CERKL, a Retinal Dystrophy Gene, Regulates Mitochondrial Transport and Dynamics in Hippocampal Neurons.

Mutations in the Ceramide Kinase-like (CERKL) gene cause retinal dystrophies, characterized by progressive degeneration of retinal neurons, which eventually lead to vision loss. Among other functions, CERKL is involved in the regulation of autophagy, mitochondrial dynamics, and metabolism in the retina. However, CERKL is nearly ubiquitously expressed, and it has been recently described to play a protective role against brain injury. Here we show that Cerkl is expressed in the hippocampus, and we use mouse hippocampal neurons to explore the impact of either overexpression or depletion of CERKL on mitochondrial trafficking and dynamics along axons. We describe that a pool of CERKL localizes at mitochondria in hippocampal axons. Importantly, the depletion of CERKL in the CerklKD/KO mouse model is associated with changes in the expression of fusion/fission molecular regulators, induces mitochondrial fragmentation, and impairs axonal mitochondrial trafficking. Our findings highlight the role of CERKL, a retinal dystrophy gene, in the regulation of mitochondrial health and homeostasis in central nervous system anatomic structures other than the retina

The Alter Retina: Alternative Splicing of Retinal Genes in Health and Disease.

Alternative splicing of mRNA is an essential mechanism to regulate and increase the diversity of the transcriptome and proteome. Alternative splicing frequently occurs in a tissueor time-specific manner, contributing to differential gene expression between cell types during development. Neural tissues present extremely complex splicing programs and display the highest number of alternative splicing events. As an extension of the central nervous system, the retina constitutes an excellent system to illustrate the high diversity of neural transcripts. The retina expresses retinal specific splicing factors and produces a large number of alternative transcripts, including exclusive tissue-specific exons, which require an exquisite regulation. In fact, a current challenge in the genetic diagnosis of inherited retinal diseases stems from the lack of information regarding alternative splicing of retinal genes, as a considerable percentage of mutations alter splicing or the relative production of alternative transcripts. Modulation of alternative splicing in the retina is also instrumental in the design of novel therapeutic approaches for retinal dystrophies, since it enables precision medicine for specific mutations

Introducción al diseño de Cursos Cero para materias básicas del primer curso de Grado en Ciencias mediante enseñanza virtual

Evaluar, analizar la situación de partida de los alumnos de nuevo acceso al 1º curso de Grado en Facultades de Ciencias de la UCM (Biología, Óptica y Optometría y Química), y generar las bases para diseñar un Curso 0 de una materia básica, Química

Overexpression of cerkl protects retinal pigment epithelium mitochondria from oxidative stress effects

The precise function of CERKL, a Retinitis Pigmentosa (RP) causative gene, is not yet fully understood. There is evidence that CERKL is involved in the regulation of autophagy, stress granules, and mitochondrial metabolism, and it is considered a gene that is resilient against oxidative stress in the retina. Mutations in most RP genes affect photoreceptors, but retinal pigment epithelium (RPE) cells may be also altered. Here, we aimed to analyze the effect of CERKL overexpression and depletion in vivo and in vitro, focusing on the state of the mitochondrial network under oxidative stress conditions. Our work indicates that the depletion of CERKL increases the vulnerability of RPE mitochondria, which show a shorter size and altered shape, particularly upon sodium arsenite treatment. CERKL-depleted cells have dysfunctional mitochondrial respiration particularly upon oxidative stress conditions. The overexpression of two human CERKL isoforms (558 aa and 419 aa), which display different protein domains, shows that a pool of CERKL localizes at mitochondria in RPE cells and that CERKL protects the mitochondrial network both in size and shape against oxidative stress. Our results support CERKL being a resilient gene that regulates the mitochondrial network in RPE as in retinal neurons and suggest that RPE cell alteration contributes to particular phenotypic traits in patients carrying CERKL mutations

