@article{GrebinykPrylutskaGrebinyketal.2021, author = {Grebinyk, Anna and Prylutska, Svitlana and Grebinyk, Sergii and Evstigneev, Maxim and Krysiuk, Iryna and Skaterna, Tetiana and Horak, Iryna and Sun, Yanfang and Drobot, Liudmyla and Matyshevska, Olga and Prylutskyy, Yuriy and Ritter, Uwe and Frohme, Marcus}, title = {Antitumor efficiency of the natural alkaloid berberine complexed with C60 fullerene in Lewis lung carcinoma in vitro and in vivo}, series = {Cancer Nanotechnology}, volume = {12}, journal = {Cancer Nanotechnology}, publisher = {BioMed Central}, issn = {1868-6966}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-14749}, pages = {24}, year = {2021}, abstract = {Background Berberine (Ber) is a herbal alkaloid with pharmacological activity in general and a high anticancer potency in particular. However, due to its low bioavailability, the difficulty in reaching a target and choosing the right dose, there is a need to improve approaches of Ber use in anticancer therapy. In this study, Ber, noncovalently bound to a carbon nanostructure C60 fullerene (C60) at various molar ratios of the components, was explored against Lewis lung carcinoma (LLC). Methods C60-Ber noncovalent nanocomplexes were synthesized in 1:2, 1:1 and 2:1 molar ratios. Ber release from the nanocomplexes was studied after prolonged incubation at different pH with the liquid chromatography-mass spectrometry analysis of free Ber content. Biological effects of the free and C60-complaxated Ber were studied in vitro towards LLC cells with phase-contrast and fluorescence microscopy, flow cytometry, MTT reduction, caspase activity and wound closure assays. The treatment with C60-Ber nanocomplex was evaluated in vivo with the LLC-tumored C57Bl mice. The mice body weight, tumor size, tumor weight and tumor weight index were assessed for four groups, treated with saline, 15 mg C60/kg, 7.5 mg Ber/kg or 2:1 C60-Ber nanocomplex (15 mg C60/kg, 7.5 mg Ber/kg). Results Ber release from C60-Ber nanocomplexes was promoted with medium acidification. LLC cells treatment with C60-Ber nanocomplexes was followed by enhanced Ber intracellular uptake as compared to free Ber. The cytotoxicity of the studied agents followed the order: free Ber < 1:2 < 1:1 < 2:1 C60-Ber nanocomplex. The potency of cytotoxic effect of 2:1 C60-Ber nanocomplex was confirmed by 21.3-fold decrease of IC50 value (0.8 ± 0.3 µM) compared to IC50 for free Ber (17 ± 2 µM). C60-Ber nanocomplexes induced caspase 3/7 activation and suppressed the migration activity of LLC cells. The therapeutic potency of 2:1 C60-Ber nanocomplex was confirmed in a mouse model of LLC. The tumor growth in the group treated with 2:1 C60-Ber nanocomplex is suppressed by approximately 50\% at the end of experiment, while in the tumor-bearing group treated with free Ber no therapeutic effect was detected. Conclusions This study indicates that complexation of natural alkaloid Ber with C60 may be a novel therapeutic strategy against lung carcinoma.}, language = {en} } @article{KuhlsMoskalenkoSukiasyanetal.2021, author = {Kuhls, Katrin and Moskalenko, Olga and Sukiasyan, Anna and Manukyan, Dezdemonia and Melik-Andreasyan, Gayane and Atshemyan, Liana and Apresyan, Hripsime and Strelkova, Margarita V. and Jaeschke, Anja and Wieland, Ralf and Frohme, Marcus and Cortes, Sofia and Keshishyan, Ara}, title = {Microsatellite based molecular epidemiology of Leishmania infantum from re-emerging foci of visceral leishmaniasis in Armenia and pilot risk assessment by ecological niche modeling}, series = {PLoS Neglected Tropical Diseases}, volume = {15}, journal = {PLoS Neglected Tropical Diseases}, number = {4}, publisher = {Public Library of Science (PLoS)}, issn = {1935-2735}, doi = {10.1371/journal.pntd.0009288}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-14004}, pages = {e0009288}, year = {2021}, abstract = {Leishmaniasis is a vector-borne disease caused by protozoan parasites of the genus Leishmania. In Armenia visceral leishmaniasis (VL) is re-emerging since 1999 after a long break of 30 years, with 167 cases recorded until 2019. Molecular diagnosis of VL was implemented only in 2016, and the causative agent was identified as L. infantum. In the present study we expanded the investigation of the causative agent to a