@article{RadkeSandigLubitzetal.2017, author = {Radke, Lars and Sandig, Grit and Lubitz, Annika and Schließer, Ulrike and von Horsten, Hans Henning and Blanchard, V{\´e}ronique and Keil, Karolin and Sandig, Volker and Giese, Christoph and Hummel, Michael and Hinderlich, Stephan and Frohme, Marcus}, title = {In Vitro Evaluation of Glycoengineered RSV-F in the Human Artificial Lymph Node Reactor}, series = {Bioengineering}, volume = {4}, journal = {Bioengineering}, number = {3}, issn = {2306-5354}, doi = {10.3390/bioengineering4030070}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-9831}, year = {2017}, abstract = {Subunit vaccines often require adjuvants to elicit sustained immune activity. Here, a method is described to evaluate the efficacy of single vaccine candidates in the preclinical stage based on cytokine and gene expression analysis. As a model, the recombinant human respiratory syncytial virus (RSV) fusion protein (RSV-F) was produced in CHO cells. For comparison, wild-type and glycoengineered, afucosylated RSV-F were established. Both glycoprotein vaccines were tested in a commercial Human Artificial Lymph Node in vitro model (HuALN®). The analysis of six key cytokines in cell culture supernatants showed well-balanced immune responses for the afucosylated RSV-F, while immune response of wild-type RSV-F was more Th1 accentuated. In particular, stronger and specific secretion of interleukin-4 after each round of re-stimulation underlined higher potency and efficacy of the afucosylated vaccine candidate. Comprehensive gene expression analysis by nCounter gene expression assay confirmed the stronger onset of the immunologic reaction in stimulation experiments with the afucosylated vaccine in comparison to wild-type RSV-F and particularly revealed prominent activation of Th17 related genes, innate immunity, and comprehensive activation of humoral immunity. We, therefore, show that our method is suited to distinguish the potency of two vaccine candidates with minor structural differences.}, language = {en} } @article{BayBuschLisdatetal.2017, author = {Bay, Daniyah H. and Busch, Annika and Lisdat, Fred and Iida, Keisuke and Ikebukuro, Kazunori and Nagasawa, Kazuo and Karube, Isao and Yoshida, Wataru}, title = {Identification of G-quadruplex structures that possess transcriptional regulating functions in the Dele and Cdc6 CpG islands}, series = {BMC Molecular Biology}, volume = {18}, journal = {BMC Molecular Biology}, number = {17}, issn = {1471-2199}, doi = {10.1186/s12867-017-0094-z}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-9811}, year = {2017}, abstract = {G-quadruplex is a DNA secondary structure that has been shown to play an important role in biological systems. In a previous study, we identified 1998 G-quadruplex-forming sequences using a mouse CpG islands DNA microarray with a fluorescent-labeled G-quadruplex ligand. Among these putative G-quadruplex-forming sequences, G-quadruplex formation was verified for 10 randomly selected sequences by CD spectroscopy and DMS footprinting analysis. In this study, the biological function of the 10 G-quadruplex-forming sequences in the transcriptional regulation has been analyzed using a reporter assay.}, language = {en} } @article{HornemannSinningCortesetal.2017, author = {Hornemann, Andrea and Sinning, Denise and Cortes, Sofia and Campino, Lenea and Emmer, Peggy and Kuhls, Katrin and Ulm, Gerhard and Frohme, Marcus and Beckhoff, Burkhard}, title = {A pilot study on fingerprinting Leishmania species from the Old World using Fourier transform infrared spectroscopy}, series = {Analytical and Bioanalytical Chemistry}, journal = {Analytical and Bioanalytical Chemistry}, issn = {1432-1130}, doi = {10.1007/s00216-017-0655-5}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-10080}, year = {2017}, abstract = {Leishmania species are protozoan parasites and the causative agents of leishmaniasis, a vector borne disease that imposes a large health burden on individuals living mainly in tropical and subtropical regions. Different Leishmania species are responsible for the distinct clinical patterns, such as cutaneous, mucocutaneous, and visceral leishmaniasis, with the latter being potentially fatal if left untreated. For this reason, it is important to perform correct species identification and differentiation. Fourier transform infrared spectroscopy (FTIR) is an analytical spectroscopic technique increasingly being used as a potential tool for identification of microorganisms for diagnostic purposes. By employing mid-infrared (MIR) spectral data, it is not only possible to assess the chemical structures but also to achieve differentiation supported by multivariate statistic analysis. This work comprises a pilot study on differentiation of Leishmania species of the Old World (L. major, L. tropica, L. infantum, and L. donovani) as well as hybrids of distinct species by using vibrational spectroscopic fingerprints. Films of intact Leishmania parasites and their deoxyribonucleic acid (DNA) were characterized comparatively with respect to their biochemical nature and MIR spectral patterns. The strains' hyperspectral datasets were multivariately examined by means of variance-based principal components analysis (PCA) and distance-based hierarchical cluster analysis (HCA). With the implementation of MIR spectral datasets we show that a phenotypic differentiation of Leishmania at species and intra-species level is feasible. Thus, FTIR spectroscopy can be further exploited for building up spectral databases of Leishmania parasites in view of high-throughput analysis of clinical specimens.}, language = {en} } @article{GottschalkStollfussLietzauetal.2017, author = {Gottschalk, Josefine Johanna and Stollfuß, Carsten and Lietzau, Kai-Henning and Foitzik, Andreas and Richetta, Maria}, title = {Conceptual study for long-term monitoring of chemotherapeutic induced cell reactions by ESPI}, series = {tm - Technisches Messen}, volume = {85}, journal = {tm - Technisches Messen}, number = {2}, issn = {2196-7113}, doi = {10.1515/teme-2017-0100}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-10676}, pages = {111 -- 118}, year = {2017}, abstract = {During the last years, various approaches on an individualized drug therapy for benign cells have been researched. However, due to the complex topic a universal approach has not been found up until this point. Commonly, the effect of cytotoxic drugs on benign cells is in most cases the same compared to regular cells while the actual effect on patient still can't be predicted. In order to reduce unwanted side effects or unspecific drug reactions a test system for patients which allows to analyse the interaction between cytotoxic agents and the targeted cells is needed. Furthermore, this should also include an adequate measurement system which is capable to work in a natural environment and without any additional preparation. In terms of this work, a first proof of concept with different benign cells and cytotoxic agents is presented while monitoring the obtained displacement using electronic speckle pattern interferometry (ESPI).}, language = {en} }