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- Article (585) (remove)
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Institute
- Fachbereich Chemie und Biotechnologie (585) (remove)
Youssef Hafidi
; Hicham El Hatka
; Dominik Schmitz
; Manuel Krauss
; Jürgen Pettrak
; Markus Biel
; Najim Ittobane
Drought and water shortage are serious problems in many arid and semi-arid regions. This problem is getting worse and even continues in temperate climatic regions due to climate change. To address this problem, the use of biodegradable hydrogels is increasingly important for the application as water-retaining additives in soil. Furthermore, efficient (micro-)nutrient supply can be provided by the use of tailored hydrogels. Biodegradable polyaspartic acid (PASP) hydrogels with different available (1,6-hexamethylene diamine (HMD) and L-lysine (LYS)) and newly developed crosslinkers based on diesters of glycine (GLY) and (di-)ethylene glycol (DEG and EG, respectively) were synthesized and characterized using Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) and regarding their swelling properties (kinetic, absorbency under load (AUL)) as well as biodegradability of PASP hydrogel. Copper (II) and zinc (II), respectively, were loaded as micronutrients in two different approaches: in situ with crosslinking and subsequent loading of prepared hydrogels. The results showed successful syntheses of di-glycine-ester-based crosslinkers. Hydrogels with good water-absorbing properties were formed. Moreover, the developed crosslinking agents in combination with the specific reaction conditions resulted in higher water absorbency with increased crosslinker content used in synthesis (10% vs. 20%). The prepared hydrogels are candidates for water-storing soil additives due to the biodegradability of PASP, which is shown in an exemple. The incorporation of Cu(II) and Zn(II) ions can provide these micronutrients for plant growth.
Kevin Hoffstadt
; Marcell Nikolausz
; Simone Krafft
; Maria Bonatelli
; Vivekanantha Kumar
; Hauke Harms
; Isabel Kuperjans
Optical Fibers as Dosimeter Detectors for Mixed Proton/Neutron Fields - A Biological Dosimeter
(2023)
Jana Niedermeier
; Crystal Penner
; Samuel Usherovich
; Camille Bélanger-Champagne
; Elisabeth Paulßen
; Hoehr Cornelia
In recent years, proton therapy has gained importance as a cancer treatment modality due to its conformality with the tumor and the sparing of healthy tissue. However, in the interaction of the protons with the beam line elements and patient tissues, potentially harmful secondary neutrons are always generated. To ensure that this neutron dose is as low as possible, treatment plans could be created to also account for and minimize the neutron dose. To monitor such a treatment plan, a compact, easy to use, and inexpensive dosimeter must be developed that not only measures the physical dose, but which can also distinguish between proton and neutron contributions. To that end, plastic optical fibers with scintillation materials (Gd₂O₂S:Tb, Gd₂O₂S:Eu, and YVO₄:Eu) were irradiated with protons and neutrons. It was confirmed that sensors with different scintillation materials have different sensitivities to protons and neutrons. A combination of these three scintillators can be used to build a detector array to create a biological dosimeter.
Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems.
Svenja Trapp
; Tom Lammers
; Gokce Engudar
; Cornelia Hoehr
; Antonia G. Denkova
; Elisabeth Paulßen
; Robin M. de Kruijff
Sophia Florence Dellmann
; J. Glorius
; Yu A. Litvinov
; R. Reifarth
; Kafa Al-Khasawneh
; M. Aliotta
; L. Bott
; Benjamin Brückner
; C. G. Bruno
; Ruijiu Chen
; T. Davinson
; T. Dickel
; Iris Dillmann
; D. Dmytriev
; P. Erbacher
; D. Freire-Fernández
; Oliver Forstner
; H. Geissel
; K. Göbel
; Christopher J. Griffin
; R. Grisenti
; Alexandre Gumberidze
; Emma Haettner
; Siegbert Hagmann
; M. Heil
; R. Heß
; P.-M. Hillenbrand
; R. Joseph
; B. Jurado
; Christophor Kozhuharov
; I. Kulikov
; Bastian Löher
; Christoph Langer
; Guy Leckenby
; C. Lederer-Woods
; M. Lestinsky
; S. A. Litvinov
; B. A. Lorenz
; E. Lorenz
; J. Marsh
; Esther Babette Menz
; T. Morgenroth
; N. Petridis
; Jerome Pibernat
; U. Popp
; Athanasios Psaltis
; Shahab Sanjari
; C. Scheidenberger
; M. Sguazzin
; Ragandeep Singh Sidhu
; Uwe Spillmann
; M. Steck
; T. Stöhlker
; A. Surzhykov
; J. A. Swartz
; H. Törnqvist
; L. Varga
; Diego Vescovi
; H. Weick
; M. Weigand
; P. Woods
; Y. Xing
; Taiyo Yamaguchi
