Role of retinal pigment epithelium-derived exosomes and autophagy in new blood vessel formation

Autophagy and exosome secretion play important roles in a variety of physiological and disease states, including the development of age‐related macular degeneration. Previous studies have demonstrated that these cellular mechanisms share common pathways of activation. Low oxidative damage in ARPE‐19 cells, alters both autophagy and exosome biogenesis. Moreover, oxidative stress modifies the protein and genetic cargo of exosomes, possibly affecting the fate of surrounding cells. In order to understand the connection between these two mechanisms and their impact on angiogenesis, stressed ARPE‐19 cells were treated with a siRNA‐targeting Atg7, a key protein for the formation of autophagosomes. Subsequently, we observed the formation of multivesicular bodies and the release of exosomes. Released exosomes contained VEGFR2 as part of their cargo. This receptor for VEGF—which is critical for the development of new blood vessels—was higher in exosome populations released from stressed ARPE‐19. While stressed exosomes enhanced tube formation, exosomes became ineffective after silencing VEGFR2 in ARPE‐19 cells and were, consequently, unable to influence angiogenesis. Moreover, vessel sprouting in the presence of stressed exosomes seems to follow a VEGF‐independent pathway. We propose that abnormal vessel growth correlates with VEGFR2‐expressing exosomes release from stressed ARPE‐19 cells, and is directly linked to autophagy

Implication of RPE-derived exosomes in blood vessel formation under stress conditions

Retinal pigment epithelium (RPE) has a crucial role in the physiology and pathophysiology of the retina due to its location and metabolism. Oxidative damage has been demonstrated as a pathogenic mechanism in several retinal diseases, and reactive oxygen species (ROS) are certainly important by-products ethanol metabolism. Autophagy and exosome secretion are known to play important roles in a variety of physiological and disease states, including the development of neovascular agerelated macular degeneration (NV AMD). Previous studies have demonstrated that these cellular mechanisms share common pathways of activation. Low oxidative damage in ARPE-19, a humanRPE cell line, alters both autophagy and exosome biogenesis. Moreover, oxidative stress modifies the protein and genetic cargo of exosomes, possibly affecting the fate of surrounding cells. In order to understand the connection between these two mechanisms and their impact on angiogenesis, stressed ARPE-19 cells were treated with a siRNA targeting Atg7, a key protein for the formation of autophagosomes. Subsequently, we observed the formation of multivesicular bodies and the release of exosomes, which were found to be decreased in number. RPE-released exosomes contained VEGFR2 as part of their cargo. This receptor for VEGF - which is critical for the development of new blood vessels in NV AMD - was increased in exosome populations released from ARPE-19 cells under oxidative stress. While stressed-RPE exosomes enhanced tube formation of endothelial cells, exosomes became ineffective after silencing VEGFR2 in ARPE-19 cells and were, as a result, unable to influence angiogenesis. Moreover, vessel sprouting in the presence of stressed-RPE exosomes seems to follow a VEGF independent pathway. In view of these results, we propose that abnormal vessel growth correlates with VEGFR2-expression.MedicinaCiencias de la Salu

Mitochondrial Dynamics and Mitophagy in the 6-Hydroxydopamine Preclinical Model of Parkinson's Disease

We discuss the participation of mitochondrial dynamics and autophagy in the 6-hydroxidopamine-induced Parkinson’s disease model. The regulation of dynamic mitochondrial processes such as fusion, fission, and mitophagy has been shown to be an important mechanism controlling cellular fate. An imbalance in mitochondrial dynamics may contribute to both familial and sporadic neurodegenerative diseases including Parkinson’s disease. With special attention we address the role of second messengers as the role of reactive oxygen species and the mitochondria as the headquarters of cell death. The role of molecular signaling pathways, for instance, the participation of Dynamin-related protein 1(Drp1), will also be addressed. Furthermore evidence demonstrates the therapeutic potential of small-molecule inhibitors of mitochondrial division in Parkinson’s disease. For instance, pharmacological inhibition of Drp1, through treatment with the mitochondrial division inhibitor-1, results in the abrogation of mitochondrial fission and in a decrease of the number of autophagic cells. Deciphering the signaling cascades that underlie mitophagy triggered by 6-OHDA, as well as the mechanisms that determine the selectivity of this response, will help to better understand this process and may have impact on human treatment strategies of Parkinson’s disease

Role of retinal pigment epithelium‐derived exosomes and autophagy in new blood vessel formation

