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    Nonlinear Optical Investigation of Microbial Chromoproteins
    (Lausanne : Frontiers Media, 2020) Krekic, Szilvia; Zakar, Tomás; Gombos, Zoltán; Valkai, Sándor; Mero, Mark; Zimányi, László; Heiner, Zsuzsanna; Dér, András
    Membrane-bound or cytosolic light-sensitive proteins, playing a crucial role in energy- and signal-transduction processes of various photosynthetic microorganisms, have been optimized for sensing or harvesting light by myriads of years of evolution. Upon absorption of a photon, they undergo a usually cyclic reaction series of conformations, and the accompanying spectro-kinetic events assign robust nonlinear optical (NLO) properties for these chromoproteins. During recent years, they have attracted a considerable interest among researchers of the applied optics community as well, where finding the appropriate NLO material for a particular application is a pivotal task. Potential applications have emerged in various branches of photonics, including optical information storage and processing, higher-harmonic and white-light continuum generation, or biosensorics. In our earlier work, we also raised the possibility of using chromoproteins, such as bacteriorhodopsin (bR), as building blocks for the active elements of integrated optical (IO) circuits, where several organic and inorganic photonic materials have been considered as active components, but so far none of them has been deemed ideal for the purpose. In the current study, we investigate the linear and NLO properties of biofilms made of photoactive yellow protein (PYP) and bR. The kinetics of the photoreactions are monitored by time-resolved absorption experiments, while the refractive index of the films and its light-induced changes are measured using the Optical Waveguide Lightmode Spectroscopy (OWLS) and Z-scan techniques, respectively. The nonlinear refractive index and the refractive index change of both protein films were determined in the green spectral range in a wide range of intensities and at various laser repetition rates. The nonlinear refractive index and refractive index change of PYP were compared to those of bR, with respect to photonics applications. Our results imply that the NLO properties of these proteins make them promising candidates for utilization in applied photonics, and they should be considered as valid alternatives for active components of IO circuits. © Copyright © 2020 Krekic, Zakar, Gombos, Valkai, Mero, Zimányi, Heiner and Dér.
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    Thermal Impact on the Culturable Microbial Diversity Along the Processing Chain of Flour From Crickets (Acheta domesticus)
    (Lausanne : Frontiers Media, 2020) Fröhling, Antje; Bußler, Sara; Durek, Julia; Schlüter, Oliver K.
    The role of insects for human consumption has lately increased in interest and in order to deliver safe and high-quality raw materials and ingredients for food and feed applications, processing of insects is a major pre-requisite. For edible insects a thermal treatment and appropriate storage conditions are recommended to minimize the microbiological risk and the impact of processing methods on the microbial contamination needs to be considered and determined. Based on standard process conditions for the production of Acheta domesticus flour, different heating treatments were used to reduce the microbial load of A. domesticus. In addition, the drying temperature and drying time were varied to determine whether the required residual moisture of <5% can be achieved more quickly with consistent microbial quality. The influence of the process conditions on the microbial community of A. domesticus along the processing chain was finally investigated under optimized process conditions. The total viable count was reduced from 9.24 log10 CFU/gDM to 1.98 log10 CFU/gDM along the entire processing chain. While Bacillaceae, Enterobacteriaceae, Enterococcaceae, and yeast and molds were no longer detectable in the A. domesticus flour, Staphylococcaceae and mesophilic spore forming bacteria were still found in the flour. The results indicate that the steaming process is essential for effectively increasing microbial safety since this processing step showed the highest inactivation. It is recommended to not only evaluate the total viable count but also to monitor changes in microbial diversity during processing to ensure microbial safety of the final product. © Copyright © 2020 Fröhling, Bußler, Durek and Schlüter.
