TY - JOUR A1 - Sagu Tchewonpi, Sorel A1 - Huschek, Gerd A1 - Waldbach Braga, Tess A1 - Rackiewicz, Michal A1 - Homann, Thomas A1 - Rawel, Harshadrai Manilal T1 - Design of Experiment (DoE) for Optimization of HPLC Conditions for the Simultaneous Fractionation of Seven α-Amylase/Trypsin Inhibitors from Wheat (Triticum aestivum L.) JF - Processes : open access journal N2 - Wheat alpha-amylase/trypsin inhibitors remain a subject of interest considering the latest findings showing their implication in wheat-related non-celiac sensitivity (NCWS). Understanding their functions in such a disorder is still unclear and for further study, the need for pure ATI molecules is one of the limiting problems. In this work, a simplified approach based on the successive fractionation of ATI extracts by reverse phase and ion exchange chromatography was developed. ATIs were first extracted from wheat flour using a combination of Tris buffer and chloroform/methanol methods. The separation of the extracts on a C18 column generated two main fractions of interest F1 and F2. The response surface methodology with the Doehlert design allowed optimizing the operating parameters of the strong anion exchange chromatography. Finally, the seven major wheat ATIs namely P01083, P17314, P16850, P01085, P16851, P16159, and P83207 were recovered with purity levels (according to the targeted LC-MS/MS analysis) of 98.2 ± 0.7; 98.1 ± 0.8; 97.9 ± 0.5; 95.1 ± 0.8; 98.3 ± 0.4; 96.9 ± 0.5, and 96.2 ± 0.4%, respectively. MALDI-TOF-MS analysis revealed single peaks in each of the pure fractions and the mass analysis yielded deviations of 0.4, 1.9, 0.1, 0.2, 0.2, 0.9, and 0.1% between the theoretical and the determined masses of P01083, P17314, P16850, P01085, P16851, P16159, and P83207, respectively. Overall, the study allowed establishing an efficient purification process of the most important wheat ATIs. This paves the way for further in-depth investigation of the ATIs to gain more knowledge related to their involvement in NCWS disease and to allow the absolute quantification in wheat samples. KW - wheat KW - α-amylase/trypsin inhibitors KW - fractionation KW - purification KW - reversed-phase chromatography KW - ion-exchange chromatography KW - design of experiment KW - LC–MS/MS KW - MALDI-TOF-MS Y1 - 2022 U6 - https://doi.org/10.3390/pr10020259 SN - 2227-9717 VL - 10 SP - 1 EP - 18 PB - MDPI CY - Basel, Schweiz ET - 2 ER - TY - JOUR A1 - Sallat, Stephan T1 - Sprachstörungen in Kita und Schule BT - Alltags- und unterrichtsintegrierte Intervention und Adaption JF - Spektrum Patholinguistik 15 Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-572360 SN - 978-3-86956-542-2 SN - 1866-9433 SN - 1866-9085 IS - 15 SP - 47 EP - 70 PB - Universitätsverlag Potsdam CY - Potsdam ER - TY - JOUR A1 - Sarabadani, Jalal A1 - Metzler, Ralf A1 - Ala-Nissila, Tapio T1 - Driven polymer translocation into a channel: Isoflux tension propagation theory and Langevin dynamics simulations JF - Physical Review Research N2 - Isoflux tension propagation (IFTP) theory and Langevin dynamics (LD) simulations are employed to study the dynamics of channel-driven polymer translocation in which a polymer translocates into a narrow channel and the monomers in the channel experience a driving force fc. In the high driving force limit, regardless of the channel width, IFTP theory predicts τ ∝ f βc for the translocation time, where β = −1 is the force scaling exponent. Moreover, LD data show that for a very narrow channel fitting only a single file of monomers, the entropic force due to the subchain inside the channel does not play a significant role in the translocation dynamics and the force exponent β = −1 regardless of the force magnitude. As the channel width increases the number of possible spatial configurations of the subchain inside the channel becomes significant and the resulting entropic force causes the force exponent to drop below