TY - GEN A1 - Rosellini, Matteo A1 - Schulze, Alicia A1 - Omer, Ejlal A. A1 - Ali, Nadeen T. A1 - Marini, Federico A1 - Küpper, Jan-Heiner A1 - Efferth, Thomas T1 - The effect of plastic-related compounds on transcriptome-wide gene expression on CYP2C19-overexpressing HepG2 cells T2 - Molecules N2 - In recent years, plastic and especially microplastic in the oceans have caused huge problems to marine flora and fauna. Recently, such particles have also been detected in blood, breast milk, and placenta, underlining their ability to enter the human body, presumably via the food chain and other yet-unknown mechanisms. In addition, plastic contains plasticizers, antioxidants, or lubricants, whose impact on human health is also under investigation. At the cellular level, the most important enzymes involved in the metabolism of xenobiotic compounds are the cytochrome P450 monooxygenases (CYPs). Despite their extensive characterization in the maintenance of cellular balance, their interactions with plastic and related products are unexplored. In this study, the possible interactions between several plastic-related compounds and one of the most important cytochromes, CYP2C19, were analyzed. By applying virtual compound screening and molecular docking to more than 1000 commercially available plastic-related compounds, we identified candidates that are likely to interact with this protein. A growth inhibition assay confirmed their cytotoxic activity on a CYP2C19-transfected hepatic cell line. Subsequently, we studied the effect of the selected compounds on the transcriptome-wide gene expression level by conducting RNA sequencing. Three candidate molecules were identified, i.e., 2,2′-methylene bis(6-tert-butyl-4-methylphenol), 1,1-bis(3,5-di-tert-butyl-2-hydroxyphenyl) ethane, and 2,2′-methylene bis(6-cyclohexyl-4-methylphenol)), which bound with a high affinity to CYP2C19 in silico. They exerted a profound cytotoxicity in vitro and interacted with several metabolic pathways, of which the ‘cholesterol biosynthesis process’ was the most affected. In addition, other affected pathways involved mitosis, DNA replication, and inflammation, suggesting an increase in hepatotoxicity. These results indicate that plastic-related compounds could damage the liver by affecting several molecular pathways. KW - cytotoxicity KW - ecotoxicity KW - hepatotoxicity KW - microplastic KW - RNA sequencing Y1 - 2023 UR - https://www.mdpi.com/1420-3049/28/16/5952 U6 - https://doi.org/10.3390/molecules28165952 SN - 1420-3049 VL - 28 IS - 16 ER - TY - PAT A1 - Scheibner, Katrin A1 - Küpper, Jan-Heiner A1 - Schmidtke, Kai-Uwe A1 - Miethbauer, Sebastian A1 - Herzog, Natalie T1 - Verfahren zur Herstellung von biogenen Stoffen T1 - Method for producing biogenic substances Y1 - 2020 UR - https://patents.google.com/patent/EP3068868B1/de ER - TY - PAT A1 - Scheibner, Katrin A1 - Kiebist, Jan A1 - Schmidtke, Kai-Uwe A1 - Küpper, Jan-Heiner T1 - Verfahren zur enzymatischen Herstellung von zytostatischen 4-Hydroxy-Oxazaphosphorinen“ Y1 - 2020 UR - https://www.b-tu.de/enzymtechnologie/publikationen/patente ER - TY - PAT A1 - Scheibner, Katrin A1 - Küpper, Jan-Heiner A1 - Jung, Friedrich T1 - In vitro Arzneimittel aus Prodrugs und deren Verwendung Y1 - 2021 UR - https://www.b-tu.de/enzymtechnologie/publikationen/patente ER - TY - GEN A1 - Buchmueller, Julia A1 - Enge, Anne-Margarethe A1 - Peters, Antonia A1 - Ebmeyer, Johanna A1 - Küpper, Jan-Heiner A1 - Schäfer, Bernd A1 - Bräuning, Albert A1 - Hessel-Pras, Stefanie T1 - The chemical structure impairs the intensity of genotoxic effects promoted by 1,2-unsaturated pyrrolizidine alkaloids in vitro T2 - Food and Chemical Toxicology Y1 - 2022 U6 - https://doi.org/10.1016/j.fct.2022.113049 SN - 1873-6351 SN - 0278-6915 VL - Vol. 164 ER - TY - GEN A1 - Abdelfatah, Sara A1 - Nass, Janine A1 - Knorz, Caroline A1 - Klauck, Sabine M. A1 - Küpper, Jan-Heiner A1 - Efferth, Thomas T1 - Pyrrolizidine alkaloids cause cell cycle and DNA damage repair defects as analyzed by transcriptomics in cytochrome P450 3A4-overexpressing HepG2 clone 9 cells T2 - Cell biology and toxicology Y1 - 2022 U6 - https://doi.org/10.1007/s10565-021-09599-9 SN - 1573-6822 SN - 0742-2091 VL - 38 IS - 2 SP - 325 EP - 345 ER - TY - GEN A1 - Geburek, Ina A1 - Rutz, Lukas A1 - Gao, Lan A1 - Küpper, Jan-Heiner A1 - These, Anja A1 - Schrenk, Dieter T1 - The metabolic pattern of hepatotoxic pyrrolizidine alkaloids in liver cells T2 - Chemical Research in Toxicology N2 - Contamination with 1,2-unsaturated pyrrolizidine alkaloids (PAs) is a serious problem for certain phytomedicines, foods, and animal feeds. Several of these PAs are genotoxic and carcinogenic, primarily in the liver, upon cytochrome P450 (CYP)-catalyzed activation into reactive (pyrrolic and pyrrole-like) metabolites. Here we investigated the metabolism of selected PAs (echimidine, europine, lasiocarpine, lycopsamine, retrorsine, and senecionine) in rat hepatocytes in primary culture and in human CYP3A4-transfected HepG2 cells. The open-chained diesters echimidine and lasiocarpine and the cyclic diester senecionine were extensively metabolized in rat hepatocytes into a broad spectrum of products released into the medium. A large portion of unidentified, possibly irreversibly bound, products remained in the cells while detectable amounts of reactive and other metabolites were found in the incubation media. In HepG2-CYP3A4 cells, lasiocarpine was more extensively metabolized than echimidine and senecionine which also gave rise to the release of pyrrolic metabolites. In human cells, no pyrrolic metabolites were detected in retrorsine or lycopsamine incubations, while no such metabolites were detected from europine in both cell types. Other types of metabolic changes comprised modifications such as side chain demethylation or oxygenation reactions like the formation of N-oxides. The latter, considered as a detoxification step, was a major pathway with cyclic diesters, was less distinctive for echimidine and lycopsamine and almost negligible for lasiocarpine and europine. Our data are in agreement with previously published cyto- and genotoxicity findings and suggests that the metabolic pattern may contribute substantially to the specific toxic potency of a certain congener. In addition, marked differences were found for certain congeners between rat hepatocytes and transfected human HepG2 cells, whereby a high level of bioactivation was found for lasiocarpine, whereas a very low level of bioactivation was observed for monoesters, in particular in human cells. Y1 - 2021 UR - https://pubs.acs.org/doi/abs/10.1021/acs.chemrestox.0c00507 U6 - https://doi.org/10.1021/acs.chemrestox.0c00507 VL - 34 IS - 4 SP - 1101 EP - 1113 ER - TY - GEN A1 - Jung, Conrad H. G. A1 - Waldeck, Peter A1 - Petrick, Ingolf A1 - Braune, Steffen A1 - Küpper, Jan-Heiner A1 - Jung, Friedrich T1 - Bioreactor for the cultivation of Arthrospira platensis under controlled conditions T2 - Journal of Cellular Biotechnology N2 - A vertical flat-type bioreactor consisting of transparent polyethylene (food safe) was constructed, which is characterized by a flexible design and allows the adjustment of a light path of 1 cm – 8.5 cm and a working volume of 1.5 l – 4 l. To characterize