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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. |
- Vaivads, Andris, et al.
(författare)
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EIDOSCOPE : particle acceleration at plasma boundaries
- 2012
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Ingår i: Experimental astronomy. - : Springer Science and Business Media LLC. - 0922-6435 .- 1572-9508. ; 33:2-3, s. 491-527
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Tidskriftsartikel (refereegranskat)abstract
- We describe the mission concept of how ESA can make a major contribution to the Japanese Canadian multi-spacecraft mission SCOPE by adding one cost-effective spacecraft EIDO (Electron and Ion Dynamics Observatory), which has a comprehensive and optimized plasma payload to address the physics of particle acceleration. The combined mission EIDOSCOPE will distinguish amongst and quantify the governing processes of particle acceleration at several important plasma boundaries and their associated boundary layers: collisionless shocks, plasma jet fronts, thin current sheets and turbulent boundary layers. Particle acceleration and associated cross-scale coupling is one of the key outstanding topics to be addressed in the Plasma Universe. The very important science questions that only the combined EIDOSCOPE mission will be able to tackle are: 1) Quantitatively, what are the processes and efficiencies with which both electrons and ions are selectively injected and subsequently accelerated by collisionless shocks? 2) How does small-scale electron and ion acceleration at jet fronts due to kinetic processes couple simultaneously to large scale acceleration due to fluid (MHD) mechanisms? 3) How does multi-scale coupling govern acceleration mechanisms at electron, ion and fluid scales in thin current sheets? 4) How do particle acceleration processes inside turbulent boundary layers depend on turbulence properties at ion/electron scales? EIDO particle instruments are capable of resolving full 3D particle distribution functions in both thermal and suprathermal regimes and at high enough temporal resolution to resolve the relevant scales even in very dynamic plasma processes. The EIDO spin axis is designed to be sun-pointing, allowing EIDO to carry out the most sensitive electric field measurements ever accomplished in the outer magnetosphere. Combined with a nearby SCOPE Far Daughter satellite, EIDO will form a second pair (in addition to SCOPE Mother-Near Daughter) of closely separated satellites that provides the unique capability to measure the 3D electric field with high accuracy and sensitivity. All EIDO instrumentation are state-of-the-art technology with heritage from many recent missions. The EIDOSCOPE orbit will be close to equatorial with apogee 25-30 RE and perigee 8-10 RE. In the course of one year the orbit will cross all the major plasma boundaries in the outer magnetosphere; bow shock, magnetopause and magnetotail current sheets, jet fronts and turbulent boundary layers. EIDO offers excellent cost/benefits for ESA, as for only a fraction of an M-class mission cost ESA can become an integral part of a major multi-agency L-class level mission that addresses outstanding science questions for the benefit of the European science community.
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