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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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- Delon, Claire, et al.
(författare)
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Modelling land–atmosphere daily exchanges of NO, NH3, and CO2 in a semi-arid grazed ecosystem in Senegal
- 2019
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Ingår i: Biogeosciences. - : Copernicus GmbH. - 1726-4189. ; , s. 2049-2077
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Tidskriftsartikel (refereegranskat)abstract
- Three different models (STEP–GENDEC–NOflux, Zhang2010, and Surfatm) are used to simulate NO, CO2, and NH3 fluxes at the daily scale for 2 years (2012–2013) in a semi-arid grazed ecosystem at Dahra (15∘24′10′′ N, 15∘25′56′′ W, Senegal, Sahel). Model results are evaluated against experimental results acquired during three field campaigns. At the end of the dry season, when the first rains re-wet the dry soils, the model STEP–GENDEC–NOflux simulates the sudden mineralization of buried litter, leading to pulses in soil respiration and NO fluxes. The contribution of wet season fluxes of NO and CO2 to the annual mean is respectively 51 % and 57 %. NH3 fluxes are simulated by two models: Surfatm and Zhang2010. During the wet season, air humidity and soil moisture increase, leading to a transition between low soil NH3 emissions (which dominate during the dry months) and large NH3 deposition on vegetation during wet months. Results show a great impact of the soil emission potential, a difference in the deposition processes on the soil and the vegetation between the two models with however a close agreement of the total fluxes. The order of magnitude of NO, NH3, and CO2 fluxes is correctly represented by the models, as well as the sharp transitions between seasons, specific to the Sahel region. The role of soil moisture in flux magnitude is highlighted, whereas the role of soil temperature is less obvious. The simultaneous increase in NO and CO2 emissions and NH3 deposition at the beginning of the wet season is attributed to the availability of mineral nitrogen in the soil and also to microbial processes, which distribute the roles between respiration (CO2 emissions), nitrification (NO emissions), volatilization, and deposition (NH3 emission/deposition). The objectives of this study are to understand the origin of carbon and nitrogen compounds exchanges between the soil and the atmosphere and to quantify these exchanges on a longer timescale when only a few measurements have been performed.
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