Publikationen am Institut für Chemie und Biotechnik

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Ehrhardt, Linda; Günther, Mike; Böhme, Manfred; Köhler, Michael; Cao-Riehmer, Jialan
Three soil bacterial communities from an archaeological excavation site of an ancient coal mine near Bennstedt (Germany) characterized by 16S r-RNA sequencing. - In: Environments, ISSN 2076-3298, Bd. 9 (2022), 9, 115, S. 1-19

This metagenomics investigation of three closely adjacent sampling sites from an archaeological excavation of a pre-industrial coal mining exploration shaft provides detailed information on the composition of the local soil bacterial communities. The observed significant differences between the samples, reflected in the 16S r-RNA analyses, were consistent with the archaeologically observed situation distinguishing the coal seam, the rapidly deposited bright sediment inside an exploration shaft, and the topsoil sediment. In general, the soils were characterized by a dominance of Proteobacteria, Actinobacteria, Acidobacteria, and Archaea, whereas the coal seam was characterized by the highest proportion of Proteobacteria; the topsoil was characterized by very high proportions of Archaea - in particular, Nitrosotaleaceae - and Acidobacteria, mainly of Subgroup 2. Interestingly, the samples of the fast-deposited bright sediment showed a rank function of OTU abundances with disproportional values in the lower abundance range. This could be interpreted as a reflection of the rapid redeposition of soil material during the refilling of the exploration shaft in the composition of the soil bacterial community. This interpretation is supported by the observation of a comparatively high proportion of reads relating to bacteria known to be alkaliphilic in this soil material. In summary, these investigations confirm that metagenomic analyses of soil material from archaeological excavations can provide valuable information about the local soil bacterial communities and the historical human impacts on them.



https://doi.org/10.3390/environments9090115
Radivoievych, Alexandar; Kolp, Benjamin; Grebinyk, Sergii; Prylutska, Svitlana; Ritter, Uwe; Zolk, Oliver; Glökler, Jörn Felix; Frohme, Marcus; Grebinyk, Anna
Prestine C60 fullerene as a novel agent in sonodynamic treatment of cancer cells. - In: FEBS Open Bio, ISSN 2211-5463, Bd. 12 (2022), S. 74

https://doi.org/10.1002/2211-5463.13440
Mazétyté-Stasinskiené, Raminta; Freiberger, Emma; Täuscher, Eric; Köhler, Michael
Four-level structural hierarchy: microfluidically supported synthesis of polymer particle architectures incorporating fluorescence-labeled components and metal nanoparticles. - In: Langmuir, ISSN 1520-5827, Bd. 38 (2022), 29, S. 8794-8804

Hierarchical assemblies of functional polymer particles are promising due to their surface as well as physicochemical properties. However, hierarchical composites are complex and challenging to form due to the many steps necessary for integrating different components into one system. Highly structured four-level composite particles were formed in a four-step process. First of all, gold (Au) nanoparticles, poly(methyl methacrylate) (PMMA) nanoparticles, and poly(tripropylene glycol diacrylate) (poly-TPGDA) microparticles were individually synthesized. By applying microfluidic techniques, polymer nano- and microparticles were formed with tunable size and surface properties. Afterwards, the negatively charged gold nanoparticles and PMMA particles functionalized with a positively charged surface were mixed to form Au/PMMA assemblies. The Au/PMMA composites were mixed and incubated with poly-TPGDA microparticles to form ternary Au/PMMA/poly-TPGDA assemblies. For the formation of composite-containing microparticles, Au/PMMA/poly-TPGDA composites were dispersed in an aqueous acrylamide-methylenebisacrylamide solution. Monomer droplets were formed in a co-flow microfluidic device and photopolymerized by UV light. In this way, hierarchically structured four-level composites consisting of four different size ranges - 0.025/0.8/30/1000 μm - were obtained. By functionalizing polymer nano- and microparticles with different fluorescent dyes, it was possible to visualize the same composite particle under two different excitation modes (λex = 395-440 and λex = 510-560 nm). The Au/PMMA/poly-TPGDA composite-embedded polyacrylamide microparticles can be potentially used as a model for the creation of composite particles for sensing, catalysis, multilabeling, and biomedical applications.



