Hepatitis C virus infection of primary tupaia hepatocytes leads to selection of quasispecies variants, induction of interferon-stimulated genes and NF-kappaB nuclear translocation

Systems for in vitro culture of Hepatitis C virus (HCV) are essential tools to analyse virus-cell interactions and to investigate relevant pathophysiological aspects of HCV infection. Although the HCV replicon methodology has increased our understanding of HCV biology, this system does not reproduce the natural infection. Recently, tupaia (Tupaia belangeri chinensis) hepatocytes have been utilized for in vitro culture of HCV. In the present work, primary tupaia hepatocytes infected in vitro with HCV were used to analyse the evolution of HCV quasispecies in infected cells and the ability of the virus to influence antiviral and proinflammatory responses in cells sustaining virus replication. The results confirmed the potential of tupaia hepatocytes as a model for HCV infection, although this system is limited by rapid loss of differentiated cell phenotype in culture. These findings revealed an extraordinary plasticity of HCV quasispecies, which underwent rapid evolution to tupaia-tropic variants as early as 24 h after infection. It was also shown that HCV could activate interferon-sensitive genes, albeit modestly in comparison with other viruses such as Semliki Forest virus. Importantly, HCV activated NF-kappaB in primary hepatocytes and upregulated NF-kappaB-responsive genes including the chemokines MCP-1 and CXCL2 (MIP-2). This effect may play a role in induction of the hepatic inflammatory reaction in vivo. In summary, HCV quasispecies adapt rapidly to the specific biology of the host and HCV stimulates a blunted interferon response while inducing a proinflammatory phenotype in the infected cell

Follow-Up Study Confirms the Presence of Gastric Cancer DNA Methylation Hallmarks in High-Risk Precursor Lesions

Intestinal metaplasia confers an increased risk of progression to gastric cancer. However, some intestinal metaplasia patients do not develop cancer. The development of robust molecular biomarkers to stratify patients with advanced gastric precursor lesions at risk of cancer progression will contribute to guiding programs for prevention. Starting from a genome-wide methylation study, we have simplified the detection method regarding candidate-methylation tests to improve their applicability in the clinical environment. We identified CpG methylation at the ZNF793 and RPRM promoters as a common event in intestinal metaplasia and intestinal forms of gastric cancer. Furthermore, we also showed that Helicobacter pylori infection influences DNA methylation in early precursor lesions but not in intestinal metaplasia, suggesting that therapeutic strategies to prevent epigenome reprogramming toward a cancer signature need to be adopted early in the precursor cascade. To adopt prevention strategies in gastric cancer, it is imperative to develop robust biomarkers with acceptable costs and feasibility in clinical practice to stratified populations according to risk scores. With this aim, we applied an unbiased genome-wide CpG methylation approach to a discovery cohort composed of gastric cancer (n = 24), and non-malignant precursor lesions (n = 64). Then, candidate-methylation approaches were performed in a validation cohort of precursor lesions obtained from an observational longitudinal study (n = 264), with a 12-year follow-up to identify repression or progression cases. H. pylori stratification and histology were considered to determine their influence on the methylation dynamics. As a result, we ascertained that intestinal metaplasia partially recapitulates patterns of aberrant methylation of intestinal type of gastric cancer, independently of the H. pylori status. Two epigenetically regulated genes in cancer, RPRM and ZNF793, consistently showed increased methylation in intestinal metaplasia with respect to earlier precursor lesions. In summary, our result supports the need to investigate the practical utilities of the quantification of DNA methylation in candidate genes as a marker for disease progression. In addition, the H. pylori-dependent methylation in intestinal metaplasia suggests that pharmacological treatments aimed at H. pylori eradication in the late stages of precursor lesions do not prevent epigenome reprogramming toward a cancer signature

Recommendations for ophthalmologic practice during the easing of COVID-19 control measures

