Comparative proteomic analysis of normal and NQO1-null mouse embryo fibroblasts

Comunicaciones a congreso

H2O2-preconditioned human adipose-derived stem cells (HC016) increase theirresistance to oxidative stress byoverexpressing Nrf2 and bioenergetic adaptation

BackgroundMesenchymal stem cells, including those derived from human adipose tissue (hASCs), are currently being widely investigated for cell therapy. However, when transplanted at the site of injury, the survival and engraftment rates of hASCs are low, mainly due to the harsh microenvironment they encounter, characterized by inflammation and oxidative stress. To overcome these therapeutic limitations, cell preconditioning with low-concentration of hydrogen peroxide (H2O2) has been proposed as a plausible strategy to increase their survival and adaptation to oxidative stress. Nonetheless, the underlying mechanisms of this approach are not yet fully understood. In this study, we analyzed molecular and bioenergetic changes that take place in H2O2 preconditioned hASCs.MethodsLong-term exposure to a low concentration of H2O2 was applied to obtain preconditioned hASCs (named HC016), and then, their response to oxidative stress was analyzed. The effect of preconditioning on the expression of Nrf2 and its downstream antioxidant enzymes (HO-1, SOD-1, GPx-1, and CAT), and of NF-kappa B and its related inflammatory proteins (COX-2 and IL-1 beta), were examined by Western blot. Finally, the Seahorse XF96 Flux analysis system was used to evaluate the mitochondrial respiration and glycolytic function, along with the total ATP production.ResultsWe found that under oxidative conditions, HC016 cells increased the survival by (i) decreasing intracellular ROS levels through the overexpression of the transcription factor Nrf2 and its related antioxidant enzymes HO-1, SOD-1, GPx-1, and CAT; (ii) reducing the secretion of pro-inflammatory molecules COX-2 and IL-1 beta through the attenuation of the expression of NF-kappa B; and (iii) increasing the total ATP production rate through the adaption of their metabolism to meet the energetic demand required to survive.ConclusionsH(2)O(2) preconditioning enhances hASC survival under oxidative stress conditions by stimulating their antioxidant response and bioenergetic adaptation. Therefore, this preconditioning strategy might be considered an excellent tool for strengthening the resistance of hASCs to harmful oxidative stress.Partial funding for this project was provided by the Department of Economic Development and Competitiveness of the Basque Government, the European Regional Development Fund (PREMISE IG-2015/0000558), and the University of the Basque Country (UPV/EHU; research grants PES 17/29 and 16/37)

Hydrogen Peroxide-Preconditioned Human Adipose-Derived Stem Cells Enhance the Recovery of Oligodendrocyte-Like Cells after Oxidative Stress-Induced Damage

Oxidative stress associated with neuroinflammation is a key process involved in the pathophysiology of neurodegenerative diseases, and therefore, has been proposed as a crucial target for new therapies. Recently, the therapeutic potential of human adipose-derived stem cells (hASCs) has been investigated as a novel strategy for neuroprotection. These cells can be preconditioned by exposing them to mild stress in order to improve their response to oxidative stress. In this study, we evaluate the therapeutic potential of hASCs preconditioned with low doses of H2O2 (called HC016 cells) to overcome the deleterious effect of oxidative stress in an in vitro model of oligodendrocyte-like cells (HOGd), through two strategies: i, the culture of oxidized HOGd with HC016 cell-conditioned medium (CM), and ii, the indirect co-culture of oxidized HOGd with HC016 cells, which had or had not been exposed to oxidative stress. The results demonstrated that both strategies had reparative effects, oxidized HC016 cell co-culture being the one associated with the greatest recovery of the damaged HOGd, increasing their viability, reducing their intracellular reactive oxygen species levels and promoting their antioxidant capacity. Taken together, these findings support the view that HC016 cells, given their reparative capacity, might be considered an important breakthrough in cell-based therapies.Partial funding for this project was provided by the Department of Economic Development and Competitiveness of the Basque Government, the European Regional Development Fund (PREMISE IG-2015/0000558) and the University of the Basque Country (UPV/EHU; research grants GIU 19/088 and PES 17/29 and 16/37)

CYB5R3 overexpression preserves skeletal muscle mitochondria and autophagic signaling in aged transgenic mice

