Fingolimod phosphate protection against mitochondrial damage in neuronal cells

Background: Major role of oxidative stress in the pathogenesis of neurodegenerative diseases have been suggested, being mitochondria one of the main sources of ROS. Aim: In the present work, we have studied the antioxidant effect of fingolimod phosphate (FP) on neuronal mitochondrial function and morphology using a model of mitochondrial oxidative damage induced by menadione (Vitk3). Methods: SN4741 neuronal cells were grown (70-80% confluence) and used as control (non-treated cells) or treated cells with Vitk3 15 µM alone or in presence of FP 50 nM during 4 hours. Mitochondrial membrane potential (MMP), cytochrome c oxidase (COX) activity, mitochondrial oxygen consumption rate (OCR), mitochondrial distribution (MTG) and morphology (EM) were analysed. Statistical differences were determined using one-way ANOVA. Results: Vitk3 incubation produces a dramatical decrease in MMP compared to control (43.7 %); this can be almost totally reverted by the co-incubation of Vitk3 in presence of FP (p<0.05). A 20.7 % decrease in COX activity has been found after Vitk3 incubation, again this effect was counteracted when Vitk3 and FP are combined, restoring COX activity to control levels (p<0.05). Vitk3 incubation triggers initially an increase in OCR, decreasing dramatically (61%) after 4 hours. In experiments co-incubating Vitk3 in presence of FP, the OCR decrease found was reduced to only 17% (p<0.05). In experiments with MitoTracker™ Green, we found a change in the network pattern distribution after Vitk3 administration that partially disappears when co-incubated in presence of FP. Almost all the mitochondria treated with Vitk3 show ultrastructural alterations at the electron microscopy level while normal mitochondria can be found when Vitk3 and FP are combined. Conclusion: FP protects against the mitochondrial damage induced by Vitk3, as seen by the results obtained in mitochondrial functional markers, distribution and morphology.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. PS13/14: Study of the non-immunological mechanisms of action of Gilenya (Fingolimod) as therapeutic tool in Multiple Sclerosis and/or other neurodegenerative diseases. Novartis Farmacéutica S.A

The role of population change in the increased economic differences in mortality: a study of premature death from all causes and major groups of causes of death in Spain, 1980-2010

Background: An increase has been observed in differences in mortality between the richest and poorest areas of rich countries. This study assesses whether one of the proposed explanations, i.e., population change, might be responsible for this increase in Spain. Methods: Observational study based on average income, population change and mortality at provincial level. The premature mortality rate (ages 0-74 years) was estimated for all causes and for cancer, cardiovascular disease and external causes across the period 1980-2010. In the years analysed, provinces were grouped into tertiles based on provincial income, with the mortality rate ratio (MMR) being estimated by taking the tertile of highest-income provinces as reference. Population change was then controlled for to ascertain whether it would modify the rate ratio. Results: In all-cause mortality, the magnitude of the MRR for provinces in the poorest versus the richest tertile was 1.01 in 1980 and 1.12 in 2010; in cardiovascular mortality, the MMRs for these same years were 1.08 and 1.31 respectively; and in the case of cancer and external-cause mortality, MMR magnitude was similar in 1980 and 2010. The magnitude of the MMR remained unchanged in response to adjustment for population change, with the single exception of 1980, when it increased in all-cause and cardiovascular mortality. Conclusion: The increase in the difference in premature mortality between the richest and poorest areas in Spain is due to the increased difference in cardiovascular mortality. This increase is not accounted for by population change. In rich countries, more empirical evidence is thus needed to test other alternative explanations for the increase in economic differences in mortality.This study was conducted thanks to support from the Research Project PI12/01459 “Population change and geographical inequalities in mortality” financed by the Ministry of Science and Innovation

Tipos de colaboraciones tecnológicas y los elementos más importantes para su desarrollo

Las colaboraciones tecnológicas son acuerdos en los que se comparten conocimientos, habilidades, tecnologías, métodos de fabricación, instalaciones y hasta muestras de fabricación. Éstas se llevan realizando desde los años 80 y, debido al cambio constante de la tecnología, han aumentado considerablemente.El objetivo de este trabajo de fin de grado ha sido analizar los tipos de colaboraciones tecnológicas con diferentes tipos de socios y países, así como los elementos más importantes para llevarlas a cabo. El estudio se ha realizado con el panel de datos PITEC (2016), en el cual hemos analizado inicialmente una muestra de 4518 empresas para finalmente centrarnos solamente en las 2034 empresas que realizan colaboraciones tecnológicas. Los resultados obtenidos muestran que la mayoría de colaboraciones que se dan son con empresas del mismo país, siendo éstas con clientes o instituciones. En lo relativo a los elementos que se tienen en cuenta para realizar dichas colaboraciones, el principal factor es la escasez de recursos; sobre la fuente de información, según los resultados obtenidos, debe ser de dentro de la empresa; en cuanto a los objetivos que las empresas se ponen para realizar una cooperación, los principales son buscar una calidad mayor en bienes y servicios, y ampliar la gama de los productos ofrecidos.<br /

