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    Cell stimulation versus cell death induced by sequential treatments with pulsed electric fields and cold atmospheric pressure plasma
    (San Francisco, California, US : PLOS, 2018) Steuer, Anna; Wolff, Christina M.; von Woedtke, Thomas; Weltmann, Klaus-Dieter; Kolb, Juergen F.
    Pulsed electric fields (PEFs) and cold atmospheric pressure plasma (CAP) are currently both investigated for medical applications. The exposure of cells to PEFs can induce the formation of pores in cell membranes and consequently facilitate the uptake of molecules. In contrast, CAP mainly acts through reactive species that are generated in the liquid environment. The objective of this study was to determine, if PEFs combined with plasma-treated cell culture medium can mutually reinforce effects on viability of mammalian cells. Experiments were conducted with rat liver epithelial WB-F344 cells and their tumorigenic counterpart WB-ras for a direct comparison of non-tumorigenic and tumorigenic cells from the same origin. Viability after treatments strongly depended on cell type and applied field strength. Notably, tumorigenic WB-ras cells responded more sensitive to the respective treatments than non-tumorigenic WB-F344 cells. More cells were killed when plasma-treated medium was applied first in combination with treatments with 100-μs PEFs. For the reversed treatment order, i.e. application of PEFs first, the combination with 100-ns PEFs resulted in a stimulating effect for non-tumorigenic but not for tumorigenic cells. The results suggest that other mechanisms, besides simple pore formation, contributed to the mutually reinforcing effects of the two methods.
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    Nitrosylation vs. oxidation – How to modulate cold physical plasmas for biological applications
    (San Francisco, California, US : PLOS, 2019) Lackmann, Jan-Wilm; Bruno, Giuliana; Jablonowski, Helena; Kogelheide, Friederike; Offerhaus, Björn; Held, Julian; Schulz-von der Gathen, Volker; Stapelmann, Katharina; von Woedtke, Thomas; Wende, Kristian
    Thiol moieties are major targets for cold plasma-derived nitrogen and oxygen species, making CAPs convenient tools to modulate redox-signaling pathways in cells and tissues. The underlying biochemical pathways are currently under investigation but especially the role of CAP derived RNS is barely understood. Their potential role in protein thiol nitrosylation would be relevant in inflammatory processes such as wound healing and improving their specific production by CAP would allow for enhanced treatment options beyond the current application. The impact of a modified kINPen 09 argon plasma jet with nitrogen shielding on cysteine as a thiol-carrying model substance was investigated by FTIR spectroscopy and high-resolution mass spectrometry. The deposition of short-lived radical species was measured by electron paramagnetic resonance spectroscopy, long-lived species were quantified by ion chromatography (NO2-, NO3-) and xylenol orange assay (H2O2). Product profiles were compared to samples treated with the so-called COST jet, being introduced by a European COST initiative as a reference device, using both reference conditions as well as conditions adjusted to kINPen gas mixtures. While thiol oxidation was dominant under all tested conditions, an Ar + N2/O2 gas compositions combined with a nitrogen curtain fostered nitric oxide deposition and the desired generation of S-nitrosocysteine. Interestingly, the COST-jet revealed significant differences in its chemical properties in comparison to the kINPen by showing a more stable production of RNS with different gas admixtures, indicating a different •NO production pathway. Taken together, results indicate various chemical properties of kINPen and COST-jet as well as highlight the potential of plasma tuning not only by gas admixtures alone but by adjusting the surrounding atmosphere as well.
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    The stability of memristive multidirectional associative memory neural networks with time-varying delays in the leakage terms via sampled-data control
    (San Francisco, California, US : PLOS, 2018) Wang, Weiping; Yu, Xin; Luo, Xiong; Wang, Long; Li, Lixiang; Kurths, Jürgen; Zhao, Wenbing; Xiao, Jiuhong
    In this paper, we propose a new model of memristive multidirectional associative memory neural networks, which concludes the time-varying delays in leakage terms via sampled-data control. We use the input delay method to turn the sampling system into a continuous time-delaying system. Then we analyze the exponential stability and asymptotic stability of the equilibrium points for this model. By constructing a suitable Lyapunov function, using the Lyapunov stability theorem and some inequality techniques, some sufficient criteria for ensuring the stability of equilibrium points are obtained. Finally, numerical examples are given to demonstrate the effectiveness of our results.
