| 3. |
- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- 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. 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 vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most 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 field 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. 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. 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.
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| 4. |
- Di Lorenzo Tillborg, Adriana, et al.
(författare)
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When policies collide : Possibilities and Challenges of Practice-based Research
- 2022
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Ingår i: Proceedings of the 21st International Seminar of the ISME Commission on Policy: Culture, Media and Education Virtual Conference. ; , s. 48-55
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Konferensbidrag (refereegranskat)abstract
- This paper addresses possibilities and challenges on the methodological level during the initial phase of a practice-based research project. The research project aims to contribute to knowledge about pupils’ participation and influence in decision-making from the perspectives of teachers, leaders, and pupils in Sweden’s Art and Music Schools. A further aim is to contribute to the development of practice-based research. The project is inspired by collaborative action research in the sense that, through collaboration between the researcher(s) and the practitioner(s), action and reflection are interconnected to developing the practice. One main challenge is that, even though the research project focuses on pupils’ participation and influence, the pupils’ voices have so far been silenced. The challenge is a consequence of two policies that collide: the convention on the rights of the children, which ensures the rights of the child to participate or not; and the Ethics Review Act, which ensures protection rights. Hence, the approach to conducting practice-based research and making visible the pupils’ voices have exposed ethical challenges with possible implications for the aim of the study, the design, and the methodology. The next step of this project is to find a way to include the pupils’ voices. The challenges during this project have not discouraged me from seeing the potential of practice-based research for developing Sweden’s Art and Music Schools. Rather, they might contribute to the development of this project and the field of practice-based research. When working to improve collaboration between research and practice, we must continue to discuss and reflect on ethical and methodological challenges to develop ways of critically examining and addressing contradicting policies.
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