National Repository of Grey Literature 40 records found  1 - 10nextend  jump to record: Search took 0.00 seconds. 
New fungal fucosidases and their synthetic potential
Glozlová, Michaela ; Beránek, Martin (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Glozlová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultants: doc. RNDr. Pavla Bojarová, Ph.D., Mgr. Pavlína Nekvasilová Title of diploma thesis: New fungal fucosidases and their synthetic potential The literary research focuses on α-L-fucosidases, their occurrence, functions, and use in the synthesis of fucosylated compounds, which were also a part of the research, particularly their presence in the human body and involvement in pathological conditions. Furthermore, it explores the oligosaccharides found in human milk, their impact on infant health, and their synthesis, which can be facilitated through the use of α-L-fucosidases. The experimental section involves the analysis of a library of filamentous fungi with the aim of identifying a representative strain with α-L-fucosidase activity towards pNP-α-L-fucoside. Recombinant α-L-fucosidases were selected based on searches in genetic databases and prepared. The feasibility of their production in Pichia pastoris and Escherichia coli was tested, and the produced enzymes were purified by ion exchange chromatography, characterized and their regioselectivity and synthetic potential for the synthesis of fucosylated...
Effect of enzymes on the degradation of thermosensitive copolymers for controlled release of drugs
Dávidíková, Anna Mária ; Michlovská, Lenka (referee) ; Vojtová, Lucy (advisor)
Termocitlivé hydrogély, ktoré sa používajú ako nosiče liečiv v dnešnej dobe zažívajú obrovský záujem v spoločnosti. Používajú sa ako spôsob dopravy liečiva na predom určené miesto a po dosiahnutí určeného miesta slúžia ako dlhodobé úložisko liečiv v tele. Vlastnosti hydrogélov môžu byť upravované tak, aby dokázali naviazať rôzne druhy liečiv, aby boli citlivé na rôzne vonkajšie impulzy a aby degradovali pomalšie, čo predĺži životnosť liečiv v tele. Aby sme vedeli, ako upraviť vlastnosti hydrogélov a ako sa budú správať, musíme poznať ich degradačný proces a chovanie v rôznych degradačných médiách. Práve preto sú degradačné štúdie veľmi dôležitou súčasťou štúdií hydrogélov. Táto bakalárska práca je zameraná na sledovanie degradácie rôznych roztokov termocitlivého hydrogélu vo viacerých degradačných médiách. Teoretická časť práce zahrňuje krátky prehľad o tom, čo sú to termocitlivé hydrogély, ich charakterizácia a popis vlastností spolu s použitím hydrogélov ako nosičov liečiv. Ďalej sa teoretická časť venuje degradácií hydrogélov celkovo ako aj degradácií konkrétne použitého hydrogélu v tejto bakalárskej práci. V prvom rade bol syntetizovaný živou polymeráciou za otváraním kruhu v inertnej atmosfére termocitlivý PLGA-PEG-PLGA kopolymér, tvorený poly(mliečnou-co-glykolovou kyselinou)-b-poly(ethylen glykolom)-b-poly(mliečnou-co-glykolovou kyselinou). Následne bol kopolymér charakterizovaný na protónovej nukleárnej magnetickej rezonancii a na gélovej chromatografii. Z nasyntetizovaného kopolyméru boli pripravené tri vodné roztoky hydrogélu o rôznych koncentráciách a to 10, 15 a 20 hm %. Vzorky pripravených hydrogélov za teploty 37 , boli degradované v rôznych degradačných roztokoch (UPW (ultra čistá voda), NaCl, PBS (fosfátový pufr) a roztok PBS s enzýmom). Degradačná štúdia bola hlavným cieľom tejto bakalárskej práce. Jednotlivé vzorky boli odobrané v konkrétne stanovené časy a to po 1, 2, 3, 7, 9, 14, 16, 21, 23 a 30 dňoch. Pri vzorkách s enzýmom bola predpokladaná rýchlejšia degradácia vzoriek, tak tieto odbery boli nastavené po 1, 2 a 3 dňoch. Ďalším cieľom tejto práce bola analýza odobraných vzoriek hydrogélov po degradácií na gélovej chromatografii a analýza degradačných médií na kvapalinovej chromatografií s hmotnostným spektrometrom. Počas degradácie vzoriek bola prevedená taktiež gravimetrická analýza a pri odoberaní degradačných roztokov bolo merané ich pH. Následne boli všetky získané dáta vyhodnotené a bolo sledované, či enzým urýchľuje PLGA-PEG-PLGA hydrolytickú degradačnú kinetiku. Výsledky poukazujú na to, že enzým má nízky vplyv na rýchlosť degradácie. Výsledky tejto práce poslúžia ako základy pre budúce degradačné štúdie.
