Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Evaluación de una prueba rápida para diagnóstico de dengue en el nivel local Evaluation of a rapid dengue test at the local level

Justificación y objetivo: Analizar concordancia entre la prueba rápida Dengue Duo IgM & IgG® de la casa comercial PanBio®, aplicada en el nivel local de diferentes áreas del país y en el Centro Nacional de Referencia de Virología (CNRV), con resultados obtenidos por MACELISA del CNRV. Métodos: Resultados de 828 determinaciones de IgM dengue por Dengue Duo IgM & IgG® del nivel local y del CNRV se compararon por el índice kappa con los resultados del MAC-ELISA del CNRV. Para verificar el cumplimiento en procedimientos de la casa comercial, se realizó una visita de inspección a los laboratorios y se envió muestras incógnitas para evaluar el desempeño. Resultados: El índice kappa de la prueba rápida en el nivel local contra el MAC-ELISA fue 0,62 (IC 95% 0,55-0,69) y la concordancia de los positivos del 75% (IC 95% 71-79). El índice kappa de la prueba rápida en el CNRV fue 0,81 (IC 95% 0,88-1,74) y la concordancia de los positivos del 93% (IC 95% 90-95). El 100% de los laboratorios mostraron condiciones adecuadas para aplicar esta prueba; su personal fue capacitado, pero la atención de otros análisis dificultó la lectura en el tiempo establecido. La concordancia de las incógnitas fue del 100%. Conclusión: Se evidenció una diferencia significativa en el índice kappa y la concordancia de los positivos, al aplicar la prueba rápida en los laboratorios y en el CNRV, hallazgos que documentan que la descentralización de esta metodología requerirá un acompañamiento que identifique factores causales de discordancia para subsanarlos y promover la generación de resultados consistentes que orienten la toma de decisiones acertadas.Background and aim: To analyze concordance of results obtained using the Dengue Duo IgM &IgG® rapid test (Pan Bio®) applied at the local level in different areas across the country and the MAC ELISA used at National Reference Center for Virology (CNRV) Methodology: The results of 828 dengue tests carried out with the Duo IgM& IgG® (Pan Bio®) at local level and at the CNRV were compared by the kappa index with those obtained with a MAC ELISA test at the CNRV. An inspection visit to participating laboratories was made and unknown samples were sent to evaluate the performance of the participants and to check the fulfillment of the procedures used with the recommendations of the manufacturer. Results: The kappa index of the results of the rapid test used at local level in comparison with the MAC ELISA was 0.62 (CI 95% 90-95); the concordance of the positive samples was 75% (CI 95% 71-79). The kappa index of the rapid test used at the CNRV was 0.81 (CI 95% 0.88- 1.74) and the concordance of the positive samples was 93% (CI 95% 90-95). All laboratories showed ideal facilities to carry out the tests and the staff got the training required; however, the attention to other analyses affected the reading within the time recommended. The concordance of the unknown samples was 100%. Conclusion: A significant difference was demonstrated in the kappa index and the concordance of positive samples between the rapid tests carried out at the participant laboratories and at the CNRV. These results imply that decentralization of this methodology would require continuous support to identify correctable factors of disagreement and to promote the generation of consistent results to orient the decision-making process

Genetic evolution of influenza viruses among selected countries in Latin America, 2017-2018.

OBJECTIVE:Since the 2009 influenza pandemic, Latin American (LA) countries have strengthened their influenza surveillance systems. We analyzed influenza genetic sequence data from the 2017 through 2018 Southern Hemisphere (SH) influenza season from selected LA countries, to map the availability of influenza genetic sequence data from, and to describe, the 2017 through 2018 SH influenza seasons in LA. METHODS:We analyzed influenza A/H1pdm09, A/H3, B/Victoria and B/Yamagata hemagglutinin sequences from clinical samples from 12 National Influenza Centers (NICs) in ten countries (Argentina, Brazil, Chile, Colombia, Costa Rica, Ecuador, Mexico, Paraguay, Peru and Uruguay) with a collection date from epidemiologic week (EW) 18, 2017 through EW 43, 2018. These sequences were generated by the NIC or the WHO Collaborating Center (CC) at the U.S Centers for Disease Control and Prevention, uploaded to the Global Initiative on Sharing All Influenza Data (GISAID) platform, and used for phylogenetic reconstruction. FINDINGS:Influenza hemagglutinin sequences from the participating countries (A/H1pdm09 n = 326, A/H3 n = 636, B n = 433) were highly concordant with the genetic groups of the influenza vaccine-recommended viruses for influenza A/H1pdm09 and influenza B. For influenza A/H3, the concordance was variable. CONCLUSIONS:Considering the constant evolution of influenza viruses, high-quality surveillance data-specifically genetic sequence data, are important to allow public health decision makers to make informed decisions about prevention and control strategies, such as influenza vaccine composition. Countries that conduct influenza genetic sequencing for surveillance in LA should continue to work with the WHO CCs to produce high-quality genetic sequence data and upload those sequences to open-access databases

The epidemiological signature of influenza B virus and its B/Victoria and B/Yamagata lineages in the 21st century

We describe the epidemiological characteristics, pattern of circulation, and geographical distribution of influenza B viruses and its lineages using data from the Global Influenza B Study. We included over 1.8 million influenza cases occurred in thirty-one countries during 2000-2018. We calculated the proportion of cases caused by influenza B and its lineages; determined the timing of influenza A and B epidemics; compared the age distribution of B/Victoria and B/Yamagata cases; and evaluated the frequency of lineage-level mismatch for the trivalent vaccine. The median proportion of influenza cases caused by influenza B virus was 23.4%, with a tendency (borderline statistical significance, p = 0.060) to be higher in tropical vs. temperate countries. Influenza B was the dominant virus type in about one every seven seasons. In temperate countries, influenza B epidemics occurred on average three weeks later than influenza A epidemics; no consistent pattern emerged in the tropics. The two B lineages caused a comparable proportion of influenza B cases globally, however the B/Yamagata was more frequent in temperate countries, and the B/Victoria in the tropics (p = 0.048). B/Yamagata patients were significantly older than B/Victoria patients in almost all countries. A lineage-level vaccine mismatch was observed in over 40% of seasons in temperate countries and in 30% of seasons in the tropics. The type B virus caused a substantial proportion of influenza infections globally in the 21st century, and its two virus lineages differed in terms of age and geographical distribution of patients. These findings will help inform health policy decisions aiming to reduce disease burden associated with seasonal influenza

Erratum to: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (Autophagy, 12, 1, 1-222, 10.1080/15548627.2015.1100356

non present

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field