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Design of Stationary Machine for Threadig Plastic Sheathing on Wide Bandsaw Blades
Riedl, Jan ; Vespalec, Arnošt (referee) ; Koutný, Daniel (advisor)
This thesis deals with the design of a machine for threading plastic sheathing on single-sided band saw blades with dimensions from 4500 x 80 x 1.0 mm to 12500 x 290 x 1.85 mm. The aim of the thesis is to create an assembly model, the text is focused on a detailed description of the machine. The device was designed in SOLIDWORKS software. The main goal and partial goals of the thesis were met. The thesis is developed to the level of structural design and assembly description, along with calculations of key parts in the structural solution. The method of operating the machine is outlined. The thesis can serve as a starting point for future designs of a machine for threading protective plastic sheathing on saw bands of the stated dimensions.
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Design of Stationary Machine for Threadig Plastic Sheathing on Wide Bandsaw Blades
Riedl, Jan ; Vespalec, Arnošt (referee) ; Koutný, Daniel (advisor)
This thesis deals with the design of a machine for threading plastic sheathing on single-sided band saw blades with dimensions from 4500 x 80 x 1.0 mm to 12500 x 290 x 1.85 mm. The aim of the thesis is to create an assembly model, the text is focused on a detailed description of the machine. The device was designed in SOLIDWORKS software. The main goal and partial goals of the thesis were met. The thesis is developed to the level of structural design and assembly description, along with calculations of key parts in the structural solution. The method of operating the machine is outlined. The thesis can serve as a starting point for future designs of a machine for threading protective plastic sheathing on saw bands of the stated dimensions.
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Voltammetric Behavior of 4-Aminophthalimide Label using Hanging Mercury Drop Electrode
Ferenčíková, Z. ; Daňhel, Aleš ; Riedl, Jan ; Hocek, Michal ; Fojta, Miroslav
4-aminophthalimide (API), covalently attached to nucleotide (dCTP or dATP) has recently beendesigned as a fluorescent label for the detection of DNA-protein interaction. However API is electrochemically reducible and therefore it is possible to use alternatively the API-modified nuclotides as DNA redox labels. The electrochemical behavior of labeled nucleotides and labeled DNA was studied using cyclic voltammetry and transfer stripping cyclic voltammetry at hanging mercury drop electrode. Observed CVs showed irreversible reduction signals of 4-aminophthalimide label without production of consequently oxidizable/reducible species.
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Utilisation of enzymatic labelling with 4-aminophtalimide and 4-hydroxybenzylideneimidazolinone fluorescent derivates for monitoring of DNA-protein interaction
Orság, Petr ; Pivoňková, Hana ; Riedl, Jan ; Hocek, Michal ; Fojta, Miroslav
The 5’-substituted deoxycytosine triphosphates with conjugated solvatochromic derivates of 4-aminophtalimide (API) and derivates of the green fluorescent protein, 4-hydroxybenzylideneimidazolinone (HBI) were synthetized and successfully tested for enzymatic incorporation using primer extension assay. Site specifically labelled oligonucleotide probes were prepared and tested for interaction with p53 and SSB proteins, displaying distinct DNA-binding properties. The incorporation of multiple fluorescent labels did not interfere with natural protein binding and protein interaction leaded in both cases the to the gradual ratiometric increase of the fluorescence intensity moreover accompanied with the changes of the fluorescence emission spectra profile. Neither effect was observed after incubation with BSA, non-DNA binding protein, confirming the specificity of the interaction. Modified nucleoside triphosphates with conjugated fluorescence labels derivates of API and HBI can be used as substrates for preparation of the specific oligonucleotide labelled probes and provide the novel tool for studying and monitoring the DNA-protein interaction.
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Polymerase synthesis of base-modified DNA: New methods and new applications
Balintová, Jana ; Daďová, Jitka ; Kielkowski, Pavel ; Ménová, Petra ; Vaníková, Zuzana ; Riedl, Jan ; Raindlová, Veronika ; Fojta, Miroslav ; Hocek, Michal
Diverse base-modified oligonucleotides and double-stranded DNA molecules were prepared by polymerase incorporation of modified nucleoside triphosphates. The methods include primer extension, PCR, nicking enzyme amplification reaction and mixed incorporations. The modified nucleic acids were used in redox and fluorescence labelling and coding, as well as regulation of binding and cross-linking with proteins.
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Syntéza modifikovaných fluorescenčních nukleosidů
Riedl, Jan ; Hocek, Michal
A series of modified nucleosides bearing the fluorescent substituted biaryl labels attached to nucleobase was prepared and their luminescent properties were evaluated. The modified nucleosides show strong fluorescence at ca. 400 nm when excited at 310 nm in water depending on substitution pattern.
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