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    Femtosecond laser induced step-like structures inside transparent hydrogel due to laser induced threshold reduction
    (San Francisco, California, US : PLOS, 2019) Saerchen, Emanuel; Liedtke-Gruener, Susann; Kopp, Maximilian; Heisterkamp, Alexander; Lubatschowski, Holger; Ripken, Tammo
    In the area of laser material processing, versatile applications for cutting glasses and transparent polymers exist. However, parasitic effects such as the creation of step-like structures appear when laser cutting inside a transparent material. To date, these structures were only described empirically. This work establishes the physical and chemical mechanisms behind the observed effects and describes the influence of process and material parameters onto the creation of step-like structures in hydrogel, Dihydroxyethylmethacrylat (HEMA). By focusing laser pulses in HEMA, reduced pulse separation distance below 50 nm and rise in pulse energy enhances the creation of unintended step-like structures. Spatial resolved Raman-spectroscopy was used to measure the laser induced chemical modification, which results into a reduced breakdown threshold. The reduction in threshold influences the position of optical breakdown for the succeeding laser pulses and consequently leads to the step-like structures. Additionally, the experimental findings were supplemented with numerical simulations of the influence of reduced damage threshold onto the position of optical breakdown.
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    ColiCoords: A Python package for the analysis of bacterial fluorescence microscopy data
    (San Francisco, California, US : PLOS, 2019) Smit, Jochem H.; Li, Yichen; Warszawik, Eliza M.; Herrmann, Andreas; Cordes, Thorben; Gilestro, Giorgio F
    Single-molecule fluorescence microscopy studies of bacteria provide unique insights into the mechanisms of cellular processes and protein machineries in ways that are unrivalled by any other technique. With the cost of microscopes dropping and the availability of fully automated microscopes, the volume of microscopy data produced has increased tremendously. These developments have moved the bottleneck of throughput from image acquisition and sample preparation to data analysis. Furthermore, requirements for analysis procedures have become more stringent given the demand of various journals to make data and analysis procedures available. To address these issues we have developed a new data analysis package for analysis of fluorescence microscopy data from rod-like cells. Our software ColiCoords structures microscopy data at the single-cell level and implements a coordinate system describing each cell. This allows for the transformation of Cartesian coordinates from transmission light and fluorescence images and single-molecule localization microscopy (SMLM) data to cellular coordinates. Using this transformation, many cells can be combined to increase the statistical power of fluorescence microscopy datasets of any kind. ColiCoords is open source, implemented in the programming language Python, and is extensively documented. This allows for modifications for specific needs or to inspect and publish data analysis procedures. By providing a format that allows for easy sharing of code and associated data, we intend to promote open and reproducible research. The source code and documentation can be found via the project’s GitHub page.
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    Discovery of chitin in skeletons of non-verongiid Red Sea demosponges
    (San Francisco, California, US : PLOS, 2018) Ehrlich, Hermann; Shaala, Lamiaa A.; Youssef, Diaa T. A.; Żółtowska- Aksamitowska, Sonia; Tsurkan, Mikhail; Galli, Roberta; Meissner, Heike; Wysokowski, Marcin; Petrenko, Iaroslav; Tabachnick, Konstantin R.; Ivanenko, Viatcheslav N.; Bechmann, Nicole; Joseph, Yvonne; Jesionowski, Teofil
    Marine demosponges (Porifera: Demospongiae) are recognized as first metazoans which have developed over millions of years of evolution effective survival strategies based on unique metabolic pathways to produce both biologically active secondary metabolites and biopolymer-based stiff skeletons with 3D architecture. Up to date, among marine demosponges, only representatives of the Verongiida order have been known to synthetize biologically active substances as well as skeletons made of structural polysaccharide chitin. This work, to our knowledge, demonstrates for the first time that chitin is an important structural component within skeletons of non-verongiid demosponges Acarnus wolffgangi and Echinoclathria gibbosa collected in the Red Sea. Calcofluor white staining, FTIR and Raman analysis, ESI-MS, SEM, and fluorescence microscopy as well as a chitinase digestion assay were applied in order to confirm, with strong evidence, the finding of α-chitin in the skeleton of both species. We suggest that, the finding of chitin within these representatives of Poecilosclerida order is a promising step in the evaluation of these sponges as novel renewable sources for both biologically active metabolites and chitin, which are of prospective application for pharmacology and biomedicine.
