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

A sociedade de risco midiatizada, o movimento antivacinação e o risco do autismo

Observam-se modificações epidemiológicas de doenças infecciosas entre famílias de classe média de países industrializados por força de crenças ligadas aos riscos da vacinação. Estas se expandem globalmente por conta de redes de sites, blogs e celebridades de ampla influência. Em vista da complexidade de tal fenômeno cultural, em sua analítica são articulados conceitos contemporâneos alinhados à ideia de reflexividade na sociedade de risco, assim como o da sociedade midiatizada receptora de enunciações de perigos e proteções em mútua referência e contradição. Discute-se a frequente emergência de tensões derivadas de ciclos de enunciações e incompletudes constituídas como “biovalores” simbólicos. Enfatiza-se o efeito persistente de enunciações ameaçadoras e fraudulentas a abastecer redes sociais virtuais que, há quase três décadas, ampliam o debate acerca da ligação do autismo com as vacinas. Conclui-se que os processos de produção de sentidos interligam-se em diversos níveis nos quais circulam representações que sustentam a comunicação e a identidade dos grupos com base em referenciais histórico-culturais

Optimization of cisplatin for the treatment of hormone dependent tumoral diseases Part.1. Use of steroidal ligands

A review. Platinum complexes such as cisplatin and carboplatin are metal based drugs, which are widely used in cancer chemotherapy. However, in the current therapy of hormone-dependent breast and prostate cancer they are not established. This might be the result of intrinsic and acquired resistance of the tumors during the therapy. Therefore, several attempts were done to design platinum complexes to address these tumors. Steroidal and non-steroidal drugs were modified for the coordination to platinum. The mode of action implies a binding to the estrogen receptor, a selective accumulation in the tumor cells and a specific binding to DNA. In part 1 of this review article we describe the use of steroidal ligands to optimize cisplatin for the treatment of hormone dependent tumoral diseases

Optimization of cisplatin for the treatment of hormone-dependent tumoral diseases Part.2 Use of non-steroidal ligands

A review. Platinum complexes with carrier ligands were synthesized to overcome intrinsic and acquired resistance of the tumors during the therapy. For the treatment of breast and prostate cancer steroidal and non-steroidal ligands are available. This article is a crit. review of the attempts made during the last 3 decades to design platinum complexes with a selective mode of action against hormone-dependent tumors. In part 1 of this paper the use of steroidal carrier ligands is described while in part 2 the view is focussed on derivs. of non-steroidal drugs suitable to coordinate to platinum. Esp. the [1,2-diarylethylenediamine]platinum(II) complexes are interesting analogous of cisplatin