Role of Insulin-Growth Factor II on mitochondrial recovery in a cellular model of Parkinson's Disease

Insulin-growth factor II (IGF-II) has shown antioxidant and neuroprotective effects in some neurodegenerative disorders. ROS causes damage to cellular macromolecules affecting several cellular processes and resulting in cell death. Mitochondrial ROS damage has a critical role in the pathobiology of PD. The objective was to assess the IGF-II role in the recovery of the oxidative damage produced on mitochondrial in a cellular model of PD. SN4741 cell line was treated as follows: MPP+ alone, in presence of IGF-II and/or co-incubated BMS (Ins/IGF-I receptors antagonist) or AB (anti-IGF-II-receptor). To assess the effect of IGF-II in the recovery of MPP+ damage, this treatment was removed after 2 h and replaced during another 2 h by medium, IGF-II or IGF-II + BMS or IGF-II + AB. Cell death was analysed through annexin-V Mitochondrial structure, localization and morphology was studied by western blot/ immunochemistry of Mitofilin (Mtf) and electron microscopy; function by Mitotracker and oxygen consumption rate. IGF-II prevented MPP+ cell death. In morphological/structural studies, MPP+ treated cells showed swollen mitochondria with loss of cristae, and electron-lucent matrix, inducing a mitochondrial number reduction. IGF-II retrieved normal-shaped mitochondria with intact cristae and outer/inner membranes. Moreover, MPP+ incubation significantly reduced the expression levels of Mtf compared to the CO. This expression was found in areas that had a very weak mark, indicating mitochondrial destruction or dysfunction. IGF-II coincubation, recovered the expression of Mtf, remaining associated with healthy mitochondrial function. Additionally, the decrease in OCR levels after MPP+ administration was recovered in presence of IGF-II. The BMS-receptor blockage did not modify the IGF-II responses, and AB limited its effect. In conclusion, IGF-II recovers mitochondrial structure and function due to MPP+ damage. This improvement is carried out through the specific IGF-II receptor.Supported by M.G-F.&L.J.S. Proyectos I+D+I-Programa Operativo-FEDER Andalucía 2014-2020 (UMA18-FEDERJA-004) Junta de Andalucía. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Papel del receptor S1P sobre el estrés oxidativo mitocondrial en cultivo neuronal

Introducción: Fingolimod, fármaco inmunomodulador, presenta propiedades neuroprotectoras que podrían promover la recuperación de la función cognitiva en enfermedades neurodegenerativas. El estrés oxidativo parece tener un papel fundamental en la patogénesis de dichas enfermedades, siendo la mitocondria una de las fuentes más importantes de especies reactivas de oxigeno (ROS). Objetivo: Determinar la implicación del receptor S1P en los efectos neuroprotectores mostrados por fingolimod fosfato (FP), forma activa de fingolimod, en un modelo celular de estrés oxidativo mitocondrial inducido por menadiona (Vitk3). Material y métodos: La línea celular SN4741 (70-80 % confluencia), se utilizó como control o se trató con Vitk3 15 µM en presencia o ausencia de FP 50 nM o FP 50 nM + W123 10 µM (antagonista S1P) durante 4 horas para estudiar: niveles de ROS mitocondrial según el marcaje de la producción de anión superóxido (O2−.); activación de caspasa-3; niveles de tioles totales (TTLs); marcadores mitocondriales (potencial de membrana mitocondrial-PMM-, actividad citocromo c oxidasa-COX- y consumo de oxígeno-OCR-). Las diferencias estadísticas se determinaron usando ANOVA de un factor. Resultados: W123 revierte parcialmente el efecto protector de FP sobe muerte celular programada, desencadenada por aumento de ROS (p<0,05) y consumo de reserva de antioxidante (p<0,05). El efecto de FP sobre los marcadores mitocondriales PMM, actividad COX y OCR es abolido con W123 (p<0,05). Conclusión: El receptor S1P está implicado en gran parte de los efectos protectores de FP, indicando un papel fundamental de S1P en el mantenimiento de la homeostasis mitocondrial. Proyecto financiado por Novartis Farmacéutica SA (PS13/14).Campus de Excelencia Internacional Andalucía Tech. Proyecto financiado por Novartis Farmacéutica SA (PS13/14). Programa operativo de empleo juvenil; Junta de Andalucía and Fondo Social Europeo (EU). CTS507 and CTS156 from Consejería de Economía Innovación Ciencia y Empresa, Junta de Andalucía and Plan Propio de la Universidad de Málaga 2016

