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- 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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2. |
- Aharonian, Felix, et al.
(author)
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Measurements of resonant scattering in the Perseus Cluster core with Hitomi SXS
- 2018
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In: Publications of the Astronomical Society of Japan. - : Oxford University Press (OUP). - 0004-6264 .- 2053-051X. ; 70:2
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Journal article (peer-reviewed)abstract
- Thanks to its high spectral resolution (similar to 5 eV at 6 keV), the Soft X-ray Spectrometer (SXS) on board Hitomi enables us to measure the detailed structure of spatially resolved emission lines from highly ionized ions in galaxy clusters for the first time. In this series of papers, using the SXS we have measured the velocities of gas motions, metallicities and the multi-temperature structure of the gas in the core of the Perseus Cluster. Here, we show that when inferring physical properties from line emissivities in systems like Perseus, the resonant scattering effect should be taken into account. In the Hitomi waveband, resonant scattering mostly affects the Fe XXV He alpha line (w)-the strongest line in the spectrum. The flux measured by Hitomi in this line is suppressed by a factor of similar to 1.3 in the inner similar to 30 kpc, compared to predictions for an optically thin plasma; the suppression decreases with the distance from the center. The w line also appears slightly broader than other lines from the same ion. The observed distortions of the w line flux, shape, and distance dependence are all consistent with the expected effect of the resonant scattering in the Perseus core. By measuring the ratio of fluxes in optically thick (w) and thin (Fe XXV forbidden, He beta, Ly alpha) lines, and comparing these ratios with predictions from Monte Carlo radiative transfer simulations, the velocities of gas motions have been obtained. The results are consistent with the direct measurements of gas velocities from line broadening described elsewhere in this series, although the systematic and statistical uncertainties remain significant. Further improvements in the predictions of line emissivities in plasma models, and deeper observations with future X-ray missions offering similar or better capabilities to the Hitomi SXS, will enable resonant scattering measurements to provide powerful constraints on the amplitude and anisotropy of cluster gas motions.
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3. |
- Aharonian, Felix, et al.
(author)
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Temperature structure in the Perseus cluster core observed with Hitomi
- 2018
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In: Publications of the Astronomical Society of Japan. - : Oxford University Press (OUP). - 0004-6264 .- 2053-051X. ; 70:2
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Journal article (peer-reviewed)abstract
- The present paper explains the temperature structure of X-ray emitting plasma in the core of the Perseus cluster based on 1.8-20.0 keV data obtained with the Soft X-ray Spectrometer (SXS) on board the Hitomi Observatory. A series of four observations was carried out, with a total effective exposure time of 338 ks that covered a central region of similar to 7' in diameter. SXS was operated with an energy resolution of similar to 5 eV (full width at half maximum) at 5.9 keV. Not only fine structures of K-shell lines in He-like ions, but also transitions from higher principal quantum numbers were clearly resolved from Si through Fe. That enabled us to perform temperature diagnostics using the line ratios of Si, S, Ar, Ca, and Fe, and to provide the first direct measurement of the excitation temperature and ionization temperature in the Perseus cluster. The observed spectrum is roughly reproduced by a single-temperature thermal plasma model in collisional ionization equilibrium, but detailed line-ratio diagnostics reveal slight deviations from this approximation. In particular, the data exhibit an apparent trend of increasing ionization temperature with the atomic mass, as well as small differences between the ionization and excitation temperatures for Fe, the only element for which both temperatures could be measured. The best-fit two-temperature models suggest a combination of 3 and 5 keV gas, which is consistent with the idea that the observed small deviations from a single-temperature approximation are due to the effects of projecting the known radial temperature gradient in the cluster core along the line of sight. A comparison with the Chandra/ACIS and the XMM-Newton/RGS results, on the other hand, suggests that additional lower-temperature components are present in the intracluster medium (ICM), but not detectable with Hitomi/SXS giving its 1.8-20 keV energy band.
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5. |
- Kawamura, Akira, et al.
(author)
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Parameterization of rain cell properties using an advection-diffusion model and rain gage data
- 1996
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In: Atmospheric Research. - : Elsevier BV. - 0169-8095. ; 42:1-4, s. 67-73
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Journal article (peer-reviewed)abstract
- To reduce flooding risks and improve urban drainage management, there is a need to increase the forecasting accuracy for rainfall models on small typical urban time and space scales. Increased rainfall forecasting accuracy will in turn improve runoff prediction and thus, prevent flooding hazards, decrease pollution discharge through combined sewers, increase waste water treatment efficiency, etc. For this purpose, we analyzed the parameters of a two-dimensional stochastic advection-diffusion model including a Fourier domain method and an extended Kalman filter algorithm for investigation of motion, shape, size, and intensity distribution of convective rainfall. The resulting set of model parameters (advective velocity, apparent turbulent diffusion, and development/decay of rainfall rate) is used to study convective rainfall variability. It appears that the speed at which the rainfall cell is advected is not dependent on the cell development stage or apparent diffusion. Instead, there is a dependence between the source/sink term and apparent diffusion. This can be explained by the turbulent updraft of warm air which results in large rainfall intensity increase. This strong turbulence results in larger diffusion (and vice versa). The behavior of the model parameters is therefore physically explainable and relevant. The results can be used as first choice of parameter values when modeling convective rainfall over ungaged areas.
