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Lignite biosolubilization and bioconversion by Bacillus sp.: the collation of analytical data
(2021)
The vast metabolic potential of microbes in brown coal (lignite) processing and utilization can greatly contribute to innovative approaches to sustainable production of high-value products from coal. In this study, the multi-faceted and complex coal biosolubilization process by Bacillus sp. RKB 7 isolate from the Kazakhstan coal-mining soil is reported, and the derived products are characterized. Lignite solubilization tests performed for surface and suspension cultures testify to the formation of numerous soluble lignite-derived substances. Almost 24% of crude lignite (5% w/v) was solubilized within 14 days under slightly alkaline conditions (pH 8.2). FTIR analysis revealed various functional groups in the obtained biosolubilization products. Analyses of the lignite-derived humic products by UV-Vis and fluorescence spectrometry as well as elemental analysis yielded compatible results indicating the emerging products had a lower molecular weight and degree of aromaticity. Furthermore, XRD and SEM analyses were used to evaluate the biosolubilization processes from mineralogical and microscopic points of view. The findings not only contribute to a deeper understanding of microbe–mineral interactions in coal environments, but also contribute to knowledge of coal biosolubilization and bioconversion with regard to sustainable production of humic substances. The detailed and comprehensive analyses demonstrate the huge biotechnological potential of Bacillus sp. for agricultural productivity and environmental health.
L.setulosus is offered for creation of biopreparation «Setulin», possesing he- mostatic action, the basic reactant of biopreparation is diterpen – lagochirzin. Results under the maintenance and dynamics of diterpen lagochirzin accumula- tion in various parts of L.setulosus are presented: in roots, stalks, leaves, flowers and calyx lobes during the growing season, and also results on conditions of cultivation L.setulosus in the conditions of a typical sierozem are resulted. From the obtained data is visible, that the given species of a plant is endemic. It is established, that dynamics of accumulation of lagochirzin in phytomass accrues from the beginning to the middle of the growing season. The chemical analysis of L.setulosus on a localization of lagochirzin in various organs of a plant, has shown, that the greatest quantity of lagochirzin collects in calyx lobes of the plants. Also it is established, that L.setulosus can be cultivated in the conditions of the typical sierozem, a mineral food is necessary for the given species of plants of Lagochilus genus, except nitric fertilizers. Comparative studying of wild-growing and cultural forms of L.setulosus has shown, that in the cultivated phytomass of plants the maintenance of lagochirzin on 17-20 % higher than in the wild-growing species.
The purpose of the current study was to examine the reproducibility of fascicle length and pennation angle of gastrocnemius medialis while human walking. To the best of our knowledge, this is the first study of the reproducibility of fascicle length and pennation angle of gastrocnemius medialis in vivo during human gait. Twelve males performed 10 gait trials on a treadmill, in 2 separate days. B-mode ultrasonography, with the ultrasound probe firmly adjusted in the transverse and frontal planes using a special cast, was used to measure the fascicle length and the pennation angle of the gastrocnemius medialis (GM). A Vicon 624 system with three cameras operating at 120 Hz was also used to record the ankle and knee joint angles. The results showed that measurements of fascicle length and pennation angle showed high reproducibility during the gait cycle, both within the same day and between different days. Moreover, the root mean square differences between the repeated waveforms of both variables were very small, compared with their ranges (fascicle length: RMS = ∼3 mm, range: 38–63 mm; pennation angle: RMS = ∼1.5°, range: 22–32°). However, their reproducibility was lower compared to the joint angles. It was found that representative data have to be derived by a wide number of gait trials (fascicle length ∼six trials, pennation angle more than 10 trials), to assure the reliability of the fascicle length and pennation angle in human gait.
Achtsnicht, Stefan ; Tödter, Julia ; Niehues, Julia ; Telöken, Matthias ; Offenhäusser, Andreas ; Krause, Hans-Joachim ; Schröper, Florian
For performing point-of-care molecular diagnostics, magnetic immunoassays constitute a promising alternative to established enzyme-linked immunosorbent assays (ELISA) because they are fast, robust and sensitive. Simultaneous detection of multiple biomolecular targets from one body fluid sample is desired. The aim of this work is to show that multiplex magnetic immunodetection based on magnetic frequency mixing by means of modular immunofiltration columns prepared for different targets is feasible. By calculations of the magnetic response signal, the required spacing between the modules was determined. Immunofiltration columns were manufactured by 3D printing and antibody immobilization was performed in a batch approach. It was shown experimentally that two different target molecules in a sample solution could be individually detected in a single assaying step with magnetic measurements of the corresponding immobilization filters. The arrangement order of the filters and of a negative control did not influence the results. Thus, a simple and reliable approach to multi-target magnetic immunodetection was demonstrated.