Parental practices and problematic internet use in aclinical sample of adolescents

Introducción: Estudios previos han demostrado el papel determinante de las prácticas parentales sobre el uso problemático de Internet (UPI) pero se desconoce qué variables familiares específicas están asociadas al UPI en muestras clínicas. Además, existe poca concordancia entre la percepción de padres e hijos sobre las variables de crianza familiar y de uso de Internet. Por este motivo, en este estudio se analiza la relación entre el UPI y las prácticas parentales en una muestra clínica, controlando los problemas de conducta y la personalidad de los adolescentes, y se examina la concordancia a través de correlación intraclase entre las percepciones de padres e hijos en estas variables. Método: Se cuenta con una muestra de 92 adolescentes de dos centros de salud mental infanto-juvenil. Resultados: Se encuentra que las prácticas de disciplina inconsistente y baja implicación parental son predictoras de UPI. Además, la concordancia encontrada entre padres e hijos es moderada-baja, tanto en las prácticas parentales percibidas como en el UPI. Discusión: Este estudio permite determinar la importancia de las prácticas parentales en el UPI de los adolescentes que acuden a centros de salud mental, así como la necesidad de realizar una evaluación exhaustiva más allá de los autoinformes, dada la baja concordancia entre padres e hijosIntroduction: Previous studies have shown the determining role of parenting practices on problematic Internet use (PIU), but it is unknown which specific family variables are associated with UPI in clinical samples. In addition, there is often little concordance through intraclass correlation between the perception of parents and children on the variables of family rearing and Internet use. For this reason, the objectives of this study are to analyze the relationship between UPI and parental practices in a clinical sample, controlling for behavior problems and adolescents› personality, and to examine the concordance between the perceptions of parents and children. Method: Data was collected from a sample of 92 adolescents in two mental health centers for children and adolescents. Results: The results indicate that inconsistent discipline practices and low parental involvement are predictors of UPI. In addition, the concordance between parents and children is moderate-to -low both in parenting practices and in the UPI. Discussion: This study makes it possible to determine the importance of parenting practices in the UPI of adolescents who attend mental health care; additionally, the need for an exhaustive evaluation beyond self-reports is detected, given the low concor- dance between parents and childre

New molecular mechanisms to explain the neuroprotective effects of insulin-like growth factor II in a cellular model of Parkinson's disease

Introduction: One of the hallmarks of Parkinsońs Disease (PD) is oxidative distress, leading to mitochondrial dysfunction and neurodegeneration. Insulin-like growth factor II (IGF-II) has been proven to have antioxidant and neuroprotective effects in some neurodegenerative diseases, including PD. Consequently, there isgrowing interest in understanding the different mechanisms involved in the neuroprotective effect of this hormone. Objectives: To clarify the mechanism of action of IGF-II involved in the protective effect of this hormone. Methods: The present study was carried out on a cellular model PD based on the incubation of dopaminergic cells (SN4741) in a culture with the toxic 1-methyl-4-phenylpyridinium (MPP+), in the presence of IGF-II. This model undertakes proteomic analyses in order to understand which molecular cell pathways might be involved in the neuroprotective effect of IGF-II. The most important proteins found in the proteomic study were tested by Western blot, colorimetric enzymatic activity assay and immunocytochemistry. Along with the proteomic study, mitochondrial morphology and function were also studied by transmission electron microscopy and oxygen consumption rate. The cell cycle was also analysed using 7AAd/BrdU staining, and flow cytometry. Results: The results obtained indicate that MPP+, MPP++IGF-II treatment and IGF-II, when compared to control, modified the expression of 197, 246 proteins and 207 respectively. Some of these proteins were found to be involved in mitochondrial structure and function, and cell cycle regulation. Including IGF-II in the incubation medium prevents the cell damage induced by MPP+, recovering mitochondrial function and cell cycle dysregulation, and thereby decreasing apoptosis. Conclusion: IGF-II improves mitochondrial dynamics by promoting the association of Mitofilin with mitochondria, regaining function and redox homeostasis. It also rebalances the cell cycle, reducing the amount of apoptosis and cell death by the regulation of transcription factors, such as Checkpoint kinase 1

Cocaine detrimentally affects mitochondrial functionality and cell viability in dopaminergic neurons.