New fungal fucosidases and their synthetic potential
Glozlová, Michaela ; Beránek, Martin (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Glozlová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultants: doc. RNDr. Pavla Bojarová, Ph.D., Mgr. Pavlína Nekvasilová Title of diploma thesis: New fungal fucosidases and their synthetic potential The literary research focuses on α-L-fucosidases, their occurrence, functions, and use in the synthesis of fucosylated compounds, which were also a part of the research, particularly their presence in the human body and involvement in pathological conditions. Furthermore, it explores the oligosaccharides found in human milk, their impact on infant health, and their synthesis, which can be facilitated through the use of α-L-fucosidases. The experimental section involves the analysis of a library of filamentous fungi with the aim of identifying a representative strain with α-L-fucosidase activity towards pNP-α-L-fucoside. Recombinant α-L-fucosidases were selected based on searches in genetic databases and prepared. The feasibility of their production in Pichia pastoris and Escherichia coli was tested, and the produced enzymes were purified by ion exchange chromatography, characterized and their regioselectivity and synthetic potential for the synthesis of fucosylated...
Nanoporous Enzyme-Based Impedimetric Biosensor
Košelová, Z.
The ordered nanopores surfaces has been studied in the field of biosensors for the detection of affinity interactions such as antigen-antibody or nucleic acid hybridization. Nanopores are frequently fabricated by anodic oxidation of aluminium. This method is practically simple method of highly ordered pore formation, but is not suitable for massive production. The principle of detection is usually based on the degree of pore blockage by biomolecular interaction that allows a sensitive electrochemical response. Here we show the development and characterization of the nanopore-based enzymatic biosensor for the detection of analytes. We performed and tested the concept of the simple and versatile method of nanopores formation on the electrode using non-conductive polystyrene nanoparticles. The variability in pore size was modulated by nanoparticles diameter (40 and 80 nm). We measured response of our designed biosensor using electrochemical impedance spectroscopy in presence of redox mediator at low potential. The smaller diameter of nanopores increases charge transfer resistance. Biosensors with 80 nm nanoparticles had behaviour similar to reference without nanoparticles, therefore the pore was too wide for this design. The limit of detection from Rct showed to be at most 0.01 mM for every sample type. This proposed biosensor contributes to the basic knowledge about nanopores sensors, and it could have a potential, for example, in the development of ―point of care‖ devices.
The impact of synthetic chelators from the group of 4-acylpyrazole-5-ones on alcohol dehydrogenase
Sobolová, Zuzana ; Mladěnka, Přemysl (advisor) ; Jirkovský, Eduard (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Zuzana Sobolová Supervisor: Prof. Přemysl Mladěnka, Pharm.D., Ph.D. Consultant: Marcel Hrubša, MS.c. Title of diploma thesis: The impact of synthetic chelators from the group of 4-acylpyrazole-5-ones on alcohol dehydrogenase Inhibitors of alcohol dehydrogenase play a key role in the treatment of poisoning caused by methanol, ethanol, ethylene glycol, and their toxic metabolites. Such treatment can therefore be considered as life-saving. In clinical practice, a single substance, fomepizole, is used. This thesis is aimed at investigation of the effect of eleven experimental chelators from the 4- acylpyrazol-5-ones group on equine and yeast alcohol dehydrogenases, and the possible definition of structural features that would be the most suitable for interaction with this enzyme. Due to the ability of these substances to chelate zinc ions, it is believed that they could inhibit an enzyme that has a zinc molecule in the structure. The measurement itself is based on principle of increase in absorbance during the enzymatic reaction, which is caused by the formation of NADH. This measurement is relatively simple, efficient and, due to used kinetic method, also less susceptible to be affected by...
Acitivity of cellulase C1 and FP in commercial products and its response during hydrolysis of biomass
FOUNĚ, Jakub
The deal of the work was to determine the activity of enzyme cellulase in commercial product Onozuka R-10.
