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Takenaga, Shoko ; Schneider, Benno ; Erbay, E. ; Biselli, Manfred ; Schnitzler, Thomas ; Schöning, Michael Josef ; Wagner, Torsten
A new microfluidic assembly method for semiconductor-based biosensors using 3D-printing technologies was proposed for a rapid and cost-efficient design of new sensor systems. The microfluidic unit is designed and printed by a 3D-printer in just a few hours and assembled on a light-addressable potentiometric sensor (LAPS) chip using a photo resin. The cell growth curves obtained from culturing cells within microfluidics-based LAPS systems were compared with cell growth curves in cell culture flasks to examine biocompatibility of the 3D-printed chips. Furthermore, an optimal cell culturing within microfluidics-based LAPS chips was achieved by adjusting the fetal calf serum concentrations of the cell culture medium, an important factor for the cell proliferation.
Immel, Timo ; Grützke, Martin ; Späte, Anne-Katrin ; Groth, Ulrich ; Öhlschläger, Peter ; Huhn, Thomas
Chelate stabilization of a titanium(IV)–salan alkoxide by ligand exchange with 2,6-pyridinedicarboxylic acid (dipic) resulted in heptacoordinate complex 3 which is not redox-active, stable on silica gel and has increased aqueous stability. 3 is highly toxic in HeLa S3 and Hep G2 and has enhanced antitumor efficacy in a mouse cervical-cancer model.
Henken, F. E. ; Oosterhuis, K. ; Öhlschläger, Peter ; Bosch, L. ; Hooijberg, E. ; Haanen, J. B. A. G. ; Steenbergen, R. D. M.
Persistent infection with high-risk human papillomaviruses (hrHPV) can result in the formation of anogenital cancers. As hrHPV proteins E6 and E7 are required for cancer initiation and maintenance, they are ideal targets for immunotherapeutic interventions. Previously, we have described the development of DNA vaccines for the induction of HPV16 E6 and E7 specific T cell immunity. These vaccines consist of ‘gene-shuffled’ (SH) versions of HPV16 E6 and E7 that were fused to Tetanus Toxin Fragment C domain 1 (TTFC) and were named TTFC-E6SH and TTFC-E7SH. Gene-shuffling was performed to avoid the risk of inducing malignant transformation at the vaccination site. Here, we describe the preclinical safety evaluation of these candidate vaccines by analysis of their transforming capacity in vitro using established murine fibroblasts (NIH 3T3 cells) and primary human foreskin keratinocytes (HFKs). We demonstrate that neither ectopic expression of TTFC-E6SH and TTFC-E7SH alone or in combination enabled NIH 3T3 cells to form colonies in soft agar. In contrast, expression of HPV16 E6WT and E7WT alone or in combination resulted in effective transformation. Similarly, retroviral transduction of HFKs from three independent donors with both TTFC-E6SH and TTFC-E7SH alone or in combination did not show any signs of immortalization. In contrast, the combined expression of E6WT and E7WT induced immortalization in HFKs from all donors. Based on these results we consider it justified to proceed to clinical evaluation of DNA vaccines encoding TTFC-E6SH and TTFC-E7SH in patients with HPV16 associated (pre)malignancies.
Wiegand, Sandra ; Voigt, Birgit ; Albrecht, Dirk ; Bongaerts, Johannes ; Evers, Stefan ; Hecker, Michael ; Daniel, Rolf ; Liesegang, Heiko
Efficient FACS selection procedure for cells undergoing Flp-mediated site-specific conversions
(1998)
Transcription-promoting genomic sites in mammalia: their elucidation and architectural principles
(1998)
Bode, J. ; Bartsch, J. ; Boulikas, T. ; Iber, M. ; Mielke, C. ; Schübeler, D. ; Seibler, Jost ; Benham, C.
Feng, Yong-Qing ; Seibler, Jost ; Alami, Raouf ; Eisen, Andrew ; Westerman, Karen A. ; Leboulch, Philippe ; Fiering, Steven ; Bouhassira, Eric E.