An elevated consumption of cocaine (benzoylmethylecgonine), which causes anesthetic and stimulant effects on the central nervous system, may be associated with several neurodegenerative conditions affecting dopaminergic neurons, such as Parkinson's disease (PD). To investigate the impact of cocaine on cell viability and morphology, dopaminergic neurons from the substantia nigra (SN4741) were cultured. Analysis involved assessing cell death (LDH levels) and cell morphology (GIEMSA staining) after a 24-hour treatment period. Additionally, the effects on reactive oxygen species (ROS) generation (DH2), membrane potential (JC-1), oxygen consumption rate (OCR), and mitochondrial stress (Seahorse) were evaluated after a 6-hour treatment. The optimal concentration of cocaine for experimental use (2 mM) was identified, inducing a substantial 39.75% neuronal death. Examination of neuronal death (LDH) revealed a remarkable 280% increase following cocaine treatment. Optical analysis demonstrated heightened mortality and detrimental changes in neuronal morphology post-cocaine treatment, including a globose shape, loss of synapses, extremely thin membrane, and cell aggregation. In the "short time" experiments, mitochondrial oxidative damage was evident in SN cells treated with cocaine, leading to the demise of 75% of the cells. Furthermore, a significant 173.6% increase in reactive oxygen species (ROS) production and a 20% reduction in mitochondrial membrane potential (JC-1 assay) were observed. Cocaine treatment also resulted in a notable 60% decrease in mitochondrial oxygen consumption. In summary, a concentration of 2 mM cocaine induces a considerable rise in mitochondrial oxidative damage, subsequently causing morphological alterations and progressive death of dopaminergic neurons due to the accumulation of reactive oxygen species (ROS).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Inequalities in total mortality and by cause of death according to the level of education in Navarra: findings from a longitudinal study from 2001 to 2008

Fundamentos: Dada la ausencia de evidencia científica, el objetivo fue mostrar las desigualdades en mortalidad según el nivel de estudios en Navarra y la contribución de las principales causas de defunción a la magnitud de desigualdades en la mortalidad por todas las causas de muerte. Métodos: Todos los ciudadanos de 25 años y mayores residentes en Navarra en 2001 fueron seguidos durante 7 años para conocer su estado vital. El indicador de posición socioeconómica utilizado fue el nivel de estudios. Se estimaron las tasas de mortalidad general y por causa de muerte ajustadas por edad según la educación. Posteriormente, se calcularon la diferencia relativa (razón) y la diferencia absoluta de tasas entre las categorías más baja y más alta de nivel de estudios y la contribución de las principales causas de muerte a la diferencia absoluta. Resultados: La razón de tasas por todas las causas de muerte fue 1,37 en hombres y 1,23 en mujeres. El virus de la inmunodeficiencia humana (VIH) (25,84) y los accidentes no intencionales (3,78) presentaron las razones de tasas más altas en los hombres y la diabetes mellitus (4,92) y el VIH (4,38) en las mujeres. Las enfermedades cardiovasculares constituyeron la causa de muerte que más contribuyó a la diferencia absoluta en mortalidad: 26% en hombres y 48% en mujeres. Conclusiones: La tasa de mortalidad en la población navarra muestra un gradiente inverso con el nivel educativo, a excepción de algunas localizaciones de cáncer. Las enfermedades cardiovasculares son la causa de muerte que más contribuye a las desigualdades absolutas en mortalidad, mientras que otras causas de muerte que muestran importantes desigualdades relativas contribuyen poco a las desigualdades absolutas.Background: Due to the lack of evidence, the objective was to show the inequalities in mortality by educational level in Navarra and the contribution of the main causes of death to the magnitude of inequalities in mortality from all causes of death. Methods: All citizens aged 25 years and older residing in Spain in 2001 were followed during 7 years to determine their vital status. Level of education was used as socioeconomic status indicator. It was estimated the age-adjusted total mortality rate and mortality rate from cause-specific mortality by educational level. Then it was calculated the relative difference (ratio) and the absolute difference in rates between the lowest and highest levels of education and the contribution of the main causes of death to the absolute difference. Results: The rate ratio for all causes of death was 1.37 in men and 1.23 in women. The human immunodeficiency virus (HIV) (25.84) and unintentional injuries (3.78) are the causes of death with higher rate ratio in men and diabetes (4.92) and HIV (4.38) in women. Cardiovascular diseases were the leading causes of death that contribute most to the absolute difference in mortality: 26% in men and 48% women. Conclusions: The mortality rate in the Navarre population shows an inverse gradient with educational level, except in some cancer sites. Cardiovascular disease is the leading cause of death that contributes most to the absolute inequalities in mortality, while other causes of death that show significant relative inequalities, contribute little to the absolute inequalities.Fundación Caja Navarr