characterization at strain level and the identification of its phylogenetic position among the L. infantum genotypes circulating worldwide. This is the first study addressing genetic diversity and population structure of L. infantum in Armenia and in Transcaucasia. Armenia is an extremely interesting region due to its bio-geographic specificities e.g. the high number of different climates in this small mountainous country and the observed high diversity of sand fly species, part of which occurring in very high altitudes. Ecological niche modeling based on registered VL cases and sand fly vectors collected in active VL foci revealed that the risk of further spread of VL is very high due to climate change. Studies of this region should be expanded to enable targeted control measures.}, language = {en} } @article{WielandKuhlsLentzetal.2021, author = {Wieland, Ralf and Kuhls, Katrin and Lentz, Hartmut H.K. and Conraths, Franz and Kampen, Helge and Werner, Doreen}, title = {Combined climate and regional mosquito habitat model based on machine learning}, series = {Ecological Modelling}, volume = {452}, journal = {Ecological Modelling}, address = {Elsevier}, issn = {0304-3800}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-15649}, year = {2021}, abstract = {Besides invasive mosquito species also several native species are proven or suspected vectors of arboviruses as West Nile or Usutu virus in Western Europe. Habitat models of these native vectors can be a helpful tool for assessing the risk of autochthonous occurrence, outbreaks and spread of diseases caused by such arboviruses. Modelling native mosquitoes is complicated because of the perfect adaptation to the climatic and landscape conditions and their high abundance in contrast to invasive species. Here we present a new approach for such a habitat model for native mosquito species in Germany, which are considered as vectors of West Nile virus (WNV). Epizootic emergence of WNV was registered in Germany since 2018. The models are based on surveillance data of mosquitoes from the German citizen science project "M{\"u}ckenatlas" complemented by data from systematic trap monitoring in Germany, and on data freely available from the Deutscher Wetterdienst (DWD) and OpenStreetMap (OSM). While climatic factors still play an important role, we could show that habitat suitability is predictable only by the combination of the climate model with a regional model. Both models were based on a machine-learning approach using XGBoost. Evaluation of the accuracy of the models was done by statistical analysis, determining among others feature importances using the SHAP-Library. Final output of the combined climatic and regional models are maps showing the superposed habitat suitability which are generated through a number of steps described in detail. These maps also include the registered cases of WNV infections in the selected region of Germany.}, language = {en} } @phdthesis{Grebinyk2021, author = {Grebinyk, Anna}, title = {Synergistic Chemo- and Photodynamic Treatment of Cancer Cells with C60 Fullerene Nanocomplexes}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-14137}, school = {Technische Hochschule Wildau}, year = {2021}, abstract = {Recent progress in nanotechnology has attracted interest to a biomedical application of the carbon nanoparticle C60 fullerene (C60) due to its unique structure and versatile biological activity. In the current study the dual functionality of C60 as a photosensitizer and a drug nanocarrier was exploited to improve the efficiency of chemotherapeutic drugs towards human leukemic cells. Pristine C60 demonstrated time-dependent accumulation with predominant mitochondrial localization in leukemic cells. C60's effects on leukemic cells irradiated with high power single chip LEDs of different wavelengths were assessed to find out the most effective photoexcitation conditions. A C60-based noncovalent nanosized system as a carrier for an optimized drug delivery to the cells was evaluated in accordance to its physicochemical properties and toxic effects. Finally, nanomolar amounts of C60-drug nanocomplexes in 1:1 and 2:1 molar ratios were explored to improve the efficiency of cell treatment, complementing it with photodynamic approach. A proposed treatment