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

Role of Oregano (Origanum vulgare) Essential Oil as a Surface Fungus Inhibitor on Fermented Sausages: Evaluation of Its Effect on Microbial and Physicochemical Characteristics

Oregano essential oil (OEO) was evaluated to determine its effect on the growth of natural contaminating molds on the surface of Spanish fermented sausage, the development of the internal microbial population of the sausage, and the physicochemical properties of the sausage. Results indicated a dramatic reduction in the contaminant molds. At the end of ripening, the main endogenous fungal species in control samples were Mucor racemosus (55%), Aspergillus fumigatus (20.6%), Cladosporium sphaerospermum (11.1%), Acremonium strictum (7.9%), and Aspergillus niger (4.7%). In samples treated with OEO, M. racemosus and A. fumigatus were the only species isolated; the treatment was more effective against A. fumigatus than against M. racemosus. The use of OEO to inhibit surface fungi did not affect the sausage drying process, pH, water activity, or color changes during ripening. These parameters change in a typical pattern for fermented dry-cured sausages during ripening. At the end of ripening, OEO-treated sausages had lower hardness and greater chewiness than the control but showed similar textural properties to sausages treated with potassium sorbate. [...