The movement of magnetic beads due to a magnetic field gradient is of great interest in different application fields. In this report we present a technique based on a magnetic tweezers setup to measure the velocity factor of magnetically actuated individual superparamagnetic beads in a fluidic environment. Several beads can be tracked simultaneously in order to gain and improve statistics. Furthermore we show our results for different beads with hydrodynamic diameters between 200 and 1000 nm from diverse manufacturers. These measurement data can, for example, be used to determine design parameters for a magnetic separation system, like maximum flow rate and minimum separation time, or to select suitable beads for fixed experimental requirements.
In modern bioanalytical methods, it is often desired to detect several targets in one sample within one measurement. Immunological methods including those that use superparamagnetic beads are an important group of techniques for these applications. The goal of this work is to investigate the feasibility of simultaneously detecting different superparamagnetic beads acting as markers using the magnetic frequency mixing technique. The frequency of the magnetic excitation field is scanned while the lower driving frequency is kept constant. Due to the particles’ nonlinear magnetization, mixing frequencies are generated. To record their amplitude and phase information, a direct digitization of the pickup-coil’s signal with subsequent Fast Fourier Transformation is performed. By synchronizing both magnetic beads using frequency scanning in magnetic frequency mixing technique magnetic fields, a stable phase information is gained. In this research, it is shown that the amplitude of the dominant mixing component is proportional to the amount of superparamagnetic beads inside a sample. Additionally, it is shown that the phase does not show this behaviour. Excitation frequency scans of different bead types were performed, showing different phases, without correlation to their diverse amplitudes. Two commercially available beads were selected and a determination of their amount in a mixture is performed as a demonstration for multiplex measurements.
Sensitive and rapid detection of cholera toxin subunit B using magnetic frequency mixing detection
(2019)
Achtsnicht, Stefan ; Neuendorf, Christian ; Faßbender, Tobias ; Nölke, Greta ; Offenhäusser, Andreas ; Krause, Hans-Joachim ; Schröper, Florian
Cholera is a life-threatening disease caused by the cholera toxin (CT) as produced by some Vibrio cholerae serogroups. In this research we present a method which directly detects the toxin’s B subunit (CTB) in drinking water. For this purpose we performed a magnetic sandwich immunoassay inside a 3D immunofiltration column. We used two different commercially available antibodies to capture CTB and for binding to superparamagnetic beads. ELISA experiments were performed to select the antibody combination. The beads act as labels for the magnetic frequency mixing detection technique. We show that the limit of detection depends on the type of magnetic beads. A nonlinear Hill curve was fitted to the calibration measurements by means of a custom-written python software. We achieved a sensitive and rapid detection of CTB within a broad concentration range from 0.2 ng/ml to more
than 700 ng/ml.
