pH-triggered endosomal escape of pore-forming Listeriolysin O toxin-coated gold nanoparticles

The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (ERC grant agreement n° 338133)Background: A major bottleneck in drug delivery is the breakdown and degradation of the delivery system through the endosomal/lysosomal network of the host cell, hampering the correct delivery of the drug of interest. In nature, the bacterial pathogen Listeria monocytogenes has developed a strategy to secrete Listeriolysin O (LLO) toxin as a tool to escape the eukaryotic lysosomal system upon infection, allowing it to grow and proliferate unharmed inside the host cell. Results: As a “proof of concept”, we present here the use of purifed His-LLO H311A mutant protein and its conjuga tion on the surface of gold nanoparticles to promote the lysosomal escape of 40 nm-sized nanoparticles in mouse embryonic fbroblasts. Surface immobilization of LLO was achieved after specifc functionalization of the nanoparti cles with nitrile acetic acid, enabling the specifc binding of histidine-tagged proteins. Conclusions: Endosomal acidifcation leads to release of the LLO protein from the nanoparticle surface and its self-assembly into a 300 Å pore that perforates the endosomal/lysosomal membrane, enabling the escape of nanoparticles.Depto. de Química FísicaFac. de Ciencias QuímicasTRUEUnión Europea. FP7Ministerio de Ciencia e Innovación (MICINN)Comunidad de MadridUniversidad Complutense de Madridpu

Effectiveness of gold nanorods of different sizes in photothermal therapy to eliminate melanoma and glioblastoma cells

Gold nanorods are the most commonly used nanoparticle in photothermal therapy (PTT) due to their efficiency to convert the light into heat. This study aimed to investigate the efficacy of gold nanorods of different sizes (large and small) in eliminating two types of cancer cells: glioblastoma and melanoma cells. After the cytotoxic profiles of the nanoparticles were stablished, PTT was applied to the melanoma and glioblastoma cells with a high efficiency and mortality rate. Several methods were used to evaluate the efficiency of PTT including fluorescence, confocal or dark field microscopy, biochemical analysis, and flow cytometry. To determine cell viability and the type of cell death triggered by PTT, Calcein-propidium iodide, Annexin-V staining and dehydrogenase activity assays were performed. Our findings revealed that after PTT was applied to the cancer cells, the main cell death was apoptosis. This is advantageous as the presence of apoptotic cells can stimulate antitumoral immunity in vivo. Considering the high efficacy of these gold nanorods in PTT, large nanoparticles could be useful for biofunctionalization purposes. Large nanorods offer a greater surface area for attaching biomolecules, thereby promoting high sensitivity and specificity in recognizing target cancer cells. Additionally, large nanoparticles could also be beneficial for theragnostic applications, involving both therapy and diagnosis, due to their superior detection sensitivity

Effectiveness of gold nanorods of different sizes in photothermal therapy to eliminate melanoma and glioblastoma cells

Gold nanorods are the most commonly used nanoparticle in photothermal therapy (PTT) due to their efficiency to convert the light into heat. This study aimed to investigate the efficacy of gold nanorods of different sizes (large and small) in eliminating two types of cancer cells: glioblastoma and melanoma cells. After the cytotoxic profiles of the nanoparticles were stablished, PTT was applied to the melanoma and glioblastoma cells with a high efficiency and mortality rate. Several methods were used to evaluate the efficiency of PTT including fluorescence, confocal or dark field microscopy, biochemical analysis, and flow cytometry. To determine cell viability and the type of cell death triggered by PTT, Calcein-propidium iodide, Annexin-V staining and dehydrogenase activity assays were performed. Our findings revealed that after PTT was applied to the cancer cells, the main cell death was apoptosis. This is advantageous as the presence of apoptotic cells can stimulate antitumoral immunity in vivo. Considering the high efficacy of these gold nanorods in PTT, large nanoparticles could be useful for biofunctionalization purposes. Large nanorods offer a greater surface area for attaching biomolecules, thereby promoting high sensitivity and specificity in recognizing target cancer cells. Additionally, large nanoparticles could also be beneficial for theragnostic applications, involving both therapy and diagnosis, due to their superior detection sensitivity

