| 2. |
- 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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| 3. |
- Cheng, Cheng, et al.
(author)
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Deep H i Mapping of Stephan's Quintet and Its Neighborhood
- 2023
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In: Astrophysical Journal. - 0004-637X. ; 954:1
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Journal article (peer-reviewed)abstract
- We carried out deep mapping observations of the atomic hydrogen (H i) 21 cm line emission in a field centered on the famous galaxy group Stephan's Quintet (SQ), using the Five-hundred-meter Aperture Spherical Telescope (FAST) equipped with a 19-beam receiver. The final data cube reaches an H i column density sensitivity of 5σ = 2.1 × 1017 cm-2 per 20 km s-1 channel with an angular resolution of 4.′0. The discovery of a large diffuse feature of the H i emission in the outskirts of the intragroup medium of SQ was reported in a previous paper (Xu et al.). Here we present a new study of the total H i emission of SQ and the detection of several neighboring galaxies, exploiting the high sensitivity and the large sky coverage of the FAST observations. A total H i mass of M H I = 3.48 ± 0.35 × 1010 M ☉ is found for SQ, which is significantly higher than previous measurements in the literature. This indicates that, contrary to earlier claims, SQ is not H i deficient. The excessive H i gas is mainly found in the velocity ranges of 6200-6400 km s-1 and 6800-7000 km s-1, which were undetected in previous observations that are less sensitive than ours. Our results suggest that the "missing H i"in compact groups may be hidden in the low-density diffuse neutral gas instead of in the ionized gas.
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