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Epitachophoretic separation and concentration of large volume samples
Voráčová, Ivona ; Foret, František ; Datinská, Vladimíra ; Novotný, Jakub ; Gheibi, P. ; Berka, J. ; Astier, Y.
There has been a growing interest in developing isotachophoretic protocols for concentration and purification of DNA as an alternative to the solid phase extraction protocols of DNA preparation. Also concentration of DNA from large volume samples is required. We have developed a new device suitable for separation, concentration and collection of ions from several milliliter sample volumes into microliter fractions. We have used a discontinuous electrolyte system comprising of the leading and terminating electrolytes utilized for concentration of samples containing DNA fragments. The method is simple, fast with high concentration factor and without extensive surface interactions.
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A New evaluation of combined antitumor effects of natural and synthetic nuclear retinoid receptor ligands in human breast carcinoma cells
Strouhalová, Dana ; Laštovičková, Markéta ; Bobál, P. ; Macejová, D. ; Mosná, B. ; Brtko, J. ; Bobálová, Janette
This work represents the study of the effects of biologically active ligands of nuclear retinoid X receptors, triorganotin compounds, on proteomic profile of MDA-MB-231 cells. We focused primarily on tributyl/phenyltin derivatives that suggest the combined antitumor effect when they are used with natural nuclear retinoid receptor ligand all-trans retinoic acid (ATRA). Proteomic strategies connected to PDQuest evaluation were applied in this study. Some specific proteins with tumor process association were chosen for more detail study. Our findings indicate that selected triorganotins derivatives resulted to significantly reduced level of studied proteins belonging to metabolic pathway or to other cellular processes as apoptosis or epithelial–mesenchymal transition. In the case of vimentin, the key protein of EMT process, the silencing was set up by triorganotin compounds and significantly enhanced by combination with ATRA. This additive effect on downregulation can result from the potential engagement of RXR/RAR heterodimer, considering co-addition of both partner ligands of mentioned heterodimer to the treatment.
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Rapid fabrication of microextraction devices for direct determination of basic drugs in dried blood spots
Ryšavá, Lenka ; Přikryl, Jan ; Dvořák, Miloš ; Malá, Zdeňka ; Kubáň, Pavel
A 3D-printed cutter was designed for rapid fabrication of supported liquid membrane (SLM) extraction devices compatible with injection system of commercial capillary electrophoresis (CE) instrument. Low cost and single use microextraction devices were fabricated from polypropylene micropipette tips with high precision (dimension variances ≤ 3% RSD). Practical applicability of the microextraction devices was examined by in-line coupling of SLM extractions to CE for direct analysis of four model basic drugs in dried blood spot extracts. Excellent repeatability of the hyphenated technique was achieved (RSD ≤ 7.6%), transfer of analytes across the SLM ranged from 23 to 73% and limits of detection were as low as 0.07 μg/mL.
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A Microfluidic devide for monitoring cell migration
Ledvina, Vojtěch ; Klepárník, Karel ; Legartová, Soňa ; Bártová, Eva
A polydimethylsiloxane microfluidic device for monitoring directed cell migration was developed. The device comprises a main channel barred by pillar arrays with decreasing spacing and utilizes chemotactic movement of cells in the direction of increasing gradient of fetal bovine serum. HeLa cell line transfected with Premo FUCCI Cell Cycle Sensor was used as a model organism. The cells were imaged for several days using a scanning confocal microscope and the rate of migration in different cell cycle phases was evaluated.
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