Transcriptomic analysis of Streptomyces coelicolor differentiation in solid sporulating cultures: first compartmentalized and second multinucleated mycelia have different and distinctive transcriptomes

[EN] Streptomycetes are very important industrial bacteria, which produce two thirds of all clinically relevant secondary metabolites. They have a complex developmental-cycle in which an early compartmentalized mycelium (MI) differentiates to a multinucleated mycelium (MII) that grows inside the culture medium (substrate mycelium) until it starts to growth into the air (aerial mycelium) and ends up forming spores. Streptomyces developmental studies have focused mainly on the later stages of MII differentiation (aerial mycelium and sporulation), with regulation of pre-sporulation stages (MI/MII transition) essentially unknown. This work represents the first study of the Streptomyces MI transcriptome, analyzing how it differs from the MII transcriptome. We have used a very conservative experimental approach to fractionate MI from MII and quantify gene expressions. The expression of well characterized key developmental/metabolic genes involved in bioactive compound production (actinorhodin, undecylprodigiosin, calcium-dependent antibiotic, cpk, geosmin) or hydrophobic cover formation-sporulation (bld, whi, wbl, rdl, chp, ram) was correlated with MII differentiation. Additionally, 122 genes conserved in the Streptomyces genus, whose biological function had not been previously characterized, were found to be differentially expressed (more than 4-fold) in MI or MII. These genes encoded for putative regulatory proteins (transcriptional regulators, kinases), as well as hypothetical proteins. Knowledge about differences between the MI (vegetative) and MII (reproductive) transcriptomes represents a huge advance in Streptomyces biology that will make future experiments possible aimed at characterizing the biochemical pathways controlling pre-sporulation developmental stages and activation of secondary metabolism in StreptomycesSIThis research was funded by grant BIO2010-16303 from the Subdirección General de Proyectos de Investigación, (DGI), Ministry of Science and Innovation (MICINN), Spain; and by an ERC Starting Grant (Strp-differentiation 280304). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Lecciones básicas de Derecho e instituciones de la Unión Europea

Los materiales docentes que aquí se presentan pretenden ser un texto de ayuda y consulta para aquellos que se acercan al Derecho de la Unión Europea; en ellos no sólo se contienen los conceptos básicos de la materia, sino que se han añadido a éstos, una serie de actividades didácticas y formativas específicamente orientadas al estudiante, y que le han de servir como instrumento para poder asentar sus conocimientos mediante la realización de prácticas y autoevaluaciones.The teaching materials presented here are intended as a help text and consultation for those who come to the European Union law; they not only contain the basic concepts of matter, but that have been added to these, a series of educational activities and training specifically geared toward the student, and that you have to serve as the instrument to be able to hone their knowledge through practical work and self-evaluations

Hereditary Leiomyomatosis and Renal Cell Cancer Syndrome in Spain: Clinical and Genetic Characterization

Simple Summary Hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome is a very rare hereditary disorder characterized by cutaneous leiomyomas (CLMs), uterine leiomyomas (ULMs), renal cysts (RCys) and renal cell cancer (RCC), with no data on its prevalence worldwide. No genotype-phenotype associations have been described. The aim of our study was to describe the genotypic and phenotypic features of the largest series of patients with HLRCC from Spain reported to date. Of 27 FH germline pathogenic variants, 12 were not previously reported in databases. Patients with missense pathogenic variants showed higher frequencies of CLMs, ULMs and RCys, than those with loss-of-function variants. The frequency of RCCs (10.9%) was lower than those reported in the previously published series. Hereditary leiomyomatosis and renal cell cancer syndrome (HLRCC) is a very rare hereditary disorder characterized by cutaneous leiomyomas (CLMs), uterine leiomyomas (ULMs), renal cysts (RCys) and renal cell cancers (RCCs). We aimed to describe the genetics, clinical features and potential genotype-phenotype associations in the largest cohort of fumarate hydratase enzyme mutation carriers known from Spain using a multicentre, retrospective study of individuals with a genetic or clinical diagnosis of HLRCC. We collected clinical information from medical records, analysed genetic variants and looked for genotype-phenotype associations. Analyses were performed using R 3.6.0. software. We included 197 individuals: 74 index cases and 123 relatives. CLMs were diagnosed in 65% of patients, ULMs in 90% of women, RCys in 37% and RCC in 10.9%. Twenty-seven different pathogenic variants were detected, 12 (44%) of them not reported previously. Patients with missense pathogenic variants showed higher frequencies of CLMs, ULMs and RCys, than those with loss-of-function variants (p = 0.0380, p = 0.0015 and p = 0.024, respectively). This is the first report of patients with HLRCC from Spain. The frequency of RCCs was lower than those reported in the previously published series. Individuals with missense pathogenic variants had higher frequencies of CLMs, ULMs and RCys