strategy was developed for C60 nanocomplexes with the common chemotherapeutic drug Doxorubicin, whose intracellular accumulation and localization, cytotoxicity and mechanism of action were investigated. The developed strategy was revealed to be transferable to an alternative potent anticancer drug - the herbal alkaloid Berberine. Hereafter, a strong synergy of treatments arising from the combination of C60-mediated drug delivery and C60 photoexcitation was revealed. Presented data indicate that a combination of chemo- and photodynamic treatments with C60-drug nanoformulations could provide a promising synergetic approach for cancer treatment.}, subject = {cancer}, language = {en} } @article{PetrilloFabbriKagklietal.2021, author = {Petrillo, Mauro and Fabbri, M and Kagkli, Dafni Maria and Querci, Maddalena and Van den Eede, Guy and Alm, Erik and Aytan-Aktug, Derya and Capella-Gutierrez, Salvador and Carrillo, Catherine and Cestaro, Alessandro and Chan, Kok-Gan and Coque, Teresa and Endrullat, Christoph and Gut, Ivo and Hammer, Paul and Kay, Gemma L. and Madec, Jean-Yves and Mather, Alison E. and McHardy, Alice Carolyn and Naas, Thierry and Paracchini, Valentina and Peter, Silke and Pightling, Arthur and Raffael, Barbara and Rossen, John and Rupp{\´e}, Etienne and Schlaberg, Robert and Vanneste, Kevin and Weber, Lukas M. and Westh, Henrik and Angers-Loustau, Alexandre}, title = {A roadmap for the generation of benchmarking resources for antimicrobial resistance detection using next generation sequencing [version 1; peer review: 2 approved with reservations]}, series = {F1000Research}, journal = {F1000Research}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-15699}, year = {2021}, abstract = {Next Generation Sequencing technologies significantly impact the field of Antimicrobial Resistance (AMR) detection and monitoring, with immediate uses in diagnosis and risk assessment. For this application and in general, considerable challenges remain in demonstrating sufficient trust to act upon the meaningful information produced from raw data, partly because of the reliance on bioinformatics pipelines, which can produce different results and therefore lead to different interpretations. With the constant evolution of the field, it is difficult to identify, harmonise and recommend specific methods for large-scale implementations over time. In this article, we propose to address this challenge through establishing a transparent, performance-based, evaluation approach to provide flexibility in the bioinformatics tools of choice, while demonstrating proficiency in meeting common performance standards. The approach is two-fold: first, a community-driven effort to establish and maintain "live" (dynamic) benchmarking platforms to provide relevant performance metrics, based on different use-cases, that would evolve together with the AMR field; second, agreed and defined datasets to allow the pipelines' implementation, validation, and quality-control over time. Following previous discussions on the main challenges linked to this approach, we provide concrete recommendations and future steps, related to different aspects of the design of benchmarks, such as the selection and the characteristics of the datasets (quality, choice of pathogens and resistances, etc.), the evaluation criteria of the pipelines, and the way these resources should be deployed in the community.}, language = {en} } @article{GloeklerLimIdaetal.2021, author = {Gl{\"o}kler, J{\"o}rn and Lim, Theam Soon and Ida, Jeunice and Frohme, Marcus}, title = {Isothermal amplifications - a comprehensive review on current methods}, series = {Critical Reviews in Biochemistry and Molecular Biology}, journal = {Critical Reviews in Biochemistry and Molecular Biology}, publisher = {Taylor \& Francis}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-15154}, pages = {1 -- 44}, year = {2021}, abstract = {AbstractThe introduction of nucleic acid amplification techniques has revolutionized the field of medical diagnostics in the last decade. The advent of PCR catalyzed the increasing application of DNA, not just for molecular cloning but also for molecular based diagnostics. Since the introduction of PCR, a deeper understanding of molecular mechanisms and enzymes involved in DNA/RNA