Producción y caracterización de microesferas de alginato funcionalizadas con colágeno y ácido hialurónico como soporte tridimensional para el cultivo de células de mieloma múltiple

[ES] El mieloma múltiple (MM) es una neoplasia hematológica caracterizada por una expansión clonal de las células plasmáticas (CPs). Los cultivos 2D convencionales se ha visto que no consiguen reproducir el desarrollo de la enfermedad y actualmente se están llevando a cabo diseños y desarrollos de modelos de cultivo 3D que permitan reproducir “in vitro” el comportamiento de las CPs y su resistencia a fármacos mediada por el microambiente medular. La progresión a una medicina personalizada sería un hecho realista con un modelo 3D que permita el cultivo de células ex vivo de pacientes. El objetivo principal del siguiente trabajo ha consistido en desarrollar un modelo de cultivo 3D basado en microgel, que mediante el recubrimiento de las microesferas con distintas biomoléculas procedentes de la médula ósea, como son el colágeno tipo I y el ácido hialurónico (HA), permita evaluar la influencia de estas biomoléculas en la proliferación celular de las CPs. El microgel ha sido producido mediante un sistema de microfluídica, permitiendo obtener microesferas de alginato de un tamaño comprendido 170-190 µm, y con un alto rendimiento. Las microesferas fueron funcionalizadas con Colágeno Tipo I y ácido hialurónico por el método Layer-by-Layer. La presencia de Colágeno y HA en las microesferas se confirmó de forma cualitativa por FTIR, mientras que, por medio de BCA se evaluó de forma cuantitativa la presencia de Colágeno Tipo I. Posteriormente, las células de la línea RPMI 8226 se cultivaron en suspensión agitada y entre las microesferas del microgel. La proliferación celular se evaluó mediante el ensayo PicoGreen. Se observaron diferencias significativas de proliferación dependiendo de las condiciones de cultivo: células en suspensión agitada, microgel sin funcionalizar y microgel funcionalizados con HA, Col y ambas biomoléculas. En definitiva, el desarrollo de un cultivo 3D basado en microgel presenta diferencias significativas en cuanto a la proliferación de las CPs con respecto a células en suspensión, observándose mayores diferencias en los microgeles recubiertos con HA. Esto corroboraría la importancia de desarrollar entornos 3D que permitan reproducir “in vitro” el microambiente medular.[EN] Multiple myeloma is a hemathological neoplasm characterised by a clonal expansion of plasmatic cells (PC). Conventional 2D cultures aren’t able to reproduce the development of the disease, so, 3D culture models are currently being developed in order to allow the “in vivo” reproduction of plasmatic cells behaviour and their resistance to drugs mediated by the bone marrow (BM) microenvironment. Progression to a personalized medicine would be a realistic fact with this 3D models for cell ex vivo culture. The main objective of this work is the development of a microgel based 3D cell culture model, that due to their coating with different biomolecules originated in the bone marrow, like the type I collagen and hyaluronic acid, allows to evaluate the influence of this biomolecules in the proliferation of PC. The microgel has been produced in a microfluidic system, allowing us to obtain alginate microspheres of an average size of between 170-190 µm and optimum production throughput. Microspheres were functionalized with type I collagen and hyaluronic acid by a layer-by-layer method. The material characterization of the different coatings applied was made by the means of Fourier transformed infrared spectroscopy, while the type I collagen coating was essayed through bicinconinic acid essay (BCA). RPMI 8226 cell line has been cultivated between the microgel microspheres by maintaining the suspension in an orbital shaker. Cell proliferation was evaluated through PicoGreen assay. There were significant differences in proliferation depending on culture conditions: cells in suspension, cells with microgel and cells with different functionalized microgels. Definitely, the development of a microgel based 3D culture presents significant differences regarding the proliferation of the PC when compared to cells in suspension, observing this way major differences in microgels coated with hyaluronic acid. This fact would bear out the importance of developing 3D models that allow "in vitro” reproduction of the BM microenvironment.Díaz Benito, B. (2019). Producción y caracterización de microesferas de alginato funcionalizadas con colágeno y ácido hialurónico como soporte tridimensional para el cultivo de células de mieloma múltiple. http://hdl.handle.net/10251/124733TFG

Letter in response to article in journal of infection: “High SARS-CoV-2 antibody prevalence among healthcare workers exposed to COVID-19 patients”

Letter to editorUnidad Docente de Inmunología, Oftalmología y ORLFac. de Óptica y OptometríaTRUEpu

Quality of Life and Autonomy in Patients with Intermittent Bladder Catheterization Trained by Specialized Nurses

