TALEN mediated gene editing in a mouse model of Fanconi anemia

The promising ability to genetically modify hematopoietic stem and progenitor cells by precise gene editing remains challenging due to their sensitivity to in vitro manipulations and poor efficiencies of homologous recombination. This study represents the first evidence of implementing a gene editing strategy in a murine safe harbor locus site that phenotypically corrects primary cells from a mouse model of Fanconi anemia A. By means of the co-delivery of transcription activator-like effector nucleases and a donor therapeutic FANCA template to the Mbs85 locus, we achieved efficient gene targeting (23%) in mFA-A fibroblasts. This resulted in the phenotypic correction of these cells, as revealed by the reduced sensitivity of these cells to mitomycin C. Moreover, robust evidence of targeted integration was observed in murine wild type and FA-A hematopoietic progenitor cells, reaching mean targeted integration values of 21% and 16% respectively, that were associated with the phenotypic correction of these cells. Overall, our results demonstrate the feasibility of implementing a therapeutic targeted integration strategy into the mMbs85 locus, ortholog to the well-validated hAAVS1, constituting the first study of gene editing in mHSC with TALEN, that sets the basis for the use of a new safe harbor locus in mice.The authors would like to thank Miguel A. Martin for the careful maintenance of NSG mice and Omaira Alberquilla for her technical assistance in flow cytometry. The authors also thank the Fundación Botín for promoting translational research at the Hematopoietic Innovative Therapies Division of the CIEMAT. CIBERER is an initiative of the “Instituto de Salud Carlos III” and “Fondo Europeo de Desarrollo Regional (FEDER)”. Finally, although this work was entirely performed with mouse cells, the authors feel deeply grateful to the FA patients and families, clinicians and to Aurora de la Cal as the secretary of the Spanish FA network for always instilling motivation and providing an example for work in this disease. This work was supported by grants from the “7th Framework Program European Commission (HEALTH-F5-2012-305421; EUROFANCOLEN)”, “Ministerio de Sanidad, Servicios Sociales e Igualdad” (EC11/060 and EC11/550), “Ministerio de Economía, Comercio y Competitividad y Fondo Europeo de Desarrollo Regional (FEDER)” (SAF2015-68073-R), “Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III” (RD12/0019/0023) Consejería de Educación, Juventud y Deporte de la Comunidad de Madrid (AvanCell Project; B2017/BMD3692) and the German Federal Ministry of Education and Research (BMBF-01EO0803)S

TVIII - Arquitectura y Ciudad - AR301 - 202101

1Descripción: El Taller VIII, Arquitectura y Ciudad, es el taller en donde el alumno desarrolla un proyecto arquitectónico a partir de un contacto directo con el contexto urbano existente, por lo tanto, el grado de complejidad se incrementan al tener ahora otra gran consideración: LA CIUDAD. Se basa en una Intervención urbana y arquitectónica en un lugar de la ciudad (nivel barrial o nivel sectorial) en donde se han identificado previamente las posibilidades y necesidades del sitio. El análisis del lugar hace énfasis en: Espacio público, viabilidad, conectividad, identidad, sostenibilidad, vitalidad y dinámicas urbanas. Una vez identificada la zona, hecha la investigación correspondiente y obtener conclusiones sobre la problemática, carencias, virtudes, posibilidades y necesidades, se realiza una propuesta urbana grupal para el área mencionada, planteando soluciones a nivel macro (nivel barrial o nivel sectorial) y micro (infraestructura necesaria / proyecto específico). Propósito: Desarrollar propuestas arquitectónicas en las que la atención del diseño tome en consideración la problemática urbana de un sector de la ciudad, para y resolverla de manera que se extienda a la renovación y el embellecimiento del entorno urbano. El curso de TVIII ¿ Arquitectura y Ciudad ha sido diseñado con el propósito de introducir al estudiante en la visión de la ciudad como conjunto urbano. Se plantea propuestas urbano-arquitectónicas para el desarrollo sostenido y ordenado del hábitat. La asignatura del Taller VIII contribuye directamente al desarrollo de la competencia específica de la carrera: Diseño Fundamentado (que corresponde a los criterios NAAB : (PC2, PC3, PC8, PC5, SC3, SC5). Competencias en el nivel de logro 03. Tiene como rrequisito la asignatura de Taller VII ¿ Taller de Integración

