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

Effects of Once-Weekly Exenatide on Cardiovascular Outcomes in Type 2 Diabetes.

Abstract BACKGROUND: The cardiovascular effects of adding once-weekly treatment with exenatide to usual care in patients with type 2 diabetes are unknown. METHODS: We randomly assigned patients with type 2 diabetes, with or without previous cardiovascular disease, to receive subcutaneous injections of extended-release exenatide at a dose of 2 mg or matching placebo once weekly. The primary composite outcome was the first occurrence of death from cardiovascular causes, nonfatal myocardial infarction, or nonfatal stroke. The coprimary hypotheses were that exenatide, administered once weekly, would be noninferior to placebo with respect to safety and superior to placebo with respect to efficacy. RESULTS: In all, 14,752 patients (of whom 10,782 [73.1%] had previous cardiovascular disease) were followed for a median of 3.2 years (interquartile range, 2.2 to 4.4). A primary composite outcome event occurred in 839 of 7356 patients (11.4%; 3.7 events per 100 person-years) in the exenatide group and in 905 of 7396 patients (12.2%; 4.0 events per 100 person-years) in the placebo group (hazard ratio, 0.91; 95% confidence interval [CI], 0.83 to 1.00), with the intention-to-treat analysis indicating that exenatide, administered once weekly, was noninferior to placebo with respect to safety (P<0.001 for noninferiority) but was not superior to placebo with respect to efficacy (P=0.06 for superiority). The rates of death from cardiovascular causes, fatal or nonfatal myocardial infarction, fatal or nonfatal stroke, hospitalization for heart failure, and hospitalization for acute coronary syndrome, and the incidence of acute pancreatitis, pancreatic cancer, medullary thyroid carcinoma, and serious adverse events did not differ significantly between the two groups. CONCLUSIONS: Among patients with type 2 diabetes with or without previous cardiovascular disease, the incidence of major adverse cardiovascular events did not differ significantly between patients who received exenatide and those who received placebo. (Funded by Amylin Pharmaceuticals; EXSCEL ClinicalTrials.gov number, NCT01144338 .)

Gift politics: Exposure and surveillance in the anthropocene

This article discusses the role of gift relations in the Anthropocene. We reinterpret Mauss’s original concept of the gift to understand its application and transformation in a social context that increasingly sees human behavior as a resource for the realization of governmental and corporate objectives. Contemporary gift rela- tions focus on reciprocity through personal data instead of physical artifacts, and on promoting control and consumerism instead of forging moral and personal obligations. In our analysis, we distinguish two important elements. First, gifts are used to elicit voluntary exposure of personal data by individuals. In exchange for personal data, people are granted material or immaterial rewards. Second, gift relations have a pervasive element of surveillance that aims to influence behavior through p

Balance or Trade-off? Online Security Technologies and Fundamental Rights

In this contribution I argue that the image of the balance is often used to defend the idea of a trade-off. To understand the drawbacks of this line of thought I will explore the relationship between online security technologies and fundamental rights, notably privacy, non-discrimination, freedom of speech and due process. After discriminating between three types of online security technologies I will trace the reconfiguration of the notion of privacy in the era of smart environments. This will lead to an inquiry into the metaphor of the scale, building on the triple test regarding the justification of the limitation of fundamental rights such as privacy. The conclusion will be that in the case of a trade-off, infringing measures will have to be balanced by effective safeguards. No trade-off without balanc

Does cloud computing have a silver lining?

Despite the language of immateriality and weightlessness, network communications have a significant materiality and weight when considered from the standpoint of production, consumption and recycling. In this paper, we concentrate on the energy signatures of the server industry on which internet communications depend, with special consideration of Google server farms. The move towards thin clients and cloud computing raises the stakes for the server farm business, and makes it more urgent to confront the finite environment in which information circulates. We assess recent and near-future growth in server traffic, and suggest that business models and regulatory systems have yet to recognise the challenges they pose. <br/

The Biopolitical Public Domain: the Legal Construction of the Surveillance Economy

Within the political economy of informational capitalism, commercial surveillance practices are tools for resource extraction. That process requires an enabling legal construct, which this essay identifies and explores. Contemporary practices of personal information processing constitute a new type of public domain — a repository of raw materials that are there for the taking and that are framed as inputs to particular types of productive activity. As a legal construct, the biopolitical public domain shapes practices of appropriation and use of personal information in two complementary and interrelated ways. First, it constitutes personal information as available and potentially valuable: as a pool of materials that may be freely appropriated as inputs to economic production. That framing supports the reorganization of sociotechnical activity in ways directed toward extraction and appropriation. Second, the biopolitical public domain constitutes the personal information harvested within networked information environments as raw. That framing creates the backdrop for culturally-situated techniques of knowledge production and for the logic that designates those techniques as sites of legal privilege

Political public relations and mediatization: The strategies of news management

In highly mediatized democracies, political parties and other political actors have to manage the news and adapt to the fact that news media have become largely independent and highly influential. In this chapter, the authors bridge theory and research on strategic party behavior, mediatization, and political public relations to analyze why and how political parties seek to manage the news to further their strategic goals. Based on a review of the literature, the authors suggest a typology of strategies and tactics that political parties can use to manage the news. One conclusion is that adaption to news media need to follow from the strategic goals of the party. Otherwise adapting to news media is not strategic but rather surrendering to news media and news media logic