unity. Y1 - 2022 U6 - https://doi.org/10.1103/PhysRevResearch.4.033003 SN - 2643-1564 VL - 4 SP - 033003-1 EP - 033003-14 PB - American Physical Society CY - College Park, Maryland, USA ET - 3 ER - TY - JOUR A1 - Schjeide, Brit-Maren A1 - Schenke, Maren A1 - Seeger, Bettina A1 - Püschel, Gerhard T1 - Validation of a novel double control quantitative copy number PCR method to quantify off-target transgene integration after CRISPR-induced DNA modification JF - Methods and protocols : M&Ps N2 - In order to improve a recently established cell-based assay to assess the potency of botulinum neurotoxin, neuroblastoma-derived SiMa cells and induced pluripotent stem-cells (iPSC) were modified to incorporate the coding sequence of a reporter luciferase into a genetic safe harbor utilizing CRISPR/Cas9. A novel method, the double-control quantitative copy number PCR (dc-qcnPCR), was developed to detect off-target integrations of donor DNA. The donor DNA insertion success rate and targeted insertion success rate were analyzed in clones of each cell type. The dc-qcnPCR reliably quantified the copy number in both cell lines. The probability of incorrect donor DNA integration was significantly increased in SiMa cells in comparison to the iPSCs. This can possibly be explained by the lower bundled relative gene expression of a number of double-strand repair genes (BRCA1, DNA2, EXO1, MCPH1, MRE11, and RAD51) in SiMa clones than in iPSC clones. The dc-qcnPCR offers an efficient and cost-effective method to detect off-target CRISPR/Cas9-induced donor DNA integrations. KW - CRISPR editing validation KW - copy number analyses KW - homology-directed repair KW - homologous recombination deficiency Y1 - 2022 U6 - https://doi.org/10.3390/mps5030043 SN - 2409-9279 VL - 5 IS - 3 SP - 1 EP - 14 PB - MDPI CY - Basel, Schweiz ER - TY - JOUR A1 - Schlör, Anja A1 - Hirschberg, Stefan A1 - Ben Amor, Ghada A1 - Meister, Toni Luise A1 - Arora, Prerna A1 - Pöhlmann, Stefan A1 - Hoffmann, Markus A1 - Pfänder, Stephanie A1 - Eddin, Omar Kamal A1 - Kamhieh-Milz, Julian A1 - Hanack, Katja T1 - SARS-CoV-2 neutralizing camelid heavy-chain-only antibodies as powerful tools for diagnostic and therapeutic applications JF - Frontiers in Immunology N2 - Introduction: The ongoing COVID-19 pandemic situation caused by SARS-CoV-2 and variants of concern such as B.1.617.2 (Delta) and recently, B.1.1.529 (Omicron) is posing multiple challenges to humanity. The rapid evolution of the virus requires adaptation of diagnostic and therapeutic applications. Objectives: In this study, we describe camelid heavy-chain-only antibodies (hcAb) as useful tools for novel in vitro diagnostic assays and for therapeutic applications due to their neutralizing capacity. Methods: Five antibody candidates were selected out of a naïve camelid library by phage display and expressed as full length IgG2 antibodies. The antibodies were characterized by Western blot, enzyme-linked immunosorbent assays, surface plasmon resonance with regard to their specificity to the recombinant SARS-CoV-2 Spike protein and to SARS-CoV-2 virus-like particles. Neutralization assays were performed with authentic SARS-CoV-2 and pseudotyped viruses (wildtype and Omicron). Results: All antibodies efficiently detect recombinant SARS-CoV-2 Spike protein and SARS-CoV-2 virus-like particles in different ELISA setups. The best combination was shown with hcAb B10 as catcher antibody and HRP-conjugated hcAb A7.2 as the detection antibody. Further, four out of five antibodies potently neutralized authentic wildtype SARS-CoV-2 and particles pseudotyped with the SARS-CoV-2 Spike proteins of the wildtype and Omicron variant, sublineage BA.1 at concentrations between 0.1 and 0.35 ng/mL (ND50). Conclusion: Collectively, we report novel camelid hcAbs suitable for diagnostics and potential therapy. KW - camelid heavy-chain-only antibodies KW - single domain antibodies KW - nanobodies