Experimental determination of the cross sections of proton capture on radioactive nuclei is extremely difficult. Therefore, it is of substantial interest for the understanding of the production of the p-nuclei. For the first time, a direct measurement of proton-capture cross sections on stored, radioactive ions became possible in an energy range of interest for nuclear astrophysics. The experiment was performed at the Experimental Storage Ring (ESR) at GSI by making use of a sensitive method to measure (p,γ) and (p,n) reactions in inverse kinematics. These reaction channels are of high relevance for the nucleosyn-thesis processes in supernovae, which are among the most violent explosions in the universe and are not yet well understood. The cross section of the ¹¹⁸Te(p,γ) reaction has been measured at energies of 6 MeV/u and 7 MeV/u. The heavy ions interacted with a hydrogen gas jet target. The radiative recombination process of the fully stripped ¹¹⁸Te ions and electrons from the hydrogen target was used as a luminosity monitor. An overview of the experimental method and preliminary results from the ongoing analysis will be presented.
Omnia El Bergui
; Aziz Abouabdillah
; Mohamed Bourioug
; Dominik Schmitz
; Markus Biel
; Abdellah Aboudrare
; Manuel Krauss
; Ahlem Jomaa
; Sebastian Romuli
; Joachim Müller
; Mustapha Fagroud
; Rachid Bouabid
Throughout the last decade, and particularly in 2022, water scarcity has become a critical concern in Morocco and other Mediterranean countries. The lack of rainfall during spring was worsened by a succession of heat waves during the summer. To address this drought, innovative solutions, including the use of new technologies such as hydrogels, will be essential to transform agriculture. This paper presents the findings of a study that evaluated the impact of hydrogel application on onion (Allium cepa) cultivation in Meknes, Morocco. The treatments investigated in this study comprised two different types of hydrogel-based soil additives (Arbovit® polyacrylate and Huminsorb® polyacrylate), applied at two rates (30 and 20 kg/ha), and irrigated at two levels of water supply (100% and 50% of daily crop evapotranspiration; ETc). Two control treatments were included, without hydrogel application and with both water amounts. The experiment was conducted in an open field using a completely randomized design. The results indicated a significant impact of both hydrogel-type dose and water dose on onion plant growth, as evidenced by various vegetation parameters. Among the hydrogels tested, Huminsorb® Polyacrylate produced the most favorable outcomes, with treatment T9 (100%, HP, 30 kg/ha) yielding 70.55 t/ha; this represented an increase of 11 t/ha as compared to the 100% ETc treatment without hydrogel application. Moreover, the combination of hydrogel application with 50% ETc water stress showed promising results, with treatment T4 (HP, 30 kg, 50%) producing almost the same yield as the 100% ETc treatment without hydrogel while saving 208 mm of water.
Nuclear magnetic resonance (NMR) spectrometric methods for the quantitative analysis of pure heparin in crude heparin is proposed. For quantification, a two-step routine was developed using a USP heparin reference sample for calibration and benzoic acid as an internal standard. The method was successfully validated for its accuracy, reproducibility, and precision. The methodology was used to analyze 20 authentic porcine heparinoid samples having heparin content between 4.25 w/w % and 64.4 w/w %. The characterization of crude heparin products was further extended to a simultaneous analysis of these common ions: sodium, calcium, acetate and chloride. A significant, linear dependence was found between anticoagulant activity and assayed heparin content for thirteen heparinoids samples, for which reference data were available. A Diffused-ordered NMR experiment (DOSY) can be used for qualitative analysis of specific glycosaminoglycans (GAGs) in heparinoid matrices and, potentially, for quantitative prediction of molecular weight of GAGs. NMR spectrometry therefore represents a unique analytical method suitable for the simultaneous quantitative control of organic and inorganic composition of crude heparin samples (especially heparin content) as well as an estimation of other physical and quality parameters (molecular weight, animal origin and activity).
The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates.
An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates.