Utilización del cultivo de tejidos vegetales in vitro para el estudio de la respuesta de los cítricos al estrés salino

Este trabajo describe la puesta a punto de un sistema in vitro para el estudio de la toxicidad del NaCl en tres genotipos de cítricos, evitando el filtro de iones que constituye la raíz. Los cultivos se establecieron a partir de plantas de mandarino Cleopatra, citrange Carrizo y citrumelo CPB4475 cultivadas en invernadero. Se ensayaron diferentes concentraciones de NaCl y se seleccionó 60 mM como tratamiento salino para los diferentes experimentos. Los brotes de todos los genotipos estudiados acumularon concentraciones similares de iones cloruro cuando se cultivaron desprovistos del sistema radicular y mostraron los mismos daños foliares. No se observó incremento en la concentración de malondialdehido (como indicador del daño oxidativo) en ninguno de los genotipos y todos ellos mostraron patrones similares de señalización hormonal, con independencia de su tolerancia o sensibilidad cuando se cultivan en campo. El sistema in vitro descrito se postula como una herramienta útil para el estudio de los procesos bioquímicos implicados en la respuesta de los cítricos al estrés salino.In this work, an in vitro experimental system has been developed to study the toxic effect of NaCl on three citrus genotypes, avoiding the ion filter that represents the root system. Cultures were established from greenhouse growing plants of Cleopatra mandarin, Carrizo citrange and citrumelo CPB4475. Several salt concentrations were tested and 60 mM NaCl was selected as saline treatment. Shoots from all studied genotypes accumulated similar levels of chloride when cultured without roots and exhibited similar leaf damage. No increases in malondialdehyde levels (as a measure of oxidative stress) were observed in any genotype and similar patterns of hormonal signalling were exhibited in the three genotypes, despite their different tolerance under field conditions. The in vitro culture system has been proved as a useful tool to study biochemical processes involved in the response of citrus to salt stress.Este estudio fue financiado por el Ministerio de Ciencia e Innovación y la Fundació Bancaixa/Universitat Jaume I a través de la concesión de los proyectos AGL2010-22195-C03-01/AGR y P1 1B2009-01, respectivament

Clinical Heterogeneity of Pulmonary Arterial Hypertension Associated With Variants in TBX4

Background: The knowledge of hereditary predisposition has changed our understanding of Pulmonary Arterial Hypertension. Genetic testing has been widely extended and the application of Pulmonary Arterial Hypertension specific gene panels has allowed its inclusion in the diagnostic workup and increase the diagnostic ratio compared to the traditional sequencing techniques. This is particularly important in the differential diagnosis between Pulmonary Arterial Hypertension and Pulmonary Venoocclusive Disease. Methods: Since November 2011, genetic testing is offered to all patients with idiopathic, hereditable and associated forms of Pulmonary Arterial Hypertension or Pulmonary Venoocclusive Disease included in the Spanish Registry of Pulmonary Arterial Hypertension. Herein, we present the clinical phenotype and prognosis of all Pulmonary Arterial Hypertension patients with disease-associated variants in TBX4. Results: Out of 579 adults and 45 children, we found in eight patients from seven families, disease-causing associated variants in TBX4. All adult patients had a moderate-severe reduction in diffusion capacity. However, we observed a wide spectrum of clinical presentations, including Pulmonary Venoocclusive Disease suspicion, interstitial lung disease, pulmonary vascular abnormalities and congenital heart disease. Conclusions: Genetic testing is now essential for a correct diagnosis work-up in Pulmonary Arterial Hypertension. TBX4-associated Pulmonary Arterial Hypertension has marked clinical heterogeneity. In this regard, a genetic study is extremely useful to obtain an accurate diagnosis and provide appropriate management.This project was founded by Project "Bases Gene´tico Moleculares de la Medicina de Precisio´n en la Hipertensio´n Arterial Pulmonar". Funder: Instituto Carlos III. Ministerio de Economı´a y Competitividad. https://www.isciii.es/Paginas/Inicio.aspx Award number: PI 18/01233 Grant Recipient: P E-

La metamorfosis de Europa ¿Triunfo de los mercados?

Curso de veran

Saturated fatty acid-enriched small extracellular vesicles mediate a crosstalk inducing liver inflammation and hepatocyte insulin resistance

