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Examination of specific IgGs with the ELISA method
ŠANTRŮČKOVÁ, Kamila
The aim of this study is to determine the levels of specific IgG anti-SARS-CoV-2 by ELISA in selected individuals and to evaluate whether the QuantiVac ELISA (IgG) kit is suitable for the determination. In the theoretical part, the bachelor thesis focuses on immunochemical methods. Immunochemical methods are based on the interaction between antigen and antibody. Thanks to immunochemical methods we can determine the presence of specific antibodies. One of the important immunochemical methods is ELISA or Enzyme-linked immunosorbent assay. ELISA is a special type of enzyme immunoassay. The basic principle is the interaction of antigen and antibody to form an immunocomplex. In order for this immunocomplex to be measurable, a conjugate must be added to cause a colour reaction. The practical part is focused on the testing of specific IgG antibodies against SARS-CoV-2. Sixteen human serum samples were examined. The serum samples were from people of different age groups. The sera were from vaccinated persons, vaccinated persons with a history of Covid 19 disease, unvaccinated persons with a history of Covid 19 disease, or unvaccinated persons without a history of Covid 19 disease. In addition, six calibrators, a positive and a negative control were measured. A total of 24 microtiter wells were occupied. The Anti-SARS-CoV-2 QuantiVac ELISA kit from EUROIMMUN was used. All measurements were performed using a DYNAREAD photometer and processed using AlaDYN software. The results were different levels of antibodies. Out of 16 serum samples, 2 samples came out negative, one sample came out borderline and 13 samples came out positive.
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The preparation of tablets by selective laser sintering.
Ficek, Lukáš ; Mužíková, Jitka (advisor) ; Šnejdrová, Eva (referee)
Charles University, Faculty of pharmacy in Hradec Králové Department of: Pharmaceutical Technology Mentor: doc. PharmDr Jitka Mužíková, Ph. D Consultant: Mgr. Thao Tranová Student: Lukáš Ficek Title of Thesis: The preparation of tablets by laser sintering This work focuses on the preparation of orally dispersible tablets by selective laser sintering (SLS). The present project is a pilot study conducted on the 3D printer Sintratec Kit at the Department of Pharmaceutical Technology. SLS is a 3D printing method based on the fusion of powder particles using a laser beam that sinters the individual particles together in the layers of material.1-2 The matrix forming polymer used in work was Kollidon® VA64 which was combined with two pigments namely Candurin® Gold Sheen and NTX Ruby Red. In addition, mixtures of the polymer with a co-processed dry binder Prosolv® ODT G2 and a physical mixture of mannitol, silicified microcrystalline cellulose, and crospovidone at the concentrations of 20%, 40%, and 60% were used. The influence of printing parameters, especially laser speed, on tablet quality was tested. NTX Ruby Red pigment was selected for final tablet printing. The best powder formulation for optimal printing progress and high-quality properties of the orally dispersible tablets was the mixture of...
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Test benches for microcontroller kits
Žaludek, Ivo ; Arm, Jakub ; Bradáč, Zdeněk
This paper deals with the design of a test bench for modules used in educational labs. The first section outlines the requirements for the tester, followed by a description of the printed circuit board design process. Implementation details of the software are then examined, including a graphical user interface based on Model-View-Controller (MVC) architecture that allows for easy result printing. The obtained results are presented, and further work is discussed.
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Comparison of similarities of mass spectra and structures of small molecules
Malíčková, Viktorie ; Galgonek, Jakub (advisor) ; Škrhák, Vít (referee)
Methods for measuring the similarity of mass spectra and the structures of small molecules are crucial for advancements in medicinal chemistry, pharmacology, and metabolomics. One commonly used method for comparing the mass spectra of molecules is cosine similarity. This measures the similarity between two non-zero vectors by calculating the cosine of the angle between them. Comparing the mass spectra of molecules enables searching in molecular databases, clustering of spectra, and exploration of spectral libraries. Structural similarity is measured based on various molecular fingerprints, such as Daylight, RDKit, Atom-Pair, Topological Torsion, Extended-Connectivity fingerprints, and others. These fingerprints are compared using similarity coefficients. The methods for comparing structures and mass spectra of molecules mentioned can be applied using bioinformatic libraries such as RDKit and CDK for generating and analyzing structural fingerprints, and the MatchMS library for comparing mass spectra. The work provides a theoretical overview of molecular descriptors, including various types of molecular fingerprints and techniques for measuring structural similarity, as well as the principles of mass spectrometry and approaches to comparing mass spectra. The practical part of the work focuses on...
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Investigation of changes in epigenetic modofications in rat brain induced by morphine treatment
Hronová, Anna ; Novotný, Jiří (advisor) ; Roubalová, Lenka (referee)
This study addressed the impact of varying lengths of abstinence periods on the epitranscriptomic modification of m6 A and the enzymes involved in this modification as well as on transcription factors in the brains of morphine-dependent rats. The methods used included a colorimetric kit to determine m6 A levels and Western blotting to quantify the levels of enzymes and transcription factors. The results showed that different lengths of morphine abstinence did not have a significant effect on m6 A modification, suggesting that this pathway was likely not influenced by morphine abstinence. However, significant changes were observed in the levels of enzymes involved in the regulation of m6 A, which were localized and time-specific, without a clear trend. Examination of transcriptional regulators ∆FosB and MeCP2 also revealed significant changes only in certain parts of the brain and at specific times, but without a clear trend in changes in levels. The outcome of this study is that the dynamics of transcriptional regulators, N6 -methyl-adenosine, and enzymes involved in m6 A modification are localized and time-specific, opening new questions regarding the mechanisms that govern neurobiological processes during the abstinence period.
