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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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- Beal, Jacob, et al.
(author)
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Robust estimation of bacterial cell count from optical density
- 2020
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In: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Journal article (peer-reviewed)abstract
- Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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- Ablikim, M., et al.
(author)
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Branching fraction measurement of J/ψ→KSKL and search for J/ψ→KSKS
- 2017
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In: Physical Review D. - 2470-0010 .- 2470-0029. ; 96:11
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Journal article (peer-reviewed)abstract
- Using a sample of 1.31 x 10(9) J/Psi events collected with the BESIII detector at the BEPCII collider, we study the decays of J/Psi -> KSKL and KSKS. The branching fraction of J/Psi -> KSKL is determined to be B(J/Psi -> KSKL) = (1.93 +/- 0.01 (stat) +/- 0.05 (syst)) x 10(-4), which significantly improves on previous measurements. No clear signal is observed for the J/Psi -> KSKS process, and the upper limit at the 95% confidence level for its branching fraction is determined to be B(J/Psi -> KSKS) < 1.4 x 10(-8), which improves on the previous searches by 2 orders in magnitude and reaches the order of the Einstein-Podolsky-Rosen expectation.
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- Ablikim, M., et al.
(author)
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Evidence for e(+)e(-)->gamma chi c1,2 at center-of-mass energies from 4.009 to 4.360 GeV
- 2015
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In: Chinese Physics C. - : IOP Publishing. - 1674-1137 .- 2058-6132. ; 39:4
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Journal article (peer-reviewed)abstract
- Using data samples collected at center-of-mass energies of root s=4.009, 4.230, 4.260, and 4.360 GeV with the BESIII detector operating at the BEPCII collider, we perform a search for the process e(+)e(-)->gamma chi(cJ) (J=0, 1, 2) and find evidence for e(+)e(-)->gamma chi(c1) and e(+)e(-)->gamma chi(c2) with statistical significances of 3.0 sigma and 3.4 sigma, respectively. The Born cross sections sigma(B)(e(+)e(-)->gamma chi(cJ)), as well as their upper limits at the 90% confidence level (C.L.) are determined at each center-of-mass energy.
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- Ablikim, M., et al.
(author)
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Evidence for e+e−→γηc(1S) at center-of-mass energies between 4.01 and 4.60 GeV
- 2017
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In: Physical Review D. - 2470-0010 .- 2470-0029. ; 96:5
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Journal article (peer-reviewed)abstract
- We present first evidence for the process e(+)e(-) -> gamma eta(c)(1S) at six center-of-mass energies between 4.01 and 4.60 GeV using data collected by the BESIII experiment operating at BEPCII. We measure the Born cross section at each energy using a combination of twelve eta(c)(1S) decay channels. We also combine all six energies under various assumptions for the energy-dependence of the cross section. If the process is assumed to proceed via the Y(4260), we measure a peak Born cross section sigma(peak)(e(+)e(-) -> gamma eta(c)(1S)) = 2.11 +/- 0.49 (stat.) +/- 0.36 (syst.) pb with a statistical significance of 4.2 sigma.
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- Ablikim, M., et al.
(author)
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Improved measurements of two-photon widths of the chi(cJ) states and helicity analysis for chi(c2) -> gamma gamma
- 2017
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In: Physical Review D. - : AMER PHYSICAL SOC. - 2470-0010 .- 2470-0029. ; 96:9
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Journal article (peer-reviewed)abstract
- Based on 448.1 x 10(6) Psi(3686) events collected with the BESIII detector, the decays Psi(3686) -> gamma chi(cJ), chi(cJ) -> gamma gamma(J = 0, 1, 2) are studied. The decay branching fractions of chi(c0,2) -> gamma gamma are measured to be B(chi(c0) -> gamma gamma) = (1.93 +/- 0.08 +/- 0.05 +/- 0.05) x 10(-4) and B(chi(c2) -> gamma gamma) = (3.10 +/- 0.09 +/- 0.07 +/- 0.11) x 10(-4) which correspond to two-photon decay widths of Gamma(gamma gamma)(chi(c0)) = 2.03 +/- 0.08 +/- 0.06 +/- 0.13 keV and Gamma(gamma gamma)(chi(c2)) = 0.60 +/- 0.02 +/- 0.01 +/- 0.04 keV with a ratio of R = Gamma(gamma gamma)(chi(c2))/Gamma(gamma gamma)(chi(c0)) = 0.295 +/- 0.014 +/- 0.007 +/- 0.027, where the uncertainties are statistical, systematic and associated with the uncertainties of B(Psi(3686) -> gamma chi(c0,2)) and the total widths Gamma(chi(c0,2)), respectively. For the forbidden decay of chi(c1) -> gamma gamma, no signal is observed, and an upper limit on the two-photon width is obtained to be Gamma(gamma gamma)(chi(c1)) < 5.3 eV at the 90% confidence level. The ratio of the two-photon widths between helicity-zero and helicity-two components in the decay chi(c2) -> gamma gamma is also measured to be f(0/2) = Gamma(lambda=0)(gamma gamma) (chi(c2))/Gamma(lambda=2)(gamma gamma) (chi(c2)) = (0.0 +/- 0.6 +/- 1.2) x 10(-2), where the uncertainties are statistical and systematic, respectively.
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- Ablikim, M., et al.
(author)
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Improved measurements of X-cJ -> Sigma(+) (Sigma)over-bar(-) and Sigma(0)(Sigma)over-bar(0) decays
- 2018
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In: Physical Review D. - : American Physical Society. - 2470-0010 .- 2470-0029. ; 97:5
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Journal article (peer-reviewed)abstract
- Using a data sample of (448.1 +/- 2.9) x 10(6) psi (3686) events collected with the BESIII detector at the BEPCII collider, we present measurements of branching fractions for the decays X-cJ -> Sigma(+) (Sigma) over bar (-) and Sigma(0) (Sigma) over bar (0) The decays X-c1.2 -> Sigma(+) (Sigma) over bar (-) and Sigma (Sigma) over bar (0) are observed for the first time, and the branching fractions for X-c0 -> Sigma(+) (Sigma) over bar (-) and Sigma(0) (Sigma) over bar (0) decays are measured with improved precision. The branching fraction ratios between the charged and neutral modes are consistent with the prediction of isospin symmetry.
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10. |
- Ablikim, M., et al.
(author)
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Measurement of the matrix elements for the decays eta' -> eta pi(+) pi(-) and eta' -> eta pi(0)pi(0)
- 2018
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In: Physical Review D. - : AMER PHYSICAL SOC. - 2470-0010 .- 2470-0029. ; 97:1
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Journal article (peer-reviewed)abstract
- Based on a sample of 1.31 x 10(9) J/psi events collected with the BESIII detector, the matrix elements for the decays eta' -> eta pi(+) pi(-) and eta' -> eta pi(0)pi(0) are determined using 351,016 eta' -> (eta -> gamma gamma)pi' pi(-) and 56,249 eta' -> (eta -> gamma gamma)pi(0) pi(0) events with background levels less than 1%. Two commonly used representations are used to describe the Dalitz plot density. We find that an assumption of a linear amplitude does not describe the data well. A small deviation of the obtained matrix elements between eta' -> eta pi(+) pi(-) and eta' -> eta pi(0)pi(0) is probably caused by the mass difference between charged and neutral pions or radiative corrections. No cusp structure in eta' -> eta pi(0)pi(0) is observed.
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