The transgeneticist's toolbox: novel methods for the targeted modification of eukaryotic genomes
(2000)
Bode, Jürgen ; Schlake, Thomas ; Iber, Michaela ; Schübeler, Dirk ; Seibler, Jost ; Snezhkov, Evgeney ; Nikolaev, Lev
Cesari, Francesca ; Rennekampff, Verena ; Vintersten, Kristina ; Vuong, Lam Giang ; Seibler, Jost ; Bode, Jürgen ; Wiebel, Franziska F. ; Nordheim, Alfred
Goetze, Sandra ; Baer, Alexandra ; Winkelmann, Silke ; Nehlsen, Kristina ; Seibler, Jost ; Maass, Karin ; Bode, Jürgen
Seibler, Jost ; Zevnik, Branko ; Küter-Luks, Birgit ; Andreas, Susanne ; Kern, Heidrun ; Hennek, Thomas ; Rode, Anja ; Heimann, Cornelia ; Faust, Nicole ; Kauselmann, Gunther ; Schoor, Michael ; Jaenisch, Rudolf ; Rajewsky, Klaus ; Kühn, Ralf ; Schwenk, Frieder
Seibler, Jost ; Küter-Luks, Birgit ; Kern, Heidrun ; Streu, Sandra ; Plum, Leona ; Maurer, Jan ; Kühn, Ralf ; Brüning, Jens C. ; Schwenk, Frieder
Plum, Leona ; Ma, Xiaosong ; Hampel, Brigitte ; Balthasar, Nina ; Coppari, Roberto ; Münzberg, Heike ; Shanabrough, Marya ; Burdakov, Denis ; Rother, Eva ; Janoschek, Ruth ; Alber, Jens ; Belgardt, Bengt F. ; Koch, Linda ; Seibler, Jost ; Schenk, Frieder ; Fekete, Csaba ; Suzuki, Akira ; Mak, Tak W. ; Krone, Wilhelm ; Horvath, Tamas L. ; Ashcroft, Frances M. ; Brüning, Jens C.
Seibler, Jost ; Kleinridders, Andre ; Küter-Luks, Birgit ; Niehaves, Sandra ; Brüning, Jens C. ; Schwenk, Frieder
Investigation of TRPV1 loss-of-function phenotypes in transgenic shRNA expressing and knockout mice
(2008)
Christoph, Thomas ; Bahrenberg, Gregor ; Vry, Jean de ; Englberger, Werner ; Erdmann, Volker A. ; Frech, Moritz ; Kögel, Babette ; Röhl, Thomas ; Schiene, Klaus ; Schröder, Wolfgang ; Seibler, Jost ; Kurreck, Jens
Koch, Linda ; Wunderlich, F. Thomas ; Seibler, Jost ; Könner, A. Christine ; Hampel, Brigitte ; Irlenbusch, Sigrid ; Brabant, Georg ; Kahn, C. Ronald ; Schwenk, Frieder ; Brüning, Jens C.
Kotnik, Katarina ; Popova, Elena ; Todiras, Mihail ; Mori, Marcelo A. ; Alenina, Natalia ; Seibler, Jost ; Bader, Michael
Glaser, Stefan ; Lubitz, Sandra ; Loveland, Kate L. ; Ohbo, Kazu ; Robb, Lorraine ; Schwenk, Frieder ; Seibler, Jost ; Roellig, Daniela ; Kranz, Andrea ; Anastassiadis, Konstantinos ; Stewart, A. Francis
Jordan, Sabine D. ; Krüger, Markus ; Willmes, Diana M. ; Redemann, Nora ; Wunderlich, F. Thomas ; Brönneke, Hella S. ; Merkwirth, Carsten ; Kashkar, Hamid ; Olkkonen, Vesa M. ; Böttger, Thomas ; Braun, Thomas ; Seibler, Jost ; Brüning, Jens C.