A perspectiva de gênero nas opiniões e discursos das mulheres sobre o cuidado

Objetivo: Explorar el significado que las mujeres cuidadoras dan a sus prácticas de cuidado y analizar la relación de dichas prácticas con las trayectorias laborales. Método: Estudio de corte cualitativo con diseño fenomenológico, realizado en Sevilla, España con cuidadoras familiares. La estrategia de obtención de datos ha sido la entrevista semi-estructurada. Los datos fueron analizados mediante análisis de contenido, se utilizó el software informático Atlas-ti GmbH versión 5.0®. Las categorías definidas son: historia cuidadora e historia laboral y significado del cuidado en sus vidas. Resultados: Participaron del estudio veinte cuidadoras. Las mujeres cuidadoras narran unas historias de cuidados extensas, cuidan a distintas personas de la unidad familiar en distintos momentos temporales. Los cuidados que desempeñan son muy diversos, contemplando la dimensión física y social. Se observa una contradicción entre el trabajo de cuidado y la incorporación y/o mantenimiento en el mercado laboral. Conclusión: Las mujeres expresan un concepto de cuidados integral que contempla la responsabilidad, la disponibilidad, el acompañamiento, así como estados afectivos, que generan un gran impacto en su salud. Las mujeres entrevistadas ajustan la participación en el mercado laboral a las necesidades de cuidados de la familia. Así, la perspectiva de género debe ser incorporada por los profesionales sanitarios que atienden los problemas de salud de las cuidadoras.Objective: To investigate the meanings given by women caregivers to their care practices and to analyze the relationship between such practices and their work history. Method: This was a phenomenological qualitative cohort study conducted in Seville, Spain, with family caregivers. Data were collected using semi-structured interviews and submitted to content analysis, using Atlas-ti GmbH software version 5.0®. The following categories emerged: caregiving history and work history, and the meaning of care in their lives. Results: Twenty caregivers participated in the study. Women caregivers told stories of wide-ranging care, including several individuals in their family at different moments. They performed different forms of care, encompassing physical and social dimensions. Their discourse expressed the contradiction between their caregiving role and entering and/or maintaining their place in the labor market. Conclusion: The women expressed a concept of comprehensive care that includes responsibility, availability, and companionship, as well as emotional states, which greatly impact health. They tailored their participation in the labor market according to their family’s care needs. Thus, the gender perspective must be incorporated by health care providers who work with the health of caregivers.Objetivo: Explorar o significado que as mulheres cuidadoras dão às suas práticas de cuidado e analisar a relação dessas práticas com as trajetórias de trabalho. Método: Estudo qualitativo, com desenho fenomenológico de desígnio, realizado em Sevilha, Espanha com cuidadoras familiares. A estratégia de obtenção de dados foi a entrevista semiestruturada. Os dados foram analisados com a metodologia de análise de conteúdo, por meio do software Atlas-ti GmbH, versão 5.0®. As categorias definidas são: história de cuidados e história laboral e o significado do cuidado em suas vidas. Resultados: Participaram do estudo 20 cuidadoras. As mulheres cuidadoras narram algumas histórias de cuidados extensas e cuidam de diferentes pessoas da unidade familiar em diferentes momentos temporais. Os cuidados que desempenham são muito diversos, contemplando a dimensão física e social. Observa-se uma contradição entre o trabalho de cuidado e a incorporação e/ou manutenção no mercado de trabalho. Conclusão: As mulheres expressam um conceito integrante de cuidados que abarca a responsabilidade, a disponibilidade, o acompanhamento, como também estados afetivos que geram um grande impacto em sua saúde. As mulheres entrevistadas ajustam a participação no mercado de trabalho com as necessidades de cuidados da família. Deste modo, a perspectiva de gênero deve ser incorporada pelos profissionais da saúde que atendem aos problemas de saúde das cuidadoras

A perspectiva de gênero nas opiniões e discursos das mulheres sobre o cuidado

Objective: To investigate the meanings given by women caregivers to their care practices and to analyze the relationship between such practices and their work history. Method: This was a phenomenological qualitative cohort study conducted in Seville, Spain, with family caregivers. Data were collected using semi-structured interviews and submitted to content analysis, using Atlas-ti GmbH software version 5.0®. The following categories emerged: caregiving history and work history, and the meaning of care in their lives. Results: Twenty caregivers participated in the study. Women caregivers told stories of wide-ranging care, including several individuals in their family at different moments. They performed different forms of care, encompassing physical and social dimensions. Their discourse expressed the contradiction between their caregiving role and entering and/or maintaining their place in the labor market. Conclusion: The women expressed a concept of comprehensive care that includes responsibility, availability, and companionship, as well as emotional states, which greatly impact health. They tailored their participation in the labor market according to their family’s care needs. Thus, the gender perspective must be incorporated by health care providers who work with the health of caregivers

The CRISPR/Cas9 system efficiently reverts the tumorigenic ability of BCR/ABL in vitro and in a xenograft model of chronic myeloid leukemia

