CRISPR/CasRx proof-of-concept for RNA degradation: A future tool against RNA viruses?

Producción CientíficaInfluenza viruses provide a great threat for the human population, causing highly contagious respiratory infections that can lead to serious clinical complications. There are a limited variety of influenza antivirals, and these antivirals are subjected to the constant emergence of resistances. Therefore, the development of new antiviral strategies to combat influenza viruses and other RNA viruses must be promoted. In this work, we design a proof-of-concept of a recently described CRISPR/Cas tool that has been proposed as a possible future RNA virus antiviral, named CRISPR/CasRx. For this, we verified the efficiency of the CasRx endonuclease in the degradation of the eGFP mRNA reporter gene and we established the best conditions for, and the efficient performance of, the CRISPR/CasRx system. The results were measured by fluorescence microscopy, flow cytometry, and qRT-PCR. The analyses demonstrated a reduction in fluorescence, regardless of the amount of eGFP reporter plasmid transfected. The analyses showed an 86–90% reduction in fluorescence by flow cytometry and a 51–80% reduction in mRNA expression by qRT-PCR. Our results demonstrate that the CasRx endonuclease is an efficient tool for eGFP mRNA knockdown. Therefore, subsequent experiments could be useful for the development of a new antiviral tool

Magneto-mechanical system to reproduce and quantify complex strain patterns in biological materials

Biological cells and tissues are continuously subjected to mechanical stress and strain cues from their surrounding substrate. How these forces modulate cell and tissue behavior is a major question in mechanobiology. To conduct studies under controlled varying physiological strain scenarios, a new virtually-assisted experimental system is proposed allowing for non-invasive and real-time control of complex deformation modes within the substrates. This approach is based on the use of extremely soft magneto-active polymers, which mimic the stiffness of biological materials. Thus, the system enables the untethered control of biological substrates providing reversible mechanical changes and controlling heterogeneous patterns. Motivated on a deep magneto-mechanical characterization across scales, a multi-physics and multi-scale in silico framework was developed to guide the experimental stimulation setup. The versatility and viability of the system have been demonstrated through its ability to reproduce complex mechanical scenarios simulating local strain patterns in brain tissue during a head impact, and its capability to transmit physiologically relevant mechanical forces to dermal fibroblasts. The proposed framework opens the way to understanding the mechanobiological processes that occur during complex and dynamic deformation states, e.g., in traumatic brain injury, pathological skin scarring or fibrotic heart remodeling during myocardial infarction.The authors thank Denis Wirtz (Johns Hopkins University) and Jean-Christophe Olivo-Marin (Institute Pasteur) for relevant discussion. The authors acknowledge support from the European Research Council (ERC) under the European Union's Horizon 2020 Research and Innovation Program (Grant agreement No. 947723, project: 4D-BIOMAP), and from Programa de Apoyo a la Realizacion de Proyectos Interdiscisplinares de I+D para Jovenes Investigadores de la Universidad Carlos III de Madrid and Comunidad de Madrid (project: BIOMASKIN). MAMM and CGC acknowledges support from the Ministerio de Ciencia, Innovacion y Universidades, Spain (FPU19/03874 and FPU20/01459) and DGG acknowledges support from the Talent Attraction grant (CM 2018 - 2018-T2/IND-9992) from the Comunidad de Madrid

PPARγ as an indicator of vascular function in an experimental model of metabolic syndrome in rabbits

Background and aims: Underlying mechanisms associated with vascular dysfunction in metabolic syndrome (MetS) remain unclear and can even vary from one vascular bed to another. Methods: In this study, MetS was induced by a high-fat, high-sucrose diet, and after 28 weeks, aorta and renal arteries were removed and used for isometric recording of tension in organ baths, protein expression by Western blot, and histological analysis to assess the presence of atherosclerosis. Results: MetS induced a mild hypertension, pre-diabetes, central obesity and dyslipidaemia. Our results indicated that MetS did not change the contractile response in either the aorta or renal artery. Conversely, vasodilation was affected in both arteries in a different way. The aorta from MetS showed vascular dysfunction, including lower response to acetylcholine and sodium nitroprusside, while the renal artery from MetS presented a preserved relaxation to acetylcholine and an increased sensitivity to sodium nitroprusside. We did not find vascular oxidative stress in the aorta from MetS, but we found a significant decrease in PPARγ, phospho-Akt (p-Akt) and phospho-eNOS (p-eNOS) protein expression. On the other hand, we found oxidative stress in the renal artery from MetS, and PPARγ, Akt and p-Akt were overexpressed. No evidence of atherosclerosis was found in arteries from MetS. Conclusions: MetS affects vascular function differently depending on the vessel. In the aorta, it decreases both the vasodilation and the expression of the PPARγ/Akt/eNOS pathway, while in the renal artery, it increases the expression of PPARγ/Akt signalling pathway without decreasing the vasodilation

Tuberculosis prophylaxis with levofloxacin in liver transplant patients is associated with a high incidence of tenosynovitis: safety analysis of a multicenter randomized trial