Experiencias de Innovación docente en los Estudios Jurídicos: una visión práctica

Esta publicación se enmarca dentro de las actividades del Grupo de Investigación de la Universidad de Extremadura Fiscalitas & Iuris.Este trabajo surge con el objetivo principal de dar visibilidad y publicidad a las nuevas técnicas docentes en el seno de la Facultad de Derecho de la UEx. Como se sabe, se ha producido un innegable y significativo avance en el uso de nuevas técnicas docentes y también de las TICs aplicadas a la docencia en la Facultad de Derecho, no obstante, aún es necesario profundizar en el uso de las mismas y extenderlas entre todos los miembros del claustro de profesores, y fundamentalmente entre aquellos que llevan más años ejerciendo la docencia a través de la colaboración y la coordinación con los profesores noveles, que son quienes principalmente se sirven en mayor medida de tales instrumentos docentes. De otra parte, también era necesario que los docentes más experimentados pudieran encontrar un foro en el que transmitir y compartir con los noveles cuales son las técnicas e instrumentos docentes que ellos han venido utilizando durante el ejercicio de su magisterio, de modo que, en el marco de una relación sinalagmática, se produjera una interacción entre uno u otro grupo de docentes, a fin de fomentar el necesario debate y el intercambio de experiencias e instrumentos docentes, y en su caso el desarrollo y perfeccionamiento de los mismos; algo que hemos pretendido realizar con este trabajo, y que en buena medida hemos logrado. Las finalidades y objetivos concretos que perseguíamos, en atención a la situación expuesta eran fundamentalmente tres: • En primer lugar, la implementación de un proyecto de innovación docente integrado por una diversidad de actividades coordinadas, cada uno de ellas bajo la directa coordinación de un profesor o profesora de la UEx, aplicado a una o varias asignaturas impartidas en la Facultad de Derecho. • En segundo lugar, el establecimiento en la Facultad de Derecho de un foro de coordinación e intercambio de buenas prácticas docentes sobre la base de cada uno de las actividades coordinadas, en el que pudieran participar profesores noveles y veteranos. Para ello se desarrolló espacio virtual de innovación docente en estudios jurídicos, a través del Campus Virtual de la UEx, en el que los Profesores noveles y veteranos pudieron y puede compartir recursos e informaciones sobre prácticas de innovación. • Y, en tercer lugar, la difusión y consolidación de instrumentos de innovación docente directamente aplicadas a la docencia de los estudios jurídicos, mediante la transferencia de los resultados y la publicación de los mismos; a fin de que esta transferencia sirva de base a futuras profundizaciones en el campo de la innovación docente en los estudios jurídicos.Proyecto “Desarrollo, profundización e intercambio de buenas prácticas de innovación docente en la Facultad de Derecho” (UEx 2015-2016

Endoscopic Submucosal Dissection. Sociedad Española de Endoscopia Digestiva (SEED) clinical guideline

The reason to write this guideline is to familiarize Spanish endoscopists and gastroenterologists not only with the general indications of the procedure and possible complications but also the dedicated tools

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Nucleoredoxin downregulation reduces β-Catenin levels and shifts hematopoietic differentiation towards myeloid lineage in vitro