the performance of the bioreactor, cultivation experiments were performed with the cyanobacterium Arthrospira platensis (AP). The growth was assessed continuously by monitoring optical density and intermittently by measuring the dry weight of the AP biomass. An on-line measurement technique for estimating biomass production rate in a photosynthetic microalgae culture was developed. The oxygen produced by AP in the culture medium was flushed out sparging using a mixture of air and CO2 (1%). Factors which might influence the AP growth were monitored: pH, temperature, oxygen concentration and the filling level were corrected automatically to compensate evaporation losses. As an example, the huge influence of the light intensity on the AP growth was tested. The increase of the photon flux density of 15 to 1200μmol/(l · d) led to a 22-fold increase of the productivity and a 3.1-fold shorter doubling time. Using an online measurement technique - together with the control of the growth process via a wireless local area network (WLAN) router and virtual private network - allows monitoring the growth of Arthrospira platensis remotely. Y1 - 2021 UR - https://content.iospress.com/articles/journal-of-cellular-biotechnology/jcb210032 U6 - https://doi.org/10.3233/JCB-210032 VL - 7 IS - 1 SP - 35 EP - 40 ER - TY - GEN A1 - Braune, Steffen A1 - Krüger-Genge, Anne A1 - Kammerer, Sarah A1 - Jung, Friedrich A1 - Küpper, Jan-Heiner T1 - Phycocyanin from Arthrospira platensis as Potential Anti-Cancer Drug: Review of In Vitro and In Vivo Studies T2 - Life Y1 - 2021 UR - https://www.mdpi.com/2075-1729/11/2/91 U6 - https://doi.org/10.3390/life11020091 VL - 11 IS - 2 SP - 91 ER - TY - GEN A1 - Schulz, Christian A1 - Jung, Friedrich A1 - Küpper, Jan-Heiner T1 - Inhibition of phase-1 biotransformation and cytostatic effects of diphenyleneiodonium on hepatoblastoma cell line HepG2 and a CYP3A4-overexpressing HepG2 cell clone T2 - Clinical Hemorheology and Microcirculation N2 - Cell-based in vitro liver models are an important tool in the development and evaluation of new drugs in pharmacological and toxicological drug assessment. Hepatic microsomal enzyme complexes, consisting of cytochrome P450 oxidoreductase (CPR) and cytochrome P450 monooxygenases (CYPs), play a decisive role in catalysing phase-1 biotransformation of pharmaceuticals and xenobiotics. For a comprehensive understanding of the phase-1 biotransformation of drugs, the availability of well-characterized substances for the targeted modulation of in vitro liver models is essential. In this study, we investigated diphenyleneiodonium (DPI) for its ability to inhibit phase-1 enzyme activity and further its toxicological profile in an in vitro HepG2 cell model with and without recombinant expression of the most important drug metabolization enzyme CYP3A4. Aim of the study was to identify effective DPI concentrations for CPR/CYP activity modulation and potentially associated dose and time dependent hepatotoxic effects. The cells were treated with DPI doses up to 5,000nM (versus vehicle control) for a maximum of 48 h and subsequently examined for CYP3A4 activity as well as various toxicological relevant parameters such as cell morphology, integrity and viability, intracellular ATP level, and proliferation. Concluding, the experiments revealed a time- and concentration-dependent DPI mediated partial and complete inhibition of CYP3A4 activity in CYP3A4 overexpressing HepG2-cells (HepG2-CYP3A4). Other cell functions, including ATP synthesis and consequently the proliferation were negatively affected in both in vitro cell models. Since neither cell integrity