This work was supported by the Ayudas para el fomento de la investigacion clinica independiente [EC 10-120] and Programa Intramural Consorcio de Apoyo a la Investigación Biomédica en Red 2010. Other funding sources: National R&D&I Plan 2008–2011 and the Instituto de Salud Carlos III (ISCIII), Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía y Competitividad, Spanish Network for Research in Infectious Diseases [RD06/0008, RD12/0015] - co-financed by European Development Regional Fund “A way to achieve Europe” ERDF. Consorcio de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas is financed by the ISCIII.Background: It is necessary to develop a safe alternative to isoniazid for tuberculosis prophylaxis in liver transplant recipients. This study was designed to investigate the efficacy and safety of levofloxacin. Methods: An open-label, prospective, multicenter, randomized study was conducted to compare the efficacy and safety of levofloxacin (500 mg q24h for 9 months) initiated in patients awaiting liver transplantation and isoniazid (300 mg q24h for 9 months) initiated post-transplant when liver function was stabilized. Efficacy was measured by tuberculosis incidence at 18 months after transplantation. All adverse events related to the medication were recorded. Results: CONSORT guidelines were followed in order to present the results. The safety committee suspended the study through a safety analysis when 64 patients had been included (31 in the isoniazid arm and 33 in the levofloxacin arm). The reason for suspension was an unexpected incidence of severe tenosynovitis in the levofloxacin arm (18.2%). Although the clinical course was favorable in all cases, tenosynovitis persisted for 7 weeks in some patients. No patients treated with isoniazid, developed tenosynovitis. Only 32.2% of patients randomized to isoniazid (10/31) and 54.5% of patients randomized to levofloxacin (18/33, P = .094) completed prophylaxis. No patient developed tuberculosis during the study follow-up (median 270 days). Conclusions: Levofloxacin prophylaxis of tuberculosis in liver transplant candidates is associated with a high incidence of tenosynovitis that limits its potential utility.Ayudas para el fomento de la investigación clínica independiente [EC 10-120]Programa Intramural Consorcio de Apoyo a la Investigación Biomédica en Red 2010National R&D&I Plan 2008–2011Instituto de Salud Carlos III (ISCIII)Ministerio de Economía y Competitividad RD06/0008, RD12/0015European Development Regional Fun

Cribado de la depresión mayor en la infancia y adolescencia (Parte 2)

El síndrome depresivo que aparece durante la infancia o la adolescencia tiende a seguir un curso crónico, con alta probabilidad de permanecer en la edad adulta. Su sintomatología repercute en la vida diaria del menor y, en el caso de la depresión mayor, supone un riesgo para la aparición de conducta suicida. Dada la variabilidad de sus manifestaciones clínicas y las dificultades en el diagnóstico, se plantea la posible utilidad del uso de cuestionarios de cribado en la población infantil y adolescente, a partir de un análisis del rendimiento de estos y de la eficacia de las medidas terapéuticas actuales. El presente trabajo se ha dividido en dos partes, y al final de este se recopilan las recomendaciones de distintos grupos de trabajo y tras el análisis de la evidencia, las recomendaciones que el grupo PrevInfad ha consensuado para la consulta de los pediatras en Atención Primaria

Etnobotánica, aprovechamiento tradicional de plantas y patrimonio cultural como instrumento de innovación para el desarrollo sostenible

Memoria ID-0147. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2017-2018

Percepción, valores y actitudes sobre el medio ambiente y el tratamiento de la problemática ambiental en los medios de comunicación

Memoria ID-0142. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015

Importance of Timely Treatment Initiation in Infantile-Onset Pompe Disease, a Single-Centre Experience

Abstract Classic infantile Pompe disease (IPD) is a rare lysosomal storage disorder characterized by severe hypertrophic cardiomyopathy and profound muscle weakness. Without treatment, death occurs within the first 2 years of life. Although enzyme replacement therapy (ERT) with alglucosidase alfa has improved survival, treatment outcome is not good in many cases and is largely dependent on age at initiation. The objective of the study was (a) to analyse the different stages in the diagnosis and specific treatment initiation procedure in IPD patients, and (b) to compare clinical and biochemical outcomes depending on age at ERT initiation (<1 month of age vs. <3 months of age). Here, we show satisfactory clinical and biochemical outcomes in two IPD patients after early treatment initiation before 3 months of life with immunomodulatory therapy in the ERT-naïve setting, with a high ERT dose from the beginning. Despite the overall good evolution, the patient who initiated treatment <1 month of life presented even better outcomes than the patient who started treatment <3 months of life, with an earlier normalization of hypertrophic cardiomyopathy, along with CK normalization, highlighting the importance of early treatment initiation in this progressive disease before irreversible muscle damage has occurred.This work was partially funded by the Basque Department of Education (IT1281-19)

La enseñanza en el máster de abogacía a través del análisis de casos reales de carácter multidisciplinar. Aprendizaje "a partir del estudio de razonamientos parciales temáticos"

Generar blog para publicaciones de alumnos del Máster Abogacía, con "entradas" dirigidas. Habría análisis desde varias disciplinas añadiéndose estudios con enfoque basado en otras áreas de conocimiento, generando interdisciplinariedad

Planificación y diseño de recursos para poner en marcha la versión on-line del Máster oficial las TIC en Educación

Memoria ID-0180. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015