Filters

2017 - 201922020 - 20223

More filters

2020 - 20235
Reset filters

Settings

Subject: Screening ×

Search Results

Now showing 1 - 5 of 5
  • Thumbnail Image
    Item
    Identification of two kinase inhibitors with synergistic toxicity with low-dose hydrogen peroxide in colorectal cancer cells in vitro
    (Basel : MDPI AG, 2020) Freund, Eric; Liedtke, Kim-Rouven; Miebach, Lea; Wende, Kristian; Heidecke, Amanda; Kaushik, Nagendra Kumar; Choi, Eun Ha; Partecke, Lars-Ivo; Bekeschus, Sander
    Colorectal carcinoma is among the most common types of cancers. With this disease, diffuse scattering in the abdominal area (peritoneal carcinosis) often occurs before diagnosis, making surgical removal of the entire malignant tissue impossible due to a large number of tumor nodules. Previous treatment options include radiation and its combination with intraperitoneal heat-induced chemotherapy (HIPEC). Both options have strong side effects and are often poor in therapeutic efficacy. Tumor cells often grow and proliferate dysregulated, with enzymes of the protein kinase family often playing a crucial role. The present study investigated whether a combination of protein kinase inhibitors and low-dose induction of oxidative stress (using hydrogen peroxide, H2O2) has an additive cytotoxic effect on murine, colorectal tumor cells (CT26). Protein kinase inhibitors from a library of 80 substances were used to investigate colorectal cancer cells for their activity, morphology, and immunogenicity (immunogenic cancer cell death, ICD) upon mono or combination. Toxic compounds identified in 2D cultures were confirmed in 3D cultures, and additive cytotoxicity was identified for the substances lavendustin A, GF109203X, and rapamycin. Toxicity was concomitant with cell cycle arrest, but except HMGB1, no increased expression of immunogenic markers was identified with the combination treatment. The results were validated for GF109203X and rapamycin but not lavendustin A in the 3D model of different colorectal (HT29, SW480) and pancreatic cancer cell lines (MiaPaca, Panc01). In conclusion, our in vitro data suggest that combining oxidative stress with chemotherapy would be conceivable to enhance antitumor efficacy in HIPEC. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
  • Thumbnail Image
    Item
    Polymer-based controlled-release fed-batch microtiter plate - diminishing the gap between early process development and production conditions
    (Berlin ; Heidelberg : Springer, 2019) Keil, T.; Dittrich, B.; Lattermann, C.; Habicher, T.; Büchs, J.
    Background: Fed-batch conditions are advantageous for industrial cultivations as they avoid unfavorable phenomena appearing in batch cultivations. Those are for example the formation of overflow metabolites, catabolite repression, oxygen limitation or inhibition due to elevated osmotic concentrations. For both, the early bioprocess development and the optimization of existing bioprocesses, small-scale reaction vessels are applied to ensure high throughput, low costs and prompt results. However, most conventional small-scale procedures work in batch operation mode, which stands in contrast to fed-batch conditions in large-scale bioprocesses. Extensive expenditure for installations and operation accompany almost all cultivation systems in the market allowing fed-batch conditions in small-scale. An alternative, more cost efficient enzymatic glucose release system is strongly influenced by environmental conditions. To overcome these issues, this study investigates a polymer-based fed-batch system for controlled substrate release in microtiter plates. Results: Immobilizing a solid silicone matrix with embedded glucose crystals at the bottom of each well of a microtiter plate is a suitable technique for implementing fed-batch conditions in microtiter plates. The results showed that the glucose release rate depends on the osmotic concentration, the pH and the temperature of the medium. Moreover, the applied nitrogen source proved to influence the glucose release rate. A new developed mathematical tool predicts the glucose release for various media conditions. The two model organisms E. coli and H. polymorpha were cultivated in the fed-batch microtiter plate to investigate the general applicability for microbial systems. Online monitoring of the oxygen transfer rate and offline analysis of substrate, product, biomass and pH confirmed that fed-batch conditions are comparable to large-scale cultivations. Furthermore, due to fed-batch conditions in microtiter plates, product formation could be enhanced by the factor 245 compared to batch cultivations. Conclusions: The polymer-based fed-batch microtiter plate represents a sophisticated and cost efficient system to mimic typical industrial fed-batch conditions in small-scale. Thus, a more reliable strain screening and early process development can be performed. A systematical scale-down with low expenditure of work, time and money is possible. © 2019 The Author(s).