nor cell viability were reduced, the effect of DPI in HepG2 can be assessed as cytostatic rather than cytotoxic. KW - Phase-1 KW - biotransformation KW - CYP KW - cytochrome P450 monooxygenase KW - CYP3A4 KW - diphenyleneiodonium KW - DPI KW - HepG2 KW - HepG2-CYP3A4 KW - hepatocytes KW - NADPH-cytochrome P450 oxidoreductase KW - POR KW - CPR Y1 - 2021 UR - https://content.iospress.com/articles/clinical-hemorheology-and-microcirculation/ch219117 U6 - https://doi.org/10.3233/CH-219117 SN - 1875-8622 SN - 1386-0291 VL - 79 IS - 1 SP - 231 EP - 243 ER - TY - GEN A1 - Krüger-Genge, Anne A1 - Jung, Conrad H. G. A1 - Braune, Steffen A1 - Harb, K. A1 - Westphal, S. A1 - Klöpzig, Stefanie A1 - Küpper, Jan-Heiner A1 - Jung, Friedrich T1 - Effect of Arthrospira powders from different producers on the formation of endothelial cell monolayers T2 - Clinical Hemorheology and Microcirculation N2 - Arthrospira platensis (AP) and some of its derived products have well-established biological activities as antioxidants or as agents to reduce cardiovascular disease risk factors. Furthermore, AP products have gained increasing importance as potential anti-cancer agents. However, the ingredients of the available products vary greatly with the origin, the type of production and processing, which could have significant consequences for their biological effects. Therefore, the composition and biological influence of five distinct AP powders, which were acquired commercially or produced at a public biotechnology institute, were investigated in regard to their endothelialization capacity using a cell impedance- (CI) based measurement method. The study revealed that the AP composition and especially the influence on HUVEC proliferation differed significantly between the five AP powders up to 109%. Thus, it could be shown that the method used allows the reliable detection of quantitative differences in biological effects of different AP preparations. KW - Arthrospira platensis KW - spirulina KW - endothelial cells Y1 - 2021 UR - https://content.iospress.com/articles/clinical-hemorheology-and-microcirculation/ch219200 U6 - https://doi.org/10.3233/CH-219200 SN - 1875-8622 SN - 1386-0291 VL - 79 IS - 1 SP - 193 EP - 203 ER - TY - GEN A1 - Krüger-Genge, Anne A1 - Steinbrecht, Susanne A1 - Küpper, Jan-Heiner A1 - Lendlein, Andreas A1 - Jung, Friedrich T1 - Evidence for cytostatic effect of cyclophosphamide on human vein endothelial cells in cancer therapy: preliminary in vitro results. T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189125 SN - 1875-8622 SN - 1386-0291 VL - 69 IS - 1-2 SP - 267 EP - 276 ER - TY - GEN A1 - Jung, Friedrich A1 - Jung, Conrad H. G. A1 - Krüger-Genge, Anne A1 - Waldeck, Peter A1 - Küpper, Jan-Heiner T1 - Factors influencing the growth of Spirulina platensis in closed photobioreactors under CO₂ – O₂ conversion T2 - Journal of Cellular Biotechnology N2 - Since there is growing interest throughout the world in photosynthetic microbes as a potential source of food or food supplements, an assessment of factors which influence the biomass obtained in bioreactors, protein contents and constituents is important. This work reviews the autotrophic cultivation conditions of Spirulina platensis especially the dependency on the strain, the composition of the nutrient solution, pH, temperature of the medium, light intensity and color as well as exposure rhythm, the flow rate and composition of the aerating gas mixture and the bubble size, the content of oxygen, CO₂ and HCO₃ in the medium and last but not least from the optical density of the spirulina suspension during growth. KW - spirulina