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

Electronic state modification in laser deposited amorphous carbon films by the inclusion of nitrogen

In this study, we investigate the effect of the inclusion of nitrogen in amorphous carbon thin films deposited by pulsed laser deposition, which results in stress induced modifications to the band structure and the concomitant changes to the electronic transport properties. The microstructural changes due to nitrogen incorporation were examined using electron energy-loss spectroscopy and Raman scattering. The band structure was investigated using spectroscopic ellipsometry data in the range of. 1.5-5 eV, which was fitted to the Tauc Lorentz model parametrization and optical transmittance measurements. The dielectric constant evaluated using optical techniques was compared to that obtained with electrical measurements, assuming a Poole-Frenkel type conduction process based on the best fits to data. The electrical conduction mechanism is discussed for both low and high electric fields, in the context of the shape of the band density of states. By relating a wide range of measurement techniques, a detailed relationship between the microstructure, and the optical and the electrical structures of a-CNx films is obtained. From these measurements, it was found that, primarily, the change in density of the film, with increasing nitrogen pressure, affects the band structure of the amorphous carbon nitride. This is due to the fact that the density affects the stress in the film, which also impacts the localized states in the band gap. These results are supported by density of states measurements using scanning tunneling spectroscopy.</p