Národní úložiště šedé literatury Nalezeno 32 záznamů.  1 - 10dalšíkonec  přejít na záznam: Hledání trvalo 0.01 vteřin. 
Enzymatic Incorporation of Biotin into DNA for DNA Hybridization Analysis and for Sensitive Detection of PCR-Amplified DNA
Špaček, Jan ; Zenka, Martin ; Haroniková, Lucia ; Havran, Luděk ; Fojta, Miroslav
We present two enzymatical electrochemical assays for DNA analysis. For hybridization analysis we used probes with biotin-14-dC introduced to 3' OH end by terminal transferase. For detection of PCR products we used Deep Vent polymerase to incorporate biotin-14-dCduring PCR. In both cases streptavidin-alkaline phosphatase conjugate was subsequently attached to the incorporated biotins and was used to catalyze dephosphorylation of 1-naphthyl phosphate to 1-naphthol, the electrochemical signal of which was utilized for detection. Compared to the former method, biotin incorporation during PCR offers lower molar detection limits, whereas application of the biotin-tailed probe can provide us with more selective detection.
Primer Extension Reaction at the Gold Electrode Surface
Vidláková, Pavlína ; Havran, Luděk ; Fojta, Miroslav
One option of covalent DNA labeling is incorporation of chemically modified nucleotides using corresponding deoxynucleotide triphosphates into DNA molecules using polymerase-catalyzed primer extension. Primer extension is usually performed in solution and then the product is analyzed, often after a suitable separation. In our study we tested the possibility to prepare oligonucleotides with enzymatically incorporated anthraquinone labeled nucleotides using primer extension on templates immobilize at a gold electrode surface.
Natural and Synthetic Components of Nucleic Acids as Reactive Sites for DNA Modification by Osmium Tetroxide Complexes
Havran, Luděk ; Špaček, Jan ; Rozkosna, Jana ; Fojta, Miroslav
Natural DNA electroactivity find wide use in electrochemical analysis of its interactions and damage. For some applications is useful applied redox active tag to improve specificity of the analysis. One from utilized labels in this field are complexes of osmium tetroxide with nitrogen ligands (Os, L), which produce with DNA electroactive covalent adducts. Prime reaction site for Os, L is C=C double bond in pyrimidine nucleobases. If is 2,2'-bipyridine used as ligand, reaction is specific for single strand DNA. In this contribution will be presented results of electrochemical analysis of Os, bipy adducts with different ODN containing chemically modified purine bases.
Electrochemical Reduction and Oxidation of Nucleic Acids Bases and their Analogues: a Brief Overview
Fojta, Miroslav ; Špaček, Jan ; Dudová, Zdenka ; Pivoňková, Hana ; Daňhel, Aleš ; Fojt, Lukáš ; Havran, Luděk
Nucleic acids are known as electroactive biomacromolecules containing electrochemically reducible or oxidizable constituents. Nucleobases cytosine, 5-methylcytosine, adenine and guanine can be reduced in aqueous media on mercury or silver amalgam electrodes. Oxidation of all natural nucleic acids bases (in addition to the above mentioned ones, also uracil and thymine) was demonstrated using various types of carbon electrodes. Some of synthetic nucleobases or nucleotide analogues (e g., 7-deazapurines, cytidine analogues used as epigenetic modulators, etc.) exhibit specific electrochemical properties that differ from those of the parent bases and can be utilized to determine the given substance in the presence of natural nucleic acids or their components.
Voltammetry of Metallothioneins on Mercury Meniscus Modified Silver Solid Amalgam Electrode
Šestáková, Ivana ; Křivská, D. ; Josypčuk, Bohdan ; Nováková, Kateřina ; Navrátil, Tomáš
Peak of AgMT is formed on m-AgSAE from MT in alkaline media. The height of this peak is proportional to the MT concentration in range 37- to 301 ng /ml and can be increased with accumulation. In ammonia solution with Co(III) salt, apart from usual Brdička response at -1400 mV another, more positive catalytic system is observed, when formation of AgMT is allowed. This phenomenon has been confirmed by elimination voltammetry.