[EN]The importance of dissecting signaling pathways governing cell differentiation is based on their relevance not only for understanding basic biological phenomena but also for better comprehending the underlying mechanisms of pathologic alterations such as cancer. A paradigm of cell differentiation processes is hematopoiesis, where a single stem cell gives rise to multiple, fully differentiated, cell lineages. Nucleoredoxin (Nrx), a member of the thioredoxin family, is an important redox-sensitive modulator of Wnt/ -catenin signaling, a key pathway for the control of hematopoiesis. In this work, the relevance of Nrx for the differentiation of mouse hematopoietic progenitor cells has been analyzed in vitro. Nrx silencing leads to a dramatic reduction in the size of the Lin and LSK progenitor populations. Moreover, there is also a remarkable decrease in CD3+ cells and an enhancement in the percentage of CD11b+Gr1 myeloid cells. This myeloid bias would agree with the inhibition of the Wnt/ -catenin pathway. Interestingly, a reduction in -catenin at the protein level was observed upon Nrx silencing. Our results strongly support the importance of Nrx for hematopoietic differentiation, which could be mediated by the regulation of the Wnt/ -catenin pathway

CX3CR1-deficient microglia shows impaired signalling of the transcription factor NRF2: Implications in tauopathies

TAU protein aggregation is the main characteristic of neurodegenerative diseases known as tauopathies. Low-grade chronic inflammation is also another hallmark that indicates crosstalk between damaged neurons and glial cells. Previously, we have demonstrated that neurons overexpressing TAUP301L release CX3CL1, which activates the transcription factor NRF2 signalling to limit over-activation in microglial cells in vitro and in vivo. However, the connection between CX3CL1/CX3CR1 and NRF2 system and its functional implications in microglia are poorly described. We evaluated CX3CR1/NRF2 axis in the context of tauopathies and its implication in neuroinflammation. Regarding the molecular mechanisms that connect CX3CL1/CX3CR1 and NRF2 systems, we observed that in primary microglia from Cx3cr1-/- mice the mRNA levels of Nrf2 and its related genes were significantly decreased, establishing a direct linking between both systems. To determine functional relevance of CX3CR1, migration and phagocytosis assays were evaluated. CX3CR1-deficient microglia showed impaired cell migration and deficiency of phagocytosis, as previously described for NRF2-deficient microglia, reinforcing the idea of the relevance of the CX3CL1/CX3CR1 axis in these events. The importance of these findings was evident in a tauopathy mouse model where the effects of sulforaphane (SFN), an NRF2 inducer, were examined on neuroinflammation in Cx3cr1+/+ and Cx3cr1-/- mice. Interestingly, the treatment with SFN was able to modulate astrogliosis but failed to reduce microgliosis in Cx3cr1-/- mice. These findings suggest an essential role of the CX3CR1/NRF2 axis in microglial function and in tauopathies. Therefore, polymorphisms with loss of function in CX3CR1 or NRF2 have to be taken into account for the development of therapeutic strategies. Keywords: Inflammation, Neurodegeneration, TAU, Migration, TAM receptors, AXL, Microgliosis, Sulforaphan

Nucleoredoxin Downregulation Reduces β-Catenin Levels and Shifts Hematopoietic Differentiation towards Myeloid Lineage In Vitro

The importance of dissecting signaling pathways governing cell differentiation is based on their relevance not only for understanding basic biological phenomena but also for better comprehending the underlying mechanisms of pathologic alterations such as cancer. A paradigm of cell differentiation processes is hematopoiesis, where a single stem cell gives rise to multiple, fully differentiated, cell lineages. Nucleoredoxin (Nrx), a member of the thioredoxin family, is an important redox-sensitive modulator of Wnt/β-catenin signaling, a key pathway for the control of hematopoiesis. In this work, the relevance of Nrx for the differentiation of mouse hematopoietic progenitor cells has been analyzed in vitro. Nrx silencing leads to a dramatic reduction in the size of the Lin− and LSK progenitor populations. Moreover, there is also a remarkable decrease in CD3+ cells and an enhancement in the percentage of CD11b+Gr1− myeloid cells. This myeloid bias would agree with the inhibition of the Wnt/β-catenin pathway. Interestingly, a reduction in β-catenin at the protein level was observed upon Nrx silencing. Our results strongly support the importance of Nrx for hematopoietic differentiation, which could be mediated by the regulation of the Wnt/β-catenin pathway