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Laboratory diagnosis of microRNA in different diagnostic groups
Hrachovcová, Eliška ; Urbánková Rathouská, Jana (advisor) ; Tripská, Katarína (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Student: Bc. Eliška Hrachovcová Supervisor: PharmDr. Jana Rathouská, Ph.D. Consultant: doc. RNDr. Pavlína Kušnierová, Ph.D. Title of diploma thesis: Laboratory diagnostics of microRNAs in different diagnostic groups Background: Identification of potential miRNA targets for differential diagnosis of patients with atherosclerosis and dementia. Comparison of miRNA expression and uptake in serum samples with cerebrospinal fluid samples. Selection of appropriate miRNAs for differential diagnosis in patients with atherosclerosis and dementia. Furthermore, the selection of miRNA targets with the most significant fold-change and also the lowest p-value for subsequent validation studiues. Methods: To identify potential miRNAs, the received samples were centrifuged, aliquoted and stored at - 80C. miRNAs were isolated using the iCatcher Circulating cfRNA 1000 kit and the CatchGene isolator. Reverse transcription and amplification with detection were then performed. Data were processed in Excel and then statistically processed in GenEx using Mann-Whitney test. Results: The results were evaluated for 2 groups of patients: group A (samples of patients with Parkinson's disease and dementia) and group B (with...
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Complex molecular diagnostics of gastrointestinal stromal tumors
Kalfusová, Alena ; Kodet, Roman (advisor) ; Mandys, Václav (referee) ; Vaněček, Tomáš (referee)
Complex molecular diagnostics of gastrointestinal stromal tumors ABSTRACT Most of gastrointestinal stromal tumors (GISTs) are characterized by activating mutations in the KIT and PDGFRA genes (80 - 85 %). Mutations in the BRAF, KRAS, defects in the SDH complex or FGFR1::TACC1, ETV6::NTRK3 fusions were detected in the group of KIT/PDGFRA non-mutated GISTs (10 - 15 %). Targeted therapy with Imatinib mesylate (IM) marked a significant breakthrough in treating GISTs. A significant problem with targeted therapy is the formation of primary or secondary resistance. The biological behavior of the GIST is unpredictable. Determining the activity of selected markers for cell proliferation and senescence will enable the objectification risk of tumor behavior's aggressivness. The aim of the first part of our work was the mutational analysis of the KIT and PDGFRA genes and the detection of secondary mutations in patients with disease progression. We detected primary mutations in KIT and PDGFRA genes in 83.5 % of samples. We also demonstrated the presence of secondary mutations in 16 patients with progression of disease (in 29 progressive lesions - 69 %). We confirmed significant intratumor and also intertumor heterogeneity of secondary mutations. The aim of the second part of the thesis was the analysis of alterations in...
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Thrombin generation in pathophysiological conditions
Malíková, Ivana
Introduction: Thrombophilic conditions may be accompanied by increased thrombin generation (TG), especially during pregnancy when the increase in thrombin generation results from a series of changes in the haemostatic system. During administration of anticoagulant treatment, it is desirable to reduce the increase in thrombin generation. Aims of the study: The first aim of our study was to compare TG in individuals with a proven factor V Leiden mutation and individuals with elevated levels of factor VIII. Another aim was to compare patients with VTE and also with or without proven FV Leiden mutation that are taking different anticoagulation treatments. The final aim was to assess thrombin generation in pregnant women and their response to LMWH administration during severe thrombophilic conditions. Group of patients and methods: There were included 170 healthy blood donors in the control group, the congenital and acquired thrombophilia group included 44 individuals with FV Leiden mutation and 38 individuals with factor VIII level >150 % without other proven thrombophilia. 347 subjects with VTE taking direct coagulation inhibitors were included in the anticoagulation group. Sixty women with a prophylactic dose of LMWH were included in the group of pregnant women. Kit Technothrombin® TGA RC Low and RC...
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Mobilní kamera realizovaná prostředky ROS2
Onderka, Daniel ; Peringer, Petr (referee) ; Janoušek, Vladimír (advisor)
This thesis focuses on the middleware Robot Operating System 2. Specifically, it aims to explore and demonstrate its capabilities. It utilizes an Adeept AWR 4WD robot as the foundational hardware platform, onto which the ROS2 system is deployed. The system offers both manual control and a wandering mode equipped with obstacle avoidance functionality. In addition to its primary goal, the thesis explores various related subjects. One of them is the integration of the developed system with the Gazebo simulator. All functionality implemented on the real physical robot has been duplicated in the simulation environment. Furthermore, the original Adeept AWR kit was extended with additional hardware. This extension allowed for the use of slam_toolbox and Nav2 libraries for mapping and navigation purposes. The final extension involves the use of ros2_control for controlling robot motion.
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Sensor for bars of mallet percussion instruments
Pribol, Sebastián ; Balík, Miroslav (referee) ; Jirásek, Ondřej (advisor)
This thesis addresses the issue of capturing melodic percussion instruments using dynamic sensors that can be attached to a musical instrument. It includes theoretical knowledge, analysis, and selection of appropriate sensing methods, and the actual implementation, which consists of testing, integration with MIDI, manufacturing of the device itself, and final calibration and adjustments for proper functionality. The device uses an ESP32-S3 Dev Kit N8R8 microcontroller, which was programmed in the Arduino IDE for proper operation. The device consists of twelve piezoelectric sensors that capture the dynamics of playing the musical instrument and send MIDI information to a DAW via a Bluetooth module.
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