Understanding underlying mechanisms in Nemaline Myopathy: the role of cardiolipin.

Motivation: Nemaline myopathy (NM) is one of the most common forms of congenital myopathies, a group of heterogeneous genetic muscle disorders. The main characteristic of NM is the presence of inclusion bodies in muscle fibers, the so-called 'nemaline body' or rods. These rod-like inclusions are mostly composed of actin, alpha-actin, and other Z-band filaments. Actin Alpha 1 (ACTA1) and Nebulin (NEB) gene mutations are the main cause of NM. A recent study, employing dermal fibroblasts from individuals with NM mutations demonstrated that incorrect actin filament polymerization is associated with mitochondrial dysfunction; In addition, L-carnitine and linoleic acid, two mitochondrial-boosting compounds, enhanced actin filament polymerization and restore mitochondrial bioenergetics. However, the underlying molecular mechanisms of these compounds are unknown. This study aims to bridge this gap by analyzing the effect of these compounds on cardiolipin (CL) concentrations, due to its significant role in the structure and the functioning of the mitochondria. Cardiolipin is a tetra-acylated diphosphatidylglycerol lipid that is predominantly localized in the inner mitochondrial membrane (IMM).Methods: Four NM patients participated in this research, two patients with ACTA1 mutations and, two patients with NEB mutations. Variations in the expression of proteina involved in cardiolipin synthesis were analyzed by Western Blotting: cardiolipin synthase 1 (CLS1), which synthesized immature CL on the stromal side of the IMM and phospholipid-lysophospholipid transacylase Tafazzin (TAZ), which is involved in the remodeling phase to produce mature CL. Also, the concentration of cardiolipin in fibroblast was analyzed by fluorometric assay. Fibroblasts from two patients carrying one of these mutations were treated with L-carnitine and linoleic acid for a week.Results: Mutant fibroblasts derived from NM patients presented lower expression levels of TAZ protein. In addition, CL concentration was decreased in patients' fibroblasts compared to untreated fibroblasts and controls.Conclusions: Due to the mitochondrial dysfunction in mutant fibroblasts, cardiolipin is not maturing properly. Therefore, CL levels are decreased. The increased cardiolipin concentration by L-carnitine and linoleic acid treatment suggests that this may be one of the mechanisms of action by which these compounds improve mitochondrial function and actin polymerization

Didáctica a través del cuento. Validación cupic. / Didactic through the cuento. Validación cupic.

La literatura infantil ha sido considerado como un recurso pedagógico para ofrecer conocimiento a los niños. En este documento nos vamos a centrar en los cuentos, más concretamente en los cuentos infantiles.El principal objetivo de este documento, es conocer la idea que se tiene sobre el cuento por ello se ha realizado un cuestionario para conocer las consideraciones que se tienen sobre el uso del cuento y sus formas.El cuento, está considerado como un fuerte recurso educativo, por ello se ha considerado necesario realizar una labor de investigación a través de diferentes autores para corroborar esta afirmación. Por ello, nos hemos aproximado a una conceptualización del cuento, su clasificación y las características de cada uno de ellos, el trabajo de emociones a partir de cuentos y el valor educativo propio de los estos en la etapa de educación infantil.Dada la forma en la que se organiza la metodología en Infantil, el cuento, dadas sus características es un recurso que va ayudar a docentes a conseguir los propósitos educativos propios de la etapa, promoviendo un aprendizaje significativo e integral. Lo más importante para que un cuento provoque un aprendizaje es, la elección de los mismos y la forma en la que se utilice, de ahí la importancia en conocerlos y saber de los cuentos.Este proceso de revisión minuciosa dio como resultado la duda —no resuelta en artículos de investigación presentados en los últimos años— acerca de si los docentes —parte fundamental del proceso de E-A— son conscientes o no de la importancia subyacente al uso de los cuentos como herramienta didáctica. Es por ello que se procedió a la confección y validación de un cuestionario (CUPIC) que, sin lugar a duda, servirá como base para futuras investigaciones

HIV-1 Tropism Testing in Subjects Achieving Undetectable HIV-1 RNA : Diagnostic Accuracy, Viral Evolution and Compartmentalization