In the context of the COVID-19 pandemic, this paper provides recommendations for medical eye care during the easing of control measures after lockdown. The guidelines presented are based on a literature review and consensus among all Spanish Ophthalmology Societies regarding protection measures recommended for the ophthalmologic care of patients with or without confirmed COVID-19 in outpatient, inpatient, emergency and surgery settings. We recommend that all measures be adapted to the circumstances and availability of personal protective equipment at each centre and also highlight the need to periodically update recommendations as we may need to readopt more restrictive measures depending on the local epidemiology of the virus. These guidelines are designed to avoid the transmission of SARS-CoV-2 among both patients and healthcare staff as we gradually return to normal medical practice, to prevent postoperative complications and try to reduce possible deficiencies in the diagnosis, treatment and follow-up of the ophthalmic diseases. With this update (5th) the Spanish Society of Ophthalmology is placed as one of the major ophthalmology societies providing periodic and systematized recommendations for ophthalmic care during the COVID-19 pandemic

Next-generation sequencing of bile cell-free DNA for the early detection of patients with malignant biliary strictures

Objective: despite significant progresses in imaging and pathological evaluation, early differentiation between benign and malignant biliary strictures remains challenging. Endoscopic retrograde cholangiopancreatography (ERCP) is used to investigate biliary strictures, enabling the collection of bile. We tested the diagnostic potential of next-generation sequencing (NGS) mutational analysis of bile cell-free DNA (cfDNA). Design: a prospective cohort of patients with suspicious biliary strictures (n=68) was studied. The performance of initial pathological diagnosis was compared with that of the mutational analysis of bile cfDNA collected at the time of first ERCP using an NGS panel open to clinical laboratory implementation, the Oncomine Pan-Cancer Cell-Free assay. Results: an initial pathological diagnosis classified these strictures as of benign (n=26), indeterminate (n=9) or malignant (n=33) origin. Sensitivity and specificity of this diagnosis were 60% and 100%, respectively, as on follow-up 14 of the 26 and eight of the nine initially benign or indeterminate strictures resulted malignant. Sensitivity and specificity for malignancy of our NGS assay, herein named Bilemut, were 96.4% and 69.2%, respectively. Importantly, one of the four Bilemut false positives developed pancreatic cancer after extended follow-up. Remarkably, the sensitivity for malignancy of Bilemut was 100% in patients with an initial diagnosis of benign or indeterminate strictures. Analysis of 30 paired bile and tissue samples also demonstrated the superior performance of Bilemut. Conclusion: implementation of Bilemut at the initial diagnostic stage for biliary strictures can significantly improve detection of malignancy, reduce delays in the clinical management of patients and assist in selecting patients for targeted therapies.Funding: we thank the financial support of CIBERehd; grants PI16/01126 and PI19/00163 from Instituto de Salud Carlos III (ISCIII) cofinanced by ’Fondo Europeo de Desarrollo Regional’ (FEDER) ’Una manera de hacer Europa’; grants 58/2017 and 55/2018 from Gobierno de Navarra Salud; grant 0011-1411-2020-000010 from AGATA Strategic Project from Gobierno de Navarra; grant 2020/101 from Euroregion Nouvelle Aquitaine-Euskadi-Navarra; Fundación Eugenio Rodríguez Pascual; Fundación Mario Losantos, Fundación M Torres; grant 2018/117 from AMMF, the Cholangiocarcinoma Charity; the COST Action CA181122 Euro-cholangio-Net; POSTD18014AREC postdoctoral fellowship from AECC to MA; and Ramón y Cajal Program contracts RYC-2014-15242 and RYC-2018-024475-1 to FJC and MGFB

CiTiEs (Ciudades: Tiempo + Espacio). Implementación de itinerarios didácticos para la enseñanza virtual y presencial del Patrimonio cultural de Madrid a través del aprendizaje cooperativo

Retomando los itinerarios didácticos diseñados en un PID anterior (2019/20, nº 363), se implementarán materiales digitales para la Educación patrimonial de Madrid. Se utilizarán técnicas de aprendizaje cooperativo y recursos para la enseñanza virtual.Depto. de Didáctica de las Ciencias Experimentales , Sociales y MatemáticasFac. de EducaciónFALSEsubmitte

