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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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- Badziak, J., et al.
(författare)
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Application of ion diagnostics to control the laser-induced removal of surface layer of a carbon substrate
- 2006
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Ingår i: Plasma 2005. - : AIP. - 073540304X ; , s. 295-298
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Konferensbidrag (refereegranskat)abstract
- Among other methods of detritiation of in-vessel tokamak components the application of lasers for removal of fuel trapped in co-deposited layers is under investigation. The paper presents preparation and tests of ion diagnostic methods for on-line measurement of the amount and characteristics of ablated carbon, hydrogen/deuterium and contaminant species from the graphite target (plate) of the main toroidal limiter of the TEXTOR tokamak. For removal of the surface layer from the graphite limiter plate Nd:YAG laser was used. Determination of the characteristics of laser-produced ions has been performed by means of ion collectors and an electrostatic ion-energy analyser. The main ion stream parameters were measured depending on the number of laser shots and the laser power density on the target surface. The properties of modified carbon sample surface were determined with the use of optical methods and compared with the results of the ion measurements.
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- Gasior, P., et al.
(författare)
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Characterization of laser-irradiated co-deposited layers on plasma facing components from a tokamak
- 2006
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Ingår i: Physica Scripta. - 0031-8949 .- 1402-4896. ; T123, s. 99-103
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Tidskriftsartikel (refereegranskat)abstract
- An experimental setup and ion diagnostic method for laser-induced fuel removal and decomposition of co-deposited layers on plasma facing components from tokamaks are described. Nd:YAG 3.5 ns pulse laser with a repetition rate of 10 Hz and single-pulse energy of up to 0.8J at 1.06 mu m has been used for irradiation of a graphite limiter tile from the TEXTOR tokamak. Comparative studies have been performed for a pure graphite plate as a reference target. Energy of emitted ions has been measured using a time-of-flight method. Early results show that laser pulses efficiently ablate the co-deposit removing both fuel species and heavy components such as Si, Ni, Cr, Fe and W present in the layers. Surface topography of the irradiated targets is also presented.
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- Gasior, P., et al.
(författare)
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Effective laser-induced removal of co-deposited layers from plasma-facing components in a tokamak
- 2006
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Ingår i: Czechoslovak Journal of Physics. - : Springer Science and Business Media LLC. - 0011-4626 .- 1572-9486. ; 56, s. B67-B72
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Tidskriftsartikel (refereegranskat)abstract
- An experimental set-up and spectroscopy diagnostic method for laser-induced fuel removal and decomposition of co-deposited layers on plasma-facing components from tokamaks are described. For irradiation of a graphite limiter tile from the TEXTOR tokamak Nd:YAG 3.5-ns pulse laser with a repetition rate of 10 Hz and single pulse energy of up to 0,8 J at 1,06 mu m has been used. The spectroscopy system allowed recording of spectra in the visible wavelength range including CII and D alpha spectral lines. The evolution of CII and Da spectral lines was observed pulse-by-pulse during the co-deposit removal. The efficient ablation of the 45 mu m thick co-deposit occured after approximately 50 laser pulses.
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- Torrisi, Lorenzo, et al.
(författare)
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Single crystal silicon carbide detector of emitted ions and soft x rays from power laser-generated plasmas
- 2009
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Ingår i: Journal of Applied Physics. - : American Institute of Physics (AIP). - 0021-8979 .- 1089-7550. ; 105, s. 123304-1-123304-7
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Tidskriftsartikel (refereegranskat)abstract
- A single-crystal silicon carbide SiC detector was used for measurements of soft x rays, electrons, and ion emission from laser-generated plasma obtained with the use of the Prague Asterix Laser System PALS at intensities of the order of 1016 W/cm2 and pulse duration of 300 ps. Measurements were performed by varying the laser intensity and the nature of the irradiated target. The spectra obtained by using the SiC detector show not only the photopeak due to UV and softx-ray detection, but also various peaks due to the detection of energetic charged particles. Time-of-flight technique was employed to determine the ion kinetic energy of particles emitted from the plasma and to perform a comparison between SiC and traditional ion collectors. The detector was also employed by inserting absorber films of different thickness in front of the SiC surface inorder to determine, as a first approximation, the mean energy of the soft x-ray emission from the plasma.
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- Van Parys, A., et al.
(författare)
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Assessment of Dietary Choline Intake, Contributing Food Items, and Associations with One-Carbon and Lipid Metabolites in Middle-Aged and Elderly Adults: The Hordaland Health Study
- 2022
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Ingår i: Journal of Nutrition. - : Elsevier BV. - 0022-3166. ; 152:2, s. 513-524
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Tidskriftsartikel (refereegranskat)abstract
- Background: Choline is an essential nutrient for humans and is involved in various physiologic functions. Through its metabolite betaine, it is closely connected to the one-carbon metabolism, and the fat-soluble choline form phosphatidylcholine is essential for VLDL synthesis and secretion in the liver connecting choline to the lipid metabolism. Dietary recommendations for choline are not available in the Nordic countries primarily due to data scarcity. Objectives: The aim of this study was to investigate the dietary intake of total choline and individual choline forms, dietary sources, and the association of total choline intake with circulating one-carbon metabolites and lipids. Methods: We included 5746 participants in the Hordaland Health Study, a survey including community-dwelling adults born in 1925-1927 (mean age 72 y, 55% women) and 1950-1951 (mean age 48 y, 57% women). Dietary data were obtained using a 169-item FFQ, and choline content was calculated using the USDA Database for Choline Content of Common Foods, release 2. Metabolites of the one-carbon and lipid metabolism were measured in a nonfasting blood sample obtained at baseline, and the association with total choline intake was assessed using polynomial splines. Results: The geometric mean (95% prediction interval) energy-adjusted total choline intake was 260 (170, 389) mg/d, with phosphatidylcholine being the main form (44%). The major food items providing dietary choline were eggs, low-fat milk, potatoes, and leafy vegetables. Dietary total choline was inversely associated with circulating concentrations of total homocysteine, glycine, and serine and positively associated with choline, methionine, cystathionine, cysteine, trimethyllysine, trimethylamine-N-oxide, and dimethylglycine. A weak association was observed between choline intake and serum lipids. Conclusions: Phosphatidylcholine was the most consumed choline form in community-dwelling adults in Norway. Our findings suggest that choline intake is associated with the concentration of most metabolites involved in the one-carbon and lipid metabolism. © 2021 The Author(s). Published by Oxford University Press on behalf of the American Society for Nutrition.
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