Die qualitative und quantitative Detektion von Zielsubstanzen innerhalb einer wässrigen Probe ist für viele Fragestellungen von Interesse, etwa bei der Detektion von Kontaminationen in Trinkwasser in Krisensituationen. Hierbei ist es nicht nur wichtig, dass Pathogene möglichst sensitiv detektiert werden können, sondern auch, dass die Analyse schnell erfolgt, um Betroffenen im Katastrophenfall zügig sicheres Trinkwasser zu Verfügung stellen zu können. Da bei einem solchen Szenario nicht von einer in der Nähe befindlichen funktionierenden Laborinfrastruktur ausgegangen werden kann, ist es wichtig, dass die Messung direkt vor Ort erfolgen kann. Im Rahmen dieser Arbeit wurde untersucht, ob eine derartige Schnellanalytik mithilfe von superparamagnetischen Beads (MBs) und der magnetischen Frequenzmischtechnik möglich ist. Dabei werden die MBs mit Hilfe von primären Antikörpern an die Zielsubstanz gebunden und mit sekundären Antikörpern an die Poren-Oberfläche eines Polyethylen-Filters fixiert (Sandwich-Immunoassay). So kann die Quantifizierung der Zielsubstanz auf eine magnetische Messung der immobilisierten MB-Marker zurückgeführt werden. Die magnetische Frequenzmischtechnik basiert auf der Anregung der Probe mit Magnetfeldern zweier verschiedener Frequenzen. Die durch die nichtlineare Magnetisierungsform der superparamagnetischen MBs entstehenden Mischfrequenzen werden typischerweise mithilfe einer zweistufigen Lock-in-Detektion analysiert (analoge Demodulation), die in einem Magnetreader als Handheldgerät realisiert wurde. Zusätzlich zu dieser Technik wurde das Prinzip der direkten Digitalisierung des gesamten Antwortsignals mit anschließender Fourier-Analyse der erzeugten Mischfrequenzen experimentell umgesetzt, um die Amplituden und Phasen mehrerer Mischfrequenzen simultan zu erfassen. Eine Möglichkeit zur Sensitivitätssteigerung ist die magnetische Aufkonzentration, indem vor der magnetischen Analyse eine Separation der MBs aus einem größeren Probenvolumen mittels magnetischem Feldgradienten durchgeführt wird. Zur Charakterisierung verschiedener kommerzieller MBs hinsichtlich ihrer magnetischen Separierbarkeit wurde ein Aufbau zur Messung ihrer magnetophoretischen Beweglichkeiten realisiert und ihre Geschwindigkeiten im Gradientenfeld mikroskopisch gemessen.Da eine Probe oftmals nicht nur auf eine einzige Zielsubstanz, sondern simultan auf mehrere verschiedene Pathogene hin untersucht werden soll, wurden verschiedene Ansätze entwickelt und getestet, die einen solchen multiparametrischen magnetischen Immunoassay ermöglichen. Einerseits wurde eine räumliche Separation der Bindungsbereiche für verschiedene Zielsubstanzen realisiert, die sequentiell ausgewertet werden können. Andererseits wurde die Unterscheidung von verschiedenen Zielsubstanzen anhand der Charakteristika der an sie gebundenen, verschieden funktionalisierten MB-Typen untersucht. Für eine solche Unterscheidung wurde zum einen die Anregefrequenz der magnetischen Frequenzmischtechnik während einer Messung variiert. Damit konnte gezeigt werden, dass sich verschiedene MB-Sorten anhand der Phase ihrer Frequenzmischsignale voneinander unterscheiden lassen. Weiterhin wurde gezeigt, dass sich der Signalverlauf einer binären Mischung zweier verschiedener MB-Typen als gradueller Übergang der Verläufe der beiden reinen MB-Lösungen ergibt. Eine weitere Analysemethode für einen multiparametrischen Immunoassay besteht darin, ein zusätzliches einstellbares statisches magnetisches Offsetfeld zu verwenden. Hierfür wurden mehrere Aufbauten auf Basis von Permanent- und Elektromagneten simuliert, konstruiert und charakterisiert. Mithilfe von Simulationen konnte gezeigt werden, dass eine auf diesem Verfahren beruhende Unterscheidung für MBs mit unterschiedlichen magnetischen Partikelmomenten möglich ist. Als direkte Anwendung des hier entwickelten Magnetreaders in Zusammenspiel mit der digitalen Demodulation wurde ein magnetischer Assay gegen die B-Untereinheit des Choleratoxins in Trinkwasser mit einem niedrigen Detektionslimit von 0,2 ng/ml demonstriert.
Achten, Richard ; Bauer, Andreas ; Bertram, Walter ; Cremer, Markus ; Daemen, Jos ; Dehnhardt, Markus ; Fleischer, Manfred ; Kirchner, Peter ; Leyendecker, Marco ; Pietrzyk, Uwe ; Schmitz, Jakob ; Ziemons, Karl ; Zilles, Karl
Die vorliegende Erfindung betrifft eine Vorrichtung zum Halten eines lebenden Objektes bei physiologischen Messungen mit einem Basiselement und Mitteln zum Arretieren des lebenden Objektes, die atraumatische Ohrenhalter sowie ein Mundstück aufweisen, das mit einer Aussparung für die Zähne des Lebewesens versehen ist, mit einem Masseanteil von wenigstens 95% an glasfaserverstärktem Kunststoff.