The contribution of altered post-transcriptional gene silencing to the development of insulin resistance and type 2 diabetes mellitus so far remains elusive. Here, we demonstrate that expression of microRNA (miR)-143 and 145 is upregulated in the liver of genetic and dietary mouse models of obesity. Induced transgenic overexpression of miR-143, but not miR-145, impairs insulin-stimulated AKT activation and glucose homeostasis. Conversely, mice deficient for the miR-143–145 cluster are protected from the development of obesity-associated insulin resistance. Quantitative-mass-spectrometry-based analysis of hepatic protein expression in miR-143-overexpressing mice revealed miR-143-dependent downregulation of oxysterol-binding-protein-related protein (ORP) 8. Reduced ORP8 expression in cultured liver cells impairs the ability of insulin to induce AKT activation, revealing an ORP8-dependent mechanism of AKT regulation. Our experiments provide direct evidence that dysregulated post-transcriptional gene silencing contributes to the development of obesity-induced insulin resistance, and characterize the miR-143–ORP8 pathway as a potential target for the treatment of obesity-associated diabetes.
Hasegawa, Maki ; Kapelyukh, Yury ; Tahara, Harunobu ; Seibler, Jost ; Rode, Anja ; Krueger, Sylvia ; Lee, Dongtao N. ; Wolf, C. Roland ; Scheer, Nico
Raab, Monika ; Kappel, Sven ; Krämer, Andrea ; Sanhaji, Mourad ; Matthess, Yves ; Kurunci-Csacsko, Elisabeth ; Calzada-Wack, Julia ; Rathkolb, Birgit ; Rosman, Jan ; Adler, Thure ; Busch, Dirk H. ; Esposito, Irene ; Fuchs, Helmut ; Gailus-Durner, Valérie ; Klingenspor, Martin ; Wolf, Eckhard ; Sänger, Nicole ; Prinz, Florian ; Hrabe de Angelis, Martin ; Seibler, Jost ; Yuan, Juping ; Bergmann, Martin ; Knecht, Rainald ; Kreft, Bertolt ; Strebhardt, Klaus
Meiotic functions of RAD18
(2011)
Inagaki, Akiko ; Sleddens-Linkels, Esther ; Wassenaar, Evelyne ; Ooms, Marja ; Cappellen, Wiggert A. van ; Hoeijmakers, Jan H. J. ; Seibler, Jost ; Vogt, Thomas F. ; Shin, Myung K. ; Grootegoed, J. Anton ; Baarends, Willy M.
Obesity-induced overexpression of miR-802 impairs glucose metabolism through silencing of Hnf1b
(2013)
Kornfeld, Jan-Wilhelm ; Baitzel, Catherina ; Könner, A. Christine ; Nicholls, Hayley T. ; Vogt, Merly C. ; Herrmanns, Karolin ; Scheja, Ludger ; Haumaitre, Cécile ; Wolf, Anna M. ; Knippschild, Uwe ; Seibler, Jost ; Cereghini, Silvia ; Heeren, Joerg ; Stoffel, Markus ; Brüning, Jens C.
Gebeshuber, Christoph A. ; Kornauth, Christoph ; Dong, Lihua ; Sierig, Ralph ; Seibler, Jost ; Reiss, Martina ; Tauber, Stefanie ; Bilban, Martin ; Wang, Shijun ; Kain, Renate ; Böhmig, Georg A. ; Moeller, Marcus J. ; Gröne, Hermann-Josef ; Englert, Christoph ; Martinez, Javier ; Kerjaschki, Dontscho
Michalak, Ewa Malgorzata ; Nacerddine, Karim ; Pietersen, Alexandra ; Beuger, Vincent ; Pawlitzky, Inka ; Cornelissen-Steijger, Paulien ; Wientjens, Ellen ; Tanger, Ellen ; Seibler, Jost ; Lohuizen, Maarten van ; Jonkers, Jos
A High-Throughput Functional Complementation Assay for Classification of BRCA1 Missense Variants
(2013)
Bouwman, Peter ; Gulden, Hanneke van der ; Heijden, Ingrid van der ; Drost, Rinske ; Klijn, Christiaan N. ; Prasetyanti, Pramudita ; Pieterse, Mark ; Wientjens, Ellen ; Seibler, Jost ; Hogervorst, Frank B. L. ; Jonkers, Jos
Breuer, Lars ; Raue, Markus ; Kirschbaum, M. ; Mang, Thomas ; Schöning, Michael Josef ; Thoelen, R. ; Wagner, Torsten
Poly(N-isopropylacrylamide) (PNIPAAm) hydrogel films with incorporated graphene oxide (GO) were developed and tested as light-stimulated actuators. GO dispersions were synthesized via Hummers method and characterized toward their optical properties and photothermal energy conversion. The hydrogels were prepared by means of photopolymerization. In addition, the influence of GO within the hydrogel network on the lower critical solution temperature (LCST) was investigated by differential scanning calorimetry (DSC). The optical absorbance and the response to illumination were determined as a function of GO concentration for thin hydrogel films. A proof of principle for the stimulation with light was performed.
