Fakultät Naturwissenschaften
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Publication 3D chromatin maps of a brown alga reveal U/V sex chromosome spatial organization(2024) Liu, Pengfei; Vigneau, Jeromine; Craig, Rory J.; Barrera-Redondo, Josué; Avdievich, Elena; Martinho, Claudia; Borg, Michael; Haas, Fabian B.; Liu, Chang; Coelho, Susana M.; Liu, Pengfei; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Vigneau, Jeromine; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Craig, Rory J.; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Barrera-Redondo, Josué; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Avdievich, Elena; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Martinho, Claudia; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Borg, Michael; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Haas, Fabian B.; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, Germany; Liu, Chang; Institute of Biology, University of Hohenheim, Stuttgart, Germany; Coelho, Susana M.; Department of Algal Development and Evolution, Max Planck Institute for Biology Tübingen, Tübingen, GermanyNuclear three dimensional (3D) folding of chromatin structure has been linked to gene expression regulation and correct developmental programs, but little is known about the 3D architecture of sex chromosomes within the nucleus, and how that impacts their role in sex determination. Here, we determine the sex-specific 3D organization of the model brown alga Ectocarpus chromosomes at 2 kb resolution, by mapping long-range chromosomal interactions using Hi-C coupled with Oxford Nanopore long reads. We report that Ectocarpus interphase chromatin exhibits a non-Rabl conformation, with strong contacts among telomeres and among centromeres, which feature centromere-specific LTR retrotransposons. The Ectocarpus chromosomes do not contain large local interactive domains that resemble TADs described in animals, but their 3D genome organization is largely shaped by post-translational modifications of histone proteins. We show that the sex determining region (SDR) within the U and V chromosomes are insulated and span the centromeres and we link sex-specific chromatin dynamics and gene expression levels to the 3D chromatin structure of the U and V chromosomes. Finally, we uncover the unique conformation of a large genomic region on chromosome 6 harboring an endogenous viral element, providing insights regarding the impact of a latent giant dsDNA virus on the host genome’s 3D chromosomal folding.Publication Analysis of secondary inorganic aerosols over the greater Athens area using the EPISODE–CityChem source dispersion and photochemistry model(2024) Myriokefalitakis, Stelios; Karl, Matthias; Weiss, Kim A.; Karagiannis, Dimitris; Athanasopoulou, Eleni; Kakouri, Anastasia; Bougiatioti, Aikaterini; Liakakou, Eleni; Stavroulas, Iasonas; Papangelis, Georgios; Grivas, Georgios; Paraskevopoulou, Despina; Speyer, Orestis; Mihalopoulos, Nikolaos; Gerasopoulos, EvangelosSecondary inorganic aerosols (SIAs) are major components of fine particulate matter (PM 2.5), having substantial implications for climate and air quality in an urban environment. In this study, a state-of-the-art thermodynamic model has been coupled to the source dispersion and photochemistry city-scale chemistry transport model EPISODE–CityChem, which is able to simulate pollutants at a horizontal resolution of 100m×100m, to determine the equilibrium between the inorganic gas and aerosol phases over the greater Athens area, Greece, for the year 2019. In agreement with in situ observations, sulfate ( SO42-) is calculated to have the highest annual mean surface concentration (2.15 ± 0.88 µgm-3) among SIAs in the model domain, followed by ammonium ( NH4+; 0.58 ± 0.14 µgm-3) and fine nitrate ( NO3-; 0.24 ± 0.22 µgm-3). Simulations denote that NO3-formation strongly depends on the local nitrogen oxide emissions, along with the ambient temperature, the relative humidity, and the photochemical activity. Additionally, we show that anthropogenic combustion sources may have an important impact on the NO3-formation in an urban area. During the cold period, the combined effect of decreased temperature in the presence of non-sea-salt potassium favors the partitioning of HNO3in the aerosol phase in the model, raising the NO3-formation in the area. Overall, this work highlights the significance of atmospheric composition and the local meteorological conditions for the equilibrium distribution of nitrogen-containing semi-volatile compounds and the acidity of inorganic aerosols, especially in urban areas where atmospheric trace elements from natural and anthropogenic sources coexist.Publication The antiviral activity of polyphenols(2025) Burkard, Markus; Piotrowsky, Alban; Leischner, Christian; Detert, Katja; Venturelli, Sascha; Marongiu, Luigi; Burkard, Markus; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, Germany; Piotrowsky, Alban; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, Germany; Leischner, Christian; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, Germany; Detert, Katja; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, Germany; Venturelli, Sascha; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, Germany; Marongiu, Luigi; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, GermanyPolyphenols are secondary metabolites produced by a large variety of plants. These compounds that comprise the class of phenolic acids, stilbenes, lignans, coumarins, flavonoids, and tannins have a wide range of employment, from food production to medical usages. Among the beneficial applications of polyphenols, their antiviral activity is gaining importance due to the increased prevalence of drug‐resistant viruses such as herpes and hepatitis B viruses. In the present review, we provide an overview of the most promising or commonly used antiviral polyphenols and their mechanisms of action focusing on their effects on enveloped viruses of clinical importance (double‐stranded linear or partially double‐stranded circular DNA viruses, negative sense single‐stranded RNA viruses with nonsegmented or segmented genomes, and positive sense single‐stranded RNA viruses). The present work emphasizes the relevance of polyphenols, in particular epigallocatechin‐3‐gallate and resveratrol, as alternative or supportive antivirals. Polyphenols could interfere with virtually all steps of viral infection, from the adsorption to the release of viral particles. The activity of polyphenols against viruses is especially relevant given the risk of widespread outbreaks associated with viruses, remarked by the recent COVID‐19 pandemic.Publication Application of AprX from Pseudomonas paralactis for the improvement of the emulsifying properties of milk, plant and insect protein and estimation of their hydrolysate’s bitter potential(2025) Volk, Veronika; Ewert, Jacob; Longhi, Miriam; Stressler, Timo; Fischer, LutzProtein properties can be modified by selective enzymatic hydrolysis. In this study, the alkaline metalloendopeptidase AprX (Serralysin; EC 3.4.24.40) from Pseudomonas paralactis was used for the tailored hydrolysis of different food proteins resulting in the production of protein hydrolysates with improved emulsifying properties. Sodium caseinate, wheat gluten and buffalo worm protein were used for AprX hydrolysis at 40 °C and pH 8 to cover a spectrum of different protein sources. A maximum degree of hydrolysis (DH) of 13.1 ± 0.2%, 14.2 ± 0.1% and 20.7 ± 0.1% was reached for sodium caseinate, wheat gluten and the worm protein, respectively. The corresponding hydrolysate properties were analyzed regarding their particle size, peptide composition, solubility, viscosity, surface hydrophobicity and interfacial tension. The emulsifying properties were investigated by the oil-droplet size, ζ-potential and stability of emulsions prepared from the hydrolysates. Using partially hydrolyzed sodium caseinate (DH = 10.6%) as an emulsifier lead to an eightfold increase of the emulsion stability (t1/2 = 180 ± 0 min) compared to unhydrolyzed sodium caseinate. The emulsion stability using wheat gluten hydrolysates (DH = 11.9%) was increased 30-fold (t1/2 = 45 ± 5 min). Simultaneously, the solubility of gluten was increased by 60%. Buffalo worm hydrolysates (DH = 14.6%) had a twofold (t1/2 = 85 ± 5 min) increased emulsion stability. In conclusion, AprX can be used to improve the solubility and emulsifying properties of food proteins at a relatively high DH.Publication Assessing the capabilities of 2D fluorescence monitoring in microtiter plates with data-driven modeling for secondary substrate limitation experiments of Hansenula polymorpha(2023) Berg, Christoph; Herbst, Laura; Gremm, Lisa; Ihling, Nina; Paquet-Durand, Olivier; Hitzmann, Bernd; Büchs, JochenBackground: Non-invasive online fluorescence monitoring in high-throughput microbioreactors is a well-established method to accelerate early-stage bioprocess development. Recently, single-wavelength fluorescence monitoring in microtiter plates was extended to measurements of highly resolved 2D fluorescence spectra, by introducing charge-coupled device (CCD) detectors. Although introductory experiments demonstrated a high potential of the new monitoring technology, an assessment of the capabilities and limits for practical applications is yet to be provided. Results: In this study, three experimental sets introducing secondary substrate limitations of magnesium, potassium, and phosphate to cultivations of a GFP-expressing H. polymorpha strain were conducted. This increased the complexity of the spectral dynamics, which were determined by 2D fluorescence measurements. The metabolic responses upon growth limiting conditions were assessed by monitoring of the oxygen transfer rate and extensive offline sampling. Using only the spectral data, subsequently, partial least-square (PLS) regression models for the key parameters of glycerol, cell dry weight, and pH value were generated. For model calibration, spectral data of only two cultivation conditions were combined with sparse offline sampling data. Applying the models to spectral data of six cultures not used for calibration, resulted in an average relative root-mean-square error (RMSE) of prediction between 6.8 and 6.0%. Thus, while demanding only sparse offline data, the models allowed the estimation of biomass accumulation and glycerol consumption, even in the presence of more or less pronounced secondary substrate limitation. Conclusion: For the secondary substrate limitation experiments of this study, the generation of data-driven models allowed a considerable reduction in sampling efforts while also providing process information for unsampled cultures. Therefore, the practical experiments of this study strongly affirm the previously claimed advantages of 2D fluorescence spectroscopy in microtiter plates.Publication Bioaccessibility and anti-inflammatory activity in Caco-2 cells of phytochemicals from industrial by-products of coffee (Coffea arabica L.)(2025) Jiménez-Gutiérrez, Milena; Zielinski, Christian; Esquivel, Patricia; Frank, Jan; Irías-Mata, Andrea; Jiménez-Aspee, FelipeCoffee by-products are rich in nutrients and bioactive compounds in free soluble form and bound to cell wall components. These compounds undergo chemical changes during gastrointestinal digestion, affecting their bioaccessibility and bioactivity. This study is the first to investigate coffee by-products from industrial wet processing to evaluate the impact of simulated gastrointestinal digestion on their phytochemical composition and subsequent anti-inflammatory activity in Caco-2 cells. Digestion significantly reduced the stability and solubility of main compounds; however, digested bioaccessible by-products still exhibited anti-inflammatory properties, reducing IL-6, IL-8, and TNF-α levels. Correlation analysis identified rutin, quercetin-3-glycoside, caffeine and 5-caffeoylquinic acid as strongly linked to cytokine suppression, suggesting key roles and possible synergies. These results highlight the potential of coffee by-products as functional ingredients targeting intestinal inflammation. Future work should confirm in vivo efficacy, optimize extraction at scale, and address regulatory requirements to support industrial application and promote circular economy benefits.Publication Capsaicin attenuates the effect of inflammatory cytokines in a HaCaT cell model for basal keratinocytes(2024) Cervantes Recalde, Maria Fernanda; Schmidt, Jana; Girardi, Cristina; Massironi, Marco; Rechl, Markus Leo; Hans, Joachim; Stuhlmann, Dominik; Somoza, Veronika; Lieder, BarbaraIntroduction: The resolution of the skin’s inflammatory response is only possible if its barrier function is restored. TRPV1 channel activation plays an important role during inflammation but the effect of this activation on the skin barrier under inflammatory conditions has not been clarified. We hypothesize that it could potentially aid the keratinocyte barrier by reducing inflammatory cytokine release and promoting tight junction development. Methods: To explore the role of TRPV1 activation in inflammation, we designed and optimized an in vitro model of keratinocytes with basal epidermal layer characteristics using HaCaT cells and TNFα to induce inflammation. Results: TNFα increased the gene expression of tight junction protein claudin 1 (CLDN1) by at least 2.60 ± 0.16-fold, in a concentration-dependent manner, over a 48 h period. The administration of a capsaicin pre-treatment reduced the CLDN1 expression to 1.51 ± 0.16-fold during the first 6 h after TNFα induction, whereas IL-8 cytokine release was reduced 0.64 ± 0.17-fold. After 48 h, CLDN1 protein levels increased by a factor of 6.57 ± 1.39 compared to cells only treated with TNFα. Discussion: These results suggest that activation of TRPV1 by capsaicin can potentiate the increase in CLDN1 expression and CLDN1 protein synthesis induced by TNFα in cultured keratinocytes, while reducing the release of IL-8.Publication Characterization of the major odor-active compounds in fresh rhizomes and leaves of Houttuynia cordata by comparative aroma extract dilution analysis(2025) Xu, Zhenli; Liu, Jing; Kreissl, Johanna; Oellig, Claudia; Vetter, Walter; Steinhaus, Martin; Frank, Stephanie; Rodov, VictorHouttuynia cordata is a culinary herb from Asia. Its edible rhizomes and leaves have a fishy aroma, the molecular background of which was unknown. A comparative aroma extract dilution analysis applied to fresh rhizomes and leaves resulted in 44 and 41 odorants, respectively, 38 of which were present with FD factors ≥1 in both samples. The odorant with the highest FD factors, whether in the rhizomes or leaves, was identified as metallic, soapy, fishy smelling 3-oxododecanal. Toward clarifying its tautomeric composition, quantum calculations suggested a predominance of the enol forms in the plant. However, the form perceived at the sniffing port during GC–O remained unclear.Publication Chlorinated paraffins in hinges of kitchen appliances(2021) Sprengel, Jannik; Vetter, WalterChlorinated paraffins (CPs) are anthropogenic pollutants of growing environmental concern. These highly complex mixtures of thousands of homologs and congeners are usually applied as additives in lubricants or as flame retardants and plasticizers in polymers and paints. Recent studies indicated the presence of high amounts of CPs in the kitchen environment whose sources could not be unequivocally identified. One option was the use of CPs as or in lubricants of hinges. To test this hypothesis, we performed wipe tests on lubricants on 29 hinges of different types of kitchen appliances (refrigerators, baking ovens, dishwashers, freezers, microwave oven, pasta machine, food processor, steam cooker) and analyzed them for short-chain CPs (SCCPs) and medium-chain CPs (MCCPs). CPs were detected in 21 samples (72%). Per wipe, SCCP concentrations ranged between 0.02 and 10 µg (median 0.23 µg), while MCCPs ranged from 0.09 to 750 µg (median 1.0 µg). Highest MCCP amounts (380 and 750 µg per wipe, respectively) were determined in new and unused appliances. A medium correlation between SCCP content and appliance age was observed, but no additional statistic correlation between SCCP/MCCP amount and appliance type or manufacturer could be observed. CPs released from hinges by volatilization, abrasion, and cleaning processes could enter the environment and come in contact with persons living in the corresponding households.Publication Comparison of binding properties of a laccase-treated pea protein-sugar beet pectin mixture with methylcellulose in a bacon-type meat analogue(2022) Moll, Pascal; Salminen, Hanna; Stadtmueller, Lucie; Schmitt, Christophe; Weiss, JochenA bacon-type meat analogue consists of different structural layers, such as textured protein and a fat mimetic. To obtain a coherent and appealing product, a suitable binder must glue those elements together. A mixture based on pea protein and sugar beet pectin (r = 2:1, 25% w/w solids, pH 6) with and without laccase addition and a methylcellulose hydrogel (6% w/w) serving as benchmark were applied as binder between textured protein and a fat mimetic. A tensile strength test, during which the layers were torn apart, was performed to measure the binding ability. The pea protein–sugar beet pectin mixture without laccase was viscoelastic and had medium and low binding strength at 25 °C (F ≤ 3.5 N) and 70 °C (F ≈ 1.0 N), respectively. The addition of laccase solidified the mixture and increased binding strength at 25 °C (F ≥ 4.0 N) and 70 °C (F ≈ 2.0 N), due to covalent bonds within the binder and between the binder and the textured protein or the fat mimetic layers. Generally, the binding strength was higher when two textured protein layers were glued together. The binding properties of methylcellulose hydrogel was low (F ≤ 2.0 N), except when two fat mimetic layers were bound due to hydrophobic interactions becoming dominant. The investigated mixed pectin–pea protein system is able serve as a clean-label binder in bacon-type meat analogues, and the application in other products seems promising.Publication Comparison of whole genomes of tick-borne encephalitis virus from mountainous alpine regions and regions with a lower