Cytochrome b5 reductase 3 (CYB5R3) overexpression activates respiratory metabolism and exerts prolongevity effects in transgenic mice, mimicking some of the salutary effects of calorie restriction. The aim of our study was to understand how CYB5R3 overexpression targets key pathways that modulate the rate of aging in skeletal muscle, a postmitotic tissue with a greater contribution to resting energy expenditure. Mitochondrial function, autophagy and mitophagy markers were evaluated in mouse hind limb skeletal muscles from young-adult (7 months old) and old (24 months old) males of wild-type and CYB5R3-overexpressing genotypes. Ultrastructure of subsarcolemmal and intermyofibrillar mitochondria was studied by electron microscopy in red gastrocnemius. CYB5R3, which was efficiently overexpressed and targeted to skeletal muscle mitochondria regardless of age, increased the abundance of complexes I, II, and IV in old mice and prevented the age-related decrease of complexes I, III, IV, and V and the mitofusin MFN-2. ATP was significantly decreased by aging, which was prevented by CYB5R3 overexpression. Coenzyme Q and the mitochondrial biogenesis markers TFAM and NRF-1 were also significantly diminished by aging, but CYB5R3 overexpression did not protect against these declines. Both aging and CYB5R3 overexpression upregulated SIRT3 and the mitochondrial fission markers FIS1 and DRP-1, although with different outcomes on mitochondrial ultrastructure: old wild-type mice exhibited mitochondrial fragmentation whereas CYB5R3 overexpression increased mitochondrial size in old transgenic mice concomitant with an improvement of autophagic recycling. Interventions aimed at stimulating CYB5R3 could represent a valuable strategy to counteract the deleterious effects of aging in skeletal muscle

Conditioned Medium from H2O2-Preconditioned Human Adipose-Derived Stem Cells Ameliorates UVB-Induced Damage to Human Dermal Fibroblasts

Human skin exposure to ultraviolet B (UVB) radiation can result in acute photodamage through oxidative modifications of cellular components and biomolecules involved in the metabolism of dermal cells. Recently, the therapeutic potential of human adipose-derived stem cells (hASCs) has been investigated as a novel strategy for photoprotection due to their pro-angiogenic properties, protective activity against oxidative stress and paracrine effect on dermal cells. To enhance these therapeutic properties, hASCs can be preconditioned by exposing them to sublethal cellular stressors. In this study, we first analyzed response capacity against UVB-induced oxidative stress in H2O2-preconditioned hASCs (called HC016 cells); and second, we evaluated the photoprotective effect of HC016-conditioned medium (CM) in an in vitro UVB irradiation model in cultured human foreskin fibroblasts (hFFs). The results demonstrated that HC016 cells have a greater capacity to respond efficiently to UVB-induced oxidative stress, evidenced by higher Nrf2 antioxidant system activity and enhanced viability and migration capacity. Further, HC016-CM treatment increased viability, migratory capacity and collagen type I synthesis in hFFs exposed to UVB radiation, as well as reducing their cytotoxicity, apoptosis, senescence and IL-6 secretion. Collectively, these findings support the view that HC016 cells could protect against UVB-induced photodamage via paracrine mechanisms.This research was founded by the University of the Basque Country (UPV/EHU, grant reference numbers: GIU19/088 and PES 21/50)

Análisis y desarrollo de las competencias personales y profesionales del profesor tutor de TFG

Durante este curso hemos conseguido llevar a cabo sólo una parte de los objetivos que teníamos previstos. No obstante, hemos constatado la relevancia del tema que estamos trabajando en los foros a los que hemos asistido, por lo que nos ha animado a pedir una continuación del proyecto para el próximo curso. En primer lugar, hemos llevado a cabo una revisión bibliográfica acerca de las competencias personales y profesionales que debe tener un buen profesor de educación superior. Estas competencias son el punto de partida para desempeñar con eficacia la función de tutorizar un TFG. Las conclusiones de esta revisión apuntan a la consideración de tres dimensiones fundamentales: las competencias personales, las pedagógicas y las técnicas. Sobre estas dimensiones se ha empezado a construir un cuestionario de evaluación de competencias. Durante este curso se han confeccionado los ítems y se ha pasado la primera batería a profesores expertos en dirigir TFGs con el objeto de obtener la validez de contenido del instrumento. En el futuro nos queda aplicar el cuestionario a profesores y alumnos para llevar a cabo el trabajo psicométrico, así como diseñar el protocolo de actuación que mejore la actuación docente de los profesores que dirigen un TFG

