Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques

Univerzita Karlova, Farmaceutická fakulta v Hradci Králové Katedra: Katedra analytické chemie Kandidát: Mgr. Lenka Javorská Školitel: prof. RNDr. Petr Solich, CSc. Konzultant: doc. RNDr. Lenka Kujovská Krčmová, Ph.D. Název dizertační práce: Vývoj bioanalytických metod pro stanovení diagnostických markerů a léčiv s využitím chromatografických technik Předkládaná disertační práce se zabývá vývojem a validací chromatografických metod pro stanovení glykopeptidového antibiotika vankomycinu, a látek souvisejících s metabolismem L-argininu v biologických tekutinách. Dále je v práci zkoumán potenciál neopterinu, kynureninu a tryptofanu jako biomarkerů imunitní odpovědi v reakci na operační zákrok a predikci pooperačních komplikací v souvislosti s onkologickými onemocněními. V teoretické části jsou popsány vývojové trendy v oblasti instrumentace kapalinové chromatografie, stacionárních fází a extrakčních technik. Je zde diskutována problematika retence a separace polárních látek, biologické matrice a její úpravy před analýzou. Dále jsou v této části charakterizovány zvolené analyty v souvislosti s klinickým záměrem, pro který byly uvedené chromatografické metody cíleně vyvíjeny. Také je zde uveden přehled vybraných publikovaných metod pro jejich stanovení s využitím kapalinové chromatografie. Poslední část...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...Katedra analytické chemieDepartment of Analytical ChemistryFarmaceutická fakulta v Hradci KrálovéFaculty of Pharmacy in Hradec Králov

The Redevelopment and Recultivation Work in the Area of the City of Ostrava

Import 02/09/2009Téma mé bakalářské práce zní: „Sanační a rekultivační práce na území města Ostravy“. Cílem mé bakalářské práce bylo charakterizovat sanační a rekultivační záměry na území města Ostravy a vyhodnotit průběh sanací a rekultivací. Bakalářská práce obsahuje legislativní podmínky nutné pro sanaci a rekultivaci, rozdělení rekultivací a procesní postupy sanací a rekultivací, jejž mají za následek ekologicky vyváženou krajinu, která je vhodná pro život člověka v čistém a krásném prostředí. Obsahuje také konkrétní sanační a rekultivační projekty, které se na území města zpracovávají.The theme of my thesis is about: „The redevelopment and recultivation work on the area of the city of Ostrava“. The aim of this study was characterizing of the redevelopment and recultivation intention on the area of the city of Ostrava and analysing process of the redevelopment and the recultivation. This thesis include the legislative conditions necessary for the redevelopment and the recultivation, division the recultivation and procedural process of the redevelopment and the recultivation, which result in ecological balanced landscape that is suitable for human life in a clean and beautiful environment. It also includes specific the redevelopment and recultivation projects, which occur in the territory of the city.Prezenční545 - Institut ekonomiky a systémů řízenívýborn

vulvovaginal candidiasis

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 pathoge

Detection of His-tag and FLAG-tag recombinant proteins

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..

Detection of His-tag and FLAG-tag recombinant proteins

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..

vulvovaginal candidiasis

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 pathoge

Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques

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..

Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques

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..

Comparing Reclamation Strategies of the Ostrava-Karvina Coal District

Import 04/07/2011Téma mé diplomové práce zní: „Rekultivační přístupy v OKR a jejich porovnání“. Cílem mé diplomové práce bylo charakterizovat způsoby rekultivace a porovnat technické řešení tradičních a progresivních přístupů. Teoretická část obsahuje legislativní podmínky, které jsou nutné pro rekultivaci území, rozdělení rekultivací, procesní postupy rekultivací, jež mají za výsledek ekologicky vyváženou krajinu a také jsou zde popsány způsoby financování rekultivací. Praktická část obsahuje řešení konkrétního projektu sanačně rekultivační stavby na území OKR, kde je mým úkolem porovnat a z ekonomického hlediska vyhodnotit realizaci rekultivační stavby při použití různých výplňových materiálů. Výsledkem bude z vytipovaných, na trhu dostupných materiálů, vyhodnotit ekonomicky nejvýhodnější materiál pro rekultivační stavby.The theme of my thesis is " Comparing Reclamation Strategies of the Ostrava-Karvina Coal District. " The aim of this thesis was to characterize of ways of recultivations and compare technical solutions with traditional and progressive approaches. This thesis include the legislative conditions necessary for the recultivations, division the recultivations and procedural process of the recultivations, which result in ecological balanced landscape and ways financing of recultivations. The practical part includes specific project of the Ostrava-Karvina Coal District, where I must to compare and from an economic perspective to assess the implementation of recultivations works by using different restorative materials. The result will be selected to market of the available materials, to evaluate the most economical material for reclamation works.Prezenční545 - Institut ekonomiky a systémů řízenívýborn

The Effect of 4-Methylcatechol on Platelets in Familial Hypercholesterolemic Patients Treated with Lipid Apheresis and/or Proprotein Convertase Subtilisin Kexin 9 Monoclonal Antibodies

Elevated low-density lipoprotein (LDL) cholesterol levels lead to atherosclerosis and platelet hyperaggregability, both of which are known culprits of arterial thrombosis. Normalization of LDL cholesterol in familial hypercholesterolemia (FH) is not an easy task and frequently requires specific treatment, such as regularly performed lipid apheresis and/or novel drugs such as proprotein convertase subtilisin kexin 9 monoclonal antibodies (PCSK9Ab). Moreover, a high resistance rate to the first-line antiplatelet drug acetylsalicylic acid (ASA) stimulated research of novel antiplatelet drugs. 4-methylcatechol (4-MC), a known metabolite of several dietary flavonoids, may be a suitable candidate. The aim of this study was to analyse the antiplatelet effect of 4-MC in FH patients and to compare its impact on two FH treatment modalities via whole-blood impedance aggregometry. When compared to age-matched, generally healthy controls, the antiplatelet effect of 4-MC against collagen-induced aggregation was higher in FH patients. Apheresis itself improved the effect of 4-MC on platelet aggregation and blood from patients treated with this procedure and pretreated with 4-MC had lower platelet aggregability when compared to those solely treated with PCKS9Ab. Although this study had some inherent limitations, e.g., a low number of patients and possible impact of administered drugs, it confirmed the suitability of 4-MC as a promising antiplatelet agent and also demonstrated the effect of 4-MC in patients with a genetic metabolic disease for the first time