| 1. |
- 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. |
- Chen, Yong, et al.
(författare)
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A Vps21 endocytic module regulates autophagy
- 2014
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Ingår i: Molecular Biology of the Cell. - 1939-4586. ; 25:20, s. 3166-3177
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Tidskriftsartikel (refereegranskat)abstract
- In autophagy, the double-membrane autophagosome delivers cellular components for their degradation in the lysosome. The conserved Ypt/Rab GTPases regulate all cellular trafficking pathways, including autophagy. These GTPases function in modules that include guanine-nucleotide exchange factor (GEF) activators and downstream effectors. Rab7 and its yeast homologue, Ypt7, in the context of such a module, regulate the fusion of both late endosomes and autophagosomes with the lysosome. In yeast, the Rab5-related Vps21 is known for its role in early- to late-endosome transport. Here we show an additional role for Vps21 in autophagy. First, vps21Δ mutant cells are defective in selective and nonselective autophagy. Second, fluorescence and electron microscopy analyses show that vps21Δ mutant cells accumulate clusters of autophagosomal structures outside the vacuole. Third, cells with mutations in other members of the endocytic Vps21 module, including the GEF Vps9 and factors that function downstream of Vps21, Vac1, CORVET, Pep12, and Vps45, are also defective in autophagy and accumulate clusters of autophagosomes. Finally, Vps21 localizes to PAS. We propose that the endocytic Vps21 module also regulates autophagy. These findings support the idea that the two pathways leading to the lysosome—endocytosis and autophagy—converge through the Vps21 and Ypt7 GTPase modules.
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| 3. |
- Li, Hui, et al.
(författare)
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A Simple Compact Reconfigurable Slot Antenna With a Very Wide Tuning Range
- 2010
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Ingår i: IEEE Transactions on Antennas and Propagation. - 0018-926X .- 1558-2221. ; 58:11, s. 3725-3728
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Tidskriftsartikel (refereegranskat)abstract
- A simple and compact slot antenna with a very wide tuning range is proposed. A 25 mm (roughly equal to lambda(H)/8, where lambda(H) corresponds to the highest frequency of the tuning range) open slot is etched at the edge of the ground. To achieve the tunability, only two lumped elements, namely, a PIN diode and a varactor diode are used in the structure. By switching the PIN diode placed at the open end of the slot, the slot antenna can resonate as a standard slot (when the switch is on) or a half slot (when the switch is off). Continuous tuning over a wide frequency range in those two modes can be achieved by adjusting the reverse bias (giving different capacitances) of the varactor diode loaded in the slot. Through optimal design, the tuning bands of the two modes are stitched together to form a very wide tuning range. The fabricated prototype has a tuning frequency range from 0.42 GHz to 1.48 GHz with S11 better than -10 dB, giving a frequency ratio (f(R) = f(u)/f(L)) of 3.52:1. The measured full spherical radiation patterns show consistent radiation characteristics of the proposed antenna within the whole tuning range.
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| 4. |
- Yu, Yufeng, et al.
(författare)
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An Electrically Small Frequency Reconfigurable Antenna With a Wide Tuning Range
- 2011
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Ingår i: IEEE Antennas and Wireless Propagation Letters. - 1536-1225 .- 1548-5757. ; 10, s. 103-106
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Tidskriftsartikel (refereegranskat)abstract
- An electrically small frequency-reconfigurable antenna with a very wide tuning band is proposed. Three varactor diodes are used to achieve the tunable capacitance. With a modified feeding structure, the measured tuning range of the fabricated antenna reaches from 457.5 to 894.5 MHz, and the tuning band enhancement is also explained through an equivalent circuit analysis.
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