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Sequential injection analysis BI-LOV - development of technique for sample treatment using solid phase extraction I
Popovská, Lenka ; Chocholouš, Petr (advisor) ; Sklenářová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Mgr. Lenka Popovská Supervisor: PharmDr. Petr Chocholouš, Ph.D. Title of diploma thesis: Sequential injection analysis BI - LOV - development of technique for sample treatment using solid phase extraction I Antioxidant activity of polyphenolic compounds can positively affect the therapy and the prevention of diseases associated with oxidative stress, e.g. cardiovascular diseases, atherosclerosis, neurodegenerative diseases (Alzheimer's and Parkinson's diseases), rheumatoid arthritis, virus diseases, neoplasms, benign prostatic hyperplasia (BHP), prostatitis etc. This diploma thesis deals with the analysis of phenolic acids, namely ferulic acid and also protocatechuic acid and o-coumaric acid. The ferulic acid was analysed within five concentrations (10,000 µg/ml, 5,000 µg/ml, 2,500 µg/ml, 1,250 µg/ml and 0,625 µg/ml), four different strong anion exchangers (TMAHP, QAE - A25, SAX - BOND and SAX - DSC), five different pH (5,0, 6,0, 7,0 a 8,5) of washing buffer and three different pH (1,5, 2,0 a 2,5) of elution buffer. Absorbance of the ferulic acid was measured at wavelengths 325 and 240 nm. In this thesis was also assessed handling with each sorbent, especially convenient use within LOV...
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Optimization of procedure for isolation of phosphorylated peptides from a peptide mixture by metal oxide affinity chromatography for mass spectrometry analysis
Popovská, Lenka ; Solich, Petr (advisor) ; Zemanová, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Lenka Popovská Supervisor: Prof. RNDr. Petr Solich, CSc. Consultant Supervisor: Mgr. Barbora Šalovská, Ph.D. Title of the rigorous thesis: Optimization of procedure for isolation of phosphorylated peptides from a peptide mixture by metal oxide affinity chromatography for mass spectrometry analysis. Protein phosphorylation is the key regulatory mechanism in cellular signalling. It is involved in most cellular processes that are regulated by a complex interplay of protein kinases and protein phosphatases and include proliferation, differentiation, and apoptosis. Defective or altered signalling pathways can lead to various diseases, and therefore it is very important to study protein phosphorylation in detail and try to understand it as much as possible. Technological development of mass spectrometry (MS) in recent years has enabled a significant increase in the number of phosphorylation sites that may be identified in a sample. Increasing scan speed together with commercial availability of high resolution MS have improved quantity and quality of the acquired data, which can be obtained in a relatively short time. However, even high-end devices would not allow to comprehensively identify the...
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Optimization of procedure for isolation of phosphorylated peptides from a peptide mixture by metal oxide affinity chromatography for mass spectrometry analysis
Popovská, Lenka ; Solich, Petr (advisor) ; Zemanová, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Lenka Popovská Supervisor: Prof. RNDr. Petr Solich, CSc. Consultant Supervisor: Mgr. Barbora Šalovská, Ph.D. Title of the rigorous thesis: Optimization of procedure for isolation of phosphorylated peptides from a peptide mixture by metal oxide affinity chromatography for mass spectrometry analysis. Protein phosphorylation is the key regulatory mechanism in cellular signalling. It is involved in most cellular processes that are regulated by a complex interplay of protein kinases and protein phosphatases and include proliferation, differentiation, and apoptosis. Defective or altered signalling pathways can lead to various diseases, and therefore it is very important to study protein phosphorylation in detail and try to understand it as much as possible. Technological development of mass spectrometry (MS) in recent years has enabled a significant increase in the number of phosphorylation sites that may be identified in a sample. Increasing scan speed together with commercial availability of high resolution MS have improved quantity and quality of the acquired data, which can be obtained in a relatively short time. However, even high-end devices would not allow to comprehensively identify the...
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Sequential injection analysis BI-LOV - development of technique for sample treatment using solid phase extraction I
Popovská, Lenka ; Chocholouš, Petr (advisor) ; Sklenářová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Mgr. Lenka Popovská Supervisor: PharmDr. Petr Chocholouš, Ph.D. Title of diploma thesis: Sequential injection analysis BI - LOV - development of technique for sample treatment using solid phase extraction I Antioxidant activity of polyphenolic compounds can positively affect the therapy and the prevention of diseases associated with oxidative stress, e.g. cardiovascular diseases, atherosclerosis, neurodegenerative diseases (Alzheimer's and Parkinson's diseases), rheumatoid arthritis, virus diseases, neoplasms, benign prostatic hyperplasia (BHP), prostatitis etc. This diploma thesis deals with the analysis of phenolic acids, namely ferulic acid and also protocatechuic acid and o-coumaric acid. The ferulic acid was analysed within five concentrations (10,000 µg/ml, 5,000 µg/ml, 2,500 µg/ml, 1,250 µg/ml and 0,625 µg/ml), four different strong anion exchangers (TMAHP, QAE - A25, SAX - BOND and SAX - DSC), five different pH (5,0, 6,0, 7,0 a 8,5) of washing buffer and three different pH (1,5, 2,0 a 2,5) of elution buffer. Absorbance of the ferulic acid was measured at wavelengths 325 and 240 nm. In this thesis was also assessed handling with each sorbent, especially convenient use within LOV...
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Cloning, expression and purification of human 17beta-HSD1
Popovská, Lenka ; Wsól, Vladimír (advisor) ; Novotná, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Popovská Supervisor: Prof. Ing. Vladimír Wsól, Ph.D. Title of diploma thesis: Cloning, expression and purification of human 17β-HSD1 17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1; SDR28C1; EC 1.1.1.62) belongs to the SDR superfamily and catalyzes the NAD(P)(H) dependent oxidoreduction of hydroxyl/keto groups at position C17 of androgens and estrogens and in this manner regulate intracellular availability of steroid hormone ligands to their nuclear receptors. This pathway constitutes a pre-receptor control mechanism. Estradiol is the most potent female sex steroid and the only one responsible for estrogen action in women. The ovary is the primary source of estradiol present in the circulation of premenopausal women, but circulating estrone and androgens originating from the adrenal gland are also converted to estradiol in peripheral tissues such as adipose tissue. After menopause, estrogen biosynthesis in peripheral tissues has a major role in estrogen action. For production of recombinant protein 17β-HSD1 bacterial strain E. coli DH10B with inserted vector pOTB7, which contained coding sequence of the respective enzyme, was used. The sequence of the enzyme was isolated from...
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