replication has spurred the development of novel methods devoid of temperature cycling. Isothermal amplification methods have since been introduced utilizing different mechanisms, enzymes, and conditions. The ease with which isothermal amplification methods have allowed nucleic acid amplification to be carried out has had a profound impact on the way molecular diagnostics are being designed after the turn of the millennium. With all the advantages isothermal amplification brings, the issues or complications surrounding each method are heterogeneous making it difficult to identify the best approach for an end-user. This review pays special attention to the various isothermal amplification methods by classifying them based on the mechanistic characteristics which include reaction formats, amplification information, promoter, strand break, and refolding mechanisms. We would also compare the efficiencies and usefulness of each method while highlighting the potential applications and detection methods involved. This review will serve as an overall outlook on the journey and development of isothermal amplification methods as a whole.}, language = {en} } @inproceedings{RadutnyNechyporenkoAlekseevaetal.2021, author = {Radutny, Radiy and Nechyporenko, Alina and Alekseeva, Victoriia and Yurevych, Nadiia and Lupyr, Andrii and Gargin, Vitaliy}, title = {Complex Automatic Evaluation of the Medical Images of the Paranasal Sinuses}, series = {Proceedings of the 4th International Conference on Informatics \& Data-Driven Medicine, Valencia, Spain, November 19 - 21, 2021}, booktitle = {Proceedings of the 4th International Conference on Informatics \& Data-Driven Medicine, Valencia, Spain, November 19 - 21, 2021}, editor = {Shakhovska, Nataliya and Salazar, Addisson and Izonin, Ivan and Campos, Jaime}, publisher = {RWTH Aachen}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-19010}, pages = {182 -- 189}, year = {2021}, abstract = {Evaluation of medical images is of key importance in the work of medical staff today. Especially this problem pays important role in otolaryngology. The aim of our work was to develop an automatic complex method for assessing the state of the human paranasal sinuses. Our research included 10 people of different sex and age, who were divided into groups, taking into account the recommendations of the WHO for 2019-2021. The structure of the mucous membrane of the maxillary sinus were calculated and compared. In the course of our research, an algorithm was developed for the automatic assessment of the state of the mucous membrane of the maxillary sinus and its bone walls according to the data of the spiral computed tomography. The difference between obtained data in the manual and automatic mode is minimal.}, language = {en} } @article{EngelsSchlegelJacobsetal.2021, author = {Engels, Andreas and Schlegel, Volker and Jacobs, Hannes and Großelindemann, Niklas and Morshed, Md. Niaz and Stenglein, Antonia Sigrid and Bonaiuto, Vincenzo and Foitzik, Andreas}, title = {3d Printer Heads for Extrusion of Biologicals Gels}, series = {Materials Science Forum}, volume = {1016}, journal = {Materials Science Forum}, publisher = {Trans Tech Publications}, issn = {0255-5476}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-18773}, pages = {435 -- 439}, year = {2021}, abstract = {Processing biological materials with 3-dimensional (3D) printers has attracted increasing interest in several research areas. At the same time, off-the-shelf solutions are increasingly available in a wider variety to meet the current market demand. However, most of the available bioprinters are closed source, thus, modifications are quite challenging or require an avoidable consultation process with the manufacturer. Furthermore, the entry prices for basic machines amount to several thousands of euros. Whereas, high-end 3D bio printers with a vast array of features are available for several hundred thousand euros. Due to the immense potential of this tool in the field of biotechnology it is important to extend the availability of this technology for research purposes in terms of adaptability and price. This ongoing work focuses on open-source 3D printer heads with the ability to extrude biological materials. The print heads include several techniques to process low as well as high viscose biomaterials such as agar and gelatin. Additionally, obstacles such as continuous substrate tempering or integration into existing 3D printers are addressed. The work presented is open source and thus freely adaptable to any user's specific needs. Our goal is to process a diverse range of biomaterials with different print techniques.