altitude(2021) Lemhöfer, G.; Chitimia-Dobler, L.; Dobler, G.; Bestehorn-Willmann, MalenaTick-borne encephalitis (TBE) has been a notifiable disease in Germany since 2001. Its causative agent, the TBE virus (TBEV), is the most important arbovirus in Europe and Northern Asia. The illness, caused by the European Subtype usually displays flu-like symptoms, but can result in sequelae and, in 2 % of all cases, in death. Over the last few decades, the virus has spread into new habitats, such as higher altitudes in the Alpine region. For this study, it was hypothesized that the environmental challenges that the virus might be exposed to at such altitudes could lead to the selection of viral strains with a higher resilience to such environmental factors. To determine whether strains identified at higher altitudes possessed different genetic traits compared to viruses from lower altitudes, an analysis of viral genomes from higher Alpine altitudes (> 500 m above sea level) (n = 5) and lower altitudes (< 500 m above sea level) (n = 4) was performed. No common phylogenetic ancestry or shared amino acid substitutions could be identified that differentiated the alpine from the lowland viral strains. These findings support the idea of many individual introductions of TBEV into the alpine region and the establishment of foci due to non-viral specific factors such as favorable conditions for vector species and host animals due to climate change.Publication Consumption of antioxidant-rich “Cerrado” cashew pseudofruit affects hepatic gene expression in obese C57BL/6J high fat-fed mice(2022) Egea, Mariana Buranelo; Pierce, Gavin; Park, Si-Hong; Lee, Sang-In; Heger, Fabienne; Shay, NeilThe pseudofruit of A. othonianum Rizzini, “Cerrado” cashew pulp, has been described as rich in flavonoids, phenolic compounds, and vitamin C. The objective of this work was to evaluate the beneficial health effects seen with the addition of “Cerrado” cashew pulp (CP) to an obesogenic high fat diet provided to C57BL/6J male mice. In week 9, the HF-fed group had a significantly higher baseline glucose concentration than the LF- or HF+CP-fed groups. In RNAseq analysis, 4669 of 5520 genes were found to be differentially expressed. Among the genes most upregulated with the ingestion of the CP compared to HF were Ph1da1, SLc6a9, Clec4f, and Ica1 which are related to glucose homeostasis; Mt2 that may be involved steroid biosynthetic process; and Ciart which has a role in the regulation of circadian rhythm. Although “Cerrado” CP intake did not cause changes in the food intake or body weight of fed mice with HF diet, carbohydrate metabolism appeared to be improved based on the observed changes in gene expression.Publication Consumption of yeast-fermented wheat and rye breads increases colitis and mortality in a mouse model of colitis(2022) Zimmermann, Julia; De Fazio, Luigia; Kaden-Volynets, Valentina; Hitzmann, Bernd; Bischoff, Stephan C.Background: Cereals are known to trigger for wheat allergy, celiac disease and non-celiac wheat sensitivity (NCWS). Inflammatory processes and intestinal barrier impairment are suspected to be involved in NCWS, although the molecular triggers are unclear. Aims: We were interested if different bread types influence inflammatory processes and intestinal barrier function in a mouse model of inflammatory bowel disease. Methods: Epithelial caspase-8 gene knockout (Casp8 ΔIEC ) and control (Casp8 fl ) mice were randomized to eight groups, respectively. The groups received different diets for 28 days (gluten-free diet, gluten-rich diet 5 g%, or different types of bread at 50 g%). Breads varied regarding grain, milling and fermentation. All diets were isocaloric. Results: Regardless of the diet, Casp8 ΔIEC mice showed pronounced inflammation in colon compared to ileum, whereas Casp8 fl mice were hardly inflamed. Casp8 fl mice could tolerate all bread types. Especially yeast fermented rye and wheat bread from superfine flour but not pure gluten challenge increased colitis and mortality in Casp8 ΔIEC mice. Hepatic expression of lipopolysaccharide-binding protein and colonic expression of tumor necrosis factor-α genes were inversely related to survival. The bread diets, but not the gluten-rich diet, also decreased colonic tight junction expression to variable degrees, without clear association to survival and inflammation. Conclusions: Bread components, especially those from yeast-fermented breads from wheat and rye, increase colitis and mortality in Casp8 ΔIEC mice highly susceptible to intestinal inflammation, whereas control mice can tolerate