Technically, HIV-1 tropism can be evaluated in plasma or peripheral blood mononuclear cells (PBMCs). However, only tropism testing of plasma HIV-1 has been validated as a tool to predict virological response to CCR5 antagonists in clinical trials. The preferable tropism testing strategy in subjects with undetectable HIV-1 viremia, in whom plasma tropism testing is not feasible, remains uncertain. We designed a proof-of-concept study including 30 chronically HIV-1-infected individuals who achieved HIV-1 RNA <50 copies/mL during at least 2 years after first-line ART initiation. First, we determined the diagnostic accuracy of 454 and population sequencing of gp120 V3-loops in plasma and PBMCs, as well as of MT-2 assays before ART initiation. The Enhanced Sensitivity Trofile Assay (ESTA) was used as the technical reference standard. 454 sequencing of plasma viruses provided the highest agreement with ESTA. The accuracy of 454 sequencing decreased in PBMCs due to reduced specificity. Population sequencing in plasma and PBMCs was slightly less accurate than plasma 454 sequencing, being less sensitive but more specific. MT-2 assays had low sensitivity but 100% specificity. Then, we used optimized 454 sequence data to investigate viral evolution in PBMCs during viremia suppression and only found evolution of R5 viruses in one subject. No de novo CXCR4-using HIV-1 production was observed over time. Finally, Slatkin-Maddison tests suggested that plasma and cell-associated V3 forms were sometimes compartmentalized. The absence of tropism shifts during viremia suppression suggests that, when available, testing of stored plasma samples is generally safe and informative, provided that HIV-1 suppression is maintained. Tropism testing in PBMCs may not necessarily produce equivalent biological results to plasma, because the structure of viral populations and the diagnostic performance of tropism assays may sometimes vary between compartments. Thereby, proviral DNA tropism testing should be specifically validated in clinical trials before it can be applied to routine clinical decision-making

Usos del e-learning en las Universidades Andaluzas: Estado de la situación y análisis de buenas prácticas. (Versión completa)

Secretaría General de Universidades, Investigación y Tecnología P07-SE-J.0267

Unraveling the effect of silent, intronic and missense mutations on VWF splicing : contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to the identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7437G>A) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of three mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that four of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease. identifier:02869074

Rate of severe and fatal infections in a cohort of patients with interstitial lung disease associated with rheumatoid arthritis: a multicenter prospective study

ObjectiveTo describe severe infection, foci of infection, microorganisms, associated factors, and impact on mortality in patients with rheumatoid arthritis–associated interstitial lung disease (RA-ILD).Patients and methodsThe study was based on a multicenter prospective cohort of patients with RA-ILD followed up from 2015 to 2023. The main outcome measures were incident severe infection and fatal infection. We evaluated infectious foci, etiologic agents, vaccination status, variables associated with lung function, and clinical-therapeutic variables in RA. The incidence rate (IR) for infection and mortality was calculated per 100 person-years, and 3 multivariate models were constructed to explore factors associated with infection.ResultsWe followed up 148 patients with RA-ILD for a median 56.7 months (699.3 person-years). During this period, 142 patients (96%) had at least 1 infection. A total of 368 infectious episodes were recorded, with an IR of 52.6 per 100 person-years. Of the 48 patients who died, 65% did so from infection. Respiratory infections were the most common first infection (74%), infection overall (74%), and fatal infection (80%) and were caused mostly by SARS CoV-2, Streptococcus pneumoniae, Pseudomonas aeruginosa, and influenza A virus. The factors associated with an increased risk of infection and death in patients with RA-ILD were age, inflammatory activity, and therapy with corticosteroids and immunosuppressants.ConclusionPatients with RA-ILD have a high risk of serious infection, especially respiratory infection. Infection develops early, is recurrent, and is frequently fatal. The presence of associated factors such as advanced age, joint inflammation, and treatment highlight the importance of integrated and preventive medical care

Izaña Atmospheric Research Center. Activity Report 2019-2020

Editors: Emilio Cuevas, Celia Milford and Oksana Tarasova.[EN]The Izaña Atmospheric Research Center (IARC), which is part of the State Meteorological Agency of Spain (AEMET), is a site of excellence in atmospheric science. It manages four observatories in Tenerife including the high altitude Izaña Atmospheric Observatory. The Izaña Atmospheric Observatory was inaugurated in 1916 and since that date has carried out uninterrupted meteorological and climatological observations, contributing towards a unique 100-year record in 2016. This reports are a summary of the many activities at the Izaña Atmospheric Research Center to the broader community. The combination of operational activities, research and development in state-of-the-art measurement techniques, calibration and validation and international cooperation encompass the vision of WMO to provide world leadership in expertise and international cooperation in weather, climate, hydrology and related environmental issues.[ES]El Centro de Investigación Atmosférica de Izaña (CIAI), que forma parte de la Agencia Estatal de Meteorología de España (AEMET), representa un centro de excelencia en ciencias atmosféricas. Gestiona cuatro observatorios en Tenerife, incluido el Observatorio de Izaña de gran altitud, inaugurado en 1916 y que desde entonces ha realizado observaciones meteorológicas y climatológicas ininterrumpidas y se ha convertido en una estación centenaria de la OMM. Estos informes resumen las múltiples actividades llevadas a cabo por el Centro de Investigación Atmosférica de Izaña. El liderazgo del Centro en materia de investigación y desarrollo con respecto a las técnicas de medición, calibración y validación de última generación, así como la cooperación internacional, le han otorgado una reputación sobresaliente en lo que se refiere al tiempo, el clima, la hidrología y otros temas ambientales afines