TY - GEN A1 - Schneider, Jens A1 - Weiss, Romano A1 - Ruhe, Madeleine A1 - Jung, Tobias A1 - Roggenbuck, Dirk A1 - Stohwasser, Ralf A1 - Schierack, Peter A1 - Rödiger, Stefan T1 - Open source bioimage informatic tools for the analysis of DNA damage and associated biomarkers T2 - Journal of Laboratory and Precision Medicine KW - bioimage informatics KW - DNA damage response (DDR) KW - double-strand break (DSB) KW - immunofluorescence microscopy KW - open source software KW - proteasome activator PA28gamma Y1 - 2019 U6 - https://doi.org/10.21037/jlpm.2019.04.05 SN - 2519-9005 VL - 4 ER - TY - GEN A1 - Jurischka, Christoph A1 - Dinter, Franziska A1 - Efimova, Anastasia A1 - Weiss, Romano A1 - Schiebel, Juliane A1 - Schulz, Christian A1 - Fayziev, Bekzodjon A1 - Schierack, Peter A1 - Fischer, Thomas A1 - Rödiger, Stefan T1 - An explorative study of polymers for 3D printing of bioanalytical test systems T2 - Clinical Hemorheology and Microcirculation N2 - Background: The 3D printing is relevant as a manufacturing technology of functional models for forensic, pharmaceutical and bioanalytical applications such as drug delivery systems, sample preparation and point-of-care tests. Objective: Melting behavior and autofluorescence of materials are decisive for optimal printing and applicability of the product which are influenced by varying unknown additives. Methods: We have produced devices for bioanalytical applications from commercially available thermoplastic polymers using a melt-layer process. We characterized them by differential scanning calorimetry, fluorescence spectroscopy and functional assays (DNA capture assay, model for cell adhesion, bacterial adhesion and biofilm formation test). Results: From 14 tested colored, transparent and black materials we found only deep black acrylonitrile-butadiene-styrene (ABS) and some black polylactic acid (PLA) useable for fluorescence-based assays, with low autofluorescence only in the short-wave range of 300-400 nm. PLA was suitable for standard bioanalytical purposes due to a glass transition temperature of approximately 60°C, resistance to common laboratory chemicals and easy print processing. For temperature-critical methods, such as hybridization reactions up to 90°C, ABS was better suited. Conclusions: Autofluorescence was not a disadvantage per se but can also be used as a reference signal in assays. The rapid development of individual protocols for sample processing and analysis required the availability of a material with consistent quality over time. For fluorescence-based assays, the use of commercial standard materials did not seem to meet this requirement. KW - Thermal analysis KW - 3D Printing Y1 - 2020 UR - https://pubmed.ncbi.nlm.nih.gov/31929149/ U6 - https://doi.org/10.3233/CH-190713 SN - 1875-8622 VL - 75 IS - 1 SP - 57 EP - 84 ER - TY - GEN A1 - Schiebel, Juliane A1 - Noack, Jonas A1 - Rödiger, Stefan A1 - Kammel, Anne A1 - Menzel, Friederike A1 - Schwibbert, Karin A1 - Weise, Matthias A1 - Weiss, Romano A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Elimport, Alexey A1 - Roggenbuck, Dirk A1 - Schierack, Peter T1 - Analysis of three-dimensional biofilms on different material surfaces T2 - Biomaterials Science Y1 - 2020 U6 - https://doi.org/10.1039/D0BM00455C SN - 2047-4849 VL - 8 IS - 12 SP - 3500 EP - 3510 ER - TY - GEN A1 - Dinter, Franziska A1 - Jurischka, Christoph A1 - Weiss, Romano A1 - Rödiger, Stefan T1 - Multiparametrische Antikörperdetektion für die Diagnostik T2 - BIOspektrum Y1 - 2020 U6 - https://doi.org/10.1077/s12268-020-1375-5 SN - 1868-6249 SN - 0947-0867 VL - 26 IS - 3 SP - 265 EP - 267 ER - TY - GEN A1 - Liedtke, Victoria A1 - Schröder, Christian A1 - Roggenbuck, Dirk A1 - Weiss, Romano A1 - Stohwasser, Ralf A1 - Schierack, Peter A1 - Rödiger, Stefan A1 - Schenk, Lysann T1 - LEDGF/p75 is required for an efficient DNA damage response T2 - International Journal of Molecular Sciences KW - LEDGF KW - CRISPR/Cas9 KW - DNA damage signaling KW - gH2AX KW - ubiquitination Y1 - 2021 U6 - https://doi.org/10.3390/ijms22115866 VL - 22 IS - 11 SP - 1 EP - 16 ER - TY - GEN A1 - Chilimoniuk, Jarosław A1 - Gosiewska, Alicja A1 - Słowik, Jadwiga A1 - Weiss, Romano A1 - Deckert, P. Markus A1 - Rödiger, Stefan A1 - Burdukiewicz, Michał T1 - coutfitteR: efficient selection of count distributions to assess DNA damage T2 - Annals of Translational Medicine Y1 - 2021 U6 - https://doi.org/10.21037/atm-20-6363 SN - 2305-5847 VL - 9 IS - 7 ER - TY - GEN A1 - Liedtke, Victoria A1 - Weiss, Romano A1 - Skifov, Anastasia A1 - Rödiger, Stefan A1 - Schenk, Lysann T1 - Fully automated viability and toxicity screening - areliable all‐in‐one attempt T2 - Cancer Medicine N2 - The CRISPR/Cas9 technology is nowadays a common tool for genome editing to achieve new insights into, for example, diagnostics and therapeutics in cancer and genetic disorders. Cell proliferation and anticancer drug response studies are widely used to evaluate the impact of editing. However, these assays are often time‐consuming, expensive, and reproducibility is an issue. To overcome this, we developed a fast and cheap assay that combines a fully automated multispectral fluorescence microscopy platform with a nuclei staining and open‐source software analysis.Here, we generated different LEDGF/p75 model cell lines to validate the effect on proliferation and chemosensitivity. Therefore, a fast protocol for an optimized all‐in‐one attempt for cytotoxicity screenings and proliferation analysis of adherent cells in a 96‐well plate format was established using differential staining with two fluorescent dyes (Hoechst 33342 and propidium iodide) for live/dead cell discrimination. Subsequently, an automated cell nuclei count and analysis were performed using bioimage informatics.With the new established assay technology, up to 50,000 cells/well can be detected and analyzed in a 96‐well plate, resulting in a fast and accurate verification of viability and proliferation with consistency of 98% compared to manual counting. Our screening revealed that LEDGF depletion using CRISPR/Cas9 showed a diminished proliferation and chemosensitivity independent of cell line origin. Moreover, LEDGF depletion caused a significant increase in 𝛾H2AX foci, indicating a substantial increase in DNA double strand breaks. LEDGF/p75 overexpression enhanced proliferation and chemoresistance underlining the role of LEDGF in DNA damage response.Independent of cancer cell type, LEDGF/p75 is a central player in DNA damage repair and is implicated in chemoresistance. Moreover, our automated fluorescence biosensor technology allowed fast and reliable data acquisition without any fixation or additional washing steps. Additionally, data analysis was implemented using the modular open‐source software that can be adapted as needed. Y1 - 2024 U6 - https://doi.org/10.1002/cam4.7392 SN - 2045-7634 VL - 13 IS - 12 PB - Wiley ER - TY - GEN A1 - Weiss, Romano A1 - Liedtke, Victoria A1 - Rödiger, Stefan T1 - Cannabidiol does not cause DNA double-strand breaks in a human liver-derived cell model T2 - Journal of cannabis research N2 - Background Cannabidiol (CBD) is a non-psychoactive cannabinoid with potential therapeutic applications, including anti-inflammatory, analgesic, and anticancer effects. However, experts raised concerns about its potential to induce DNA damage and chromosomal aberrations at low concentrations. Notably, these studies used liver cell lines, which may not fully reflect the metabolic processing of CBD, potentially limiting the generalizability of their findings. This study investigated the short time effects of CBD on DNA double-strand breaks (DSBs) and proliferation in the human liver-derived cell line HepG2. Methods HepG2 cells were treated with CBD (5 – 50 , 3 - 72h incubation). To investigate potential imbalances in the expression of cannabinoid receptors 1 and 2 (CB1 / CB2) within HepG2 cells, we examined their expression using Western blot analysis. We hypothesized that such an imbalance could be associated with pathogenic processes. Double-strand breaks were then detected (5 Etoposide (ETP) served as positive control) via indirect immunofluorescence analysis using H2AX and 53BP1 antibodies, followed by quantification of DSB foci. Results Expression of CB2 but not CB1 was downregulated by 30 % in HepG2 cells after exposure to 5 CBD (24h incubation; 0.05) and 70 % downregulated after exposure to 50 CBD (24h incubation; 0.01). This effect was dose-dependent. Whilst ETP induced dose dependent DSBs, we could not confirm findings by others that CBD significantly increases the number of H2AX and 53BP1 foci between 5 and 50 (3h incubation; 0.05). Conclusion In our model, CBD stimulated the cells, as confirmed by modulation of CB2 expression as well as changes in intracellular cAMP. Our results show that CBD in ranges between 5 to 50 does not significantly increase the amount of DNA double strand breaks in HepG2 cells compared to the control. However, we did observe a significant reduction in cell proliferation and a significant increase in intracellular cAMP levels following CBD treatment. KW - Cannabidiol KW - DNA damage KW - Proliferation KW - CAMP KW - HepG2 Y1 - 2025 U6 - https://doi.org/10.1186/s42238-025-00365-w SN - 2522-5782 VL - 8 IS - 1 SP - 1 EP - 15 PB - Springer Science and Business Media LLC ER -