Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

O kañe (olhar) na cidade : práticas de embelezamento corporal na infância feminina Kaingang

Esta dissertação tem como questão central as noções de beleza feminina corporal da infância feminina Kaingang moradora da cidade. A investigação foi realizada com dezesseis meninas com idades entre quatro e doze anos residentes em uma comunidade indígena localizada no bairro Lomba do Pinheiro em Porto Alegre. A pesquisa se desenvolveu entre os anos de 2009 e 2011 e buscou investigar, analisar e discutir em que medida as representações de beleza dos corpos femininos veiculadas nos meios de comunicação funcionam como pedagogias visuais e afetam as noções de beleza corporal feminina das meninas Kaingang. Procurou ainda compreender como essas meninas Kaingang absorvem as práticas de embelezamento e da cultura da cidade ao se relacionarem em diferentes territórios urbanos. Foram analisados aspectos corporais relacionados aos marcadores de gênero, classe social, raça e cor como construções históricas, sociais e culturais. A pesquisa de campo de cunho participante foi embasada em aportes da etnografia e em referenciais teóricos dos Estudos da Infância, Estudos Culturais e da Cultura Visual. As análises foram qualitativas nas quais foram cruzados os dados gerados a partir dos dizeres, desenhos e registros fotográficos das meninas Kaingang. Os resultados demonstraram que as meninas Kaingang da cidade possuem preocupações com a aparência de seus corpos que influenciam em suas feminilidades. Pela forma de lidarem com suas peles e cabelos, suas maneiras de vestir e se comportar as análises denotaram que suas valorações quanto ao que consideram ser belo e feio nos corpos femininos são absorvidas por meio das pedagogias culturais e visuais. A investigação explicitou ainda, que as meninas Kaingang sofrem influências dos ambientes territoriais nos quais circulam. Os meios midiáticos ou mesmo sociais e culturais da cidade reverberam nos valores que atribuem à beleza dos corpos femininos e, consequentemente, em suas identidades femininas infantis.The main objective of this dissertation is clarifying notions of urban Kaingang female children‟s bodily beauty. The investigation was conducted with sixteen four- and twelve-year-old girls living in an Indigenous community at Lomba do Pinheiro in Porto Alegre (RS). The research was developed between 2009 and 2011 and sought to investigate, analyse and discuss how representations for female bodies‟ beauty provided in the media work as visual pedagogies and act on notions of Kaingang girls‟ bodily beauty. It has also sought to understand how these Kaingang girls receive cosmetic practices and the city culture as they live in different urban territories. Bodily aspects concerning gender, class, race, and colour markers were analysed. The participating field research was drawn on ethnographic issues and theoretical referential for Child Studies, Cultural Studies and Visual Culture. Analyses were qualitative and data from Kaingang girls‟ statements, drawings and photographic shots were cross-referenced. The results demonstrated that Kaingang girls are concerned about how their bodies look like, which affect their femininity. In tune to the way they treat their skin and hair, the way they dress and behave, analyses noted that the value of what they regard as beautiful and ugly in female bodies are received through cultural and visual pedagogies. The investigation has also shown that the environment act on Kaingang girls. The city media, society and culture act on values ascribed to female bodily beauty and so their female child identities

O kañe (olhar) na cidade : práticas de embelezamento corporal na infância feminina Kaingang

Esta dissertação tem como questão central as noções de beleza feminina corporal da infância feminina Kaingang moradora da cidade. A investigação foi realizada com dezesseis meninas com idades entre quatro e doze anos residentes em uma comunidade indígena localizada no bairro Lomba do Pinheiro em Porto Alegre. A pesquisa se desenvolveu entre os anos de 2009 e 2011 e buscou investigar, analisar e discutir em que medida as representações de beleza dos corpos femininos veiculadas nos meios de comunicação funcionam como pedagogias visuais e afetam as noções de beleza corporal feminina das meninas Kaingang. Procurou ainda compreender como essas meninas Kaingang absorvem as práticas de embelezamento e da cultura da cidade ao se relacionarem em diferentes territórios urbanos. Foram analisados aspectos corporais relacionados aos marcadores de gênero, classe social, raça e cor como construções históricas, sociais e culturais. A pesquisa de campo de cunho participante foi embasada em aportes da etnografia e em referenciais teóricos dos Estudos da Infância, Estudos Culturais e da Cultura Visual. As análises foram qualitativas nas quais foram cruzados os dados gerados a partir dos dizeres, desenhos e registros fotográficos das meninas Kaingang. Os resultados demonstraram que as meninas Kaingang da cidade possuem preocupações com a aparência de seus corpos que influenciam em suas feminilidades. Pela forma de lidarem com suas peles e cabelos, suas maneiras de vestir e se comportar as análises denotaram que suas valorações quanto ao que consideram ser belo e feio nos corpos femininos são absorvidas por meio das pedagogias culturais e visuais. A investigação explicitou ainda, que as meninas Kaingang sofrem influências dos ambientes territoriais nos quais circulam. Os meios midiáticos ou mesmo sociais e culturais da cidade reverberam nos valores que atribuem à beleza dos corpos femininos e, consequentemente, em suas identidades femininas infantis.The main objective of this dissertation is clarifying notions of urban Kaingang female children‟s bodily beauty. The investigation was conducted with sixteen four- and twelve-year-old girls living in an Indigenous community at Lomba do Pinheiro in Porto Alegre (RS). The research was developed between 2009 and 2011 and sought to investigate, analyse and discuss how representations for female bodies‟ beauty provided in the media work as visual pedagogies and act on notions of Kaingang girls‟ bodily beauty. It has also sought to understand how these Kaingang girls receive cosmetic practices and the city culture as they live in different urban territories. Bodily aspects concerning gender, class, race, and colour markers were analysed. The participating field research was drawn on ethnographic issues and theoretical referential for Child Studies, Cultural Studies and Visual Culture. Analyses were qualitative and data from Kaingang girls‟ statements, drawings and photographic shots were cross-referenced. The results demonstrated that Kaingang girls are concerned about how their bodies look like, which affect their femininity. In tune to the way they treat their skin and hair, the way they dress and behave, analyses noted that the value of what they regard as beautiful and ugly in female bodies are received through cultural and visual pedagogies. The investigation has also shown that the environment act on Kaingang girls. The city media, society and culture act on values ascribed to female bodily beauty and so their female child identities

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