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Detection of His-tag and FLAG-tag recombinant proteins
Javorská, Lenka ; Zemanová, Lucie (advisor) ; Novotná, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Javorská Supervisor: RNDr. Lucie Škarydová, Ph.D Title of diploma thesis: Detection of His-tag and FLAG-tag recombinant proteins Recombinant proteins are proteins produced in genetically modified organism by recombinant DNA technology that enable insert DNA sequence of target protein in expression cell. This technology allows preparation of actually any protein in any quantity without difficulties associated with demanding purification of native protein from tissues. Recombinant proteins are produced often in form of fusion proteins with suitable peptide tag (e.g. His-tag, FLAG- tag) that can help with their detection and enable simple purification. The aim of this diploma thesis was to introduce and optimize universal method for the detection of recombinant proteins labelled with His-tag (6x histidine) and FLAG-tag. The model recombinant protein with His-tag was enzyme carbonylreductase 1 (CBR1) produced by expression system Escherichia coli. For FLAG-tag labelled protein were chosen enzymes DHRS12, HSD11β1, RODH4, DHRS3, DHRS7, DHRS8 produced by insect cells (strain Sf9). SDS polyacrylamide electrophoresis for protein separation and immunochemical detection after Western...
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Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques
Javorská, Lenka ; Solich, Petr (advisor) ; Pohanka, Miroslav (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Lenka Javorská Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques The presented dissertation is focused on the development and validation of chromatographic method for the determination of glycopeptide antibiotic vancomycin, and L-arginine and its pathway related substances in biofluids. Furthermore, the potential of neopterin, kynurenine and tryptophan as biomarkers of immune response to surgical procedure and prediction of postoperative complications is investigated in connection with oncological diseases. In the theoretical part trends in instrumentation, stationary phases and extraction techniques are described. The issues of separation and retention of polar compounds and biological matrix pretreatment before analysis are also discussed there. Next part deals with selected analytes and their application to the clinical purpose for which the chromatographic methods are developed. It also provides an overview of published methods for their determination using liquid...
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Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques
Javorská, Lenka ; Solich, Petr (advisor) ; Pohanka, Miroslav (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Lenka Javorská Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques The presented dissertation is focused on the development and validation of chromatographic method for the determination of glycopeptide antibiotic vancomycin, and L-arginine and its pathway related substances in biofluids. Furthermore, the potential of neopterin, kynurenine and tryptophan as biomarkers of immune response to surgical procedure and prediction of postoperative complications is investigated in connection with oncological diseases. In the theoretical part trends in instrumentation, stationary phases and extraction techniques are described. The issues of separation and retention of polar compounds and biological matrix pretreatment before analysis are also discussed there. Next part deals with selected analytes and their application to the clinical purpose for which the chromatographic methods are developed. It also provides an overview of published methods for their determination using liquid...
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vulvovaginal candidiasis
Javorská, Lenka ; Křivčíková, Lucie (advisor) ; Jílek, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lenka Javorská Supervisor: Ing. Lucie Křivčíková Title of bachelor thesis: Vulvovaginal candidiasis My Bachelor thesis deals with the disease known as vulvovaginal candidiasis. It is a fungal disease caused by vaginal yeast genus Candida, which is important because a high percentage of female population in the emerging and often relapse. The work is divided into three parts. The first part describes the characteristics of the genus Candida, the second part deals with the characteristics of of candidiasis as a class of diseases and the inclusion of vulvovaginal candidiasis in this group. Third and most extensive part deals with a comprehensive view on the issue of vulvovaginal candidiasis, which includes disease characteristics, a description of the predisposing factors and clinical manifestations, laboratory diagnosis and treatment. The work has also a questionnaire study, which examines the incidence of the disease, predisposing factors and women's attitude to treatment. Key words: candidiasis, vulvovaginal candidiasis, Candida sp,. vaginal mucosa, predisposing factor, opportunistic pathogen
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Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques
Javorská, Lenka ; Solich, Petr (advisor) ; Pohanka, Miroslav (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Lenka Javorská Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques The presented dissertation is focused on the development and validation of chromatographic method for the determination of glycopeptide antibiotic vancomycin, and L-arginine and its pathway related substances in biofluids. Furthermore, the potential of neopterin, kynurenine and tryptophan as biomarkers of immune response to surgical procedure and prediction of postoperative complications is investigated in connection with oncological diseases. In the theoretical part trends in instrumentation, stationary phases and extraction techniques are described. The issues of separation and retention of polar compounds and biological matrix pretreatment before analysis are also discussed there. Next part deals with selected analytes and their application to the clinical purpose for which the chromatographic methods are developed. It also provides an overview of published methods for their determination using liquid...