all types of bread without inflammation. Yet unidentified bread components other than gluten seem to play the major role.Publication CortexVR: Immersive analysis and training of cognitive executive functions of soccer players using virtual reality and machine learning(2022) Krupitzer, Christian; Naber, Jens; Stauffert, Jan-Philipp; Mayer, Jan; Spielmann, Jan; Ehmann, Paul; Boci, Noel; Bürkle, Maurice; Ho, André; Komorek, Clemens; Heinickel, Felix; Kounev, Samuel; Becker, Christian; Latoschik, Marc ErichGoal: This paper presents an immersive Virtual Reality (VR) system to analyze and train Executive Functions (EFs) of soccer players. EFs are important cognitive functions for athletes. They are a relevant quality that distinguishes amateurs from professionals. Method: The system is based on immersive technology, hence, the user interacts naturally and experiences a training session in a virtual world. The proposed system has a modular design supporting the extension of various so-called game modes. Game modes combine selected game mechanics with specific simulation content to target particular training aspects. The system architecture decouples selection/parameterization and analysis of training sessions via a coaching app from an Unity3D-based VR simulation core. Monitoring of user performance and progress is recorded by a database that sends the necessary feedback to the coaching app for analysis. Results: The system is tested for VR-critical performance criteria to reveal the usefulness of a new interaction paradigm in the cognitive training and analysis of EFs. Subjective ratings for overall usability show that the design as VR application enhances the user experience compared to a traditional desktop app; whereas the new, unfamiliar interaction paradigm does not negatively impact the effort for using the application. Conclusion: The system can provide immersive training of EF in a fully virtual environment, eliminating potential distraction. It further provides an easy-to-use analyzes tool to compare user but also an automatic, adaptive training mode.Publication Decoding the geography of natural TBEV microfoci in German: a geostatistical approach based on land-use patterns and climatological conditions(2022) Borde, Johannes P.; Glaser, Rüdiger; Braun, Klaus; Riach, Nils; Hologa, Rafael; Kaier, Klaus; Chitimia-Dobler, Lidia; Dobler, GerhardBackground: Tickborne-encephalitis (TBE) is a potentially life-threating neurological disease that is mainly transmitted by ticks. The goal of the present study is to analyze the potential uniform environmental patterns of the identified TBEV microfoci in Germany. The results are used to calculate probabilities for the present distribution of TBEV microfoci in Germany based on a geostatistical model. Methods: We aim to consider the specification of environmental characteristics of locations of TBEV microfoci detected in Germany using open access epidemiological, geographical and climatological data sources. We use a two-step geostatistical approach, where in a first step, the characteristics of a broad set of environmental variables between the 56 TBEV microfoci and a control or comparator set of 3575 sampling points covering Germany are compared using Fisher’s Exact Test. In the second step, we select the most important variables, which are then used in a MaxEnt distribution model to calculate a high resolution (400 × 400 m) probability map for the presence of TBEV covering the entire area of Germany. Results: The findings from the MaxEnt prediction model indicate that multi annual actual evapotranspiration (27.0%) and multi annual hot days (22.5%) have the highest contribution to our model. These two variables are followed by four additional variables with a lower, but still important, explanatory influence: Land cover classes (19.6%), multi annual minimum air temperature (14.9%), multi annual sunshine duration (9.0%), and distance to coniferous and mixed forest border (7.0%). Conclusions: Our findings are based on defined TBEV microfoci with known histories of infection and the repeated confirmation of the virus in the last years, resulting in an in-depth high-resolution model/map of TBEV microfoci in Germany. Multi annual actual evapotranspiration (27%) and multi annual hot days (22.5%) have the most explanatory power in our model. The results may be used to tailor specific regional preventive measures and investigations.Publication Development and validation of a novel enzyme-linked immunosorbent assay for the differentiation of tick-borne encephalitis infections caused