[Background & Aims]: Lipotoxicity triggers non-alcoholic fatty liver disease (NAFLD) progression owing to the accumulation of toxic lipids in hepatocytes including saturated fatty acids (SFAs), which activate pro-inflammatory pathways. We investigated the impact of hepatocyte- or circulating-derived small extracellular vesicles (sEV) secreted under NAFLD conditions on liver inflammation and hepatocyte insulin signalling. [Methods]: sEV released by primary mouse hepatocytes, characterised and analysed by lipidomics, were added to mouse macrophages/Kupffer cells (KC) to monitor internalisation and inflammatory responses. Insulin signalling was analysed in hepatocytes exposed to conditioned media from sEV-loaded macrophages/KC. Mice were i.v. injected sEV to study liver inflammation and insulin signalling. Circulating sEV from mice and humans with NAFLD were used to evaluate macrophage–hepatocyte crosstalk. [Results]: Numbers of sEV released by hepatocytes increased under NAFLD conditions. Lipotoxic sEV were internalised by macrophages through the endosomal pathway and induced pro-inflammatory responses that were ameliorated by pharmacological inhibition or deletion of Toll-like receptor-4 (TLR4). Hepatocyte insulin signalling was impaired upon treatment with conditioned media from macrophages/KC loaded with lipotoxic sEV. Both hepatocyte-released lipotoxic sEV and the recipient macrophages/KC were enriched in palmitic (C16:0) and stearic (C18:0) SFAs, well-known TLR4 activators. Upon injection, lipotoxic sEV rapidly reached KC, triggering a pro-inflammatory response in the liver monitored by Jun N-terminal kinase (JNK) phosphorylation, NF-κB nuclear translocation, pro-inflammatory cytokine expression, and infiltration of immune cells into the liver parenchyma. sEV-mediated liver inflammation was attenuated by pharmacological inhibition or deletion of TLR4 in myeloid cells. Macrophage inflammation and subsequent hepatocyte insulin resistance were also induced by circulating sEV from mice and humans with NAFLD. [Conclusions]: We identified hepatocyte-derived sEV as SFA transporters targeting macrophages/KC and activating a TLR4-mediated pro-inflammatory response enough to induce hepatocyte insulin resistance.This work was supported by grants PID2021-122766OB-I00 (AMV), PID2019-105989RB-I00 (JB), PID2020-113238RB-I00 (LB), PID2019-106581RB-I00 (MAM), PID2020-114148RB-I00 (MI), PID2019-107036RB-I00 (RF), and RD21/0006/0001 (ISCIII) (IL) funded by Ministerio de Ciencia e Innovación/Agencia Estatal de Investigación/10.13039/501100011033 and ERDF ‘A way of making Europe’ by the European Union (MICINN/AEI/FEDER, EU), grant EFSD/Boehringer Ingelheim European Research Programme on ‘Multi-System Challenges in Diabetes’ from the European Foundation for the Study of Diabetes (AMV), P2022/BMD-7227 (Comunidad de Madrid, Spain) (AMV), Fundación Ramón Areces (Spain) (AMV), CIBERdem (AMV and JB), CIBERhed (RF), and CIBERcv (LB) (ISCIII, Spain). LB and AMV belong to the Spanish National Research Council’s (CSIC’s) Cancer Hub. We also acknowledge the Spanish Ministry of Economy and Competitiveness (MINECO) postdoctoral contract IJCI-2015-24758 to IGM and the Spanish Ministry of Education, Culture and Sport (MECD) FPU17/02786 grant to RA

Virtual reality and augmented reality in medical education: an umbrella review

ObjectiveThis umbrella review aims to ascertain the extent to which immersive Virtual Reality (VR) and Augmented Reality (AR) technologies improve specific competencies in healthcare professionals within medical education and training, in contrast to traditional educational methods or no intervention.MethodsAdhering to PRISMA guidelines and the PICOS approach, a systematic literature search was conducted across major databases to identify studies examining the use of VR and AR in medical education. Eligible studies were screened and categorized based on the PICOS criteria. Descriptive statistics and chi-square tests were employed to analyze the data, supplemented by the Fisher test for small sample sizes or specific conditions.AnalysisThe analysis involved cross-tabulating the stages of work (Development and Testing, Results, Evaluated) and variables of interest (Performance, Engagement, Performance and Engagement, Effectiveness, no evaluated) against the types of technologies used. Chi-square tests assessed the associations between these categorical variables.ResultsA total of 28 studies were included, with the majority reporting increased or positive effects from the use of immersive technologies. VR was the most frequently studied technology, particularly in the “Performance” and “Results” stages. The chi-square analysis, with a Pearson value close to significance (p = 0.052), suggested a non-significant trend toward the association of VR with improved outcomes.ConclusionsThe results indicate that VR is a prevalent tool in the research landscape of medical education technologies, with a positive trend toward enhancing educational outcomes. However, the statistical analysis did not reveal a significant association, suggesting the need for further research with larger sample sizes. This review underscores the potential of immersive technologies to enhance medical training yet calls for more rigorous studies to establish definitive evidence of their efficacy

Dronedarone produces early regression of myocardial remodelling in structural heart disease.

Left ventricular hypertrophy (LVH) in hypertension is associated with a greater risk of sustained supraventricular/atrial arrhythmias. Dronedarone is an antiarrhythmic agent that was recently approved for the treatment of atrial fibrillation. However, its effect on early regression of LVH has not been reported. We tested the hypothesis that short-term administration of dronedarone induces early regression of LVH in spontaneously hypertensive rats (SHRs).Ten-month-old male SHRs were randomly assigned to an intervention group (SHR-D), where animals received dronedarone treatment (100 mg/kg) for a period of 14 days, or to a control group (SHR) where rats were given vehicle. A third group with normotensive control rats (WKY) was also added. At the end of the treatment with dronedarone we studied the cardiac anatomy and function in all the rats using transthoracic echocardiogram, cardiac metabolism using the PET/CT study (2-deoxy-2[18F]fluoro-D-glucose) and cardiac structure by histological analysis of myocyte size and collagen content.The hypertensive vehicle treated SHR rats developed the classic cardiac pattern of hypertensive cardiomyopathy as expected for the experimental model, with increases in left ventricular wall thickness, a metabolic shift towards an increase in glucose use and increases in myocyte and collagen content. However, the SHR-D rats showed statistically significant lower values in comparison to SHR group for septal wall thickness, posterior wall thickness, ventricular mass, glucose myocardial uptake, size of left ventricular cardiomyocytes and collagen content. All these values obtained in SHR-D rats were similar to the values measured in the normotensive WKY control group.The results suggest by three alternative and complementary ways (analysis of anatomy and cardiac function, metabolism and histological structure) that dronedarone has the potential to reverse the LVH induced by arterial hypertension in the SHR model of compensated ventricular hypertrophy

Limited genomic reconstruction of SARS-CoV-2 transmission history within local epidemiological clusters

Financiado para publicación en acceso aberto: Universidade de Vigo/CISUGA detailed understanding of how and when SARS-CoV-2 transmission occurs is crucial for designing effective prevention measures. Other than contact tracing, genome sequencing provides information to help infer who infected whom. However, the effectiveness of the genomic approach in this context depends on both (high enough) mutation and (low enough) transmission rates. Today, the level of resolution that we can obtain when describing SARS-CoV-2 outbreaks using just genomic information alone remains unclear. In order to answer this question, we sequenced 49 SARS-CoV-2 patient samples from ten local clusters in NW Spain for which partial epidemiological information was available, and inferred transmission history using genomic variants. Importantly, we obtained high-quality genomic data, sequencing each sample twice and using unique barcodes to exclude cross-sample contamination. Phylogenetic and cluster analyses showed that consensus genomes were generally sufficient to discriminate among independent transmission clusters. However, levels of intrahost variation were low, which prevented in most cases the unambiguous identification of direct transmission events. After filtering out recurrent variants across clusters, the genomic data were generally compatible with the epidemiological information but did not support specific transmission events over possible alternatives. We estimated the effective transmission bottleneck size to be 1-2 viral particles for sample pairs whose donor-recipient relationship was likely. Our analyses suggest that intrahost genomic variation in SARS-CoV-2 might be generally limited and that homoplasy and recurrent errors complicate identifying shared intrahost variants. Reliable reconstruction of direct SARS-CoV-2 transmission based solely on genomic data seems hindered by a slow mutation rate, potential convergent events, and technical artifacts. Detailed contact tracing seems essential in most cases to study SARS-CoV-2 transmission at high resolution.Xunta de Galicia | Ref. ED431C2018/54Banco SantanderConsejo Superior de Investigaciones CientíficasConferencia de Rectores de las Universidades EspañolasServizo Galego de Saúd

Cardiac glucose metabolism obtained by positron emission tomography.

<p>PET/CT images of myocardial 18 F-FDG uptake from a Wistar-Kyoto rat treated with vehicle (WKY), a spontaneously hypertensive rat treated with vehicle (SHR) and a spontaneously hypertensive rat treated with dronedarone (SHR-D) (A). Dronedarone produced a marked decrease in the glucose metabolism of the hypertrophied ventricle. There were no significant differences in standardized uptake value (SUV_heart) in either SHR-D or WKY. Statistically significant differences between WKY, SHR, and SHR-D are shown (*P<0.05 vs. WKY; ^†P<0.05 vs. SHR; ^†††P<0.001 vs. SHR). Values are given as mean ± SEM (n = 6 rats per group) (B).</p

Left ventricular geometry obtained by echocardiography.

<p>Examples of M-mode echocardiograms from a Wistar-Kyoto rat treated with vehicle (WKY), a spontaneously hypertensive rat treated with vehicle (SHR) and a spontaneously hypertensive rat treated with dronedarone (SHR-D).</p

Examples of histological sections of the left ventricle.

<p>Examples of histological sections of the subepicardial region of the left ventricle from a Wistar-Kyoto rat treated with vehicle (WKY), a spontaneously hypertensive rat treated with vehicle (SHR) and a spontaneously hypertensive rat treated with dronedarone (SHR-D), H&E (x40 objective; scale bar = 20 μm)(A) and Pricosirius red (CA, coronary artery; SE, subepicardium) (x4 objective; scale bar = 150 μm)(B).</p