Information on Several Interesting Case Reports of Liquid Mercury Intoxication
Navrátil, Tomáš ; Vlčková, Š. ; Mrázová, K. ; Nováková, Kateřina ; Zakharov, S. ; Honsová, Š. ; Pelclová, D.
This contribution reports on several mercury non-professional intoxications. It can be summarized that it is very important to differentiate among forms of mercury which can (acute or chronic) endanger human health. Contrarily to the wide spread fears, liquid mercury represents low danger for men and for environment. Solid amalgams are probably the less toxic forms of mercury compounds. On the other hand, widely spread daily consumer goods can be very dangerous and toxic (saving balls, fluorescent tubes, etc.). Moreover, these goods are not marked as “toxic mercury containing”. The article discusses the instruction for removing mercury from our environment. Most of the reported data are based on database of the Czech Toxicological Information Centre (TIC) (from the years 1995 – 2015).
Electrochemical Biosensors Based on Silver Solid Amalgam for Analysis in Flow Systems
Josypčuk, Oksana ; Barek, J. ; Josypčuk, Bohdan
In this contribution, the construction and preparation of 3 types of flow amperometric enzymatic biosensors based on silver solid amalgam (AgSA) were presented. Two of them consist of the tubular detector and enzymatic reactor of porous or powdered AgSA. The third biosensor was constructed using polished AgSA electrode. Reactors and polished AgSA electrodes were modified with thiols or chitosan and different enzymes were covalently immobilized by suitable techniques. Prepared biosensors were used for determination of ascorbic acid, glucose, hydrogen peroxide, catechol, pyrogallol, dopamine and sarcosine in model samples and in same cases in the real samples.
Unique Properties of Amalgam Electrodes
Josypčuk, Bohdan ; Josypčuk, Oksana
Depending on metal-mercury ratio and on the method of preparation amalgam can be: 1) solid compact; 2) paste; 3) liquid; 4) monocrystalline; 5) porous; 6) powdered. High potential of hydrogen overvoltage is observed on amalgam electrodes and it is comparable with one on Hg-electrodes. Another important benefit is the possibility to prepare amalgam electrode, detector or reactor of required size and shape. From amalgams have been prepared and tested many types of electrodes for batch and flow-through systems. The diversity of amalgam materials provides some unique electrochemical properties of electrodes, the most important of which are described in this paper.
Transport of Phytochelatin PC2 across Model Phospholipid Membrane
Šestáková, Ivana ; Nováková, Kateřina ; Josypčuk, Bohdan ; Navrátil, Tomáš
Using electrochemical cell with two compartments separated by phospholipid membrane on polycarbonate support, transport of phytochelatin PC2 by means of cell penetrating peptide TP10 and in lesser extent with Mastoparan X was confirmed with voltammetry. Formation of the membrane and transport was followed by electrochemical impedance spectroscopy.
Methanol Outbreak in the Czech Republic in the year 2012 – Almost Two Years Later
Navrátil, Tomáš ; Zakharov, S. ; Pelclová, D. ; Mrázová, K.
Almost two years ago, in autumn 2012, a large methanol outbreak was registered in the Czech Republic (CR). The first case was registered in September 2012 in Moravian city Havířov. The source was connected with illegal production and sale of adulterated spirits. About 50 dead and more than 150 cases of methanol intoxications have been reported since September 2012. Some cases of intoxications, connected with old methanol sources distributed in 2012, have been registered up to now. Not all of total amount of prepared methanolic drinks has been found and hundreds liters of toxic methanol have remained in stores or in households. n the other hand, many people intoxicated during this outbreak and classified as “fully recovered” by their hospital discharge, have suffered by sequels of their intoxication at present (visual and nervous system disturbances) and their number has been increasing.

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