https://doi.org/10.1021/acs.langmuir.2c00686
Konkin, Alexander; Ritter, Uwe; Konkin, Aleksei A.; Knauer, Andrea; Krinichnyi, Victor I.; Klochkov, Vladimir; Aganov, Albert; Gafurov, Marat; Wendler, Frank; Scharff, Peter
PPDN and NTCDA radical anions formation in EMIM-DCA, BMIM-BF4 EMIM-Ac ionic liquid solutions under the steady state UV and Vis light illumination: a combined X-, K-band EPR and DFT study. - In: Journal of molecular liquids, ISSN 1873-3166, Bd. 362 (2022), 119631

The radical anion of Pyrazino[2,3-f][1,10]phenanthroline-2,3-dicarbonitrile (PPDN) in blends with imidazolium based room temperature ionic liquids (RTIL): EMIM-DCA, BMIM-BF4, EMIM-Ac has been detected by X-band continues wave (CW) electron paramagnetic resonance (EPR) under steady state Xe-lamp illumination in the temperature interval from 190 to 340 K. The radical anion of 1,4,5,8-Naphthalenetetracarboxylic dianhydride (NTCDA) was registered by X- and K-band CW EPR at room temperature under the visible light CW diode laser operated at 532 nm, and Xe-lamp as well. The experimental hyperfine coupling data of both anion radicals were confirmed by DFT calculation. The formation of PPDN•- NTCDA•- and fullerene derivative (FD) radical anions is attributed to the photoelectron transfer from an IL anion to PPDN, NTCDA and FD electron acceptors. Here, the electron transfer leads to an irreversibility of these reactions due to photo-induced decomposition of the IL anions in the presence of an effective electron acceptor and is supported in the above RTILs solutions by means of EPR. For the indirect confirmation of the EMIM-DCA, EMIM-AC, BMIM-BF4 anion degradation in solutions with PPDN and NTCDA up to the transient radical state, similar data of acetate anion [OCOCH3]- decomposition, under CW Xe-Lamp photolysis resulting in •CH3 formation and its stabilization at 77 K in EMIM-Ac suspension with some FD dissolved in DCB are introduced as well. However, the main goal of this study is dedicated to the features of rotational and translational diffusion kinetics of PPDN and NTCDA radical anions in IL solutions as well to the evaluation of their application as a spin probes in ILs study in liquid phase.



https://doi.org/10.1016/j.molliq.2022.119631
Henkel, Thomas; Mayer, Günter; Hampl, Jörg; Cao-Riehmer, Jialan; Ehrhardt, Linda; Schober, Andreas; Groß, Gregor Alexander
From microtiter plates to droplets - there and back again. - In: Micromachines, ISSN 2072-666X, Bd. 13 (2022), 7, 1022, S. 1-13

Droplet-based microfluidic screening techniques can benefit from interfacing established microtiter plate-based screening and sample management workflows. Interfacing tools are required both for loading preconfigured microtiter-plate (MTP)-based sample collections into droplets and for dispensing the used droplets samples back into MTPs for subsequent storage or further processing. Here, we present a collection of Digital Microfluidic Pipetting Tips (DMPTs) with integrated facilities for droplet generation and manipulation together with a robotic system for its operation. This combination serves as a bidirectional sampling interface for sample transfer from wells into droplets (w2d) and vice versa droplets into wells (d2w). The DMPT were designed to fit into 96-deep-well MTPs and prepared from glass by means of microsystems technology. The aspirated samples are converted into the channel-confined droplets’ sequences separated by an immiscible carrier medium. To comply with the demands of dose-response assays, up to three additional assay compound solutions can be added to the sample droplets. To enable different procedural assay protocols, four different DMPT variants were made. In this way, droplet series with gradually changing composition can be generated for, e.g., 2D screening purposes. The developed DMPT and their common fluidic connector are described here. To handle the opposite transfer d2w, a robotic transfer system was set up and is described briefly.



https://doi.org/10.3390/mi13071022
Cao-Riehmer, Jialan; Chande, Charmi; Köhler, Michael
Microtoxicology by microfluidic instrumentation: a review. - In: Lab on a chip, ISSN 1473-0189, Bd. 22 (2022), 14, S. 2600-2623