Prevalencia de Sífilis mediante VDRL, prueba treponémica y su correlación con niveles de CD4 y CV en personas viviendo con virus de la inmunodeficiencia humana, que se atienden en el Hospital General Docente Ambato en el período comprendido entre Enero -

El estudio de las enfermedades de transmisión sexual en Ecuador, es un tema de alta relevancia en el país, sobre todo cuando existen cifras considerables de personas que mantienen cualesquiera de estas enfermedades en todo el territorio. El Virus de inmunodeficiencia humana, tal y como su nombre lo indica, tiene la característica de destruir gradualmente el sistema inmunitario al atacar y destruir las células CD4, haciendo que las personas que padecen de esta afección crónica se encuentren en riesgo de contraer infecciones causadas por otro de enfermedades como la sífilis, otra enfermedad de transmisión sexual que al contacto con el VIH puede mejorar su adquisición o transmisión. Por ello, este estudio tiene el tema de “Prevalencia de sífilis mediante VDRL, prueba treponémica y su correlación con niveles de CD4 y CV en personas viviendo con virus de la inmunodeficiencia humana, que se atienden en el Hospital General Docente Ambato en el período comprendido entre Enero - Diciembre 2022”, mismo que busca determinar la prevalencia del sífilis en el VIH, en los pacientes del Hospital General Docente Ambato durante el año 2022. En cuanto a metodología, se aplicó un estudio descriptivo transversal, utilizando una muestra que abarcó la totalidad de pacientes infectados por VIH, esto es, 609 personas. Dando como resultado la evidente existencia de una alta prevalencia en la Coinfección VIH-Sífilis; sin embargo, en el hospital se constató un número importante de pacientes que no se han podido realizar este examen por su falta de disponibilidad constante

Diseño de un protocolo de gamificación en el ámbito docente universitario: aplicación en el fomento del proceso de aprendizaje y la adquisición dinámica de competencias en el contexto de la Tecnología Farmacéutica

¿Se puede diseñar un protocolo de gamificación que implique a distintas personas de la comunidad universitaria, que se pueda adaptar a los perfiles de la nueva generación de estudiantes y a los aprendizajes o competencias a adquirir?

Learning analytics en Farmacia siguiendo el "rastro" de estudiantes de Tecnología Farmacéutica: primera experiencia para indagar en su potencial transformador, relacionado con el aprendizaje personalizado y adaptativo a una generación Y-Z