platensis KW - influencing factors KW - temperature KW - light intensity KW - wave length Y1 - 2019 U6 - https://doi.org/10.3233/JCB-199004 SN - 2352-3697 SN - 2352-3689 VL - 5 IS - 2 SP - 125 EP - 134 ER - TY - GEN A1 - Steinbrecht, Susanne A1 - König, Rosalie A1 - Schmidtke, Kai-Uwe A1 - Herzog, Natalie A1 - Scheibner, Katrin A1 - Krüger-Genge, Anne A1 - Jung, Friedrich A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - Metabolic activity testing can underestimate acute drug cytotoxicity as revealed by HepG2 cell clones overexpressing cytochrome P450 2C19 and 3A4 T2 - Toxicology Y1 - 2018 U6 - https://doi.org/10.1016/j.tox.2018.11.008 SN - 0300-483X VL - 412 SP - 37 EP - 47 ER - TY - GEN A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - Optimized protocol for induction of cytochrome P450 enzymes 1A2 and 3A4 in human primary-like hepatocyte cell strain HepaFH3 to study in vitro toxicology T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189321 SN - 1386-0291 SN - 1875-8622 VL - 70 IS - 4 SP - 563 EP - 571 ER - TY - GEN A1 - Krüger-Genge, Anne A1 - Jung, Friedrich A1 - Küpper, Jan-Heiner A1 - Lehmann, Christian A1 - Franke, Ralf-Peter T1 - Actin type and distribution in erythrocytes T2 - Journal of Cellular Biotechnology N2 - Erythrocytes transport oxygen from the lungs to the tissues. The excess surface area together with the elasticity of the erythrocyte cell membrane provides the flexibility needed to pass through the microvasculature where the oxygen exchange occurs. Although the architecture of the red cell and its membrane-associated cytoskeletal network is known in general, the factors that control the characteristic shape change during echinocyte formation are poorly understood. In this short report we show that in echinocytes a completely reorganized membrane cytoskeleton with a box-like structure of actin filaments prevailed indicating the importance of the actin cytoskeleton during echinocyte formation. KW - Erythrocytes KW - echinocytes KW - submembraneous cytoskeleton KW - actin Y1 - 2018 UR - https://content.iospress.com/articles/journal-of-cellular-biotechnology/jcb179014 U6 - https://doi.org/10.3233/JCB-179014 SN - 2352-3697 SN - 2352-3689 VL - 3 IS - 2 SP - 81 EP - 83 ER - TY - GEN A1 - Krüger-Genge, Anne A1 - Braune, Steffen A1 - Walter, Maria A1 - Kratz, Karl A1 - Küpper, Jan-Heiner A1 - Krengel, M. A1 - Lendlein, Andreas A1 - Jung, Friedrich T1 - Influence of surface treatments of poly(n-butyl acrylate) networks on fibroblasts adhesion, morphology and viability T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189130 SN - 1875-8622 SN - 1386-0291 VL - 69 IS - 1-2 SP - 305 EP - 316 ER - TY - GEN A1 - Nowak, Elisabeth A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - ATP-based cell viability assay is superior to trypan blue exclusion and XTT assay in measuring cytotoxicity of anticancer drugs Taxol and Imatinib, and proteasome inhibitor MG-132 on human hepatoma cell line HepG2. T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189120 SN - 1386-0291 SN - 1875-8622 VL - 69 IS - 1-2 SP - 327 EP - 336 ER - TY - GEN A1 - Uhlig, Katja A1 - Gehre, Christian P. A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner A1 - Coleman, D. A1 - Püschel, G. A1 - Duschl, Claus T1 - Real-time monitoring of oxygen consumption of hepatocytes in a microbioreactor T2 - Toxicology Letters Y1 - 2018 U6 - https://doi.org/10.1016/j.toxlet.2018.06.652 SN - 0378-4274 VL - 295 IS - Supplement 1 SP - S115 ER - TY - PAT A1 - Küpper, Jan-Heiner T1 - Vorrichtung und Verfahren zur Sequestrierung von atmosphärischen Kohlendioxid Y1 - 2017 N1 - EU-Patentanmeldung ER -