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Procalcitonin as an early marker of bacterial infection
Mikulová, Lenka ; Beránek, Martin (advisor) ; Drastíková, Monika (referee)
Introduction: Procalcitonin (PCT) is a 13 kD protein with a chain of 116 amino acids. It consists of parafollicularcells of the thyroid gland. Inflammatory procalcitonin is made up of 114 amino acids. Procalcitonin from plasma in the course of inflammatory process, does not consistof C-cells of the thyroid gland but it is generatedin the liver, and, to a lesser extent, in the lungs, kidneys, testes, prostate and small intestine. Objectives: 1. To compare the levels and the dynamics of C-reactive protein (CRP) and procalcitonin in the inflammatory process. in three patients with different diagnoses. To compare the number of examined PCT according to indicated VFN (General University Hospital) departments in the years 2009, 2012 and 2015. 3. To study the dynamics of the number of examined PCT in the years 2001- 2016. Methods: The levels of CRP and PCT were measured in the ÚLBLD laboratory of the General University Hospital in Prague in the period from May 20to June 17, 2016. The CRP was measured by immunoturbidimetricanalyzerUnicelDxC 880i. The PCT was measured by theCobas analyser using the electrochemiluminescence technology. In order to evaluate the data, graphs were processed with levels of bothanalytesfor 3 patients. Further, data needed for the evaluation of the above mentioned objectives no. 2...
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Final exam software projects
Beránek, Martin ; Prokýšek, Miloš (advisor) ; Štípek, Jiří (referee)
The thesis deals with evaluation of quality of high school software projects. Examining the sources, the text describes differences between the projects on a model school and between real world needs. Based on this, it suggests an intervention, in the form of passing materials to members of an action group composed of teachers leading the software projects. The materials supply topics that were not found in the model school's education plan. A methodology for evaluating the projects is suggested, drawing from qualitative indicators used in the real world. An impact of this intervention on the students' projects is evaluated afterwards. The impacts at the model school were not significant. Powered by TCPDF (www.tcpdf.org)
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Molecular biology investigation of somatostatin receptors in diagnostics of pituitary tumors
Drastíková, Monika ; Beránek, Martin (advisor) ; Fryšák, Zdeněk (referee) ; Patočka, Jiří (referee)
Molecular biology investigation of somatostatin receptors in diagnostics of pituitary tumors The overwhelming majority of pituitary tumors are benign, with 25-30% of them representing clinically non-functioning pituitary adenomas (NFA). The treatment of choice for NFA is transsphenoidal neurosurgery. However, the procedure is often not completely successful and frequently tumor remnants remain, resulting in regrow over the long-term follow-up period. This fact has led to the development of new therapy strategies using pharmacological treatment: somatostatin analogs (SA) and estrogen receptor modulators. SA are effectively used in the treatment of acromegaly, neuroendocrine tumors and Cushing's disease. In contrast, pharmacological treatment of NFA has for the most part been unsuccessful. Low SA effectiveness could be associated with the variable expression of target receptors on the adenomas. The aim of the study was to determine the somatostatin (SSTR subtypes 1-5) and estrogen receptor 1 (ER1) expression profile. To obtain more complex receptor profile expression in the pituitary, we also investigated dopamin receptor 2 (D2R). Methods The group of patients was made up of 105 men and 101 women (20-87 years old; median 61). 144 samples were NFA, 44 growth hormone secreting (GHomas), 9 corticotrophin...
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Alpha-1-antitrypsin deficiency analysis using real-time PCR
Blažková, Petra ; Beránek, Martin (advisor) ; Šimůnek, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Petra Blažková Supervisor: Doc. PharmDr. Martin Beránek Ph.D. Title of diploma thesis: Alpha-1-antitrypsin deficiency analysis using real-time PCR This diploma thesis focuses on the validation of the method PCR in real-time (real-time PCR) to investigate the Z and S mutations in the gene SERPINA1, located at the long arm of chromosome 14 (14q32.13) and provides instructions for making a protein named alpha-1- antitrypsin (A1AT). A1AT is a serine protease inhibitor that protects tissues from degradation by neutrophil elastase. Deficiency, which is most commonly caused by these mutations, can cause children and adult's liver and lung disease. A method of real-time PCR was applied after successful validation to a set of 46 clinical samples of DNA extracted from blood samples and 30 samples of the DNA from cells of the buccal mucosa. DNA isolation was performed by kit extraction QIAamp ® DNA Mini Kit by QIAGEN. I used a set of primers and hybridization probes according to Snyder (2006) for genotyping. Melting curve analysis was carried out in the thermocycler LightCycler 1.2. Results of DNA samples obtained from blood were compared with the results obtained through an accredited method...
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Hereditary causes of thromboembolic disease and their laboratory diagnostics
Mihlová, Radka ; Šimůnek, Tomáš (advisor) ; Beránek, Martin (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Autor: Radka Mihlová Supervisor: Doc. PharmDr. Tomáš Šimůnek, PhD. Name of Bachelor's Thesis: Hereditary causes of thromboembolic diseases and their laboratory diagnosis. The term thrombophilia is used to designation hereditary and acquired prothrombotic states in arteries, veins and microcirculation. A thrombus formation may lead to partial or complete closure of the blood vessels - deep vein thrombosis. Then, a gore can be released and subsequently travels through blood vessels and a heart, after passing close one of pulmonary arteries, resulting in a pulmonary embolism. This state is known as venous thromboembolic disease. Then, the gore can be released and subsequently travels through blood vessels and after passing through a heart a gore may result either in a pulmonary arteries, resulting in a pulmonary embolism. The aim of the thesis is to describe methods used for a diagnostics of thrombophilia and genetically dependent causes of thromboembolic disease. Early and accurate diagnostics of thrombophilia plays an important role in a prevention of thromboembolic events and complications associated with them. Currently, a large variety of molecular genetic markers is used with clearly...