  • Thumbnail Image
    Item
    Glycolytic flux control by drugging phosphoglycolate phosphatase
    ([London] : Nature Publishing Group UK, 2022) Jeanclos, Elisabeth; Schlötzer, Jan; Hadamek, Kerstin; Yuan-Chen, Natalia; Alwahsh, Mohammad; Hollmann, Robert; Fratz, Stefanie; Yesilyurt-Gerhards, Dilan; Frankenbach, Tina; Engelmann, Daria; Keller, Angelika; Kaestner, Alexandra; Schmitz, Werner; Neuenschwander, Martin; Hergenröder, Roland; Sotriffer, Christoph; von Kries, Jens Peter; Schindelin, Hermann; Gohla, Antje
    Targeting the intrinsic metabolism of immune or tumor cells is a therapeutic strategy in autoimmunity, chronic inflammation or cancer. Metabolite repair enzymes may represent an alternative target class for selective metabolic inhibition, but pharmacological tools to test this concept are needed. Here, we demonstrate that phosphoglycolate phosphatase (PGP), a prototypical metabolite repair enzyme in glycolysis, is a pharmacologically actionable target. Using a combination of small molecule screening, protein crystallography, molecular dynamics simulations and NMR metabolomics, we discover and analyze a compound (CP1) that inhibits PGP with high selectivity and submicromolar potency. CP1 locks the phosphatase in a catalytically inactive conformation, dampens glycolytic flux, and phenocopies effects of cellular PGP-deficiency. This study provides key insights into effective and precise PGP targeting, at the same time validating an allosteric approach to control glycolysis that could advance discoveries of innovative therapeutic candidates.
  • Thumbnail Image
    Item
    Continuous electroosmotic sorting of particles in grooved microchannels
    (London : Royal Soc. of Chemistry, 2017) Dubov, Alexander L.; Molotilin, Taras Y.; Vinogradova, Olga I.
    We propose a novel microfluidic fractionation concept suitable for neutrally buoyant micron-sized particles. This approach takes advantage of the ability of grooved channel walls oriented at an angle to the direction of an external electric field to generate a transverse electroosmotic flow. Using computer simulations, we first demonstrate that the velocity of this secondary transverse flow depends on the distance from the wall, so neutrally buoyant particles, depending on their size and initial location, will experience different lateral displacements. We then optimize the geometry and orientation of the surface texture of the channel walls to maximize the efficiency of particle fractionation. Our method is illustrated in a full scale computer experiment where we mimic the typical microchannel with a bottom grooved wall and a source of polydisperse particles that are carried along the channel by the forward electroosmotic flow. Our simulations show that the particle dispersion can be efficiently separated by size even in a channel that is only a few texture periods long. These results can guide the design of novel microfluidic devices for efficient sorting of microparticles.
  • Thumbnail Image
    Item
    Slowness curve surface acoustic wave transducers for optimized acoustic streaming
    (Cambridge : Royal Society of Chemistry, 2020) O'Rorke, R.; Winkler, A.; Collins, D.; Ai, Y.
    Surface acoustic waves can induce force gradients on the length scales of micro- and nanoparticles, allowing precise manipulation for particle capture, alignment and sorting activities. These waves typically occupy a spatial region much larger than a single particle, resulting in batch manipulation. Circular arc transducers can focus a SAW into a narrow beam on the order of the particle diameter for highly localised, single-particle manipulation by exciting wavelets which propagate to a common focal point. The anisotropic nature of SAW substrates, however, elongates and shifts the focal region. Acousto-microfluidic applications are highly dependent on the morphology of the underlying substrate displacement and, thus, become dependent on the microchannel position relative to the circular arc transducer. This requires either direct measurement or computational modelling of the SAW displacement field. We show that the directly measured elongation and shift in the focal region are recapitulated by an analytical model of beam steering, derived from a simulated slowness curve for 128° Y-cut lithium niobate. We show how the negative effects of beam steering can be negated by adjusting the curvature of arced transducers according to the slowness curve of the substrate, for which we present a simple function for convenient implementation in computational design software. Slowness-curve adjusted transducers do not require direct measurement of the SAW displacement field for microchannel placement and can capture smaller particles within the streaming vortices than can circular arc IDTs.