Some Research Questions and Results of UC3M in the E-Madrid Excellence Network

32 slides.-- Contributed to: 2010 IEEE Global Engineering Education Conference (EDUCON), Madrid, Spain, 14-16 April, 2010.-- Presented by C. Delgado Kloos.Proceedings of: 2010 IEEE Global Engineering Education Conference (EDUCON), Madrid, Spain, 14-16 April, 2010Universidad Carlos III de Madrid is one of the six main participating institutions in the eMadrid excellence network, as well as its coordinating partner. In this paper, the network is presented together with some of the main research lines carried out by UC3M. The remaining papers in this session present the work carried out by the other five universities in the consortium.The Excellence Network eMadrid, “Investigación y Desarrollo de Tecnologías para el e-Learning en la Comunidad de Madrid” is being funded by the Madrid Regional Government under grant No. S2009/TIC-1650. In addition, we acknowledge funding from the following research projects: iCoper: “Interoperable Content for Performance in a Competency-driven Society” (eContentPlus Best Practice Network No. ECP-2007-EDU-417007), Learn3: Hacia el Aprendizaje en la 3ª Fase (“Plan Nacional de I+D+I” TIN2008-05163/ TSI), Flexo: “Desarrollo de aprendizaje adaptativo y accesible en sistemas de código abierto” (AVANZA I+D, TSI-020301- 2008-19), España Virtual (CDTI, Ingenio 2010, CENIT, Deimos Space), SOLITE (CYTED 508AC0341), and “Integración vertical de servicios telemáticos de apoyo al aprendizaje en entornos residenciales” (Programa de creación y consolidación de grupos de investigación de la Universidad Carlos III de Madrid).Publicad

Sustained Cytotoxic Response of Peripheral Blood Mononuclear Cells from Unvaccinated Individuals Admitted to the ICU Due to Critical COVID-19 Is Essential to Avoid a Fatal Outcome

The main objective of this study was to determine the influence of the cytotoxic activity of peripheral blood mononuclear cells (PBMCs) on the outcome of unvaccinated individuals with critical COVID-19 admitted to the ICU. Blood samples from 23 individuals were collected upon admission and then every 2 weeks for 13 weeks until death (Exitus group) (n = 13) or discharge (Survival group) (n = 10). We did not find significant differences between groups in sociodemographic, clinical, or biochemical data that may influence the fatal outcome. However, direct cellular cytotoxicity of PBMCs from individuals of the Exitus group against pseudotyped SARS-CoV-2-infected Vero E6 cells was significantly reduced upon admission (−2.69-fold; p = 0.0234) and after 4 weeks at the ICU (−5.58-fold; p = 0.0290), in comparison with individuals who survived, and it did not improve during hospitalization. In vitro treatment with IL-15 of these cells did not restore an effective cytotoxicity at any time point until the fatal outcome, and an increased expression of immune exhaustion markers was observed in NKT, CD4+, and CD8+ T cells. However, IL-15 treatment of PBMCs from individuals of the Survival group significantly increased cytotoxicity at Week 4 (6.18-fold; p = 0.0303). Consequently, immunomodulatory treatments that may overcome immune exhaustion and induce sustained, efficient cytotoxic activity could be essential for survival during hospitalization due to critical COVID-19.This work was supported by the Coordinated Research Activities at the National Center of Microbiology (CNM, Instituto de Salud Carlos III) (COV20_00679) to promote an integrated response against SARS-CoV-2 in Spain (Spanish Ministry of Science and Innovation) that is coordinated by Dr Inmaculada Casas (WHO National Influenza Center of the CNM); a generous donation provided by Chiesi España, S.A.U. (Barcelona, Spain); the Spanish Ministry of Science and Innovation (PID2019-110275RB-I00). The work of Guiomar Casado is financed by CIBERINFEC, co-financed by the European Regional Development Fund (FEDER) “A way to make Europe”. The work of Montserrat Torres is supported by Instituto de Salud Carlos III (COV20_00679). The work of Fernando Ramos Martín is financed by the Spanish Ministry of Science and Innovation (PID2019-110275RB-I00). The work of Mario Manzanares is supported by a pre-doctoral grant from Instituto de Salud Carlos III (ISCIII-PFIS FI20CIII/00021). The work of Lorena Vigón is supported by a pre-doctoral grant from Instituto de Salud Carlos III (FIS PI16CIII/00034-ISCIII-FEDER). The work of Sara Rodríguez-Mora is financed by NIH grant R01AI143567.N