Intermittent bladder catheterization (IBC) involves regular urine draining using a catheter, which is removed immediately after urinary elimination. It allows for the patient's urological health to be managed and their renal function to be preserved, and it promotes autonomy. Compliance with the prescribed number of daily catheterizations, which must be conducted by the patient, and infection prevention measures are crucial. To identify the patients requiring IBC, and to determine their adherence (whether they followed the prescribed guidelines and their difficulty in carrying out the procedure, as well as to assess how the IBC influences their quality of life and state of mind after receiving self-care training from a specialized nurse), we carried out a prospective, multicenter observational study in 24 Spanish hospitals with one month of monitoring and a sample of 99 patients. The sources of information were the patients' clinical records, the King's Health Questionnaire, the Mini-Mental State Examination (MMSE), and the hospital anxiety and depression scale (HADS). Descriptive and bivariate statistics were used to analyses the paired data. After recruitment (n = 99), 79 patients completed the questionnaire at a mean age of 35.2 years (SD = 20.5 years). In total, 53.5% (53) of the sample consisted of men and 32.3% (32) had neurological damage as the reason for prescription; 67% (67.7) performed self-catheterization and 86.7% adhered to the IBC. After one month of monitoring, a statistically significant improvement in quality of life was observed in all criteria, with the exception of personal relationships (p < 0.005), as well as an improvement in anxiety and depression levels (p < 0.001). Patients who require IBC show good adherence to the IBC with a significant percentage of self-catheterization. After one month of IBC, a significant improvement in the patients' quality of life and mood was observed. These results could be attributed to adequate patient training and adequate personalization of the IBC materials by the specialized nurses

Impact of operatoŕs experience on peri-procedural outcomes with Watchman FLX: Insights from the FLX-SPA registry

Background: The Watchman FLX is a device upgrade of the Watchman 2.5 that incorporates several design enhancements intended to simplify left atrial appendage occlusion (LAAO) and improve procedural outcomes. This study compares peri-procedural results of LAAO with Watchman FLX (Boston Scientific, Marlborough, Massachusetts) in centers with varying degrees of experience with the Watchman 2.5 and Watchman FLX. Methods: Prospective, multicenter, 'real-world' registry including consecutive patients undergoing LAAO with the Watchman FLX at 26 Spanish sites (FLX-SPA registry). Implanting centers were classified according to the center's prior experience with the Watchman 2.5. A further division of centers according to whether or not they had performed ≤ 10 or > 10Watchman FLX implants was prespecified at the beginning of the study. Procedural outcomes of institutions stratified according to their experience with the Watchman 2.5 and FLX devices were compared. Results: 359 patients [mean age 75.5 (SD8.1), CHA2DS2-VASc 4.4 (SD1.4), HAS-BLED 3.8(SD0.9)] were included. Global success rate was 98.6%, successful LAAO with the first selected device size was achieved in 95.5% patients and the device was implanted at first attempt in 78.6% cases. There were only 9(2.5%) major peri-procedural complications. No differences in efficacy or safety results according to the centeŕs previous experience with Watchman 2.5 and procedural volume with Watchman FLX existed. Conclusions: The Watchman FLX attains high procedural success rates with complete LAA sealing in unselected, real-world patients, along with a low incidence of peri-procedural complications, regardless of operatoŕs experience with its previous device iteration or the number of Watchman FLX devices implanted

Implantación de un sistema integral de gestión del conocimiento para los procesos de innovación docente de la Universidad de Salamanca

Memoria ID-0312. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015

Implantación de un sistema integral de gestión del conocimiento para los procesos de innovación docente de la Universidad de Salamanca

Memoria ID-0312. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Lectura, educación y bibliotecas: ideas para crear buenos lectores

Some strategies of reading promotion are described and explaine

Biomimetic 3D Environment Based on Microgels as a Model for the Generation of Drug Resistance in Multiple Myeloma

The development of three-dimensional environments to mimic the in vivo cellular response is a problem in the building of disease models. This study aimed to synthesize and validate three-dimensional support for culturing monoclonal plasma cells (mPCs) as a disease model for multiple myeloma. The three-dimensional environment is a biomimetic microgel formed by alginate microspheres and produced on a microfluidic device whose surface has been functionalized by a layer-by-layer process with components of the bone marrow’s extracellular matrix, which will interact with mPC. As a proof of concept, RPMI 8226 cell line cells were cultured in our 3D culture platform. We proved that hyaluronic acid significantly increased cell proliferation and corroborated its role in inducing resistance to dexamethasone. Despite collagen type I having no effect on proliferation, it generated significant resistance to dexamethasone. Additionally, it was evidenced that both biomolecules were unable to induce resistance to bortezomib. These results validate the functionalized microgels as a 3D culture system that emulates the interaction between tumoral cells and the bone marrow extracellular matrix. This 3D environment could be a valuable culture system to test antitumoral drugs efficiency in multiple myeloma.Marin Paya, JC.; Díaz Benito, B.; Amaro Martins, L.; Clara Trujillo, S.; Cordón, L.; Lanceros-Mendez, S.; Gallego Ferrer, G.... (2021). Biomimetic 3D Environment Based on Microgels as a Model for the Generation of Drug Resistance in Multiple Myeloma. Universitat Politècnica de València. https://doi.org/10.4995/Dataset/10251/17831