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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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- Laouali, D., et al.
(författare)
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Source contributions in precipitation chemistry and analysis of atmospheric nitrogen deposition in a Sahelian dry savanna site in West Africa
- 2021
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Ingår i: Atmospheric Research. - : Elsevier BV. - 0169-8095. ; 251
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Tidskriftsartikel (refereegranskat)abstract
- Experimental data on precipitation chemistry were collected at a semi-arid savanna in Senegal (Dahra) in 2013, 2014 and 2017. The chemical composition of precipitation was analyzed for inorganic and organic ions, using ionic chromatography. The pH values of precipitation range from 4.50 to 8.50 with 89% of the samples having basic pH. The composition of precipitation was controlled by four source contributions: marine, terrigenous, biogenic, and organic acids emissions from vegetation. The terrigenous contribution was the highest accounting for 42% of the total annual Volume Weighted Mean ionic concentrations, due to the proximity of the Saharan desert, followed by the marine source representing 36%, due to the location of Dahra close to the Atlantic Ocean. Nitrogenous (N) contribution represents 16% of the mean annual total ionic charge, from biogenic sources and livestock crossing the site all year round. Finally, the lowest contribution is from organic acidity (5%), due to the low density of vegetation especially during the dry season. Wet deposition fluxes in Dahra for all compounds show larger values than at other Sahelian savanna sites. Dry N deposition in Dahra was also estimated by inferential method using gas concentration measurements and modeled dry deposition velocities. The total N deposition fluxes (wet plus dry) range from 3.80 to 4.81 kgN ha−1 yr−1, comparable to fluxes at other semi-arid savannas in Niger and Mali. Wet deposition contributed with 37–53% of the total N flux, suggesting that wet N deposition is equally important to dry deposition fluxes for direct N loading to savanna ecosystems in the Sahel, with a large contribution of reduced compounds. This study shows that Dahra presents a precipitation chemistry composition with characteristics close to those from other Sahelian sites, with however the specificity of being more influenced by the proximity of the Atlantic ocean and the presence of livestock year round.
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