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

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

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Introduction : The Study of Gender, Diplomacy and Negotiation

The introduction takes off from the inspirational question raised by Cynthia Enloe (1990) over two decades ago: “Where are the women?” In the field of diplomacy, this problematic has received scant attention. The introduction provides for an elaborated discussion of the state-of-the-art in the study of gender, diplomacy and international negotiation. Three core research questions guide the advancement of the conceptual framework and empirical mapping. First, where are the women located and positioned in contemporary diplomacy and international negotiation? Second, to what extent are diplomatic norms and practices of negotiations gendered? Third, to what extent have the practices of diplomacy and negotiation changed with broader and more diverse groups of diplomats

Conclusion : The Quest for Gender Justice in Diplomacy

In this last chapter, a series of more general propositions about contemporary gender and diplomacy are identified from the contributions to this book. First, it provides a historical narrative on the exclusions and inclusions in diplomacy over time, suggesting the contours of general trends in the quest to end formal barriers and bring about gender parity. Second, it critically analyses the gendered diplomatic infrastructure and how it still tends towards masculinised norms, homo-social environments and gendered division of labour. The chapter concludes with a discussion about the quest for gender justice in diplomacy and a note on some positive indicators that have been triggered by external as well as internal push-factors

Feminist IR in Europe Knowledge Production in Academic Institutions

Intro -- Contents -- List of Contributors -- List of Figures -- 1 Introduction -- References -- 2 Disparities and Diversification: Feminists in Europe Study War and/or Militaries -- Introduction -- Vital Caveats: 'IR Scholarship,' 'On War and Militaries,' 'Within Europe'? -- Militaries -- Militarism and Militarization -- Military Masculinities -- Fighting Women -- Military Families -- War -- Everyday War -- Counterinsurgency -- Conflict-Related Sexual Violence -- Concluding Remarks -- References -- 3 Feminist Security Studies in Europe: Beyond Western Academics' Club -- Introduction -- What, Where, and Who Is Feminist Security Studies? -- What is Missing? -- European Feminist Security Studies: Theories, Methodologies, Issues -- Issues and Silences -- Theories and Methods -- Concluding Thoughts -- References -- 4 Feminist Peace Research in Europe: A Snapshot -- Introduction -- Feminist Engagements with/for Peace -- A Mapping of Feminist Peace Research in Europe -- Centers of Excellence and the Production of Feminist Peace Research -- Leveraging the WPS Agenda in Peacebuilding: Negotiations, Sexual Violence, and Hybridity -- Conclusion -- References -- 5 Feminist Scholarship in Europe on the Politics of International Migration -- Introduction -- Critical Feminist Interventions in Migration Studies -- Border (In)securities and Sovereignty -- Conflict and Displacement: Determining Inclusive Refugeehood -- The Politics of Intimacy and Belonging -- Global Relations of Labor and Care -- Conclusion -- References -- 6 Feminist International Political Economy in Europe -- Introduction -- Capitalism and Neoliberalism: Connecting Discourses, Policies, and Everyday Implications -- Analyzing International Development Discourses and Organizations -- Reflexive Exercises on the Governance of Neoliberalism and Development and FeministsInstitutional Locations, Positionalities, and New Directions -- Bibliography -- 7 Feminist Foreign Policy Studies in Europe -- Introduction -- Overview -- Gender and State Foreign Policy -- EU Foreign Policy -- Securitization/Militarization of Foreign Policy -- What's Been Overlooked in European Feminist IR Scholarship on Foreign Policy and EU External Relations? -- Conclusion -- Bibliography -- 8 Feminist Knowledge Production in Europe on Diplomacy and International Negotiation -- Introduction -- Mapping Feminist Scholarship on Diplomacy and International Negotiation Across Europe -- Mapping Across States -- Mapping Across Institutional Sites -- University Sites -- Non-University Research Centers -- Diplomatic Academies and Research Centers Attached to the Ministries of Foreign Affairs (MFAs) -- Mapping Key Thinkers on Gender and Diplomacy in Europe -- Feminist Scholarship in Europe on Diplomacy and International Negotiation-Theoretical Approaches, Methodological Challenges, and Avenues for Future Research -- Conclusions -- References -- 9 Decentering Political Authority and Power: Feminist Global Governance Studies in Europe -- Introduction -- Overview: Feminist Analyses of Global Governance -- Reflections -- Theorizing the State in Global Governance -- Knowledge and Expertise as Technologies of Power -- Conclusion -- Bibliography -- IndexDescription based on publisher supplied metadata and other sources.Electronic reproduction. Ann Arbor, Michigan : ProQuest Ebook Central, YYYY. Available via World Wide Web. Access may be limited to ProQuest Ebook Central affiliated libraries

The Rankings Game: A Relational Approach to Country Performance Indicators

As the number of international rankings has risen dramatically since the 1990s, a large body of scholarship has emerged to examine and understand them. The aim of this article is to provide an overview of this body of work and to chart out fruitful directions for future research. In short, prior scholarship has been surprisingly quiet on the relations among multiple actors and their economic dimensions at the core of country performance indicator (CPI) activities. To foreground crucial socioeconomic relations, we develop a relational heuristic based upon a sports analogy: the actors involved in the creation and maintenance of CPIs can fruitfully be approached as a complex of players, referees, coaches, and audiences. Such an account helps us better understand how CPIs emerge and are sustained, even when they rely on dodgy data and their effects are perverse. We use nation brand rankings—overlooked in international relations research—as empirical illustration