[EN]CRISPR/Cas9 technology was used to abrogate p210 oncoprotein expression in the Boff-p210 cell line, a pro-B line derived from interlukin-3-dependent Baf/3, that shows IL-3-independence arising from the constitutive expression of BCR-ABL p210. Using this approach, pools of Boff-p210-edited cells and single edited cell-derived clones were obtained and functionally studied in vitro. The loss of p210 expression in Boff-p210 cells resulted in the loss of ability to grow in the absence of IL-3, as the Baf/3 parental line, showing significantly increased apoptosis levels. Notably, in a single edited cell-derived clone carrying a frame-shift mutation that prevents p210 oncoprotein expression, the effects were even more drastic, resulting in cell death. These edited cells were injected subcutaneously in immunosuppressed mice and tumor growth was followed for three weeks. BCR/ABL-edited cells developed smaller tumors than those originating from unedited Boff-p210 parental cells. Interestingly, the single edited cell-derived clone was unable to develop tumors, similar to what is observed with the parental Baf/3 cell line. CRISPR/Cas9 genomic editing technology allows the ablation of the BCR/ ABL fusion gene, causing an absence of oncoprotein expression, and blocking its tumorigenic effects in vitro and in the in vivo xenograft model of CML. The future application of this approach in in vivo models of CML will allow us to more accurately assess the value of CRISPR/Cas9 technology as a new therapeutic tool that overcomes resistance to the usual treatments for CML patients

Evaluation of biostimulation, bioaugmentation, and organic amendments application on the bioremediation of recalcitrant hydrocarbons of soil

In the present work, the operational conditions for improving the degradation rates of Total Petroleum Hydrocarbons (TPHs) in contaminated soil from a machinery park were optimized at a microcosms scale along a 90- days incubation period. In this study, bioremediation strategies and an organic amendment have been tested to verify the remediation of soil contaminated with different hydrocarbons, mineral oils, and heavy metals. Specifically, designed biostimulation and bioaugmentation strategies were compared with and without adding vermicompost. The polluted soil harboring multiple contaminants, partially attenuated for years, was used. The initial profile showed enrichment in heavy linear alkanes, suggesting a previous moderate weathering. The application of vermicompost increased five and two times the amounts of available phosphorus (P) and exchangeable potassium (K), respectively, as a direct consequence of the organic amendment addition. The microbial activity increased due to soil acidification, which influenced the solubility of P and other micronutrients. It also impacted the predominance and variability of the different microbial groups and the incubation, as reflected by phospholipid fatty acid (PLFA) results. An increase in the alkaline phosphatases and proteases linked to bacterial growth was displayed. This stimulation of microbial metabolism correlated with the degradation rates since TPHs degradation’ efficiency after vermicompost addition reached 32.5% and 34.4% of the initial hydrocarbon levels for biostimulation and bioaugmentation, respectively. Although Polycyclic Aromatic Hydrocarbons (PAHs) were less abundant in this soil, results also decreased, especially for the most abundant, the phenanthrene. Despite improving the degradation rates, results revealed that recalcitrant and hydrophobic petroleum compounds remained unchanged, indicating that mobility, linked to bioavailability, probably represents the limiting step for further soil recovery.This work is funded by the GREENER project of the European Union’s Horizon 2020 research and innovation program (Grant Agreement No. 826312). S. Curiel pre-doctoral contract was funded by Junta de Castilla y Leon ´ (ORDEN EDU/1508/2020, de 15 de diciembre)

Estudio etnobotánico y evaluación citotóxica de extractos etanólicos de plantas de uso medicinal en Tlalchi, Ixhuacán de los Reyes, Veracruz, México

Las plantas medicinales son especies vegetales que contienen sustancias empleadas con propósitos terapéuticos y pueden servir de precursores para la síntesis de nuevos fármacos. En el presente trabajo se realizó un estudio etnobotánico y toxicológico de plantas de uso medicinal de Tlalchi, Ixhuacán de los Reyes en el estado de Veracruz, México. Se registraron 22 especies distribuidas en 17 familias, destacando Asteraceae con el mayor número de frecuencia. Las especies seleccionadas fueron Solanum nigrum, Bidens pilosa, Tithonia diversifolia, Commelina diffusa, Justicia pectoralis, y Aldama dentata. Se realizó un análisis fitoquímico preliminar de las mismas, donde se obtuvo la presencia de alcaloides, flavonoides, triterpenos y saponinas. Solanum nigrum tuvo mayor toxicidad de 51.72 µg/mL. Esta última especie tuvo toxicidad en la línea celular HeLa, obteniendo una reducción del 50% de viabilidad celular a partir de las 24 h. S. nigrum podría ser un potencial agente anti-proliferativo en este tipo de células