Microtoxicology is concerned with the toxic effects of small amounts of substances. This review paper discusses the application of small amounts of noxious substances for toxicological investigation in small volumes. The vigorous development of miniaturized methods in microfluidics over the last two decades involves chip-based devices, micro droplet-based procedures, and the use of micro-segmented flow for microtoxicological studies. The studies have shown that the microfluidic approach is particularly valuable for highly parallelized and combinatorial dose-response screenings. Accurate dosing and mixing of effector substances in large numbers of microcompartments supplies detailed data of dose-response functions by highly concentration-resolved assays and allows evaluation of stochastic responses in case of small separated cell ensembles and single cell experiments. The investigations demonstrate that very different biological targets can be studied using miniaturized approaches, among them bacteria, eukaryotic microorganisms, cell cultures from tissues of multicellular organisms, stem cells, and early embryonic states. Cultivation and effector exposure tests can be performed in small volumes over weeks and months, confirming that the microfluicial strategy is also applicable for slow-growing organisms. Here, the state of the art of miniaturized toxicology, particularly for studying antibiotic susceptibility, drug toxicity testing in the miniaturized system like organ-on-chip, environmental toxicology, and the characterization of combinatorial effects by two and multi-dimensional screenings, is discussed. Additionally, this review points out the practical limitations of the microtoxicology platform and discusses perspectives on future opportunities and challenges.



https://doi.org/10.1039/D2LC00268J
Nolte, Oliver; Geitner, Robert; Volodin, Ivan A.; Rohland, Philip; Hager, Martin; Schubert, Ulrich Sigmar
State of charge and state of health assessment of viologens in aqueous-organic redox-flow electrolytes using in situ IR spectroscopy and multivariate curve resolution. - In: Advanced science, ISSN 2198-3844, Bd. 9 (2022), 17, 2200535, S. 1-10

Aqueous-organic redox flow batteries (RFBs) have gained considerable interest in recent years, given their potential for an economically viable energy storage at large scale. This, however, strongly depends on both the robustness of the underlying electrolyte chemistry against molecular decomposition reactions as well as the device's operation. With regard to this, the presented study focuses on the use of in situ IR spectroscopy in combination with a multivariate curve resolution approach to gain insight into both the molecular structures of the active materials present within the electrolyte as well as crucial electrolyte state parameters, represented by the electrolyte's state of charge (SOC) and state of health (SOH). To demonstrate the general applicability of the approach, methyl viologen (MV) and bis(3-trimethylammonium)propyl viologen (BTMAPV) are chosen, as viologens are frequently used as negolytes in aqueous-organic RFBs. The study's findings highlight the impact of in situ spectroscopy and spectral deconvolution tools on the precision of the obtainable SOC and SOH values. Furthermore, the study indicates the occurrence of multiple viologen dimers, which possibly influence the electrolyte lifetime and charging characteristics.



https://doi.org/10.1002/advs.202200535
Strutynska, Nataliia Yu.; Grynyuk, Iryna I.; Vasyliuk, Olga M.; Prylutska, Svitlana V.; Vovchenko, Ludmila L.; Kraievska, I. A.; Slobodyanik, Nikolai S.; Ritter, Uwe; Prylutskyy, Yury I.
Novel whitlockite/alginate/C60 fullerene composites: synthesis, characterization and properties for medical application. - In: The Arabian journal for science and engineering, ISSN 2191-4281, Bd. 47 (2022), 6, S. 7093-7104

The hybrid composite materials in form of spheres based on whitlockite-related calcium phosphate, Alginate (20, 30 or 50 wt.%) and C60 Fullerene (C60; 2 or 5 wt.%) were fabricated. According to XRD, elemental analysis and SEM data, the whitlockite-related (hexagonal system, space group R3c) calcium phosphate containing 0.42 wt.% of sodium was obtained in the form of particles with size 50-80 nm. It has been found that the addition of Alginate (20 wt.%) to prepared calcium phosphate leads to an increase in the compressive strength of composite by two times (from 137 to 358 MPa), and value of Young's modulus on 20% (from 460 to 558 MPa), while the presence of C60 in composition did not significant influence on this characteristic. The antibacterial activity of prepared composites with different composition and amounts (2.5, 5 or 10 mM) against Lactobacillus rhamnosus, Lactobacillus salivarius, Staphylococcus aureus and Pseudomonas aeruginosa was studied. All prepared samples did not effect on Lactobacillus. The addition of 5 wt.% C60 to phosphate-Alginate (30 wt.%) composite resulted in a tenfold decrease in the survival rate of the S. aureus strain at 5 and 10 mM of samples while P. aeruginosa was less sensitive to action of this sample and inhibition of bacteria growth was occurred only at its amount 10 mM. Thus, the results of mechanical properties and impact of created nanostructured hybrid composites on normal human microbiota (Lactobacillus) as well as pathogenic strain (S. aureus and P. aeruginosa) indicate the suitability of these promising materials for further biological test for bone therapy.