Sulfates as phase II metabolites of natural phenolic compounds
Hetman, Anastasiia ; Mladěnka, Přemysl (advisor) ; Siatka, Tomáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Anastasiia Hetman Supervisor: Assoc. Prof. Přemysl Mladěnka, PharmD., Ph.D. Co-supervisor: Assoc. Prof. Kateřina Valentová, Ph.D. Title of diploma thesis: Sulfates as phase II metabolites of natural phenolic compounds Natural monophenolic compounds, which both occur in plants and are formed during gut microbiota metabolism of polyphenols, are absorbed and pass through phase II metabolism. This results in the formation of various conjugated metabolites, such as sulfates, whose biological activity was not studied as yet. This work focused on the preparation and isolation of pure sulfated metabolites of phenolic compounds, namely 4- methylcatechol, protocatechuic, homoprotocatechuic, and 2,3,4-trihydroxybenzoic acid, and phloroglucinol, using the aryl sulphotransferase from Desulfitobacterium hafniense. As a result, two sulfated products of sufficient purity were prepared, namely 4- methylcatechol-O-sulfate (an inseparable mixture of 4-methylcatechol-2-O-sulfate and 4- methylcatechol-1-O-sulfate) and phloroglucinol-O-sulfate. Their antioxidantactivity was evaluated and compared with the activity of the parent compounds using the following six in vitro tests: DPPH (1-diphenyl-2-picrylhydrazyl) and...
Immobilization of protease V8 on magnetic particles for application to proteolytic cleavage of pepsin A
Čepa, Adam ; Pacáková, Věra (advisor) ; Tichá, Marie (referee)
This thesis is part of a long-standing research in the field of diagnosis of the stomach diseases, which is based on the gastric enzyme pepsin A mapping. It was found that a phosphorylation in the primary structure of this enzyme may serve as a marker of incipient stage of carcinogenesis. This thesis is focused on the immobilization of protease V8 isolated from microorganism Stafylococcus aureus to magnetic agarose beads. Protease V8 is a promising candidate for producing peptide maps of pepsin A. The influence of pH, temperature and reaction time on the enzyme to activity has been studied and the optimal conditions for hydrolytic catalysis of formation of peptide fragments of pepsin A.
Preparation of Influenza Neuraminidase and Polymerase Inhibitors
Zima, Václav
Influenza is an infectious disease caused by the influenza virus. This virus causes a severe viral infection that spreads easily from person to person in yearly pandemics. Vaccination is the most effective way to prevent the infection, however, due to the high rate in mutations of the virus, the vaccine needs to be often reformulated. Another option to combat influenza is based on administration of antiviral drugs. Clinical studies of isolated influenza strains ("avian flu" H5N1, 2004; "swine flu" H1N1, 2009) revealed resistance towards known influenza neuraminidase inhibitors (zanamivir, oseltamivir). The resistance is caused by structural changes close to the enzymatic site. This calls for the development of new neuraminidase inhibitors as well for development of inhibitors targeting different influenza enzymes. This Thesis is focused on design and synthesis of new inhibitors of influenza neuraminidase and RNA-dependent RNA polymerase, namely PA subunit and the assembly of PA-PB1 heterodimer enzymes (Scheme 1). Influenza neuraminidase inhibitors were prepared by C-5 derivatization of oseltamivir followed by subsequent extension of its structure with binders of 150-cavity. Binding potencies of new oseltamivir derivatives against two influenza strains were determined. The next part contributed to...
The impact of synthetic chelators from the group of 4-acylpyrazole-5-ones on alcohol dehydrogenase
Sobolová, Zuzana ; Mladěnka, Přemysl (advisor) ; Jirkovský, Eduard (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Zuzana Sobolová Supervisor: Prof. Přemysl Mladěnka, Pharm.D., Ph.D. Consultant: Marcel Hrubša, MS.c. Title of diploma thesis: The impact of synthetic chelators from the group of 4-acylpyrazole-5-ones on alcohol dehydrogenase Inhibitors of alcohol dehydrogenase play a key role in the treatment of poisoning caused by methanol, ethanol, ethylene glycol, and their toxic metabolites. Such treatment can therefore be considered as life-saving. In clinical practice, a single substance, fomepizole, is used. This thesis is aimed at investigation of the effect of eleven experimental chelators from the 4- acylpyrazol-5-ones group on equine and yeast alcohol dehydrogenases, and the possible definition of structural features that would be the most suitable for interaction with this enzyme. Due to the ability of these substances to chelate zinc ions, it is believed that they could inhibit an enzyme that has a zinc molecule in the structure. The measurement itself is based on principle of increase in absorbance during the enzymatic reaction, which is caused by the formation of NADH. This measurement is relatively simple, efficient and, due to used kinetic method, also less susceptible to be affected by...

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