Gradient, UC3M

En este artículo se presenta un resumen de las líneas de investigación que se realizan en el Laboratorio Gradient perteneciente al Grupo GAST (Grupo de Aplicaciones y Servicios Telemáticos) del Departamento de Ingeniería Telemática de la Universidad Carlos III de Madrid. La temática principal de investigación es la aplicación de tecnologías para la mejora de la enseñanza y el aprendizaje. El resumen se centra en tres líneas: Personalización del aprendizaje, uso de dispositivos móviles con fines educativos y aplicaciones de Realidad Virtual y Realidad Aumentada en educación.En este artículo se presenta un resumen de las líneas de investigación que se realizan en el Laboratorio Gradient perteneciente al Grupo GAST (Grupo de Aplicaciones y Servicios Telemáticos) del Departamento de Ingeniería Telemática de la Universidad Carlos III de Madrid. La temática principal de investigación es la aplicación de tecnologías para la mejora de la enseñanza y el aprendizaje. El resumen se centra en tres líneas: Personalización del aprendizaje, uso de dispositivos móviles con fines educativos y aplicaciones de Realidad Virtual y Realidad Aumentada en educación.Publicad

Impaired Cytotoxic Response in PBMCs From Patients With COVID-19 Admitted to the ICU: Biomarkers to Predict Disease Severity

Infection by novel coronavirus SARS-CoV-2 causes different presentations of COVID-19 and some patients may progress to a critical, fatal form of the disease that requires their admission to ICU and invasive mechanical ventilation. In order to predict in advance which patients could be more susceptible to develop a critical form of COVID-19, it is essential to define the most adequate biomarkers. In this study, we analyzed several parameters related to the cellular immune response in blood samples from 109 patients with different presentations of COVID-19 who were recruited in Hospitals and Primary Healthcare Centers in Madrid, Spain, during the first pandemic peak between April and June 2020. Hospitalized patients with the most severe forms of COVID-19 showed a potent inflammatory response that was not translated into an efficient immune response. Despite the high levels of effector cytotoxic cell populations such as NK, NKT and CD8+ T cells, they displayed immune exhaustion markers and poor cytotoxic functionality against target cells infected with pseudotyped SARS-CoV-2 or cells lacking MHC class I molecules. Moreover, patients with critical COVID-19 showed low levels of the highly cytotoxic TCRγδ+ CD8+ T cell subpopulation. Conversely, CD4 count was greatly reduced in association to high levels of Tregs, low plasma IL-2 and impaired Th1 differentiation. The relative importance of these immunological parameters to predict COVID-19 severity was analyzed by Random Forest algorithm and we concluded that the most important features were related to an efficient cytotoxic response. Therefore, efforts to fight against SARS-CoV-2 infection should be focused not only to decrease the disproportionate inflammatory response, but also to elicit an efficient cytotoxic response against the infected cells and to reduce viral replication.This work was supported by the Coordinated Research Activities at the Centro Nacional de Microbiología (CNM, Instituto de Salud Carlos III) (COV20_00679) to promote an integrated response against SARS-CoV-2 in Spain (Spanish Ministry of Science and Innovation) that is coordinated by Dr. Inmaculada Casas (WHO National Influenza Center of the CNM); a generous donation provided by Chiesi España, S.A.U. (Barcelona, Spain); the Spanish Ministry of Economy and Competitiveness (PID2019-110275RB-I00); the Spanish AIDS Research Network RD16CIII/0002/0001 that is included in Acciόn Estratégica en Salud, Plan Nacional de Investigaciόn Científica, Desarrollo e Innovaciόn Tecnolόgica 2016-2020, Instituto de Salud Carlos III, European Region Development Fund (ERDF). The work of ML-H and SR-M is financed by NIH grant R01AI143567. The work of LH is supported by a pre-doctoral grant from Instituto de Salud Carlos III (FIS PI16CIII/00034-ISCIII-FEDER).S