An enzyme-based multi-parameter biosensor is developed for monitoring the concentration of formate, d-lactate, and l-lactate in biological samples. The sensor is based on the specific dehydrogenation by an oxidized β-nicotinamide adenine dinucleotide (NAD+)-dependent dehydrogenase (formate dehydrogenase, d-lactic dehydrogenase, and l-lactic dehydrogenase, respectively) in combination with a diaphorase from Clostridium kluyveri (EC 1.8.1.4). The enzymes are immobilized on a platinum working electrode by cross-linking with glutaraldehyde (GA). The principle of the determination scheme in case of l-lactate is as follows: l-lactic dehydrogenase (l-LDH) converts l-lactate into pyruvate by reaction with NAD+. In the presence of hexacyanoferrate(III), the resulting reduced β-nicotinamide adenine dinucleotide (NADH) is then regenerated enzymatically by diaphorase. The electrochemical detection is based on the current generated by oxidation of hexacyanoferrate(II) at an applied potential of +0.3 V vs. an Ag/AgCl reference electrode. The biosensor will be electrochemically characterized in terms of linear working range and sensitivity. Additionally, the successful practical application of the sensor is demonstrated in an extract from maize silage.
Seifarth, Volker ; Goßmann, Matthias ; Grosse, J. O. ; Becker, C. ; Heschel, I. ; Artmann, Gerhard ; Temiz Artmann, Aysegül
Raue, Markus ; Wambach, M. ; Glöggler, S. ; Grefen, Dana ; Kaufmann, R. ; Abetz, C. ; Georgopanos, P. ; Handge, U. A. ; Mang, Thomas ; Blümich, B. ; Abetz, V.
Handtke, Stefan ; Volland, Sonja ; Methling, Karen ; Albrecht, Dirk ; Becher, Dörte ; Nehls, Jenny ; Bongaerts, Johannes ; Maurer, Karl-Heinz ; Lalk, Michael ; Liesegang, Heiko ; Voigt, Birgit ; Daniel, Rolf ; Hecker, Michael
Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC–MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed.
Frotscher, Ralf ; Goßmann, Matthias ; Raatschen, Hans-Jürgen ; Temiz Artmann, Aysegül ; Staat, Manfred
We present an electromechanically coupled Finite Element model for cardiac tissue. It bases on the mechanical model for cardiac tissue of Hunter et al. that we couple to the McAllister-Noble-Tsien electrophysiological model of purkinje fibre cells. The corresponding system of ordinary differential equations is implemented on the level of the constitutive equations in a geometrically and physically nonlinear version of the so-called edge-based smoothed FEM for plates. Mechanical material parameters are determined from our own pressure-deflection experimental setup. The main purpose of the model is to further examine the experimental results not only on mechanical but also on electrophysiological level down to ion channel gates. Moreover, we present first drug treatment simulations and validate the model with respect to the experiments.
Verfahren zur Bestimmung des enzymatischen Status humaner Haut in vitro [Offenlegungsschrift]
(2004)
Banowski, Bernhard ; Hoffmann, Daniele ; Wadle, Armin ; Siegert, Petra ; Sättler, Andrea ; Gerke, Thomas