Abouzar, Maryam H. ; Schöning, Michael Josef ; Poghossian, Arshak ; Christiaens, P. ; Williams, O. A. ; Wagner, P. ; Haenen, K.
Abouzar, Maryam H. ; Poghossian, Arshak ; Siqueira, José R. Jr. ; Oliveira, Osvaldo N. Jr. ; Moritz, Werner ; Schöning, Michael Josef
Abouzar, Maryam H. ; Poghossian, Arshak ; Razavi, Arash ; Besmehn, Astrid ; Bijnens, Nathalie ; Williams, Oliver A. ; Haenen, Ken ; Wagner, Patrick ; Schöning, Michael Josef
Abouzar, Maryam H. ; Poghossian, Arshak ; Razavi, A. ; Williams, O. A. ; Bijnens, N. ; Wagner, P. ; Schöning, Michael Josef
Abouzar, Maryam H. ; Poghossian, Arshak ; Pedraza, A. M. ; Gandhi, D. ; Ingebrandt, S. ; Moritz, W. ; Schöning, Michael Josef
Abouzar, Maryam H. ; Poghossian, Arshak ; Cherstvy, Andrey G. ; Pedraza, Angela M. ; Ingebrandt, Sven ; Schöning, Michael Josef
Label-free electrical detection of consecutive deoxyribonucleic acid (DNA) hybridization/denaturation by means of an array of individually addressable field-effect-based nanoplate silicon-on-insulator (SOI) capacitors modified with gold nanoparticles (Au-NP) is investigated. The proposed device detects charge changes on Au-NP/DNA hybrids induced by the hybridization or denaturation event. DNA hybridization was performed in a high ionic-strength solution to provide a high hybridization efficiency. On the other hand, to reduce the screening of the DNA charge by counter ions and to achieve a high sensitivity, the sensor signal induced by the hybridization and denaturation events was measured in a low ionic-strength solution. High sensor signals of about 120, 90, and 80 mV were registered after the DNA hybridization, denaturation, and re-hybridization events, respectively. Fluorescence microscopy has been applied as reference method to verify the DNA immobilization, hybridization, and denaturation processes. An electrostatic charge-plane model for potential changes at the gate surface of a nanoplate field-effect sensor induced by the DNA hybridization has been developed taking into account both the Debye length and the distance of the DNA charge from the gate surface.
Abouzar, Maryam H. ; Ingebrandt, S. ; Poghossian, Arshak ; Zhang, Y. ; Vu, X. T. ; Moritz, W. ; Schöning, Michael Josef
Abouzar, Maryam H. ; Ingebrandt, S. ; Poghossian, Arshak ; Zhang, Y. ; Moritz, W. ; Schöning, Michael Josef
We present new numerical results for shape optimization problems of interior Neumann eigenvalues. This field is not well understood from a theoretical standpoint. The existence of shape maximizers is not proven beyond the first two eigenvalues, so we study the problem numerically. We describe a method to compute the eigenvalues for a given shape that combines the boundary element method with an algorithm for nonlinear eigenvalues. As numerical optimization requires many such evaluations, we put a focus on the efficiency of the method and the implemented routine. The method is well suited for parallelization. Using the resulting fast routines and a specialized parametrization of the shapes, we found improved maxima for several eigenvalues.
Abel, Thomas ; Bonin, Dominik ; Albracht, Kirsten ; Zeller, Sebastian ; Brüggemann, Gert-Peter ; Burkett, Brendan ; Strüder, Heiko K.
Three-dimensional (3D) full-field measurements provide a comprehensive and accurate validation of finite element (FE) models. For the validation, the result of the model and measurements are compared based on two respective point-sets and this requires the point-sets to be registered in one coordinate system. Point-set registration is a non-convex optimization problem that has widely been solved by the ordinary iterative closest point algorithm. However, this approach necessitates a good initialization without which it easily returns a local optimum, i.e. an erroneous registration. The globally optimal iterative closest point (Go-ICP) algorithm has overcome this drawback and forms the basis for the presented open-source tool that can be used for the validation of FE models using 3D full-field measurements. The capability of the tool is demonstrated using an application example from the field of biomechanics. Methodological problems that arise in real-world data and the respective implemented solution approaches are discussed.