by different virus subtypes(2025) Freimane, Zane; Dobler, Gerhard; Chitimia-Dobler, Lidia; Karelis, Guntis; Girl, Philipp; Kuzmane, Sanita; Savicka, Oksana; Erber, Wilhelm; Zavadska, DaceObjectives: Tick-borne encephalitis (TBE) is an infection caused by the tick-borne encephalitis virus (TBEV) that can lead to symptoms of central nervous system inflammation. There are five subtypes of TBEV, three of which – European, Siberian and Far Eastern – occur in Europe. As it is thought that different subtype infections exhibit varying clinical courses and outcomes, serological differentiation of the virus subtypes is clearly important. However, to date, this has proved difficult to achieve. Methods: An ELISA format was developed based on TBE virus NS1 antigen against the European, Siberian and Far Eastern subtype. The three NS1 antigens were biotechnologically produced in a human cell line and used for ELISA coating. Sera from German (European subtype) and Russian (Siberian and/or Far Eastern subtypes) TBE patients with positive TBEV IgG were used to test the reactivity against these three NS1 antigens. Results: Testing of 23 German and 32 Russian TBEV IgG-positive sera showed that the ELISA was able to differentiate between TBEV European subtype and TBEV Siberian and Far Eastern subtype infections. Conclusions: In geographical areas where two or more TBEV subtype infections can occur, the NS1-IgG ELISA developed here constitutes an important diagnostic tool to differentiate between European subtype infections and Siberian/Far Eastern subtype infections and to use the new assay for epidemiological studies to clarify the importance of particular subtype infections in an area. Consequently, it may help to better describe and anticipate the clinical courses and outcomes of particular TBEV subtype infections.Publication A diamine oxidase from Glutamicibacter halophytocola for the degradation of histamine and tyramine in foods(2025) Kettner, Lucas; Freund, Alexander; Bechtel, Anna; Costa-Catala, Judit; Fischer, Lutz; Kettner, Lucas; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599 Stuttgart, Germany; Freund, Alexander; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599 Stuttgart, Germany; Bechtel, Anna; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599 Stuttgart, Germany; Costa-Catala, Judit; Departament de Nutrició, Ciències de l’Alimentació i Gastronomia, Campus de l’Alimentació de Torribera, Universitat de Barcelona, Av. Prat de la Riba 171, 08921 Santa Coloma de Gramenet, Spain; Fischer, Lutz; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599 Stuttgart, GermanyA novel diamine oxidase (DAO) was discovered in the bacterium Glutamicibacter halophytocola (DAO-GH). The gene of DAO-GH was integrated into the genome of the yeast Komagataella phaffii and recombinantly produced under control of the methanol-inducible AOX1 promoter in a bioreactor cultivation. A high DAO activity of 70.2 ± 5.2 µkat/Lculture (5.25 ± 0.22 µkat/gprotein) was yielded after 90 h of cultivation. The DAO-GH was partially purified by the polyethyleneimine precipitation of nucleic acids, fractionated ammonium sulfate precipitation and hydrophobic interaction chromatography, resulting in a specific DAO activity of 19.7 µkat/gProtein. The DAO-GH was then biochemically investigated regarding its potential for histamine and tyramine degradation in fermented foods and the human small intestine. Interestingly, the DAO-GH showed activity even at a low pH of 5 and low temperature of 6 °C. Both histamine and tyramine were effectively degraded and DAO-GH showed especially very high affinity towards tyramine (Km of 0.009 mM). The DAO-GH was shown to be capable of degrading around 20% of the initially applied histamine in tuna paste (pH 5.6) at 5 °C within 24 h and completely degraded the histamine in a simulated intestinal fluid within 1.5 h in bioconversion experiments. The DAO-GH was spray-dried for the production of a storable enzyme preparation. Only around 17% of activity were lost in this process and the DAO-GH remained stable at room temperature for at least 3 months. The discovery of this DAO with its very advantageous biochemical properties allows the preparation of histamine-reduced or -free fermented foods by a simple enzymatic treatment or the treatment of histamine intolerance symptoms as a dietary supplement or medicine.Publication Echinococcus multilocularis and other taeniid metacestodes of muskrats in Luxembourg: prevalence, risk factors, parasite reproduction, and genetic diversity(2022) Martini, Matilde; Dumendiak, Sonja; Gagliardo, Anna; Ragazzini, Francesco; La