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Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques
Javorská, Lenka ; Solich, Petr (advisor) ; Pohanka, Miroslav (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Lenka Javorská Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques The presented dissertation is focused on the development and validation of chromatographic method for the determination of glycopeptide antibiotic vancomycin, and L-arginine and its pathway related substances in biofluids. Furthermore, the potential of neopterin, kynurenine and tryptophan as biomarkers of immune response to surgical procedure and prediction of postoperative complications is investigated in connection with oncological diseases. In the theoretical part trends in instrumentation, stationary phases and extraction techniques are described. The issues of separation and retention of polar compounds and biological matrix pretreatment before analysis are also discussed there. Next part deals with selected analytes and their application to the clinical purpose for which the chromatographic methods are developed. It also provides an overview of published methods for their determination using liquid...
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Detection of His-tag and FLAG-tag recombinant proteins
Javorská, Lenka ; Zemanová, Lucie (advisor) ; Novotná, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Javorská Supervisor: RNDr. Lucie Škarydová, Ph.D Title of diploma thesis: Detection of His-tag and FLAG-tag recombinant proteins Recombinant proteins are proteins produced in genetically modified organism by recombinant DNA technology that enable insert DNA sequence of target protein in expression cell. This technology allows preparation of actually any protein in any quantity without difficulties associated with demanding purification of native protein from tissues. Recombinant proteins are produced often in form of fusion proteins with suitable peptide tag (e.g. His-tag, FLAG- tag) that can help with their detection and enable simple purification. The aim of this diploma thesis was to introduce and optimize universal method for the detection of recombinant proteins labelled with His-tag (6x histidine) and FLAG-tag. The model recombinant protein with His-tag was enzyme carbonylreductase 1 (CBR1) produced by expression system Escherichia coli. For FLAG-tag labelled protein were chosen enzymes DHRS12, HSD11β1, RODH4, DHRS3, DHRS7, DHRS8 produced by insect cells (strain Sf9). SDS polyacrylamide electrophoresis for protein separation and immunochemical detection after Western...
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vulvovaginal candidiasis
Javorská, Lenka ; Křivčíková, Lucie (advisor) ; Jílek, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lenka Javorská Supervisor: Ing. Lucie Křivčíková Title of bachelor thesis: Vulvovaginal candidiasis My Bachelor thesis deals with the disease known as vulvovaginal candidiasis. It is a fungal disease caused by vaginal yeast genus Candida, which is important because a high percentage of female population in the emerging and often relapse. The work is divided into three parts. The first part describes the characteristics of the genus Candida, the second part deals with the characteristics of of candidiasis as a class of diseases and the inclusion of vulvovaginal candidiasis in this group. Third and most extensive part deals with a comprehensive view on the issue of vulvovaginal candidiasis, which includes disease characteristics, a description of the predisposing factors and clinical manifestations, laboratory diagnosis and treatment. The work has also a questionnaire study, which examines the incidence of the disease, predisposing factors and women's attitude to treatment. Key words: candidiasis, vulvovaginal candidiasis, Candida sp,. vaginal mucosa, predisposing factor, opportunistic pathogen
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Alternative methods of financing foreign trade
Javorska, Lenka ; Sato, Alexej (advisor) ; Jelínková, Eva (referee) ; Machková, Hana (referee) ; Dudek, Miloslav (referee) ; Chvojka, Petr (referee)
Disertační práce se zabývá faktoringem a forfaitingem, jakožto metodami finacování zahraničního obchodu, které nejsou v České republice dosud využívány ve stejné míře jako v zahraničí. Podává nestranný a kritický pohled na oba nástroje, prezentuje nejen základní principy jejich využívání a jejich výhody, ale zaměřuje se na jejich slabé stránky, podmínky a překážky využívání. Součástí práce je také popis současného českého faktoringového a forfaitingového trhu, včetně možných prognóz budoucího vývoje a analýzy nabídky jednotlivých finančních institucí. This thesis focuses on factoring and forfaiting. Both factoring and forfaiting market in the Czech Republic are not as developed as in Western Europe. It offers critical and neutral survey of all aspects of these two instruments, presents not only basic principles and their advantages but analyzes also their weaknesses, preconditions and impediments to their utilization. The thesis also describes current Czech factoring and forfaiting market including assumptions on future developments and analysis of financial products offered by particular financial institutions.
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