El presente proyecto es consecuencia de la asistencia a las Jornadas organizadas por la UCM “Las TIC en la Enseñanza II: Innovación en el Aula”. Se generaron muchas inquietudes y muchas ideas. De todas ellas, el grupo de innovación docente consolidado, que plantea el presente proyecto, ha decidido explotar las herramientas (Logs, etc.) que la UCM pone a nuestra disposición, como María Vela indicó en su presentación, para aplicar por primera vez “Learning Analytics” y “sacar partido al rastro de nuestros estudiantes” de Farmacia, en concreto, de la asignatura Tecnología Farmacéutica. La selección de este tema está en línea con la iniciativa europea 2020 en cuanto al uso de Learning analytics para mejorar e innovar el aprendizaje y la enseñanza. La tecnología educativa ha dado lugar a la aparición de rastros o huellas digitales -y a su registro- que permiten tener datos cuya medida, análisis y estudio posterior pueden ser de utilidad para obtener conclusiones prácticas. Learning Analytics –LA- o analítica de aprendizaje es una herramienta novedosa - relacionada con el análisis de redes sociales (SNA) y el Big Data- que mediante el registro y estudio crítico de determinados indicadores discentes y docentes, contribuye a la personalización y adaptación del aprendizaje así como coopera en la planificación educativa con el objetivo de mejorar el desarrollo competencial y la significatividad de lo aprendido (Gutiérrez-Priego, 2015; García-Peñalvo, 2014). Es una primera experiencia de la que se esperan tener resultados preliminares que sirvan de base para seguir trabajando en este ámbito e indagar en el potencial transformador que se le atribuye, relacionado con el aprendizaje personalizado y adaptativo a una generación Y-Z que se caracteriza por ser nativos digitales, estar abiertos a los cambios, vivir el presente, buscar soluciones rápidas a los problemas que les surgen, estar siempre conectados y preferir sobretodo lo visual antes que el contenido denso. Estas nuevas generaciones necesitan aprender de otra manera: quieren trabajar de modo diferente en el aula, saber cómo extraer conocimiento relevante de la información que nos rodea, aprender de manera colaborativa, potenciar determinadas competencias y desarrollar nuevas habilidades (Postgrado en Digital Learning y Experiencias de Aprendizaje Emergente IL3-UB 2019). Para ello, es fundamental establecer tres aspectos clave: • qué variables se van a medir. • cómo se medirán, esto es, la identificación de criterios e indicadores mensurables así como la selección de herramientas y procedimientos de análisis, • cuál será la interpretación, aspecto en el que además del propio estudio estadístico va a ser muy significativa la experiencia y el bagaje (know-how) del analista (docente/instructor). Estos aspectos están vinculados a varios grupos específicos de estudiantes: los grupos de Tecnología Farmacéutica impartidos por los docentes del equipo. En distintos trabajos/publicaciones se ha presentado a LA como un instrumento relacionado con el aprendizaje continuo en continua transformación. Se puede deducir que en un aprendizaje –tanto virtual como presencial e híbrido- con carácter práctico como el que se desarrolla en Tecnología Farmacéutica, que busca ser proactivo, colaborativo e interactivo, la recopilación y el análisis crítico de datos (cuantitativo y cualitativo) puede resultar fundamental como ya se extrae de las decisiones estratégicas empresariales a través del BI (Business Intelligence). Además, se pretende aplicar Learning Analytics para identificar a los alumnos que necesitan apoyo adicional. Aunque generalmente se busca brindar apoyo a todos los alumnos, identificar aquellos con más necesidades suele ser difícil: a menudo, no son los alumnos que pasan al frente y preguntan. Al tener en cuenta la trayectoria académica, la participación online y el desempeño académico, Learning Analytics puede ayudar a identificar a los alumnos que probablemente necesiten más apoyo y a hacerlo con suficiente anticipación en sus estudios de modo que las intervenciones puedan obtener el máximo beneficio. Este proyecto pretende ser una continuidad a los llevados a cabo en convocatorias anteriores (conv. 2014: nº 175, conv. 2015 nº 349, conv. 2016: nº 215, conv. 2017: nº 244, conv 2018: nº 252). De forma que no se generan nuevos contenidos, sino que, para avanzar un poco más allá en el escenario de aprendizaje, se pretende poner en práctica las metodologías y habilidades definidas y desarrolladas en dichos proyectos de Innovación Docente como gamificación, un paquete de vídeos sobre contenidos prácticos de la asignatura que facilitan el aprendizaje en modelos de aprendizaje inverso en el laboratorio, materiales para el desarrollo de competencias transversales (herramientas informáticas, bases de datos, gestión de residuos generados en el laboratorio de prácticas), herramientas de evaluación y autoevaluación mediante exámenes test, materiales para el aprendizaje autónomo del alumno, siempre en el entorno de la docencia-aprendizaje de la Tecnología Farmacéutica. Para ello, se plantea la aplicación de Learning analyticis a partir de la construcción de un modelo de análisis de los datos que facilita la plataforma Moodle, teniendo en cuenta las distintas interacciones entre el docente, el estudiante y la materia desarrollada, de forma que esta información pueda transformarse en conocimiento y su comprensión pueda ayudar a la mejora de la práctica docente de profesores (Hernández Estrada, 2016). ¿Nos animamos a aprovechar los beneficios de Learning Analytics

The evolution of the ventilatory ratio is a prognostic factor in mechanically ventilated COVID-19 ARDS patients