Autophagy and exosome secretion play important roles in a variety of physiological and disease states, including the development of age‐related macular degeneration. Previous studies have demonstrated that these cellular mechanisms share common pathways of activation. Low oxidative damage in ARPE‐19 cells, alters both autophagy and exosome biogenesis. Moreover, oxidative stress modifies the protein and genetic cargo of exosomes, possibly affecting the fate of surrounding cells. In order to understand the connection between these two mechanisms and their impact on angiogenesis, stressed ARPE‐19 cells were treated with a siRNA‐targeting Atg7, a key protein for the formation of autophagosomes. Subsequently, we observed the formation of multivesicular bodies and the release of exosomes. Released exosomes contained VEGFR2 as part of their cargo. This receptor for VEGF—which is critical for the development of new blood vessels—was higher in exosome populations released from stressed ARPE‐19. While stressed exosomes enhanced tube formation, exosomes became ineffective after silencing VEGFR2 in ARPE‐19 cells and were, consequently, unable to influence angiogenesis. Moreover, vessel sprouting in the presence of stressed exosomes seems to follow a VEGF‐independent pathway. We propose that abnormal vessel growth correlates with VEGFR2‐expressing exosomes release from stressed ARPE‐19 cells, and is directly linked to autophagy.Sin financiación4.658 JCR (2018) Q1, 31/136 Medicine, Research & Experimental; Q2, 60/193 Cell Biology1.439 SJR (2018) Q1, 41/177 Molecular Medicine; Q2, 95/299 Cell BiologyNo data IDR 2018UE

Implementation of a game-based practical workshop to increase the learning and motivation in the study of facial muscles

Fisiología del Sistema Estomatognático es una asignatura de carácter básico en el plan de estudios del grado en Odontología y es imprescindible para que los estudiantes comprendan, entre otros conceptos, las funciones de los tejidos y músculos orofaciales. Sin embargo, esta asignatura puede resultar compleja y con una alta cantidad de contenidos para los estudiantes, lo que puede afectar su motivación e interés por la materia. Para incrementar el aprendizaje y motivación de los estudiantes, se realizó un taller práctico basado en el juego, durante una sesión práctica, consistentes en el reconocimiento de los músculos faciales involucrados en la masticación, la deglución y la articulación del habla. Al finalizar la actividad, los estudiantes respondieron a una encuesta de valoración, dando su opinión sobre la repercusión del taller en su aprendizaje y motivación. Como resultado, los estudiantes valoraron positivamente el taller práctico indicando que éste les permitió entender mejor la función de los músculos faciales y recomendaron la realización de esta actividad a otros estudiantes. Asimismo, el taller les pareció interesante, divertido y gratificante, a la vez que fomentó un ambiente de aprendizaje colaborativo al realizarse en parejas.Physiology of the Stomatognathic System is a basic subject in the curriculum of the degree in Dentistry and is essential for students to understand, among other concepts, the functions of orofacial tissues and muscles. However, this subject can be complex and with a high amount of content for students, which can affect their motivation and interest in the subject. In order to increase the learning and motivation of the students, a practical workshop based on games was carried out in a practical session, consisting of the recognition of the facial muscles involved in mastication, swallowing and speech articulation. At the end of the activity, students responded to an evaluation survey, giving their opinion on the impact of the workshop on their learning and motivation. As a result, the students rated the practical workshop positively, indicating that it gave them a better understanding of the function of the facial muscles and recommended the activity to other students. They also found the workshop interesting, fun and rewarding, as well as fostering a collaborative learning environment by working in pairs.Sin financiaciónSPI 2022UE

Digital escape room as an instrument for the acquisition of specific knowledge in basic biomedical sciences

Background: The COVID-19 pandemic turned the world towards digitized learning. In the present day, the increased demand for online education produces an even greater request for quality courses and well-established processes for teaching relevant contents. Simultaneously, modern pedagogical trends promote the application of gamification as a tool for improving motivation, developing new skills and strengthening knowledge retention. In this context, Digital Escape Rooms have become an educational tool of great interest, in terms of new approaches to game-based learning. Objective: The general objective is to evaluate the effectiveness of a Digital Escape Room to facilitate the acquisition and the maintenance of students' knowledge applied in Biomedical Sciences. Methods: An educational game, the Genetics Escape Room, will be created with Genially. It will be based on 4 contents: genome, mutations, polymorphism and genetically transmitted diseases. The participants will consist of ca. 240 first year students enrolled in “Cellular Biology and Human genetics” in Dentistry Degree. They will be divided into an experimental group that will play Escape Room, and a control group that will visualize a virtual master class about the same 4 contents of the Escape Room. To evaluate students´ knowledge, all participants will complete a 20-question test (5 questions for each content) performed at three different time points. Firstly, two weeks before the experiment as pre-test, to assess students’ previous knowledge, secondly at the end of the gamification or the virtual class (post-test 1), and finally in two weeks’ time to evaluate knowledge retention when compared the two methods (post-test 2). To evaluate students’ opinion about the experience, a perception survey will be completed. Hypothesis: The following hypotheses are going to be considered: - We expect that the use of the escape room may well be equally as effective as a master class in increasing student knowledge and understanding. - A higher degree of retention of knowledge for lifelong learning is expected among students who play escape room. - We expect a higher motivation and an increase in the appreciation of Human Genetics subject among students who take part in the escape room.2021/UEM22No data 2021UE