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    Biogas residue parameterization for soil organic matter modeling
    (San Francisco, California, US : PLOS, 2018-10-12) Prays, Nadia; Dominik, Peter; Sänger, Anja; Franko, Uwe
    A variety of biogas residues (BGRs) have been used as organic fertilizer in agriculture. The use of these residues affects the storage of soil organic matter (SOM). In most cases, SOM changes can only be determined in long-term observations. Therefore, predictive modeling can be an efficient alternative, provided that the parameters required by the model are known for the considered BGRs. This study was conducted as a first approach to estimating the organic matter (OM) turnover parameters of BGRs for process modeling. We used carbon mineralization data from six BGRs from an incubation experiment, representing a range of substrate inputs, to calculate a turnover coefficient k controlling the velocity of fresh organic matter (FOM) decay and a synthesis coefficient describing the SOM creation from FOM. An SOM turnover model was applied in inverse mode to identify both parameters. In a second step, we related the parameters k and to chemical properties of the corresponding BGRs using a linear regression model and applied them to a long-term scenario simulation. According to the results of the incubation experiment, the k values ranged between 0.28 and 0.58 d-1 depending on the chemical composition of the FOM. The estimated values ranged between 0.8 and 0.89. The best linear relationship of k was found to occur with pH (R2 = 0.863). Parameter is related to the Ct/Norg ratio (R2 = 0.696). Long-term scenario simulations emphasized the necessity of specific k and values related to the chemical properties for each BGR. However, further research is needed to validate and improve these preliminary results. © 2018 Prays et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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    Side effects by oral application of atmospheric pressure plasma on the mucosa in mice
    (San Francisco, California, US : PLOS, 2019) Jablonowski, Lukasz; Kocher, Thomas; Schindler, Axel; Müller, Karolina; Dombrowski, Frank; von Woedtke, Thomas; Arnold, Thomas; Lehmann, Antje; Rupf, Stefan; Evert, Matthias; Evert, Katja
    Cold atmospheric pressure plasma (CAP) has been investigated with promising results for peri-implant diseases treatment. However, prior to in-vivo applications of CAP sources in humans, short-term harmful mucosal damage or other unwanted side effects have to be reviewed. 180 male mice (B6C3F1) were divided into twelve treatment groups (n = 15). The right buccal cheek mucosa was treated with CAP. The first and second group each received continuous 10 sec irradiation with 2 different plasma sources (kINPen09, PS-MWM). The third group was treated with the kINPen09 for one minute. Control groups were treated with a corresponding dose of ultraviolet light for 8 seconds or 48 seconds and the other one was left untreated. The animals were weighed before and after treatment. The animals were sacrificed one day or one week after exposure. Stained tissue samples were histologically examined for tissue damage independently by two experienced pathologists. One day after CAP treatment histological analysis showed focal mucosal erosion with superficial ulceration and necrosis accompanied by a mild inflammatory reaction. One week after CAP treatment, the mucosal defects were completely re-epithelialized, associated with remnants of granulation tissue in the stroma irrespective of treatment duration. Furthermore, no cytological atypia was found and no severe weight loss occurred. The control groups did not show any alterations at all. CAP treatment led to a superficial mucosal damage that healed within few days. Nonetheless, further long-term experiments are necessary to exclude undesirable side effects after longer observation time. Particularly, potential carcinogenic effects must be ruled out prior to the application of CAP treatment in daily dental practice.
