• European Marine Science
• METIS Research Information System close

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. 17 1

Despite its well-established negative impacts on society and biodiversity, eutrophication continues to be one of the most pervasive anthropogenic influence along the freshwater to marine continuum. The interaction between eutrophication and climate change, particularly climate warming, was explicitly focused upon a decade ago in the paper by Moss et al. (2011), which called for an integrated response to both problems, given their apparent synergy. In this review, we summarise advances in the theoretical framework and empirical research on this issue and analyse the current understanding of the major drivers and mechanisms by which climate change can enhance eutophication, and vice versa, with a particular focus on shallow lakes. Climate change can affect nutrient loading, through changes at the catchment and landscape levels by affecting hydrological patterns and fire frequency, and through temperature effects on nutrient cycling. Biotic communities and their interactions can also be directly and indirectly affected by climate change, leading to an overall weakening of resilience to eutrophication impacts. Increasing empirical evidence now indicates several mechanisms by which eutrophying aquatic systems can increasingly act as important sources of greenhouse gases to the atmosphere, particularly methane. We also highlight potential feedbacks between eutrophication, cyanobacterial blooms, and climate change. Facing both challenges at the same time is more pressing than ever. Meaningful and strong measures at the landscape and water body levels are therefore required if we are to ensure ecosystem resilience and safe water supply, conserving biodiversity, and decreasing the carbon footprint of freshwaters. European Union’s Horizon 2020- grant agreement No 869296 The PONDERFUL Project