Curvas de Intensidad – Duración – Frecuencia (IDF) de la estación meteorológica del CIM–FICH. Serie 1986–2016

The study of the rainfall distribution and temporal evolution is interesting in hydrological design projects. The IDF curves is most useful methodology to define the design storm based on rain intensity, duration and recurrence. Its construction has often been limited by the availability of pluviographic data, in relation to the spatial coverage of the stations and the insufficient length of the records. The objective of this work is to update the curves of the IDF of the Meteorological Information Center (CIM) "Lic. Enrique B. Rodríguez", Facultad de Ingeniería y Ciencias Hídricas, Universidad Nacional del Litoral (FICH–UNL). The IDF curves was determined by Sherman methodology and was verified that the rain intensity decreases when the duration increases and, during the same duration, the rain intensity increases with the return period. The rain intensities less than and equal to 10 minutes exceed 100 mm/h for all recurrences. The IDF curves for the CIM station show a lower rain intensity and those defined for the cities of Rafaela and Paraná, with percentage differences that increase with recurrence.El estudio de la precipitación, su distribución y evolución temporal son de interés para proyectos de diseño hidrológico. Las curvas IDF es la metodología más utilizada para definir la tormenta de diseño en base a la relación entre la intensidad de la lluvia, la duración y la recurrencia. Su construcción muchas veces se ve limitada por la escasa disponibilidad de datos pluviográficos, referente a la cobertura espacial de las estaciones y a la longitud insuficiente de los registros. El presente trabajo tiene por objetivo actualizar las curvas IDF del Centro de Informaciones Meteorológicas (CIM) “Lic. Enrique B. Rodríguez”, Facultad de Ingeniería y Ciencias Hídricas, Universidad Nacional del Litoral (FICH–UNL). Las curvas IDF fueron calculadas por el método de Sherman y se verificó que la intensidad de precipitación decrece cuando aumenta la duración y para una misma duración, la intensidad aumenta con el periodo de retorno. Las intensidades menores e iguales a 10 minutos superan los 100 mm.h -1 para todas las recurrencias. Las curvas IDF determinadas para la estación del CIM presentan intensidades de lluvia menores a las definidas para las localidades de Rafaela y Paraná, con diferencias porcentuales que aumentan con la recurrencia.

Curvas de Intensidad – Duración – Frecuencia (IDF) de la estación meteorológica del CIM–FICH. Serie 1986–2016

El estudio de la precipitación, su distribución y evolución temporal son de interés para proyectos de diseño hidrológico. Las curvas IDF es la metodología más utilizada para definir la tormenta de diseño en base a la relación entre la intensidad de la lluvia, la duración y la recurrencia. Su construcción muchas veces se ve limitada por la escasa disponibilidad de datos pluviográficos, referente a la cobertura espacial de las estaciones y a la longitud insuficiente de los registros. El presente trabajo tiene por objetivo actualizar las curvas IDF del Centro de Informaciones Meteorológicas (CIM) “Lic. Enrique B. Rodríguez”, Facultad de Ingeniería y Ciencias Hídricas, Universidad Nacional del Litoral (FICH–UNL). Las curvas IDF fueron calculadas por el método de Sherman y se verificó que la intensidad de precipitación decrece cuando aumenta la duración y para una misma duración, la intensidad aumenta con el periodo de retorno. Las intensidades menores e iguales a 10 minutos superan los 100 mm.h -1 para todas las recurrencias. Las curvas IDF determinadas para la estación del CIM presentan intensidades de lluvia menores a las definidas para las localidades de Rafaela y Paraná, con diferencias porcentuales que aumentan con la recurrencia.Fil: Marcus, Ramiro. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hidricas. Santa Fe, Argentina.Fil: Morresi, María del Valle. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hidricas. Santa Fe, Argentina.Fil: Elizalde Carrillo, Esteban. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hidricas. Santa Fe, Argentina.Fil: Macor, José.Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hidricas. Santa Fe, Argentina.Fil: Cristina, Ignacio. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hidricas. Santa Fe, Argentina