KW - SARS-CoV-2 KW - neutralization KW - Omicron Y1 - 2022 U6 - https://doi.org/10.3389/fimmu.2022.930975 SN - 1664-3224 SP - 1 EP - 14 PB - Frontiers Media SA CY - Lausanne, Schweiz ER - TY - JOUR A1 - Schmidt, Johanna A1 - Eikerling, Maren A1 - Cholin, Joana T1 - Klinische Marker in der Spontansprache früh-sukzessiv bilingualer Kinder mit Deutsch als Zweitsprache JF - Spektrum Patholinguistik 15 Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-572413 SN - 978-3-86956-542-2 SN - 1866-9433 SN - 1866-9085 IS - 15 SP - 135 EP - 150 PB - Universitätsverlag Potsdam CY - Potsdam ER - TY - JOUR A1 - Schmidt, Lena Katharina A1 - Francke, Till A1 - Rottler, Erwin A1 - Blume, Theresa A1 - Schöber, Johannes A1 - Bronstert, Axel T1 - Suspended sediment and discharge dynamics in a glaciated alpine environment BT - identifying crucial areas and time periods on several spatial and temporal scales in the Ötztal, Austria JF - Earth surface dynamics N2 - Glaciated high-alpine areas are fundamentally altered by climate change, with well-known implications for hydrology, e.g., due to glacier retreat, longer snow-free periods, and more frequent and intense summer rainstorms. While knowledge on how these hydrological changes will propagate to suspended sediment dynamics is still scarce, it is needed to inform mitigation and adaptation strategies. To understand the processes and source areas most relevant to sediment dynamics, we analyzed discharge and sediment dynamics in high temporal resolution as well as their patterns on several spatial scales, which to date few studies have done. We used a nested catchment setup in the Upper Ötztal in Tyrol, Austria, where high-resolution (15 min) time series of discharge and suspended sediment concentrations are available for up to 15 years (2006–2020). The catchments of the gauges in Vent, Sölden and Tumpen range from 100 to almost 800 km2 with 10 % to 30 % glacier cover and span an elevation range of 930 to 3772 m a.s.l. We analyzed discharge and suspended sediment yields (SSY), their distribution in space, their seasonality and spatial differences therein, and the relative importance of short-term events. We complemented our analysis by linking the observations to satellite-based snow cover maps, glacier inventories, mass balances and precipitation data. Our results indicate that the areas above 2500 m a.s.l., characterized by glacier tongues and the most recently deglaciated areas, are crucial for sediment generation in all sub-catchments. This notion is supported by the synchronous spring onset of sediment export at the three gauges, which coincides with snowmelt above 2500 m but lags behind spring discharge onsets. This points at a limitation of suspended sediment supply as long as the areas above 2500 m are snow-covered. The positive correlation of annual SSY with glacier cover (among catchments) and glacier mass balances (within a catchment) further supports the importance of the glacier-dominated areas. The analysis of short-term events showed that summer precipitation events were associated with peak sediment concentrations and yields but on average accounted for only 21 % of the annual SSY in the headwaters. These results indicate that under current conditions, thermally induced sediment export (through snow and glacier melt) is dominant in the study area. Our results extend the scientific knowledge on current hydro-sedimentological conditions in glaciated high-alpine areas and provide a baseline for studies on projected future changes in hydro-sedimentological system dynamics. Y1 - 2022 U6 - https://doi.org/10.5194/esurf-10-653-2022 SN - 2196-632X SN - 2196-6311 VL - 10 IS - 3 SP - 653 EP - 669 PB - Copernicus Publications CY - Göttingen ER - TY - JOUR A1 - Schmitz-Antonischki, Dorit A1 - Heide, Judith A1 - Netzebandt, Jonka T1 - Therapie von Wortabrufstörungen mit der App LingoTalk bei einer Patientin mit Aphasie BT - eine