| 1. |
- 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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| 2. |
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| 3. |
- 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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| 5. |
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- 2019
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Journal article (peer-reviewed)
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| 6. |
- Kristan, Matej, et al.
(author)
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The Visual Object Tracking VOT2015 challenge results
- 2015
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In: Proceedings 2015 IEEE International Conference on Computer Vision Workshops ICCVW 2015. - : IEEE. - 9780769557205 ; , s. 564-586
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Conference paper (peer-reviewed)abstract
- The Visual Object Tracking challenge 2015, VOT2015, aims at comparing short-term single-object visual trackers that do not apply pre-learned models of object appearance. Results of 62 trackers are presented. The number of tested trackers makes VOT 2015 the largest benchmark on short-term tracking to date. For each participating tracker, a short description is provided in the appendix. Features of the VOT2015 challenge that go beyond its VOT2014 predecessor are: (i) a new VOT2015 dataset twice as large as in VOT2014 with full annotation of targets by rotated bounding boxes and per-frame attribute, (ii) extensions of the VOT2014 evaluation methodology by introduction of a new performance measure. The dataset, the evaluation kit as well as the results are publicly available at the challenge website(1).
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| 7. |
- Wu, Ya-Ting, et al.
(author)
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Monitoring bisphenol A and its biodegradation in water using a fluorescent molecularly imprinted chemosensor.
- 2015
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In: Chemosphere. - : Elsevier BV. - 1879-1298 .- 0045-6535. ; 119, s. 515-523
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Journal article (peer-reviewed)abstract
- In this paper, we present a simple and rapid method for monitoring bisphenol A (BPA) and its biodegradation in environmental water using a fluorescent molecularly imprinted polymer chemosensor (fMIPcs). A fluorescent molecularly imprinted polymer (fMIP) was first synthesized by precipitation polymerization method using BPA as template, dansyl methacrylate as functional monomer. Then a fMIPcs was constructed by combining the fMIP with a fluorescent microplate reader. The fMIPcs displayed selective, concentration-dependent fluorescence quenching in response to BPA in water even in the existence of interferences, thereby allowing reliable high through-put quantification of BPA via simple fluorescence measurements. The fMIPcs was able to directly quantify BPA (from 10 to 2000μgL(-1)) in different environmental water samples (distilled water, distilled water containing heavy metals and humic acid, tap water, and river water) with high accuracy, and to monitor BPA biodegradation in real-time. Using the fMIPcs, it was possible to achieve fast analytical results with lower limit of detection for BPA (3μgL(-1)) from smaller sample volume (250μL), which are superior to many relevant methods reported in the literature. Moreover, BPA levels and biodegradation rates measured by fMIPcs are comparable to the instrument-based method (HPLC). The fMIPcs developed in this work offers a new solution for simple, rapid, accurate and high through-put BPA quantification, and makes it possible to monitor BPA biodegradation in real time.
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| 8. |
- Andhikaputra, Gerry, et al.
(author)
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The impact of temperature and precipitation on all-infectious-, bacterial-, and viral-diarrheal disease in Taiwan
- 2023
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In: Science of the Total Environment. - : Elsevier BV. - 1879-1026 .- 0048-9697. ; 862
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Journal article (peer-reviewed)abstract
- BACKGROUND: The ongoing climate change will elevate the incidence of diarrheal in 2030-2050 in Asia, including Taiwan. This study investigated associations between meteorological factors (temperature, precipitation) and burden of age-cause-specific diarrheal diseases in six regions of Taiwan using 13 years of (2004-2016) population-based data.METHODS: Weekly cause-specific diarrheal and meteorological data were obtained from 2004 to 2016. We used distributed lag non-linear model to assess age (under five, all age) and cause-specific (viral, bacterial) diarrheal disease burden associated with extreme high (99th percentile) and low (5th percentile) of climate variables up to lag 8 weeks in six regions of Taiwan. Random-effects meta-analysis was used to pool these region-specific estimates.RESULTS: Extreme low temperature (15.30 °C) was associated with risks of all-infectious and viral diarrhea, with the highest risk for all-infectious diarrheal found at lag 8 weeks among all age [Relative Risk (RR): 1.44; 95 % Confidence Interval (95 % CI): 1.24-1.67]. The highest risk of viral diarrheal infection was observed at lag 2 weeks regardless the age. Extreme high temperature (30.18 °C) was associated with risk of bacterial diarrheal among all age (RR: 1.07; 95 % CI: 1.02-1.13) at lag 8 weeks. Likewise, extreme high precipitation (290 mm) was associated with all infectious diarrheal, with the highest risk observed for bacterial diarrheal among population under five years (RR: 2.77; 95 % CI: 1.60-4.79) at lag 8 weeks. Extreme low precipitation (0 mm) was associated with viral diarrheal in all age at lag 1 week (RR: 1.08; 95 % CI: 1.01-1.15)].CONCLUSION: In Taiwan, extreme low temperature is associated with an increased burden of viral diarrheal, while extreme high temperature and precipitation elevated burden of bacterial diarrheal. This distinction in cause-specific and climate-hazard specific diarrheal disease burden underscore the importance of incorporating differences in public health preparedness measures designed to enhance community resilience against climate change.
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| 9. |
- Felsberg, Michael, et al.
(author)
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The Thermal Infrared Visual Object Tracking VOT-TIR2015 Challenge Results
- 2015
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In: Proceedings of the IEEE International Conference on Computer Vision. - : Institute of Electrical and Electronics Engineers (IEEE). - 9781467383905 ; , s. 639-651
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Conference paper (peer-reviewed)abstract
- The Thermal Infrared Visual Object Tracking challenge 2015, VOTTIR2015, aims at comparing short-term single-object visual trackers that work on thermal infrared (TIR) sequences and do not apply prelearned models of object appearance. VOT-TIR2015 is the first benchmark on short-term tracking in TIR sequences. Results of 24 trackers are presented. For each participating tracker, a short description is provided in the appendix. The VOT-TIR2015 challenge is based on the VOT2013 challenge, but introduces the following novelties: (i) the newly collected LTIR (Linköping TIR) dataset is used, (ii) the VOT2013 attributes are adapted to TIR data, (iii) the evaluation is performed using insights gained during VOT2013 and VOT2014 and is similar to VOT2015.
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