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    Modeling of Individual Fruit-Bearing Capacity of Trees Is Aimed at Optimizing Fruit Quality of Malus x domestica Borkh. 'Gala'
    (Lausanne : Frontiers Media, 2021) Penzel, Martin; Herppich, Werner B.; Weltzien, Cornelia; Tsoulias, Nikos; Zude-Sasse, Manuela
    The capacity of apple trees to produce fruit of a desired diameter, i.e., fruit-bearing capacity (FBC), was investigated by considering the inter-tree variability of leaf area (LA). The LA of 996 trees in a commercial apple orchard was measured by using a terrestrial two-dimensional (2D) light detection and ranging (LiDAR) laser scanner for two consecutive years. The FBC of the trees was simulated in a carbon balance model by utilizing the LiDAR-scanned total LA of the trees, seasonal records of fruit and leaf gas exchanges, fruit growth rates, and weather data. The FBC was compared to the actual fruit size measured in a sorting line on each individual tree. The variance of FBC was similar in both years, whereas each individual tree showed different FBC in both seasons as indicated in the spatially resolved data of FBC. Considering a target mean fruit diameter of 65 mm, FBC ranged from 84 to 168 fruit per tree in 2018 and from 55 to 179 fruit per tree in 2019 depending on the total LA of the trees. The simulated FBC to produce the mean harvest fruit diameter of 65 mm and the actual number of the harvested fruit >65 mm per tree were in good agreement. Fruit quality, indicated by fruit's size and soluble solids content (SSC), showed enhanced percentages of the desired fruit quality according to the seasonally total absorbed photosynthetic energy (TAPE) of the tree per fruit. To achieve a target fruit diameter and reduce the variance in SSC at harvest, the FBC should be considered in crop load management practices. However, achieving this purpose requires annual spatial monitoring of the individual FBC of trees.
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    Plasma-Treated Water Affects Listeria monocytogenes Vitality and Biofilm Structure
    (Lausanne : Frontiers Media, 2021) Handorf, Oliver; Pauker, Viktoria Isabella; Weihe, Thomas; Schäfer, Jan; Freund, Eric; Schnabel, Uta; Bekeschus, Sander; Riedel, Katharina; Ehlbeck, Jörg
    Background: Plasma-generated compounds (PGCs) such as plasma-processed air (PPA) or plasma-treated water (PTW) offer an increasingly important alternative for the control of microorganisms in hard-to-reach areas found in several industrial applications including the food industry. To this end, we studied the antimicrobial capacity of PTW on the vitality and biofilm formation of Listeria monocytogenes, a common foodborne pathogen. Results: Using a microwave plasma (MidiPLexc), 10 ml of deionized water was treated for 100, 300, and 900 s (pre-treatment time), after which the bacterial biofilm was exposed to the PTW for 1, 3, and 5 min (post-treatment time) for each pre-treatment time, separately. Colony-forming units (CFU) were significantly reduced by 4.7 log10 ± 0.29 log10, as well as the metabolic activity decreased by 47.9 ± 9.47% and the cell vitality by 69.5 ± 2.1%, compared to the control biofilms. LIVE/DEAD staining and fluorescence microscopy showed a positive correlation between treatment and incubation times, as well as reduction in vitality. Atomic force microscopy (AFM) indicated changes in the structure quality of the bacterial biofilm. Conclusion: These results indicate a promising antimicrobial impact of plasma-treated water on Listeria monocytogenes, which may lead to more targeted applications of plasma decontamination in the food industry in the future.
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    Enhancement of Intracellular Calcium Ion Mobilization by Moderately but Not Highly Positive Material Surface Charges
    (Lausanne : Frontiers Media, 2020) Gruening, Martina; Neuber, Sven; Nestler, Peter; Lehnfeld, Jutta; Dubs, Manuela; Fricke, Katja; Schnabelrauch, Matthias; Helm, Christiane A.; Müller, Rainer; Staehlke, Susanne; Nebe, J. Barbara
    Electrostatic forces at the cell interface affect the nature of cell adhesion and function; but there is still limited knowledge about the impact of positive or negative surface charges on cell-material interactions in regenerative medicine. Titanium surfaces with a variety of zeta potentials between −90 mV and +50 mV were generated by functionalizing them with amino polymers, extracellular matrix proteins/peptide motifs and polyelectrolyte multilayers. A significant enhancement of intracellular calcium mobilization was achieved on surfaces with a moderately positive (+1 to +10 mV) compared with a negative zeta potential (−90 to −3 mV). Dramatic losses of cell activity (membrane integrity, viability, proliferation, calcium mobilization) were observed on surfaces with a highly positive zeta potential (+50 mV). This systematic study indicates that cells do not prefer positive charges in general, merely moderately positive ones. The cell behavior of MG-63s could be correlated with the materials’ zeta potential; but not with water contact angle or surface free energy. Our findings present new insights and provide an essential knowledge for future applications in dental and orthopedic surgery. © Copyright © 2020 Gruening, Neuber, Nestler, Lehnfeld, Dubs, Fricke, Schnabelrauch, Helm, Müller, Staehlke and Nebe.
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    Understanding Beta-Lactam-Induced Lysis at the Single-Cell Level
    (Lausanne : Frontiers Media, 2021) Wong, Felix; Wilson, Sean; Helbig, Ralf; Hegde, Smitha; Aftenieva, Olha; Zheng, Hai; Liu, Chenli; Pilizota, Teuta; Garner, Ethan C.; Amir, Ariel; Renner, Lars D.