Detection of kinase domain mutations in BCR::ABL1 leukemia by ultra-deep sequencing of genomic DNA

The screening of the BCR::ABL1 kinase domain (KD) mutation has become a routine analysis in case of warning/failure for chronic myeloid leukemia (CML) and B-cell precursor acute lymphoblastic leukemia (ALL) Philadelphia (Ph)-positive patients. In this study, we present a novel DNA-based next-generation sequencing (NGS) methodology for KD ABL1 mutation detection and monitoring with a 1.0E−4 sensitivity. This approach was validated with a well-stablished RNA-based nested NGS method. The correlation of both techniques for the quantification of ABL1 mutations was high (Pearson r = 0.858, p < 0.001), offering DNA-DeepNGS a sensitivity of 92% and specificity of 82%. The clinical impact was studied in a cohort of 129 patients (n = 67 for CML and n = 62 for B-ALL patients). A total of 162 samples (n = 86 CML and n = 76 B-ALL) were studied. Of them, 27 out of 86 harbored mutations (6 in warning and 21 in failure) for CML, and 13 out of 76 (2 diagnostic and 11 relapse samples) did in B-ALL patients. In addition, in four cases were detected mutation despite BCR::ABL1 < 1%. In conclusion, we were able to detect KD ABL1 mutations with a 1.0E−4 sensitivity by NGS using DNA as starting material even in patients with low levels of disease.Tis project was funded in part by CRIS CANCER FOUNDATION

Relation between pathologic findings and cellular immune responses in sheep with naturally acquired paratuberculosis

5 páginas.Objective-To investigate the relation between lesions and cellular immune response-based tests in sheep with naturally acquired paratuberculosis and to evaluate efficacy of an interferon-gamma (IFN-gamma) assay and the intradermal skin test. Animals-110 and 88 adult sheep were used to evaluate the IFN-gamma assay and skin test, respectively. Procedure-Before euthanasia, heparinized blood was collected and incubated with avian purified protein derivative (PPD). Interferon-gamma production was assessed, using a commercial enzyme immunoassay. Skin testing in the caudal fold also was performed, using avian PPD. Sensitivity and specificity of these techniques were evaluated, using histopathologic findings as reference. Results-Of the sheep examined, 61 (55.4%) had lesions that were classified in 5 categories, according to location, extension, cellular types, and presence of acid-fast bacilli. Regarding cellular immunity lest results, 55.4% of sheep with and 6.2% without pathologic changes had positive results of the IFN-gamma assay. Skin test sensitivity (55.6%) and specificity (100%) were slightly greater. Conclusion-Close association between pathologic findings and cellular immune response was apparent. Most sheep with focal (tuberculoid) and some with diffuse lesions (borderline tuberculoid), all with scant or no acid-fast bacilli in the intestine, had positive results of the IFN-gamma, assay and skin test. Most sheep with diffuse lesions and high numbers of bacilli (borderline lepromatous) were test negative. Clinical Relevance-Cellular immune response-based and serologic tests are useful tools for seeking evidence of paratuberculosis. Each technique detects different types of immune-reacting sheep, and their simultaneous use may detect most infected animals.Peer Reviewe

Relation between serologic response and pathologic findings in sheep with naturally acquired paratuberculosis

5 páginas.Objective-To investigate the relation between lesions and serologic response in sheep with naturally acquired paratuberculosis and to evaluate the efficacy of agar gel immunodiffusion (AGID) and ELISA as diagnostic tests. Animals-134 adult sheep, culled by the owner from flocks with history of severe chronic weight loss. Procedure-Before euthanasia, blood samples were collected and serum was obtained. AGID and ELISA were performed, using a protoplasmic antigen of Mycobacterium paratuberculosis. Sensitivity and specificity of these techniques were evaluated, using histopathologic findings as a reference. Histologic studies were carried out focusing on specimens containing intestinal organized lymphoid tissue. Results-Of the sheep examined, 46.3% had lesions that were classified in 5 categories, according to their location, extension, cellular types, and presence of acid-fast bacilli. Regarding serologic test results, 37.1% of sheep with lesions had a positive AGID result, and all sheep without lesions had negative results. However, ELISA positivity was slightly greater-48.4% of sheep with lesions; however, by use of ELISA, 11.1% of sheep without lesions had positive results. Conclusion-A close relation between pathologic findings and serologic response was determined, especially between presence of acid-fast bacilli (potentially heavily shedding sheep) in the intestinal lesions and positivity to serologic tests, especially AGID. Clinical Relevance-On the basis of these results, together with its high specificity, reliability, and the difficulty of fecal specimen culture in sheep, AGID is proposed as a useful tool for seeking evidence of paratuberculosis in live sheep, and therefore, is suggested as the basis for control programs in areas in which the disease is a serious threat to sheep production.Peer Reviewe