Therapiestudie JF - Spektrum Patholinguistik 15 Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-572433 SN - 978-3-86956-542-2 SN - 1866-9433 SN - 1866-9085 IS - 15 SP - 157 EP - 171 PB - Universitätsverlag Potsdam CY - Potsdam ER - TY - JOUR A1 - Schäfer, Marjänn Helena A1 - Kakularam, Kumar Reddy A1 - Reisch, Florian A1 - Rothe, Michael A1 - Stehling, Sabine A1 - Heydeck, Dagmar A1 - Püschel, Gerhard Paul A1 - Kuhn, Hartmut T1 - Male Knock-in Mice Expressing an Arachidonic Acid Lipoxygenase 15B (Alox15B) with Humanized Reaction Specificity Are Prematurely Growth Arrested When Aging JF - Biomedicines N2 - Mammalian arachidonic acid lipoxygenases (ALOXs) have been implicated in cell differentiation and in the pathogenesis of inflammation. The mouse genome involves seven functional Alox genes and the encoded enzymes share a high degree of amino acid conservation with their human orthologs. There are, however, functional differences between mouse and human ALOX orthologs. Human ALOX15B oxygenates arachidonic acid exclusively to its 15-hydroperoxy derivative (15S-HpETE), whereas 8S-HpETE is dominantly formed by mouse Alox15b. The structural basis for this functional difference has been explored and in vitro mutagenesis humanized the reaction specificity of the mouse enzyme. To explore whether this mutagenesis strategy may also humanize the reaction specificity of mouse Alox15b in vivo, we created Alox15b knock-in mice expressing the arachidonic acid 15-lipoxygenating Tyr603Asp+His604Val double mutant instead of the 8-lipoxygenating wildtype enzyme. These mice are fertile, display slightly modified plasma oxylipidomes and develop normally up to an age of 24 weeks. At later developmental stages, male Alox15b-KI mice gain significantly less body weight than outbred wildtype controls, but this effect was not observed for female individuals. To explore the possible reasons for the observed gender-specific growth arrest, we determined the basic hematological parameters and found that aged male Alox15b-KI mice exhibited significantly attenuated red blood cell parameters (erythrocyte counts, hematocrit, hemoglobin). Here again, these differences were not observed in female individuals. These data suggest that humanization of the reaction specificity of mouse Alox15b impairs the functionality of the hematopoietic system in males, which is paralleled by a premature growth arrest. KW - eicosanoids KW - lipid peroxidation KW - oxidative stress KW - polyenoic fatty acids KW - erythropoiesis Y1 - 2022 U6 - https://doi.org/10.3390/biomedicines10061379 SN - 2227-9059 VL - 10 SP - 1 EP - 22 PB - MDPI CY - Basel, Schweiz ET - 6 ER - TY - JOUR A1 - Smirnov, Lev A. A1 - Bolotov, Maxim A1 - Bolotov, Dmitri A1 - Osipov, Grigory V. A1 - Pikovsky, Arkady T1 - Finite-density-induced motility and turbulence of chimera solitons JF - New Journal of Physics N2 - We consider a one-dimensional oscillatory medium with a coupling through a diffusive linear field. In the limit of fast diffusion this setup reduces to the classical Kuramoto–Battogtokh model. We demonstrate that for a finite diffusion stable chimera solitons, namely localized synchronous domain in an infinite asynchronous environment, are possible. The solitons are stable also for finite density of oscillators, but in this case they sway with a nearly constant speed. This finite-density-induced motility disappears in the continuum limit, as the velocity of the solitons is inverse proportional to the density. A long-wave instability of the homogeneous asynchronous state causes soliton turbulence, which appears as a sequence of soliton mergings and creations. As the instability of the asynchronous state becomes stronger, this turbulence develops into a spatio-temporal intermittency. KW - chimera KW - soliton KW - finite-size effects Y1 - 2022 U6 - https://doi.org/10.1088/1367-2630/ac63d9 SN - 1367-2630 VL - 24 PB - IOP CY - London ER -