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- Klionsky, Daniel J., et al.
(author)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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In: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Research review (peer-reviewed)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. |
- Achtert, Peggy, 1982-, et al.
(author)
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Hygroscopic growth of tropospheric particle number size distributions over the North China Plain
- 2009
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In: Journal of Geophysical Research. - 0148-0227 .- 2156-2202. ; 114, s. D00G07-
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Journal article (peer-reviewed)abstract
- The hygroscopic growth of atmospheric submicrometer particle size distributions (diameter D-p ranging from 22 to 900 nm) was studied at a rural/suburban site in the North China Plain within the framework of the international Campaigns of Air Quality Research in Beijing and Surrounding Region 2006 (CAREBeijing-2006) research project. The goal was to characterize the regional aerosol in the polluted northeastern plain in China. Size descriptive hygroscopic growth factors (DHGFs) were determined as a function of relative humidity (RH) by relating the particle number size distribution at a dry condition ( 100 nm), the DHGF are substantially higher than in the Aitken particle mode (D-p < 100 nm) as a result of different chemical composition. The size-dependent behavior of the DHGF highlights the relevance of particulate sulfate production over the North China Plain, accomplished by secondary formation from the gas phase and, potentially, liquid phase processes in convective clouds. Furthermore, all results concerning the DHGF show a significant dependency on meteorological air masses. The hygroscopic growth of accumulation mode particles correlates significantly with the PM1-mass fraction of sulfate ions determined by chemical analysis. Finally, this investigation provides a parameterization of the hygroscopic growth of 250-nm particles, which might be useful when predicting visibility and radiative forcing and performing atmospheric aerosol model validations.
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