Distribución de antígeno vírico en ovinos persistentemente infectados con pestivirus

Comunicación oral presentada a la: XXX Reunión de la Sociedad Española de Anatomía Patológica Veterinaria (SEAPV). San Pedro del Pinatar, Murcia, 13-15 junio 2018.El género Pestivirus, familia Flaviviridae incluye especies de virus RNA que afectan a los rumiantes: virus de la diarrea vírica bovina (BVVD) y el virus de la Enfermedad de la frontera. Este último, muy relacionado con el BVVD, causa infertilidad y abortos en la especie ovina. Cuando el feto ovino se infecta en el primer tercio de la gestación, con un sistema inmunitario aún no desarrollado, y sobrevive a la infección, el virus se extiende por todos sus órganos. Estos animales parecen ser inmunotolerantes al virus y pueden permanecer como animales persistentemente infectados (P I). Los ovinos P I son los principales transmisores del virus por vía horizontal y vertical. El objetivo de este estudio es caracterizar la distribución del antígeno vírico de pestivirus en ovinos P I. Los animales estudiados pertenecían a un rebaño Assaf con un historial de abortos del 13%. Ante un diagnóstico de abortos por pestivirus se realizaron diferentes pruebas laboratoriales que logaron identificar 2 ovejas adultas P I, una gestante. En estas dos ovejas P I, y en la cordera nacida de una de ellas, se realizó la necropsia y se tomaron muestras de diferentes localizaciones. Para la demostración de antígeno vírico se utilizó el anticuerpo monoclonal 15C5 frente a la proteína estructural Erns. En los ovinos adultos se observó neumonía catarral crónica, amiloidosis renal y urolitiasis, hiperplasia de la pulpa blanca del bazo, cicatrices en rumen, granulomas parasitarios en intestino, infiltrados inflamatorios de células mononucleadas en páncreas y manguitos perivasculares en encéfalo. En la cordera no se detectaron lesiones relevantes excepto coroiditis focal. En todos se detectó antígeno vírico, de forma extensa e intensa, en macrófagos medulares de nódulos linfáticos y del bazo, en menor intensidad en zonas centrales de los folículos linfoides (localización de células foliculares dendríticas) en estos órganos, tonsilas y placa de Peyer. En el timo se detectó intensa inmunotinción en células de morfología compatible con células epiteliales reticulares (ovino joven). En todos se observó intensa inmunopositividad en neuronas, células de los manguitos perivasculares y gliales en encéfalo, así como en plexos nerviosos e islotes pancreáticos. En útero (macrófagos y células musculares del miometrio) y en ovario (macrófagos del estroma, células luteínicas y oocitos, éstos en el ovino más joven) se detectó también antígeno vírico. Otras células positivas fueron: endoteliales, musculares lisas en arterias, epiteliales de túbulos renales y vejiga urinaria, epiteliales del tiroides y glándula adrenal, macrófagos y células fusiformes en submucosa del tracto gastrointestinal, macrófagos de la piel, así como macrófagos alveolares, epitelio bronquiolar y condrocitos (traqueales y bronquiales), éstos últimos en el ovino más joven. El conocimiento de la distribución del virus en estos animales podría ayudar a comprender mejor la patogenia y transmisión de esta enfermedad.Peer reviewe