TVIII - Arquitectura y Ciudad - AR301 - 202102

Descripción: El Taller VIII, Arquitectura y Ciudad, es el taller en donde el alumno desarrolla un proyecto arquitectónico a partir de un contacto directo con el contexto urbano existente, por lo tanto, el grado de complejidad se incrementan al tener ahora otra gran consideración: LA CIUDAD. Se basa en una Intervención urbana y arquitectónica en un lugar de la ciudad (nivel barrial o nivel sectorial) en donde se han identificado previamente las posibilidades y necesidades del sitio. El análisis del lugar hace énfasis en: Espacio público, viabilidad, conectividad, identidad, sostenibilidad, vitalidad y dinámicas urbanas. Una vez identificada la zona, hecha la investigación correspondiente y obtener conclusiones sobre la problemática, carencias, virtudes, posibilidades y necesidades, se realiza una propuesta urbana grupal para el área mencionada, planteando soluciones a nivel macro (nivel barrial o nivel sectorial) y micro (infraestructura necesaria / proyecto específico). Propósito: Desarrollar propuestas arquitectónicas en las que la atención del diseño tome en consideración la problemática urbana de un sector de la ciudad, para y resolverla de manera que se extienda a la renovación y el embellecimiento del entorno urbano. El curso de TVIII ¿ Arquitectura y Ciudad ha sido diseñado con el propósito de introducir al estudiante en la visión de la ciudad como conjunto urbano. Se plantea propuestas urbano-arquitectónicas para el desarrollo sostenido y ordenado del hábitat. La asignatura del Taller VIII contribuye directamente al desarrollo de la competencia específica de la carrera: Diseño Fundamentado (que corresponde a los criterios NAAB : (PC2, PC3, PC8, PC5, SC3, SC5). Competencias en el nivel de logro 03. Tiene como rrequisito la asignatura de Taller VII ¿ Taller de Integración

International Nosocomial Infection Control Consortiu (INICC) report, data summary of 43 countries for 2007-2012. Device-associated module

We report the results of an International Nosocomial Infection Control Consortium (INICC) surveillance study from January 2007-December 2012 in 503 intensive care units (ICUs) in Latin America, Asia, Africa, and Europe. During the 6-year study using the Centers for Disease Control and Prevention's (CDC) U.S. National Healthcare Safety Network (NHSN) definitions for device-associated health care–associated infection (DA-HAI), we collected prospective data from 605,310 patients hospitalized in the INICC's ICUs for an aggregate of 3,338,396 days. Although device utilization in the INICC's ICUs was similar to that reported from ICUs in the U.S. in the CDC's NHSN, rates of device-associated nosocomial infection were higher in the ICUs of the INICC hospitals: the pooled rate of central line–associated bloodstream infection in the INICC's ICUs, 4.9 per 1,000 central line days, is nearly 5-fold higher than the 0.9 per 1,000 central line days reported from comparable U.S. ICUs. The overall rate of ventilator-associated pneumonia was also higher (16.8 vs 1.1 per 1,000 ventilator days) as was the rate of catheter-associated urinary tract infection (5.5 vs 1.3 per 1,000 catheter days). Frequencies of resistance of Pseudomonas isolates to amikacin (42.8% vs 10%) and imipenem (42.4% vs 26.1%) and Klebsiella pneumoniae isolates to ceftazidime (71.2% vs 28.8%) and imipenem (19.6% vs 12.8%) were also higher in the INICC's ICUs compared with the ICUs of the CDC's NHSN

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)