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

The Changing Landscape for Stroke\ua0Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non\u2013vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase 1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients\u2019 baseline characteristics. Atrial fibrillation disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age  6575 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score  652; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc = 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N = 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in Asia and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701

Dissecting the Shared Genetic Architecture of Suicide Attempt, Psychiatric Disorders, and Known Risk Factors

Background Suicide is a leading cause of death worldwide, and nonfatal suicide attempts, which occur far more frequently, are a major source of disability and social and economic burden. Both have substantial genetic etiology, which is partially shared and partially distinct from that of related psychiatric disorders. Methods We conducted a genome-wide association study (GWAS) of 29,782 suicide attempt (SA) cases and 519,961 controls in the International Suicide Genetics Consortium (ISGC). The GWAS of SA was conditioned on psychiatric disorders using GWAS summary statistics via multitrait-based conditional and joint analysis, to remove genetic effects on SA mediated by psychiatric disorders. We investigated the shared and divergent genetic architectures of SA, psychiatric disorders, and other known risk factors. Results Two loci reached genome-wide significance for SA: the major histocompatibility complex and an intergenic locus on chromosome 7, the latter of which remained associated with SA after conditioning on psychiatric disorders and replicated in an independent cohort from the Million Veteran Program. This locus has been implicated in risk-taking behavior, smoking, and insomnia. SA showed strong genetic correlation with psychiatric disorders, particularly major depression, and also with smoking, pain, risk-taking behavior, sleep disturbances, lower educational attainment, reproductive traits, lower socioeconomic status, and poorer general health. After conditioning on psychiatric disorders, the genetic correlations between SA and psychiatric disorders decreased, whereas those with nonpsychiatric traits remained largely unchanged. Conclusions Our results identify a risk locus that contributes more strongly to SA than other phenotypes and suggest a shared underlying biology between SA and known risk factors that is not mediated by psychiatric disorders.Peer reviewe

Cause related marketing in the UK: a comparative analysis of corporate behaviour in the financial and retail sectors

The purpose of cause-related marketing (CRM) is to publicise and capitalise on a firm's corporate social performance (CSP) to enhance its legitimacy. More specifically, CRM strategies focus on how firms should design their corporate social responsibility (CSR) strategies how and communicate the associated CSP in ways that will enhance a firm's reputation with its key stakeholders thereby improving long-run profitability and stakeholder relationships. Under the CRM perspective, corporate managers view CSR-related expenditures as investments tools. There successes are judged in relation to other potential investments that mayor may not be on the CSR agenda. The purpose of this research is to identify the CRM strategies used by UK firms operating in the finance and retail industries. It is primarily an empirical based project that identifies the motives and returns underlying CRM activities in two important sectors of the economy. The process oriented conceptual framework for CRM is developed and subsequently tested through both quantitative and qualitative methods. The purpose of this exercise is to gain an understanding of firms' CRM behavioural patterns and managers' underlying motives when making CRM related decisions. Simple statistics tests and semi-structured interviews are used to analyse the data. More concretely, T- tests are used to compare the means of corporate activities categorised into four types of CRM behaviours (duration of the campaign timeframe, geographic coverage, causes selection, and CRM strategies selection). This is followed by 116 semi- structured interviews with corporate and non-profit executives. The London Stock Exchange database is used as a sampling frame and the findings are presented in four empirical core chapters titled according to these four types of CRM behaviours. The primary fmding of this thesis posits that corporate managers design CRM strategies in order to demonstrate firm's legitimacy value using effective communication strategy. Since companies within the same industry are subject to more or less the same social expectations, we find that there are industry-wide similarities in the design of CRM strategies. Moreover, the thesis argues that different industries are subject to different stakeholder pressures thereby creating industry-specific CRM strategies. The analyses highlight some of these differences in the retail and financial services sector. Amongst these are the finding that i) organizational structures and the extent to which decision-making is centralised influences the type of causes and CRM strategies that are selected in the retail and financial services industries; ii) the trade-off between corporate demand for 'legitimacy' enhancing schemes and flexibility affects managers' decision regarding the duration of the campaign, iii) symbols and images are at the hearth of cause selection but these will differ substantially across sectors, iv) the industry-specific characteristics such as product types, management expertise and others affects managers' choice ofCRM strategies. This thesis explores corporate rationales and motives underlying managerial strategic decisions to invest in CRM-related activities. It offers academics and both corporate and non-profit managers insights about the role of CRM within an organisation. 3 ! I 11EThOS - Electronic Theses Online ServiceGBUnited Kingdo

The Role of Relationally Embedded Network Ties in Resource Acquisition of British Nonprofit Organizations

As nonprofit and charity organizations face increasing competition, there have been growing interests in how nonprofit organizations conduct commercial activities to raise funds as well as grow their business. However, there is lack of prior research about market-oriented and/or commercial activities in the context of nonprofit business. This study examines the process of how nonprofit organizations use relationally embedded network ties to acquire financial, human, and human capital resources to fulfill their social mission and achieve business growth. The study investigates commercial activity of three U.K.-based nonprofit organizations using the case study method. The findings contribute to insights into components of network ties for acquiring three different network resources of financial, human, and human capital. Nonprofit organizations leverage social mission to improve their ability to acquire network resources. The findings also suggest the charity and social mission of nonprofit business enhance trustworthiness in relationally embedded network ties for resource acquisition