}, language = {en} } @article{HerkenhoffBovolentaBroedeletal.2021, author = {Herkenhoff, Marcos Edgar and Bovolenta, Luiz A. and Br{\"o}del, Oliver and dos Santos, Lucilene D. and de Oliveira, Arthur C. and Chuffa, Luiz G.A. and Ribeiro, Amanda O. and Lupi, Luiz A. and Dias, Marco A.D. and Hilsdorf, Alexandre W.S. and Frohme, Marcus and Pinhal, Danillo}, title = {Variant expression signatures of microRNAs and protein related to growth in a crossbreed between two strains of Nile tilapia (Oreochromis niloticus)}, series = {Genomics}, volume = {113}, journal = {Genomics}, number = {6}, publisher = {Elsevier}, issn = {0888-7543}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-15571}, pages = {4303 -- 4312}, year = {2021}, abstract = {Nile tilapia (Oreochromis niloticus) is a species of worldwide importance for aquaculture. A crossbred lineage was developed through introgressive backcross breeding techniques and combines the high growth performance of the Chitralada (CHIT) lwith attractive reddish color of the Red Stirling (REDS) strains. Since the crossbreed has an unknown genetically improved background, the objective of this work was to characterize expression signatures that portray the advantageous phenotype of the crossbreeds. We characterized the microRNA transcriptome by high throughput sequencing (RNA-seq) and the proteome through mass spectrometry (ESI-Q-TOF-MS) and applied bioinformatics for the comparative analysis of such molecular data on the three strains. Crossbreed expressed a distinct set of miRNAs and proteins compared to the parents. They comprised several microRNAs regulate traits of economic interest. Proteomic profiles revealed differences between parental and crossbreed in expression of proteins associated with glycolisis. Distinctive miRNA and protein signatures contribute to the phenotype of crossbreed.}, language = {en} } @article{GossingRadkeBieringetal.2021, author = {Gossing, Wilhelm and Radke, Lars and Biering, Henrik and Diederich, Sven and Mai, Knut and Frohme, Marcus}, title = {The ElonginB/C-Cullin5-SOCS-Box-Complex Is a Potential Biomarker for Growth Hormone Disorders}, series = {Biomedicines}, volume = {9}, journal = {Biomedicines}, number = {2}, publisher = {MDPI}, issn = {2227-9059}, doi = {10.3390/biomedicines9020201}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-13802}, year = {2021}, abstract = {Insulin-like growth factor 1 (IGF-1) is the standard biochemical marker for the diagnosis and treatment control of acromegaly and growth hormone deficiency (GHD). However, its limitations necessitate the screening for new specific and sensitive biomarkers. The elonginB/C-cullin5-SOCS-box-complex (ECS-complex) (an intracellular five-protein complex) is stimulated by circulating growth hormone (GH) and regulates GH receptor levels through a negative feedback loop. It mediates the cells' sensitivity for GH and therefore, represents a potent new biomarker for those diseases. In this study, individual ECS-complex proteins were measured in whole blood samples of patients with acromegaly (n = 32) or GHD (n = 12) via ELISA and compared to controls. Hierarchical clustering of the results revealed that by combining the three ECS-complex proteins suppressor of cytokine signaling 2 (SOCS2), cullin-5 and ring-box protein 2 (Rbx-2), 93\% of patient samples could be separated from controls, despite many patients having a normal IGF-1 or not receiving medical treatment. SOCS2 showed the best individual diagnostic performance with an overall accuracy of 0.93, while the combination of the three proteins correctly identified all patients and controls. This resulted in perfect sensitivity and specificity for all patient groups, which demonstrates potential benefits of the ECS-complex proteins as clinical biomarkers for the diagnostics of GH-related diseases and substantiates their important role in GH metabolism.}, language = {en} }