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6. |
- Kawamura, Akira, et al.
(author)
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Real-time tracking of convective rainfall properties using a two-dimensional advection-diffusion model
- 1997
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In: Journal of Hydrology. - 0022-1694. ; 203:1-4, s. 109-118
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Journal article (peer-reviewed)abstract
- There is a need to improve rainfall forecasting capabilities for small ungaged urban catchments to reduce flooding hazards and pollution release. For this purpose, information is required on small-scale and short-term convective cell behavior. We use a two-dimensional stochastic advection-diffusion model to parameterize the space-time rainfall intensity from convective rainfall. The rainfall intensity resulting from different separable components of the rain cell, such as apparent turbulent diffusion and development/decay of rainfall intensity, is quantified for 10 observed and, for southern Sweden, representative high-intensity rainfall events. This is done following a Lagrangian approach. It is shown the used model was able to respond to rapid changes in observed rainfall intensity in both space and time, thus giving a small average root-mean-square error for all 10 events (0.06 mm min-1). When dividing the total rainfall intensity into apparent turbulent diffusion and development/decay terms, respectively, it was shown that D(y,center) and γ(center) contribute approximately equally to the observed rainfall intensity. The D(x,center) is usually only half the value of D(y,center), thus indicating less intensity contribution from this term and that the general elliptical shape of rain cells are elongated in the direction of movement. The observations indicate that the cumulus stage represents half and the dissipating stage half of the total cell development, respectively. The results can be used as first choice of parameter values when modeling rain cell movement over ungaged areas and the presented methodology can be used to study the effects of different cell components on total rainfall intensity.
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7. |
- Sasaki, Takaaki, et al.
(author)
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Elevated Intraocular Pressure, Optic Nerve Atrophy, and Impaired Retinal Development in ODAG Transgenic Mice
- 2009
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In: Investigative Ophthalmology and Visual Science. - : Association for Research in Vision and Ophthalmology (ARVO). - 0146-0404 .- 1552-5783. ; 50:1, s. 242-248
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Journal article (peer-reviewed)abstract
- PURPOSE. In an earlier study, a cDNA was cloned that showed abundant expression in the eye at postnatal day (P) 2 but was downregulated at P10; it was named ODAG (ocular development-associated gene). Its biological function was examined by generating and analyzing transgenic mice overexpressing ODAG (ODAG Tg) in the eye and by identifying ODAG-binding proteins. METHODS. Transgenic mice were generated by using the mouse Crx promoter. EGFP was designed to be coexpressed with transgenic ODAG, to identify transgene-expressing cells. Overexpression of ODAG was confirmed by Northern and Western blot analysis. IOP was measured with a microneedle technique. The eyes were macroscopically examined and histologically analyzed. EGFP expression was detected by confocal microscope. Proteins associated with ODAG were isolated by pull-down assay in conjugation with mass spectrometry. RESULTS. Macroscopically, ODAG Tg exhibited gradual protrusion of the eyeballs. The mean IOP of ODAG Tg was significantly higher than that of wild-type (WT) littermates. Histologic analysis exhibited optic nerve atrophy and impaired retinal development in the ODAG Tg eye. EGFP was expressed highly in the presumptive outer nuclear layer and weakly in the presumptive inner nuclear layer in the ODAG Tg retina. Rab6-GTPase-activating protein (Rab6-GAP) and its substrate, Rab6, were identified as ODAG-binding proteins. CONCLUSIONS. Deregulated expression of ODAG in the eye induces elevated intraocular pressure and optic nerve atrophy and impairs retinal development, possibly by interfering with the Rab6/Rab6-GAP-mediated signaling pathway. These results provide new insights into the mechanisms regulating ocular development, and ODAG Tg would be a novel animal model for human diseases caused by ocular hypertension.
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8. |
- Wannigama, Dhammika Leshan, et al.
(author)
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Tracing the transmission of mpox through wastewater surveillance in Southeast Asia
- 2023
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In: JOURNAL OF TRAVEL MEDICINE. - 1195-1982 .- 1708-8305. ; 30:5
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Journal article (peer-reviewed)abstract
- High population density and tourism in Southeast Asia increase the risk of mpox due to frequent interpersonal contacts. Our wastewater surveillance in six Southeast Asian countries revealed positive signals for Monkeypox virus (MPXV) DNA, indicating local transmission. This alerts clinicians and helps allocate resources like testing, vaccines and therapeutics in resource-limited countries.
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