Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Mechanical, Tribological, and Corrosive Properties of NbCrC_x and NbCrC_xN_y Coatings with Various Nitrogen and Carbon Contents

CrC and NbC carbide coatings both have good mechanical properties, wear resistance, and corrosion resistance. The present study seeks to combine the two coating systems in order to further enhance their properties. NbCrCx and NbCrCxNy coatings (where x and y denote the atomic percentages of carbon and nitrogen, respectively) were deposited on SKH51 substrates using a radio-frequency unbalanced magnetron sputtering system. The mechanical, tribological, and corrosive properties of the coatings were investigated and compared. Among the NbCrCx coatings, the NbCrC61 coating showed high levels of hardness, excellent adhesion strength, and good wear resistance. Among the NbCrCxNy coatings, the NbCrC55N5 coating showed high adhesion strength and hardness and excellent tribological properties. However, for nitrogen contents greater than 16 at%, the adhesion strength was dramatically reduced, resulting in poor tribological performance. Among all of the coatings, the NbCrC49 coating showed the best corrosion resistance due to its enhanced crystallinity, high adhesion strength, moderate surface roughness, and high sp3 C-C bonding ratio

Antimicrobial activities of cefoperazone-sulbactam in comparison to cefoperazone against clinical organisms from medical centers in Taiwan

Background: The multidrug-resistant Gram-negative bacteria (MDRGNBs) have emerged as important pathogens recently. Cefoperazone-sulbactam is active against a great proportion of those MDRGNBs. However, the susceptibilities data of cefoperazone-sulbactam are lacking in Taiwan. Object: This study was conducted to evaluate the susceptibilities data of cefoperazone-sulbactam aganist commonly encountered clinical pathogens in Taiwan. Materials and Methods: 2272 isolates were collected from various clinical specimens from five centers in Taiwan in 2012. The agar dilution method was used to evaluate the susceptibility of the isolated pathogens to cefoperazone and cefoperazone-sulbactam. Result: cefoperazone-sulbactam showed better activity against various GNBs, including MDRGNBs and part of carbapenem-resistant isolates tested compared to cefoperazone alone. Conclusion: Cefoperazone-sulbactam is active against most commonly encountered clinical pathogens, including MDRGNBs and part of carbapenem-esistant A. baumannii complex. It can be a potentially therapeutic agent for treating infections caused by these pathogens in Taiwan

Exploring the localization and home business strategies of the professional sports team -Formosa Dreamer case study

[[abstract]]Professional sport is one of the essential core industries for the development of the sports industry, divided into two categories: team and individual sports. The main development feature of professional basketball is to compete through the home-and-away system’s operation and establish a good emotional foundation with the residents through the home operation, which in turn creates recognition and support for the team and enhances the profits of the basketball organization. In recent years, Southeast Asia has developed a multinational professional basketball league, which covers nine countries. The “Formosa Dreamers” professional basketball team, a joint venture of Chinese entrepreneurs and has become the first professional basketball team in Taiwan, has chosen Changhua County as the home court. This report’s objective was to explore the localization and home business strategies of professional basketball games and to summarize further and analyze the characteristics of the home business strategy of Formosa Dreamers to establish the foundation for the future development of home business sports events in Taiwan.[[notice]]補正完