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    Parameterization-induced uncertainties and impacts of crop management harmonization in a global gridded crop model ensemble
    (San Francisco, California, US : PLOS, 2019) Folberth, Christian; Elliott, Joshua; Müller, Christoph; Balkovič, Juraj; Chryssanthacopoulos, James; Izaurralde, Roberto C.; Jones, Curtis D.; Khabarov, Nikolay; Liu, Wenfeng; Reddy, Ashwan; Schmid, Erwin; Skalský, Rastislav; Yang, Hong; Arneth, Almut; Ciais, Philippe; Deryng, Delphine; Lawrence, Peter J.; Olin, Stefan; Pugh, Thomas A.M.; Ruane, Alex C.; Wang, Xuhui
    Global gridded crop models (GGCMs) combine agronomic or plant growth models with gridded spatial input data to estimate spatially explicit crop yields and agricultural externalities at the global scale. Differences in GGCM outputs arise from the use of different biophysical models, setups, and input data. GGCM ensembles are frequently employed to bracket uncertainties in impact studies without investigating the causes of divergence in outputs. This study explores differences in maize yield estimates from five GGCMs based on the public domain field-scale model Environmental Policy Integrated Climate (EPIC) that participate in the AgMIP Global Gridded Crop Model Intercomparison initiative. Albeit using the same crop model, the GGCMs differ in model version, input data, management assumptions, parameterization, and selection of subroutines affecting crop yield estimates via cultivar distributions, soil attributes, and hydrology among others. The analyses reveal inter-annual yield variability and absolute yield levels in the EPIC-based GGCMs to be highly sensitive to soil parameterization and crop management. All GGCMs show an intermediate performance in reproducing reported yields with a higher skill if a static soil profile is assumed or sufficient plant nutrients are supplied. An in-depth comparison of setup domains for two EPIC-based GGCMs shows that GGCM performance and plant stress responses depend substantially on soil parameters and soil process parameterization, i.e. hydrology and nutrient turnover, indicating that these often neglected domains deserve more scrutiny. For agricultural impact assessments, employing a GGCM ensemble with its widely varying assumptions in setups appears the best solution for coping with uncertainties from lack of comprehensive global data on crop management, cultivar distributions and coefficients for agro-environmental processes. However, the underlying assumptions require systematic specifications to cover representative agricultural systems and environmental conditions. Furthermore, the interlinkage of parameter sensitivity from various domains such as soil parameters, nutrient turnover coefficients, and cultivar specifications highlights that global sensitivity analyses and calibration need to be performed in an integrated manner to avoid bias resulting from disregarded core model domains. Finally, relating evaluations of the EPIC-based GGCMs to a wider ensemble based on individual core models shows that structural differences outweigh in general differences in configurations of GGCMs based on the same model, and that the ensemble mean gains higher skill from the inclusion of structurally different GGCMs. Although the members of the wider ensemble herein do not consider crop-soil-management interactions, their sensitivity to nutrient supply indicates that findings for the EPIC-based sub-ensemble will likely become relevant for other GGCMs with the progressing inclusion of such processes.
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    Have wind turbines in Germany generated electricity as would be expected from the prevailing wind conditions in 2000-2014?
    (San Francisco, Ca. : PLOS, 2019) Germer, Sonja; Kleidon, Axel
    The planning of the energy transition from fossil fuels to renewables requires estimates for how much electricity wind turbines can generate from the prevailing atmospheric conditions. Here, we estimate monthly ideal wind energy generation from datasets of wind speeds, air density and installed wind turbines in Germany and compare these to reported actual yields. Both yields were used in a statistical model to identify and quantify factors that reduced actual compared to ideal yields. The installed capacity within the region had no significant influence. Turbine age and park size resulted in significant yield reductions. Predicted yields increased from 9.1 TWh/a in 2000 to 58.9 TWh/a in 2014 resulting from an increase in installed capacity from 5.7 GW to 37.6 GW, which agrees very well with reported estimates for Germany. The age effect, which includes turbine aging and possibly other external effects, lowered yields from 3.6 to 6.7% from 2000 to 2014. The effect of park size decreased annual yields by 1.9% throughout this period. However, actual monthly yields represent on average only 73.7% of the ideal yields, with unknown causes. We conclude that the combination of ideal yields predicted from wind conditions with observed yields is suitable to derive realistic estimates of wind energy generation as well as realistic resource potentials. © 2019 Germer, Kleidon. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.