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- 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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| 2. |
- Hara, Kyosuke, et al.
(författare)
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In-situ X-ray Transmission Observation of Carbothermic Reduction of Magnetite Powder and Macroscopic Agglomeration of Reduced Iron
- 2013
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Ingår i: ISIJ International. - : Iron and Steel Institute of Japan. - 0915-1559 .- 1347-5460. ; 53:6, s. 1010-1019
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Tidskriftsartikel (refereegranskat)abstract
- The agglomeration behaviour of reduced iron, made from magnetite powder by carbothermic reduction, was observed by using the in-situ X-ray transmission observation technique. The iron particles, above 1 mm, were clearly observed as black points. Further, the reduction speed was examined by using the thermogravimetric analysis. The bulk density of the packed powder layer and the grain size distribution of magnetite powder and carbon black powder were changed and the effects of them on the reduction speed and the agglomeration degree were examined. The agglomeration degree was evaluated with diameter of iron particles on the X-ray photographs, taken during heating, and the weight of collected iron particles after the observation experiments. Neither the bulk density of powder layer nor the grain size distribution of powder mixture affected to the reduction speed. The agglomeration degree decreased when the bulk density of the powder layer was increased by compacting. On the other hand, the agglomeration degree was increased when the grain size distribution of powder mixture was widened. Further, the height change of powder layer was also measured on the X-ray photographs and compared with the iron particles appearing behaviour to estimate the microscopic agglomeration behaviour. The mechanisms that grain size distribution affected the agglomeration degree were discussed.
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