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Contribution of Next Generation Sequencing for Laboratory Diagnostics
Votýpka, Pavel ; Beránek, Martin (advisor) ; Macháček, Miloslav (referee)
5 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Pavel Votýpka Supervisor: Doc. PharmDr. Martin Beránek, Ph.D. Consultant: Mgr. Nikola Ptáková Title of diploma thesis: Contribution of Next Generation Sequencing for Laboratory Diagnostics The endeavor to sequence the whole human genome lead not only to the knowledge acquisition regarding the human genetic information but as well to the development of new sequencing methods and technologies. In order to keep up with progress in genetic field in many clinical and research laboratories the new massive parallel sequencing equipment is being utilized. On the market are currently established four leading platforms - Illumina, Solid, Ion Torrent and 454 Life Technologies. The process of sequencing analysis can be summarized into three main steps - the sequencing library preparation, sequencing itself, variant calling and data analysis. Each part of the sequencing analysis exhibits certain specifics, we need to count with and as well its pitfalls, we need to avoid or to minimalize their impact on the analysis final result. Recently new methods termed sequencing of the 3rd generation are being developed, enabling sequence of a single DNA molecule to be determined without previous...
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Free circulating nucleic acids in plasma
Totzauerová, Kateřina ; Beránek, Martin (advisor) ; Šimůnek, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Kateřina Totzauerová Supervisor: Doc. PharmDr. Martin Beránek, Ph.D. Title of diploma thesis: Free circulating nucleic acids in blood plasma Outline: The aim of this thesis was to compare various commercially available DNA extraction kits and choose the universally optimal method for routine isolation of free circulating DNA from blood plasma samples according to their optical characteristics, the qPCR results and yield of short fragments. Methods: Four commercially available methods of extraction have been chosen to compare: QIAmp DNA Blood Mini Kit, QIAmp DSP Virus Spin Kit (both Qiagen), NucleoSpin Plasma XS (Macherey-Nagel) and Agencourt Genefind v2 (Beckman Coulter). DNA was isolated from aliquots of a pooled blood plasma of healthy individuals. Plasmatic DNA was quantified after extraction by spectrophotometry, fluorimetry and quantitative polymerase chain reaction. Fragmentation analysis on selected samples by capillary electrophoresis was also realized. Results: The best yield and purity provided the method from Qiagen QIAmp DSP Virus Spin Kit. Average value of the concentration determined qPCR was 49.95 ± 23.57 ng/mL and it gave the highest values of the fluorescence of...
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Genetic polymorphism in the NBS1 gene for diagnosis and treatment of patients with cervical carcinoma
Rataj, Michal ; Beránek, Martin (advisor) ; Šimůnek, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michal Rataj Supervisor: Doc. PharmDr. Martin Beránek Ph.D. Title of diploma thesis: Genetic polymorphism in the NBS1 gene for diagnosis and treatment of patients with cervical carcinoma The aim of this diploma thesis is to find optimal methods for screening of mutation 657del5 and estimate frequency of heterozygotes and homozygotes for the mutation 657del5 in population of the Czech republic. In the first section of the theoretical part is comprehensively pointed out the effect of factors affecting the integrity of genetic information and the formation of mutations in DNA. On the contrary, the second section devotes to the ability of cells to respond to this damage. In detail, the thesis devotes to the NBS1 gene and its product nibrin. In the complex MRE11/Rad50/NBN nibrin is an important member of the mechanisms of repair of double strand breaks NHEJ (non-homologous end joining) and HR (homologous recombination). The thesis is focused on nibrin and its functions, but also to mutations that prevent these functions and causes genetic disease Nijmegen breakage syndrome. Nibrin is translated from the sequence of the NBS1 gene. Gene NBS1 appears in population with several various...
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Genetic polymorphism in the HFE gene in the Czech population
Červinková, Barbora ; Beránek, Martin (advisor) ; Šimůnek, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Červinková Supervisor: Doc. PharmDr. Martin Beránek, Ph.D. Title of diploma work: Genetic polymorphism of the HFE gene in Czech population Outline: The main objective of the thesis was to map the incidence of the three mutations (C282Y, H63D, S65C) in the HFE gene in the Czech population. Consequently, the obtained results were compared with the reported occurrence in the world. Methods: The work involved the isolation of DNA from buccal swabs obtained from 167 donors (65 men, 102 women; average age for men was 31, for women 28). Isolated DNA strings were amplified by PCR methods using 5'-CAG ATC CTC ATC TCA CTG-3' and 5'-CTG GAT AAC CTT GGC TGT ACC CCC-3' primers for C282Y mutation, 5'-GCC ACA TCT GGC TTG AAA TT-3' and 5'-ACA TGG TTA AGG CCT GTT GC GCC ACA- 3' primers for H63D and S65C mutations. DNA samples were treated with restriction enzymes Rsa I, Bcl I and Hinf I, for C282Y, H63D and S65C mutations, respectively. Finally, the restriction fragments were separated by gel electrophoresis (2 % agarose gel). Results: C282Y mutation was present in 0 (0 %) and 19 (11,38 %) samples as homozygote or heterozygote, respectively. Moreover, C282Y mutation was present in 3 (1,80 %)...
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