Rosa, Letizia; Giunchi, Dimitri; Thielen, Frank; Romig, Thomas; Massolo, Alessandro; Wassermann, MarionMuskrats (Ondatra zibethicus) are competent intermediate hosts for Echinococcus multilocularis, are frequently infected with this zoonotic cestode, and have even been proposed as a target species to monitor endemicity levels of this parasite. However, their contribution to maintaining the parasitic lifecycle is still unclear. To obtain data on infection frequency and reproductive potential, 280 muskrats from the Grand Duchy of Luxembourg were examined for cestode larvae in the years 2013–2017. Based on morphological and molecular identification, Echinococcus multilocularis was found at a prevalence of 14.6%. Other metacestodes were Hydatigera kamiyai, with a prevalence of 45.7%, Taenia martis with 8.9%, Taenia polyacantha with 5.0%, and Versteria mustelae, which was found in 0.7% of all muskrats. More than 80% of E. multilocularis-infected muskrats contained fertile metacestodes with a mean number of >300,000 (and up to 1,609,816) protoscoleces, which is by far the highest reproductive potential known from any intermediate host species in Europe. Temporal analysis of E. multilocularis prevalence within the study period (and in comparison with earlier data) strongly indicates a robust increase in the studied area. Host age seemed to be an important risk factor for infection, as well as co-infections with Hydatigera kamiyai. A preference for the right medial lobe of the liver as the location of E. multilocularis metacestode was observed. Intraspecific genetic variation among 89 discrete E. multilocularis metacestodes was non-existent based on 300–1590 bp sections of cox1. This is a stark contrast to H. kamiyai, of which nine haplotypes were found on a short 318 bp section of cox1, resulting in genetic diversity in the small country of Luxembourg at a similar level than previously reported from large stretches of Europe and northern Asia.Publication Echinococcus multilocularis and other zoonotic helminths in red foxes (Vulpes vulpes) from a southern German hotspot for human alveolar echinococcosis(2023) Schneider, Carina; Kratzer, Wolfgang; Binzberger, Andreas; Schlingeloff, Patrycja; Baumann, Sven; Romig, Thomas; Schmidberger, JulianBackground: We describe the spatial distribution of Echinococcus multilocularis in its main definitive host, the red fox, and the distribution of human cases of alveolar echinococcosis (AE) within a highly endemic focus in southern Germany (13.7–19.9/100,000 in 1992–2018). Human cases were unequally distributed within the endemicity focus. The purpose of the study was to test whether this is reflected in the small-scale distribution of E. multilocularis in foxes. Methods: Three areas with contrasting numbers of human cases were selected within the counties of Ravensburg and Alb-Donau, Baden-Württemberg, Germany. From 2018 to 2020, a total of 240 fox carcasses were obtained from traditional hunters in these areas. Carcasses were necropsied and examined for the presence of intestinal helminths. The statistical analysis was performed with SAS version 9.4, and the geo-mapping with QGIS version 3.16.0 Hannover. Results: The prevalence of E. multilocularis in foxes was 44/106 (41.5%) in area I (commune Leutkirch and environs), 30/59 (50.8%) in area II (commune Isny and environs), and 31/75 (41.3%) in area III (commune Ehingen and environs). From 1992 to 2018, a total of nine human cases of alveolar echinococcosis were recorded in area I, five cases were recorded in study area III, and no cases were recorded in area II. No statistically significant differences between the areas were observed ( P > 0.05) for intestinal infections with E. multilocularis , and no apparent spatial correlation with the small-scale distribution of human cases was found. Concerning other zoonotic helminths, Toxocara spp. were equally common, with prevalence of 38.7%, 47.4% and 48.0%, respectively, while the frequency of Alaria alata varied among the study areas (0.0–9.4%), probably reflecting the specific habitat requirements for the establishment of its complex life cycle. Conclusions: Echinococcus multilocularis is highly prevalent in foxes in all the studied areas. The varying number of human AE cases within these areas should therefore be caused by factors other than the intensity of parasite transmission in foxes.Publication Editorial: Methods in molecular innate immunity: 2022(2025) Fritz, Jörg H.; Kufer, Thomas A.
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