Mejora de las competencias personales y profesionales del profesor tutor del Trabajo Fin de Grado (TFG)

Durante este curso hemos continuado la labor que comenzamos el curso pasado. Hemos constatado la relevancia del tema que estamos trabajando en los foros a los que hemos asistido. Después de llevar a cabo la revisión bibliográfica acerca de las competencias personales y profesionales que debe tener un buen profesor de educación superior, llegamos a la conclusión de que existen tres dimensiones fundamentales: las competencias personales, las pedagógicas y las técnicas. Sobre estas dimensiones hemos construido un cuestionario de evaluación de competencias, lo hemos cribado con la ayuda de un grupo de expertos nombrado “ad hoc” y lo hemos digitalizado . Nos encontramos en la fase de pilotaje, en que se va a aplicar el cuestionario a profesores y alumnos para llevar a cabo el trabajo psicométrico. Una vez que tengamos estos primeros resultados realizaremos el muestreo a gran escala, con el objeto de diseñar el protocolo de actuación que mejore la actuación docente de los profesores que dirigen un TFG

Experiences in flipped learning

El presente trabajo describe unas experiencias de innovación docente, de igual perfil y metodología, desarrolladas por un conjunto de profesores noveles participantes bajo la tutela experta de profesorado senior. El tipo de experiencia seleccionada ha sido el diseño y ejecución de una flipped classroom, o clase invertida. Además, se incluye la correspondiente evaluación de la acción con una valoración académica o impacto en los aprendizajes, y una valoración de la experiencia por parte del alumnado y profesorado responsable. Los resultados de cinco experiencias con impacto a mas de 270 alumnos de distinto perfil académico (tres asignaturas de titulo de Grado y dos de Máster) y las evaluaciones cuantificadas indican una valoracion muy positiva de esta metodología flipped learning (FL) por los alumnos y su deseo de aplicabilidad. Por su parte, el profesorado constata que las experiencias FL han incrementado satisfactoriamente el aprendizaje de sus alumnos y declaran una alta valoración de la experiencia para la mejora de sus competencia docentes.The present work describes experiences of teaching innovation, of the same profile and methodology, developed by a group of participating junior teachers under the expert guidance of senior teaching staff. The type of experience selected has been the design and execution of a flipped classroom, or inverted class. In addition, the corresponding evaluation of the action is included with an academic assessment or impact on learning, and an assessment of the experience by the students and teaching staff responsible. The results of five experiences with impact to more than 270 students of different academic profiles (three degrees and two Masters) and the quantified evaluations indicate a very positive assessment of this flipped learning methodology by the students and their desire for applicability. On the other hand, the teachers note that the FL experiences have successfully increased the learning of their students and declare a high evaluation of the experience for the improvement of their teaching competences

Actions for novel teacher training: design and implementation of learning activities and resources on LMS support

El presente artículo describe el desarrollo de un Proyecto formativo para la implementación de actividades y recursos de aprendizaje en la plataforma UCO-Moodle por parte de profesorado novel, bajo tutela y coordinación de profesorado con experiencia previa contrastada. Se trata de un trabajo multidisciplinar donde las acciones se han ejecutado para ocho asignaturas de cuatro Áreas de conocimiento. El impacto de las innovaciones introducidas en varias asignaturas, afectó a más de 500 alumnos pertenecientes a seis títulos de grado de la Universidad de Córdoba. El trabajo realizado se completó con una evaluación de satisfacción por parte de los alumnos de los nuevos recursos de aprendizaje así como la autoevaluación del profesorado novel sobre la experiencia desarrollada.This article describes a project for the implementation of learning activities and resources in the UCO-Moodle platform by training teachers has been developed under the supervision and coordination of senior teaching staff with proven previous experience. It is a multidisciplinary work where the actions have been executed for eight different subjects in four Areas of knowledge. The impact of the learning activities introduced in different subjects was extended to more than 500 students belonging to six undergraduate degrees from the University of Cordoba. The experience has included an evaluation of students' satisfaction with the new learning resources as well as the self-assessment of the new teaching staff on the executed actions