Background: Mortality due to COVID-19 is high, especially in patients requiring mechanical ventilation. The purpose of the study is to investigate associations between mortality and variables measured during the first three days of mechanical ventilation in patients with COVID-19 intubated at ICU admission. Methods: Multicenter, observational, cohort study includes consecutive patients with COVID-19 admitted to 44 Spanish ICUs between February 25 and July 31, 2020, who required intubation at ICU admission and mechanical ventilation for more than three days. We collected demographic and clinical data prior to admission; information about clinical evolution at days 1 and 3 of mechanical ventilation; and outcomes. Results: Of the 2,095 patients with COVID-19 admitted to the ICU, 1,118 (53.3%) were intubated at day 1 and remained under mechanical ventilation at day three. From days 1 to 3, PaO2/FiO2 increased from 115.6 [80.0-171.2] to 180.0 [135.4-227.9] mmHg and the ventilatory ratio from 1.73 [1.33-2.25] to 1.96 [1.61-2.40]. In-hospital mortality was 38.7%. A higher increase between ICU admission and day 3 in the ventilatory ratio (OR 1.04 [CI 1.01-1.07], p = 0.030) and creatinine levels (OR 1.05 [CI 1.01-1.09], p = 0.005) and a lower increase in platelet counts (OR 0.96 [CI 0.93-1.00], p = 0.037) were independently associated with a higher risk of death. No association between mortality and the PaO2/FiO2 variation was observed (OR 0.99 [CI 0.95 to 1.02], p = 0.47). Conclusions: Higher ventilatory ratio and its increase at day 3 is associated with mortality in patients with COVID-19 receiving mechanical ventilation at ICU admission. No association was found in the PaO2/FiO2 variation

Clustering COVID-19 ARDS patients through the first days of ICU admission. An analysis of the CIBERESUCICOVID Cohort

Background Acute respiratory distress syndrome (ARDS) can be classified into sub-phenotypes according to different inflammatory/clinical status. Prognostic enrichment was achieved by grouping patients into hypoinflammatory or hyperinflammatory sub-phenotypes, even though the time of analysis may change the classification according to treatment response or disease evolution. We aimed to evaluate when patients can be clustered in more than 1 group, and how they may change the clustering of patients using data of baseline or day 3, and the prognosis of patients according to their evolution by changing or not the cluster.Methods Multicenter, observational prospective, and retrospective study of patients admitted due to ARDS related to COVID-19 infection in Spain. Patients were grouped according to a clustering mixed-type data algorithm (k-prototypes) using continuous and categorical readily available variables at baseline and day 3.Results Of 6205 patients, 3743 (60%) were included in the study. According to silhouette analysis, patients were grouped in two clusters. At baseline, 1402 (37%) patients were included in cluster 1 and 2341(63%) in cluster 2. On day 3, 1557(42%) patients were included in cluster 1 and 2086 (57%) in cluster 2. The patients included in cluster 2 were older and more frequently hypertensive and had a higher prevalence of shock, organ dysfunction, inflammatory biomarkers, and worst respiratory indexes at both time points. The 90-day mortality was higher in cluster 2 at both clustering processes (43.8% [n = 1025] versus 27.3% [n = 383] at baseline, and 49% [n = 1023] versus 20.6% [n = 321] on day 3). Four hundred and fifty-eight (33%) patients clustered in the first group were clustered in the second group on day 3. In contrast, 638 (27%) patients clustered in the second group were clustered in the first group on day 3.Conclusions During the first days, patients can be clustered into two groups and the process of clustering patients may change as they continue to evolve. This means that despite a vast majority of patients remaining in the same cluster, a minority reaching 33% of patients analyzed may be re-categorized into different clusters based on their progress. Such changes can significantly impact their prognosis

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

Annual Conference on Formative Research on EFL. Practices thar inspire change.

The conference papers of the Annual Conference on Formative Research on EFL. Practices thar inspire change collect pedagogical experiences, research reports, and reflections about social issues, language teaching, teaching training, interculturality under the panorama of the Covid-19 pandemic. Each paper invites the reader to implement changes in their teaching practice through disruptive pedagogies, reflect on the social and emotional consequences of the lockdown, new paths for teacher training and different approaches for teaching interculturality. We expect to inspire new ways to train pre-service teachers and teach languages in this changing times