Does participation in therapeutic exercise programs after finishing oncology treatment still ensure an adequate health status for long-term breast cancer survivors? A ≥ 5 years follow-up study

Purpose: The aims of this study were to evaluate whether the effects of two therapeutic exercise programs are sustained over time (≥ 5 years) in long-term breast cancer survivors (LTBCS). Second, to determine the influence of the current level of physical activity (PA) performed on cancer-related fatigue (CRF) that these patients may present ≥5 years later. Methods: A prospective observational study was conducted with a cohort of 80 LTBCS in Granada during 2018. Firstly, considering their participation in one of the programs, they were allocated into two groups: usual care and therapeutic exercise program, to assess CRF, pain and pressure pain sensitivity, muscle strength, functional capacity, and quality of life. Secondly, they were also classified into 3 groups according to current level of weekly PA performed: ≤ 3, 3.1-7.4, and ≥ 7.5 (MET-hour/week) respectively, to assess its impact over CRF. Results: Although the positive effects of the programs are not sustained over time, a trend toward significance can be observed for a greater reduction in overall CRF levels, lower intensity of pain in the affected arm and cervical region, and greater functional capacity and quality of life in the group that underwent therapeutic exercise. Additionally, 66.25% of LTBCS are inactive ≥ 5 years after completion of the program and furthermore, such inactivity is accompanied by higher CRF levels (P .013 to .046). Conclusion: The positive effects of therapeutic exercise programs are not maintained over time for LTBCS. Additionally, more than half of these women (66.25%) are inactive ≥ 5 years after completion of the program, this inactivity being accompanied by higher levels of CRF.Spanish Ministry of Economy and Competitiveness (Plan Estatal de I+D+I 2013-2016)Fondo de Investigación Sanitaria del Instituto de Salud Carlos III [PI14/01627]Fondos Estructurales de la Unión Europea (FEDER)University of Granada, Plan Propio de Investigación 20163.359 Q1 JCR 20210.984 Q2 SJR 2022No data IDR 2021UE

Matching Diabetes and Alcoholism: Oxidative Stress, Inflammation, and Neurogenesis Are Commonly Involved

Diabetes and alcohol misuse are two of the major challenges in health systems worldwide. These two diseases finally affect several organs and systems including the central nervous system. Hippocampus is one of the most relevant structures due to neurogenesis and memory-related processing among other functions. The present review focuses on the common profile of diabetes and ethanol exposure in terms of oxidative stress and proinflammatory and prosurvival recruiting transcription factors affecting hippocampal neurogenesis. Some aspects around antioxidant strategies are also included. As a global conclusion, the present review points out some common hits on both diseases giving support to the relations between alcohol intake and diabetes

Oxidative stress in retinal pigment epithelium cells increases exosome secretion and promotes angiogenesis in endothelial cells.

10 páginas, 5 figurasThe retinal pigment epithelium (RPE), a monolayer located between the photoreceptors and the choroid, is constantly damaged by oxidative stress, particularly because of reactive oxygen species (ROS). As the RPE, because of its physiological functions, is essential for the survival of the retina, any sustained damage may consequently lead to loss of vision. Exosomes are small membranous vesicles released into the extracellular medium by numerous cell types, including RPE cells. Their cargo includes genetic material and proteins, making these vesicles essential for cell-to-cell communication. Exosomes may fuse with neighbouring cells influencing their fate. It has been observed that RPE cells release higher amounts of exosomes when they are under oxidative stress. Exosomes derived from cultured RPE cells were isolated by ultracentrifugation and quantified by flow cytometry. VEGF receptors (VEGFR) were analysed by both flow cytometry and Western blot. RT-PCR and qPCR were conducted to assess mRNA content of VEGFRs in exosomes. Neovascularization assays were performed after applying RPE exosomes into endothelial cell cultures. Our results showed that stressed RPE cells released a higher amount of exosomes than controls, with a higher expression of VEGFR in the membrane, and enclosed an extra cargo of VEGFR mRNA. Angiogenesis assays confirmed that endothelial cells increased their tube formation capacity when exposed to stressed RPE exosomes.This work was supported by internal funding of the university, by the local department of education, Conselleria de Educaciò, Cultura i Sport, Valencia, Spain (GVA/2014/106), and partially by other external institutions: Instituto Nacional Carlos III (PI13/00617), Red de Investigacón Cardiovascular (RD12/0042/0052).Peer reviewe