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- Beal, Jacob, et al.
(författare)
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Robust estimation of bacterial cell count from optical density
- 2020
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Tidskriftsartikel (refereegranskat)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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- 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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- 2019
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Tidskriftsartikel (refereegranskat)
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| 8. |
- Albert, Benjamin B., et al.
(författare)
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Supplementation with milk enriched with complex lipids during pregnancy : A double-blind randomized controlled trial
- 2021
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 16:2
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Tidskriftsartikel (refereegranskat)abstract
- Background Gangliosides are a class of sphingolipids that are present in the cell membranes of vertebrates. Gangliosides influence a broad range of cellular processes through effects on signal transduction, being found abundantly in the brain, and having a role in neurodevelopment.Objective We aimed to assess the effects of maternal daily consumption of ganglioside-enriched milk vs non-enriched milk and a non-supplemented group of pregnant women on maternal ganglioside levels and pregnancy outcomes.Design Double-blind parallel randomized controlled trial. Methods 1,500 women aged 20-40 years were recruited in Chongqing (China) between 11 and 14 weeks of a singleton pregnancy, and randomized into three groups: Control-received standard powdered milk formulation (>= 4 mg gangliosides/day); Complex milk lipid-enhanced (CML-E) group-same formulation enriched with complex milk lipids (>= 8 mg gangliosides/day) from milk fat globule membrane; Reference-received no milk. Serum ganglioside levels were measured in a randomly selected subsample of 250 women per group.Results CML-E milk was associated with marginally greater total gangliosides levels in maternal serum compared to Control (13.02 vs 12.69 mu g/ml; p = 0.034) but not to Reference group. CML-E milk did not affect cord blood ganglioside levels. Among the 1500 women, CML-E milk consumption was associated with a lower rate of gestational diabetes mellitus than control milk [relative risk 0.80 (95% CI 0.64, 0.99)], but which was not different to the Reference group. CML-E milk supplementation had no other effects on maternal or newborn health.Conclusions Maternal supplementation with milk fat globule membrane, as a source of gangliosides, was not associated with any adverse health outcomes, and did not increase serum gangliosides compared with the non-supplemented reference group.
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- Kristan, Matej, et al.
(författare)
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The Visual Object Tracking VOT2014 Challenge Results
- 2015
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Ingår i: COMPUTER VISION - ECCV 2014 WORKSHOPS, PT II. - Cham : Springer. - 9783319161808 - 9783319161815 ; , s. 191-217
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Konferensbidrag (refereegranskat)abstract
- The Visual Object Tracking challenge 2014, VOT2014, aims at comparing short-term single-object visual trackers that do not apply pre-learned models of object appearance. Results of 38 trackers are presented. The number of tested trackers makes VOT 2014 the largest benchmark on short-term tracking to date. For each participating tracker, a short description is provided in the appendix. Features of the VOT2014 challenge that go beyond its VOT2013 predecessor are introduced: (i) a new VOT2014 dataset with full annotation of targets by rotated bounding boxes and per-frame attribute, (ii) extensions of the VOT2013 evaluation methodology, (iii) a new unit for tracking speed assessment less dependent on the hardware and (iv) the VOT2014 evaluation toolkit that significantly speeds up execution of experiments. The dataset, the evaluation kit as well as the results are publicly available at the challenge website (http://votchallenge.net).
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| 10. |
- Qian, Li-Bing, et al.
(författare)
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Transmission of electrons through the conical glass capillary with the grounded conducting outer surface
- 2017
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Ingår i: Wuli xuebao. - : Acta Physica Sinica, Chinese Physical Society and Institute of Physics, Chinese Academy of Sciences. - 1000-3290. ; 66:12
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Tidskriftsartikel (refereegranskat)abstract
- The transmission of 1.5 keV-electrons through a conical glass capillary is reported. This study aims to understand the so-called guiding effect for the negatively charged particles (e.g. electrons). The guiding mechanism is understood quite well with positively charged particles in particular highly charged ions, but not clear with electrons, i. e., even the basic scheme mediated by the existence of negative charge patches to guide the electrons is still somewhat controversial.. The study of the charging-up dynamics causing the electrons transport inside the capillary will shed light on this issue. In order to perform this, a data acquisition system has been setup to follow the time evolution of the two-dimensional angular distribution of the transmitted electrons. The electrons are detected by the multi-channel plate (MCP) detector with a phosphor screen. The image from the phosphor screen is recorded by a charge-coupled device camera. The timing signals for the detected events are extracted from the back stack of the MCP detector and recorded by the data acquisition system, synchronized with the acquired images. The electron beam has a size of 0.5 mm x 0.5 mm and a divergence of less than 0.35.. The inner diameter of the straight part of the capillary is 1.2 mm and the exit diameter is 225 mu m. A small conducting aperture of 0.3 mm in diameter is placed at the entrance of the capillary. Two-dimensional angular distribution of the transmitted electrons through conical glass capillary and its time evolution are measured. The results show that the transmission rate decreases and reaches to a constant value for the completely discharged glass capillary with time going by. The centroid of the angular distribution moves to an asymptotic value while the width remains unchanged. These transmission characteristics are different from those indicated in our previous work (2016 Acta Phys: Si n: 65 204103). The difference originates from the different manipulations of the capillary outer surface. A conducting layer is coated on the outer surface of the capillary and grounded in this work. This isolates various discharge/charge channels and forms a new stable discharge channel. The transmission rate as a function of the tilt angle shows that the allowed transmission occurs at the tilt angle limited by the geometrical factors, i. e., the geometrical opening angle given by the aspect ratio as well as the beam divergence. The transmission characteristics suggest that most likely there are formed no negative patches to facilitate the electron transmission through the glass capillary at this selected beam energy. It is different from that of highly charged ions, where the formation of the charge patches prohibits the close collisions between the following ions and guides them out of the capillary.
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