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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. |
- Anderson, Richard I., et al.
(författare)
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VELOcities of CEpheids (VELOCE) : I. High-precision radial velocities of Cepheids
- 2024
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Ingår i: Astronomy and Astrophysics. - 0004-6361. ; 686
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Tidskriftsartikel (refereegranskat)abstract
- We present the first data release of VELOcities of CEpheids (VELOCE), dedicated to measuring the high-precision radial velocities (RVs) of Galactic classical Cepheids (henceforth, Cepheids). The first data release (VELOCE DR1) comprises 18 225 RV measurements of 258 bona fide classical Cepheids on both hemispheres collected mainly between 2010 and 2022, along with 1161 observations of 164 stars, most of which had previously been misclassified as Cepheids. The median per-observation RV uncertainty for Cepheids is 0.037 km s-1 and reaches 2 m s-1 for the brightest stars observed with Coralie. Non-variable standard stars were used to characterize RV zero-point stability and to provide a base for future cross-calibrations. We determined zero-point differences between VELOCE and 31 literature data sets using template fitting, which we also used to investigate linear period changes of 146 Cepheids. In total, 76 spectroscopic binary Cepheids and 14 candidate binary Cepheids were identified using VELOCE data alone, which are investigated in detail in a companion Paper (VELOCE II). VELOCE DR1 provides a number of new insights into the pulsational variability of Cepheids, most importantly: a) the most detailed description of the Hertzsprung progression based on RVs to date; b) the identification of double-peaked bumps in the pulsation curve; and c) clear evidence that virtually all Cepheids feature spectroscopic variability signals that lead to modulated RV variability at the level of tens to hundreds of m s-1 and that cannot be satisfactorily modeled using single-periodic Fourier series. We identified 36 stars exhibiting such modulated variability, of which 4 also exhibit orbital motion. Linear radius variations depend strongly on pulsation period and a steep increase in slope of the ΔR/p vs. log P-relation is found near 10 days. This effect, combined with significant RV amplitude differences at fixed period, challenges the existence of a tight relation between Baade-Wesselink projection factors and pulsation periods. We investigated the accuracy of RV time series measurements, Uγ, and RV amplitudes published by Gaia's third data release (Gaia DR3) and determined an offset of 0.65 ± 0.11 km s-1 relative to VELOCE. Whenever possible, we recommend adopting a single set of template correlation parameters for distinct classes of large-amplitude variable stars to avoid systematic offsets in Uγ among stars belonging to the same class. The peak-to-peak amplitudes of Gaia RVs exhibit significant (16%) dispersion. Potential differences of RV amplitudes require further inspection, notably in the context of projection factor calibration.
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