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

Determination of the furaltadone metabolite 5-methylmorpholino-3-amino-2-oxazolidinone (AMOZ) using liquid chromatography coupled to electrospray tandem mass spectrometry during the nitrofuran crisis in Portugal

Abstract The use of nitrofuran veterinary drugs as antibacterial compounds in food-producing animals has been banned in the EU since 1995. As nitrofurans are extensive and rapidly metabolized, control of their illegal use in animal production must be done in edible tissues by LC-MS/MS analysis in order to determine persistent tissue-bound metabolites. The introduction during 2002 of the multi-residue detection of nitrofuran tissue-bound metabolites by LC-MS/MS for nitrofuran control in Portuguese Residues Monitoring Plan, revealed the presence of 5-morpholinomethyl-3-amino-2-oxozolidinone (AMOZ), the bound residue of furaltadone, in a large number of samples, namely in meat poultry samples. From the 226 analysed samples in the last 4 months of 2002, 78 were non-compliant due to the presence of AMOZ (61 broilers, 11 turkeys, 5 quails and 1 pig). In this context, the aim of this paper is to describe the analytical data obtained on meat samples collected from various animal species under official Portuguese control for nitrofuran drug residues during the so-called “Portuguese nitrofuran crisis”