Diagnóstico de la calidad del agua en los viveros forestales de México

Introducción: Un vivero forestal debe contar con abundante provisión de agua de calidad. El agua de características deficientes puede contener sales, iones tóxicos, contaminantes o fitopatógenos. No obstante, en México la calidad del agua en dichos viveros ha recibido poca atención. Por ello, se planteó como objetivo realizar un diagnóstico de la calidad del agua en viveros forestales de México. Método: En 148 viveros forestales ubicados en la República Mexicana que producen planta en contenedor o en bolsa, fueron obtenidas muestras de 1 L de agua. Éstas se analizaron en el Laboratorio Central de la Universidad Autónoma Chapingo. Fueron estimados los parámetros de pH, conductividad eléctrica, concentración de bicarbonatos, calcio, boro, magnesio, sulfatos, sodio y cloro. Con los datos obtenidos se calculó la dureza total y la relación ajustada de adsorción de sodio (esta última para viveros que producen en bolsa). Resultados: Los parámetros que excedieron los límites establecidos fueron: pH con 84% (125 viveros), conductividad eléctrica con 34% (50 viveros), y concentración de bicarbonatos con 77% (114 viveros); en el resto de los parámetros medidos sólo un pequeño porcentaje está por arriba de los valores estándar. De todos los viveros, seis que producen en contenedor y dos que producen en bolsa, cumplen con los parámetros de calidad de agua, el resto tienen por lo menos un valor que no cumple con los límites establecidos. Conclusión: Con base en la proporción de viveros de cada región ecológica, la región templada sobresale con pH y bicarbonatos en altas concentraciones, y la región tropical húmeda presenta valores elevados en conductividad eléctrica. Se recomienda siempre seleccionar la ubicación del vivero entre otros factores, para contar con una fuente de agua de calidad

Maternal immune response in the placenta of sheep during recrudescence of natural congenital infection of Neospora caninum

12 páginas, 5 figuras, 2 tablas.In order to gain further insight into the pathogenesis and transmission of ovine neosporosis, the serological response of 13 naturally infected pregnant sheep was monitored. All sheep were euthanized upon the detection of a sharp increase in the level of specific antibodies against N. caninum in order to study the maternal immune response after the recrudescence of a chronic infection. Ten sheep were euthanized between 84 and 118 days of gestation, whereas the three remaining and three control not infected, pregnant sheep were euthanized at 135 days of gestation after no sharp increase in antibodies was detected. Vertical transmission was confirmed in 11 sheep by detection of N. caninum-DNA in at least one fetus, confirming recrudescence. Not all of fetuses showed pathologic microscopic lesions, however, multifocal non-purulent encephalitis was the main finding. Furthermore, nine out of the 11 vertical transmission positive sheep had lesions in placentomes (mainly multifocal necrotic foci), and the parasite was detected in eight out of 11 placentas by PCR and/or immunohistochemestry. The placentomes from sheep that suffered recrudescence showed an increased number of T lymphocytes CD3+ (CD4/CD8 < 1) and macrophages (MHC-II+), assessed by immunohistochemestry, together with an upregulation of IFN-γ, IL-10, IL-4, TNFα, IL-2 and IL-18. IL-17 was only upregulated in the three infected sheep that did not have a sharp increase in antibody levels. In the sheep that showed fetal death at the time of euthanasia (n = 3) the placental microscopic lesions were more severe, the inflammatory infiltrate was higher, and the upregulation of cytokines was greater than in those sheep carrying viable fetuses. This study suggests that, similarly to bovine neosporosis, the time of gestation when recrudescence occurs determines the viability of the fetuses and, thus, seems to be related to the severity of lesions and immune response in the placenta. These results suggest that there might be a correlation, either as cause or as a consequence, between protection against vertical transmission of the parasite and a milder maternal serological response together with a high level of transcription of IL-17 in the placenta.The present study was financed by the Ministerio de Ciencia, Innovacion y Universidades of Spain (INIA project RTA2014-00013) and the European Regional Development Fund. Daniel Gutiérrez-Exposito ´ is the recipient of a postdoctoral contract from the Ministry of Science and Innovation (grant no. FJCI-2017-32020). We wish to acknowledge María del Carmen Carro, Xavier Cortizo, María Teresa Carro and Marta Silva for their excellent technical assistance. Finally, we wish to acknowledge Dr. Luis Miguel Ortega-Mora (SALUVET group) for kindly providing the in-house polyclonal antibody against N. caninum