOS: Se confirma que las opiniones y actitudes de los médicos frente a la depresión influyen en su disposición a recomendar ejercicio físico. Además, los médicos de atención primaria con menos años de práctica profesional valoran más el papel del ejercicio físico en la salud, tanto en la depresión como en otras patologías médicas. También se observa una correlación entre la importancia que se da al ejercicio físico en la salud y la que se da a otros factores ligados al estilo de vida, especialmente la dieta.Discusión: Aunque la utilidad de recomendar ejercicio físico en la depresión va acercándose al mismo nivel de importancia que en otros problemas de salud su empleo por parte de los médicos de atención primaria es muy mejorable. Por ello, es importante insistir en la necesidad de ofrecer apoyo y programas de formación continuada a los médicos de atención primaria para ayudarles en este propósito.A sedentary lifestyle is an urgent problem in developed societies and its consequences are one of the main current problems in public health. Depression is a common reason to attend primary care in Western countries. In many cases of low and mild depression, exercise is recommended as a complement to the main therapy: Psychotherapy and/or pharmacotherapy. Nevertheless, general practitioners do not always recommend exercise to depressive patients and it is unknown why they do not. The main aim of our study was to clarify how the degree of prescribing exercise relies on a general practitioner's opinion and experience in depression treatment. A cross-sectional design was undertaken in 13 public health centers. The Depression Attitude Questionnaire (DAQ) was used to evaluate general practitioners' attitude towards depression; other variables related to the importance accorded to physical exercise; and other lifestyle aspects of physical and mental health. Our results show that general practitioners' attitude towards depression influence their willingness to recommend exercise. Moreover, less experienced general practitioners (in years) tend to appreciate the importance of exercise in health, not only in depression. A positive relationship was found between importance of exercise and importance accorded to other factors linked to lifestyle, especially diet. Although recommendation of exercise in depression is similar to other medical conditions, its prescription may be improved. Hence, it is important to point out the need for education programs for general practitioners, in order to improve their capacity to deal with their task.O problema do sedentarismo é premente nas sociedades desenvolvidas e as suas consequências para a saúde da população constituem um dos principais problemas de saúde pública actuais. A depressão é um motivo muito habitual nas consultas de cuidados primários nos países ocidentais. Foi delineada a prescrição de exercício físico como uma estratégia antidepressiva útil em depressões leves-moderadas, de forma complementar aos tratamentos de primeira escolha. Contudo, os médicos de cuidados primários nem sempre o prescrevem e desconhece-se o porquê de o fazerem. Contribuir para clarificar esta questão é o objectivo d presente trabalho. A nossa hipótese era que o grau de prescrição depende das suas opiniões e experiências no tratamento da depressão. Trata-se de um estudo descritivo transversal no qual participaram 13 centros de saúde. Utilizámos o questionário Depression Questionaire Attitude para avaliar a atitude dos médicos face à depressão e medimos outras variáveis relacionadas com a importância que atribuíam ao exercício físico e a outros aspectos do estilo de vida na saúde física e mental. Os resultados confirmam que a atitude dos médicos face à depressão influí na sua disposição para recomendar exercício físico. Adicionalmente, os médicos de cuidados primários com menos anos de prática profissional valorizam mais o papel do exercício físico na saúde, tanto na depressão como noutras patologias médicas. Também se observa uma correlação entre a importância que se dá ao exercício físico na saúde e à que se dá a outros factores ligados ao estilo de vida, especialmente à dieta. Conclui-se que apesar da utilidade de recomendar exercício físico na depressão se ir aproximando do nível de importância atribuído a outros problemas de saúde, a sua utilização por parte dos médicos de cuidados primários pode ser substancialmente melhorada. Para tal, é importante insistir na necessidade de oferecer apoio e programas de formação continuada aos médicos de cuidados primários para ajudá-los neste propósito

Estudio para la mejora de las prácticas del Grado y Máster en Trabajo Social: Bases para la modificación del curriculum académico

Este proyecto la Facultad de Trabajo Social de la UCM valora la adecuación de la estructura y los contenidos del practicum de grado de Trabajo Social, para realizar una modificación del mismo acorde a las necesidades formativas teórico-prácticas del alumnado en su futura inserción profesional

In vivo regulation and role of respiration during tomato fruit ripening

Trabajo presentado al XV Meeting of Plant Molecular Biology, celebrado online del 26 al 27 de noviembre de 2020.Peer reviewe

In vivo regulation of alternative respiration during tomato fruit ripening

Trabajo presentado a la XXIII Reunión Bianual de la Sociedad Española de Fisiología Vegetal y al XVI Congreso Hispano-Luso de Fisiología Vegetal, celebrados en Pamplona (España) del 26 al 28 de junio de 2019

Gender regimes in vulnerable households during the recession – what has changed and what not?

Depto. de Sociología: Metodología y TeoríaFac. de Trabajo SocialInstituto Complutense de Sociología para el Estudio de las Transformaciones Sociales Contemporáneas (TRANSOC)TRUEpu

Cuando lo que se hereda es la pobreza. Género, vulnerabilidad y crisis

Depto. de Sociología: Metodología y TeoríaFac. de Trabajo SocialTRUEpu

There is light in 'dark' respiration: in vivo regulation and role of alternative respiration in leaves

Trabajo presentado a la Early Career Researches’ Conference (ECRC), celebrada en Sant Feliu de Guíxols, Girona (España) del 11 al 14 de noviembre de 2019