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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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- Chen, Jing-De, et al.
(author)
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Hot-electron emission-driven energy recycling in transparent plasmonic electrode for organic solar cells
- 2022
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In: InfoMat. - : Wiley. - 2567-3165. ; 4:3
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Journal article (peer-reviewed)abstract
- Plasmonic metal electrodes with subwavelength nanostructures are promising for enhancing light harvesting in photovoltaics. However, the nonradiative damping of surface plasmon polaritons (SPPs) during coupling with sunlight results in the conversion of the excited hot-electrons to heat, which limits the absorption of light and generation of photocurrent. Herein, an energy recycling strategy driven by hot-electron emission for recycling the SPP energy trapped in the plasmonic electrodes is proposed. A transparent silver-based plasmonic metal electrode (A-PME) with a periodic hexagonal nanopore array is constructed, which is combined with a luminescent organic emitter for radiative recombination of the injected hot-electrons. Owing to the suppressed SPP energy loss via broadband hot-electron emission, the A-PME achieves an optimized optical transmission with an average transmittance of over 80% from 380 to 1200 nm. Moreover, the indium-tin-oxide-free organic solar cells yield an enhanced light harvesting with a power conversion efficiency of 16.1%.
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- Yu, Congyu, et al.
(author)
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Artificial intelligence in paleontology
- 2024
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In: Earth-Science Reviews. - 0012-8252 .- 1872-6828. ; 252
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Research review (peer-reviewed)abstract
- The accumulation of large datasets and increasing data availability have led to the emergence of data-driven paleontological studies, which reveal an unprecedented picture of evolutionary history. However, the fast-growing quantity and complication of data modalities make data processing laborious and inconsistent, while also lacking clear benchmarks to evaluate data collection and generation, and the performances of different methods on similar tasks. Recently, artificial intelligence (AI) has become widely practiced across scientific disciplines, but not so much to date in paleontology where traditionally manual workflows have been more usual. In this study, we review >70 paleontological AI studies since the 1980s, covering major tasks including micro- and macrofossil classification, image segmentation, and prediction. These studies feature a wide range of techniques such as Knowledge-Based Systems (KBS), neural networks, transfer learning, and many other machine learning methods to automate a variety of paleontological research workflows. Here, we discuss their methods, datasets, and performance and compare them with more conventional AI studies. We attribute the recent increase in paleontological AI studies most to the lowering of the entry bar in training and deployment of AI models rather than innovations in fossil data compilation and methods. We also present recently developed AI implementations such as diffusion model content generation and Large Language Models (LLMs) that may interface with paleontological research in the future. Even though AI has not yet been a significant part of the paleontologist's toolkit, successful implementation of AI is growing and shows promise for paradigm-transformative effects on paleontological research in the years to come.
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- Aad, G, et al.
(author)
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- 2015
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swepub:Mat__t
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- 2019
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Journal article (peer-reviewed)
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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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10. |
- Doughty, Tyler, 1987, et al.
(author)
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A single chromosome strain of S. cerevisiae exhibits diminished ethanol metabolism and tolerance
- 2021
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In: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 22:1
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Journal article (peer-reviewed)abstract
- Background: Eukaryotic organisms, like the model yeast S. cerevisiae, have linear chromosomes that facilitate organization and protection of nuclear DNA. A recent work described a stepwise break/repair method that enabled fusion of the 16 chromosomes of S. cerevisiae into a single large chromosome. Construction of this strain resulted in the removal of 30 of 32 telomeres, over 300 kb of subtelomeric DNA, and 107 subtelomeric ORFs. Despite these changes, characterization of the single chromosome strain uncovered modest phenotypes compared to a reference strain. Results: This study further characterized the single chromosome strain and found that it exhibited a longer lag phase, increased doubling time, and lower final biomass concentration compared with a reference strain when grown on YPD. These phenotypes were amplified when ethanol was added to the medium or used as the sole carbon source. RNAseq analysis showed poor induction of genes involved in diauxic shift, ethanol metabolism, and fatty-acid ß-oxidation during growth on ethanol compared to the reference strain. Enzyme-constrained metabolic modeling identified decreased flux through the enzymes that are encoded by these poorly induced genes as a likely cause of diminished biomass accumulation. The diminished growth on ethanol for the single chromosome strain was rescued by nicotinamide, an inhibitor of sirtuin family deacetylases, which have been shown to silence gene expression in heterochromatic regions. Conclusions: Our results indicate that sirtuin-mediated silencing in the single chromosome strain interferes with growth on non-fermentable carbon sources. We propose that the removal of subtelomeric DNA that would otherwise be bound by sirtuins leads to silencing at other loci in the single chromosome strain. Further, we hypothesize that the poorly induced genes in the single chromosome strain during ethanol growth could be silenced by sirtuins in wildtype S. cerevisiae during growth on glucose.
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