    Mechanical rupture, or lysis, of the cytoplasmic membrane is a common cell death pathway in bacteria occurring in response to β-lactam antibiotics. A better understanding of the cellular design principles governing the susceptibility and response of individual cells to lysis could indicate methods of potentiating β-lactam antibiotics and clarify relevant aspects of cellular physiology. Here, we take a single-cell approach to bacterial cell lysis to examine three cellular features-turgor pressure, mechanosensitive channels, and cell shape changes-that are expected to modulate lysis. We develop a mechanical model of bacterial cell lysis and experimentally analyze the dynamics of lysis in hundreds of single Escherichia coli cells. We find that turgor pressure is the only factor, of these three cellular features, which robustly modulates lysis. We show that mechanosensitive channels do not modulate lysis due to insufficiently fast solute outflow, and that cell shape changes result in more severe cellular lesions but do not influence the dynamics of lysis. These results inform a single-cell view of bacterial cell lysis and underscore approaches of combatting antibiotic tolerance to β-lactams aimed at targeting cellular turgor.
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    The Economic Impact of Exchanging Breeding Material: Assessing Winter Wheat Production in Germany
    (Lausanne : Frontiers Media, 2020) Lüttringhaus, Sophia; Gornott, Christoph; Wittkop, Benjamin; Noleppa, Steffen; Lotze-Campen, Hermann
    Climate change impacts imply that the stabilization and improvement of agricultural production systems using technological innovations has become vital. Improvements in plant breeding are integral to such innovations. In the context of German crop breeding programs, the economic impact of exchanging genetic material has yet to be determined. To this end, we analyze in this impact assessment the economic effects on German winter wheat production that are attributable to exchanging parental material amongst breeders in the breeding process. This exchange is supported by the breeders’ exemption, which is an integral part of the German plant variety protection legislation. It ensures that breeders can freely use licensed varieties created by other breeders for their own breeding activities and aims to speed up the development of improved varieties. For our analysis, we created a unique data set that combines variety-specific grain yield, adoption, and pedigree information of 133 winter wheat varieties. We determined the parental pedigree of each variety to see if a variety was created by interbreeding varieties that are internal or external to its specific breeder. Our study is the first that analyzes the economic impact of exchanging genetic material in German breeding programs. We found that more than 90 % of the tested varieties were bred with exchanged parental material, whereby the majority had two external parents. Also, these varieties were planted on an 8.5 times larger area than the varieties that were bred with two internal parents. Due to lower adoption, these only contributed 11 % to the overall winter wheat production in Germany, even though they yielded more. We used an economic surplus model to measure the benefits of exchanging parental breeding material on German winter wheat production. This resulted in an overall estimated economic surplus of 19.2 to 22.0 billion EUR from production year 1972 to 2018. This implies tremendous returns to using the breeder’s exemption, which, from an economic perspective, is almost cost-free for the breeder. We conclude that the exchange of breeding material contributes to improving Germany’s agricultural production and fosters the development of climate-resilient production systems and global food security. © Copyright © 2020 Lüttringhaus, Gornott, Wittkop, Noleppa and Lotze-Campen.
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    Genetic Variability of Morphological, Flowering, and Biomass Quality Traits in Hemp (Cannabis sativa L.)
    (Lausanne : Frontiers Media, 2020) Petit, Jordi; Salentijn, Elma M.J.; Paulo, Maria-Joao; Thouminot, Claire; van Dinter, Bert Jan; Magagnini, Gianmaria; Gusovius, Hans-Jörg; Tang, Kailei; Amaducci, Stefano; Wang, Shaoliang; Uhrlaub, Birgit; Müssig, Jörg; Trindade, Luisa M.