https://doi.org/10.1007/s13369-021-06552-0
Nozdrenko, Dmytro; Prylutska, Svitlana; Bogutska, Kateryna; Cherepanov, Vsevolod; Senenko, Anton; Vygovska, Oksana; Khrapatyi, Sergii; Ritter, Uwe; Prylutskyy, Yuriy; Piosik, Jacek
Analysis of biomechanical and biochemical markers of rat muscle soleus fatigue processes development during long-term use of C60 fullerene and N-acetylcysteine. - In: Nanomaterials, ISSN 2079-4991, Bd. 12 (2022), 9, 1552, S. 1-15

The development of an effective therapy aimed at restoring muscle dysfunctions in clinical and sports medicine, as well as optimizing working activity in general remains an urgent task today. Modern nanobiotechnologies are able to solve many clinical and social health problems, in particular, they offer new therapeutic approaches using biocompatible and bioavailable nanostructures with specific bioactivity. Therefore, the nanosized carbon molecule, C60 fullerene, as a powerful antioxidant, is very attractive. In this study, a comparative analysis of the dynamic of muscle soleus fatigue processes in rats was conducted using 50 Hz stimulation for 5 s with three consistent pools after intraperitoneal administration of the following antioxidants: C60 fullerene (a daily dose of 1 mg/kg one hour prior to the start of the experiment) and N-acetylcysteine (NAC; a daily dose of 150 mg/kg one hour prior to the start of the experiment) during five days. Changes in the integrated power of muscle contraction, levels of the maximum and minimum contraction force generation, time of reduction of the contraction force by 50% of its maximum value, achievement of the maximum force response, and delay of the beginning of a single contraction force response were analyzed as biomechanical markers of fatigue processes. Levels of creatinine, creatine phosphokinase, lactate, and lactate dehydrogenase, as well as pro- and antioxidant balance (thiobarbituric acid reactive substances, hydrogen peroxide, reduced glutathione, and catalase activity) in the blood of rats were analyzed as biochemical markers of fatigue processes. The obtained data indicate that applied therapeutic drugs have the most significant effects on the 2nd and especially the 3rd stimulation pools. Thus, the application of C60 fullerene has a (50-80)% stronger effect on the resumption of muscle biomechanics after the beginning of fatigue than NAC on the first day of the experiment. There is a clear trend toward a positive change in all studied biochemical parameters by about (12-15)% after therapeutic administration of NAC and by (20-25)% after using C60 fullerene throughout the experiment. These findings demonstrate the promise of using C60 fullerenes as potential therapeutic nanoagents that can reduce or adjust the pathological conditions of the muscular system that occur during fatigue processes in skeletal muscles.



https://doi.org/10.3390/nano12091552
Dorner-Reisel, Annett; Ritter, Uwe; Moje, Jens; Freiberger, Emma; Scharff, Peter
Effect of fullerene C60 thermal and tribomechanical loading on Raman signals. - In: Diamond and related materials, ISSN 0925-9635, Bd. 126 (2022), 109036, S. 1-14

Fullerene C60 powder was loaded by 1 N normal force and exposed to sliding under different frequencies for 15 min. It is shown that the velocity of the sliding movement determines the stability of the fullerene C60 powder. At slow velocity of movement with a frequency of 1 Hz under 1 N normal force, the fullerene C60 structure remains undamaged after 15 min sliding. On the contrary, high sliding velocities of 10 Hz and 50 Hz affected fragmentation of the fullerene C60, which resulted in a reduction of the coefficient of friction (COF). During sliding with 1 Hz, the friction reached the highest level with an average COF of 0.59 ± 0.03. The faster relative motion under 1 N normal force gave a lower average COF with 0.39 ± 0.03. The initial fullerene C60 powder formed a thick compressed layer in the tribomechanical loaded zone. As proven by Raman spectroscopy, operating the tribomechanical sliding test at 50 Hz stimulated the re-attraction of fresh C60 fullerene island onto the fragmented layer from outside of the loaded powder regions. The COF was increasing again up to 0.44 ± 0.04 for 1 N normal force and 50 Hz frequency. The fragmentation and decomposition of fullerene C60 with increasing sliding velocity is attributed to thermal heating up during fast relative movement. Raman spectra of the tribomechanical loaded fullerene C60 are compared with Raman spectra from slowly heated up C60 in air and with Raman spectra of laser irradiated fullerene C60.



https://doi.org/10.1016/j.diamond.2022.109036