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

Investigation of Interfacial Properties and Stability of Particle-Stabilised Emulsions in Microgravity Conditions in the Framework of the ESA-MAP FASES Programme

The poster provides and overview of the research project MAP-EASES (Fundamental and Applied Studies in Emulsion Stability) supported by the European Space Agency. The project is aimed at understanding the relationships between interfacial properties of surfactant adsorption layers at liquid-liquid interfaces and the stability and properties of the corresponding emulsions. The research programme also includes the investigation of particle-stabilised emulsions. These emulsions are widely diffuse in nature (Pickering emulsions) and have an increasing importance for different products, technologies and application subjects. Synthetic micro/nano-size particles, often in association with surfactants, are then being investigated to be utilised as new emulsion and foam additives, capable to provide long-term stability. On the other side, for some technologies it is important to understand the mechanisms underlying the particle stabilisation capability, in order to destabilise very stable emulsions, such as some crude oils. In order to respond to these fundamental and technological needs, the EASES project covers a wide spectrum of investigations and modelling: from the interaction of surfactants with particles, to droplet dynamics in emulsions, passing through the properties of single liquid-liquid interfaces and films. Benchmark experiments are planned onboard the International Space Station (ISS), exploiting the purely diffusive conditions and the absence of buoyancy, which allows for the accurate investigation of interfacial properties and of the fundamental processes involved in emulsion destabilisation. To this aim, specific innovative diagnostics have been conceived in the framework of the project, which have been realised by ESA into suitable experiment modules for the ISS. In particular, the experiment container EASES for the Fluid Science Laboratory, has been developed to produce emulsions and investigate collective properties of the emulsion, such as, destabilisation process and droplets interaction and dynamics. The module allows for the serial investigation of emulsion samples of different nature (water-in-oil, oil-in-water, surfactant-stabilised, particle stabilised) and composition (water/oil ratio, surfactant and particle concentrations) by means of two diagnostics. Optical microtomography is utilised for the investigation of transparent (dilute) emulsions, while a technique based on a Differential Scan Calorimeter allows for the investigation of opaque (concentrate) emulsions. Properties of oil-water interfaces relevant for emulsion stability, such as dilational viscoelasticity of surfactant adsorbed layers and surfactant adsorption kinetics, are instead investigated by means of the facility EASTER (Facility for Adsorption and Surface Tension), based on Capillary Pressure Tensiometry

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

International audienceIn 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