    Hemp (Cannabis sativa L.) is a bast-fiber crop well-known for the great potential to produce sustainable fibers. Nevertheless, hemp fiber quality is a complex trait, and little is known about the phenotypic variability and heritability of fiber quality traits in hemp. The aim of this study is to gain insights into the variability in fiber quality within the hemp germplasm and to estimate the genetic components, environmental components, and genotype-by-environment (G×E) interactions on fiber quality traits in hemp. To investigate these parameters, a panel of 123 hemp accessions was phenotyped for 28 traits relevant to fiber quality at three locations in Europe, corresponding to climates of northern, central, and southern Europe. In general, hemp cultivated in northern latitudes showed a larger plant vigor while earlier flowering was characteristic of plants cultivated in southern latitudes. Extensive variability between accessions was observed for all traits. Most cell wall components (contents of monosaccharides derived from cellulose and hemicellulose; and lignin content), bast fiber content, and flowering traits revealed large genetic components with low G×E interactions and high broad-sense heritability values, making these traits suitable to maximize the genetic gains of fiber quality. In contrast, contents of pectin-related monosaccharides, most agronomic traits, and several fiber traits (fineness and decortication efficiency) showed low genetic components with large G×E interactions affecting the rankings across locations. These results suggest that pectin, agronomic traits, and fiber traits are unsuitable targets in breeding programs of hemp, as their large G×E interactions might lead to unexpected phenotypes in untested locations. Furthermore, all environmental effects on the 28 traits were statistically significant, suggesting a strong adaptive behavior of fiber quality in hemp to specific environments. The high variability in fiber quality observed in the hemp panel, the broad range in heritability, and adaptability among all traits prescribe positive prospects for the development of new hemp cultivars of excellent fiber quality. © Copyright © 2020 Petit, Salentijn, Paulo, Thouminot, van Dinter, Magagnini, Gusovius, Tang, Amaducci, Wang, Uhrlaub, Müssig and Trindade.
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    Early Detection of Stripe Rust in Winter Wheat Using Deep Residual Neural Networks
    (Lausanne : Frontiers Media, 2021) Schirrmann, Michael; Landwehr, Niels; Giebel, Antje; Garz, Andreas; Dammer, Karl-Heinz
    Stripe rust (Pst) is a major disease of wheat crops leading untreated to severe yield losses. The use of fungicides is often essential to control Pst when sudden outbreaks are imminent. Sensors capable of detecting Pst in wheat crops could optimize the use of fungicides and improve disease monitoring in high-throughput field phenotyping. Now, deep learning provides new tools for image recognition and may pave the way for new camera based sensors that can identify symptoms in early stages of a disease outbreak within the field. The aim of this study was to teach an image classifier to detect Pst symptoms in winter wheat canopies based on a deep residual neural network (ResNet). For this purpose, a large annotation database was created from images taken by a standard RGB camera that was mounted on a platform at a height of 2 m. Images were acquired while the platform was moved over a randomized field experiment with Pst-inoculated and Pst-free plots of winter wheat. The image classifier was trained with 224 × 224 px patches tiled from the original, unprocessed camera images. The image classifier was tested on different stages of the disease outbreak. At patch level the image classifier reached a total accuracy of 90%. To test the image classifier on image level, the image classifier was evaluated with a sliding window using a large striding length of 224 px allowing for fast test performance. At image level, the image classifier reached a total accuracy of 77%. Even in a stage with very low disease spreading (0.5%) at the very beginning of the Pst outbreak, a detection accuracy of 57% was obtained. Still in the initial phase of the Pst outbreak with 2 to 4% of Pst disease spreading, detection accuracy with 76% could be attained. With further optimizations, the image classifier could be implemented in embedded systems and deployed on drones, vehicles or scanning systems for fast mapping of Pst outbreaks.
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    Vitronectin-based hydrogels recapitulate neuroblastoma growth conditions
    (Lausanne : Frontiers Media, 2022) Monferrer, Ezequiel; Dobre, Oana; Trujillo, Sara; González Oliva, Mariana Azevedo; Trubert-Paneli, Alexandre; Acevedo-León, Delia; Noguera, Rosa; Salmeron-Sanchez, Manuel
    The tumor microenvironment plays an important role in cancer development and the use of 3D in vitro systems that decouple different elements of this microenvironment is critical for the study of cancer progression. In neuroblastoma (NB), vitronectin (VN), an extracellular matrix protein, has been linked to poor prognosis and appears as a promising therapeutic target. Here, we developed hydrogels that incorporate VN into 3D polyethylene glycol (PEG) hydrogel networks to recapitulate the native NB microenvironment. The stiffness of the VN/PEG hydrogels was modulated to be comparable to the in vivo values reported for NB tissue samples. We used SK-N-BE (2) NB cells to demonstrate that PEGylated VN promotes cell adhesion as the native protein does. Furthermore, the PEGylation of VN allows its crosslinking into the hydrogel network, providing VN retention within the hydrogels that support viable cells in 3D. Confocal imaging and ELISA assays indicate that cells secrete VN also in the hydrogels and continue to reorganize their 3D environment. Overall, the 3D VN-based PEG hydrogels recapitulate the complexity of the native tumor extracellular matrix, showing that VN-cell interaction plays a key role in NB aggressiveness, and that VN could potentially be targeted in preclinical drug studies performed on the presented hydrogels.