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Using CRISPR-Cas9 gene editing to engineer the next generation of CAR T cells
Sabolová, Saskia ; Otáhal, Pavel (advisor) ; Heneberg, Petr (referee)
Chimeric antigen receptor (CAR T) cell therapy is currently a successful treatment for hematological malignancies and is also a rapidly evolving field of research for treating solid tumors. The potential clinical expansion of this therapy depends on overcoming many obstacles, such as the persistence of CAR T cells in the hostile tumor microenvironment, induced toxicities, or the need for the transplant to be autologous. These limitations can be mitigated by CRISPR-Cas9 gene editing, which has the potential to create CAR T cells resistant to inhibition, modulate cytokine release, decrease the risk of cytokine release syndrome or neurotoxicity, and create allogeneic CAR T cells that do not cause graft-versus-host disease. Improvements in the CRISPR-Cas9 technology field, such as the development of base and prime editors, further increase safety by bypassing the dangerous double-strand break in the genome. Although many of these modifications are still subjects of research, there are a number of ongoing or already completed clinical trials that have implemented CRISPR-Cas9 technology in their CAR T cell engineering processes.
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Speciation analysis of arsenic compounds in environmental samples
Pakosta, Lukáš ; Novotná, Marie (referee) ; Vašinová Galiová, Michaela (advisor)
Arsenic is a toxic element that exists in various species, each having different environmental and health impacts. The presence of arsenic in the environment, especially in water and soil, poses a serious risk to human health. In this bachelor's thesis, the time and temperature for the extraction of arsenic and arsenic species in soil and biota samples were optimized. For the separation of arsenic species, high-performance liquid chromatography was used, and the determination of arsenic species content and total arsenic content was carried out using a triple quadrupole mass spectrometer with inductively coupled plasma. Based on the conducted experiment, the optimal conditions were applied to soil and biota samples from Lužánky Park.
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Morphology of my bacteria III.
Procházková, Kateřina ; Mitro, Michal (referee) ; Mikyta, Svätopluk (advisor)
In my bachelor thesis titled "Morphology of my bacteria III.", I focus on a series of objects and prints depicting a parasite defined by me. Using the technique of screen printing, I portray the subject as a symbol of personal encounters with health difficulties. This results in an infinite number of growing parasites, bacteriological or virological motifs, and morphological shapes. Screen printing is primarily utilized in the work for its ability to multiply and layer, each time with a different outcome.
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Are currently used chelators of heavy metals also chelators of calcium and magnesium?
Skoupilová, Karolína ; Mladěnka, Přemysl (advisor) ; Šuťák, Róbert (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Karolína Skoupilová Supervisor: prof. Přemysl Mladěnka, Pharm.D., Ph.D. Title of diploma thesis: Are currently used chelators of heavy metals also chelators of calcium and magnesium? Calcium and magnesium are important elements for the human body. Calcium plays an important role in blood clotting and intracellular signalling, thus participating in muscle contraction and platelet aggregation, among other processes. Magnesium is a cofactor for many enzymes. A significant deficiency of both ions can manifest as disorders of heart rhythm and increased neuromuscular excitability. The aim of this study was to determine the chelating activity of currently used heavy metal chelators and to determine the stability of the indicator. First, the ideal pH for the experiment was determined. In the next step, the chelation rate of 21 known metal chelators was measured using an in vitro spectrophotometric method. Finally, the chelation of the most active calcium chelators was verified on platelet aggregation in a human blood sample. At the same time, the long-term stability of the indicator o- cresolphthalein complexone (CC) was determined. In the first step, pH 7.5 was chosen, which was the only one...
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Toxicity of selected nitramines
Papírek, Jan ; Fojt, Pavel (referee) ; Bednařík, Karel (advisor)
This Bachelor Thesis is focused on determination of toxicity of selected nitramines and nitrosoamines, which had to be synthesized soon. Out of total 10 prepared N-nitro and N- nitrosocompounds 4 were selected for testing their toxicity in test using Duckweed (Lemna Minor). Factor IC50 was determined for two compounds 3,7-dinitro-1,3,5,7-tetraazabicyclo [3.3.1]- nonane (DPT) and 3,7-dinitroso-1,3,5,7-tetraazabyciclo [3.3.1] nonane (DNPT), unfortunately factor for other compounds failed to determine, because conditions of test validity hadn’t been kept. In the end of the thesis is compared toxicity of N-nitro and N-nitrosocompounds based on results and observations.
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The study of PET bottles in terms of the content of risk elements
Kryštofová, Petra ; Řezáčová, Veronika (referee) ; Komendová, Renata (advisor)
This bachelor thesis deals with the issue of PET bottles in terms of the possible content of risk elements. The contents of antimony, copper and lead were determined in the PET packaging material and in the beverages contained in the bottles. A total of 15 samples of bottled still and infant water available in the Czech sales network were analyzed. The PET material was decomposed in an acidic medium under the action of microwave radiation under optimized conditions. Atomic absorption spectrometric methods with flame or graphite cuvette atomization were used to determine selected risk elements based on the predicted analyte concentrations. In the PET packaging material, 159.35 to 265.68 mgkg-1 of antimony were determined, the determined values of lead concentration were in the range of 10.96-20.23 mg·kg-1 and the copper concentration in the range of 0.11-7,52 mg·kg-1. In bottled water and in bottled infant water, antimony was identified only in samples labeled 6a, 8a, 9a and 15a, all lead concentrations were measured below LODPb, copper was determined only in samples 2a, 6a and 8a. In none of the samples did the concentrations given by the valid legislation be exceeded. The limits of these metals are not set in legislation in the packaging materials, only specific migration limits and they have not been exceeded. However, the high concentration of antimony in PET poses a potential risk of contamination of the contents of these packages.
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Evaluation of ecotoxicity of selected chemicals with using algal test.
Kosárová, Hedvika ; Pavlovský, Jiří (referee) ; Doležalová Weissmannová, Helena (advisor)
This bachelor thesis deals with the ecotoxicity of 2 - [(2,6-dichlorophenyl)aniline] benzene acetic acid (diclofenac), ?-methyl-4-(2-methylpropyl) benzene acetic acid (ibuprofen) and mixture of cadmium and diclofenac and studies the above using algal toxicity test in accordance with ISO 8692. The theoretical part describes the issue of pharmaceuticals in the environment, and characteristics of standards algae tests and alternative tests. Freshwater green algae Desmodesmus subspicatus were used as testing organisms in all the implemented algal toxicity tests. By the means of these tests, the IrC50 values for the standard substance potassium dichromate, the drugs diclofenac and ibuprofen and cadmium were determined. The toxicity of cadmium in mixture with diclofenac was also evaluated.
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Biotransformation of nitro aromates
Tomeček, Jaromír ; Bednařík, Karel (referee) ; Friedl, Zdeněk (advisor)
Contamination by nitroaromatic compounds, such as explosives, pesticides or other nitroaromatic compounds, is worldwide environmental problem. This text focuses on contamination by compounds from military and ammunition industry, most especially 2,4,6-trinitrotoluene (TNT), 1,3,5-trinitro-1,3,5-triazinane (RDX) and 1,3,5,7-tetranitro-1,3,5,7-tetrazocane (HMX). First goal of this work is to provide the reader with a aerobic biodegradation and transformation processes of nitroaromatic compounds named above. The second goal is to compare the toxicity of initial nitroaromates and their transformation intermediates.
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Evaluation of environmental load by perfluorinated compounds
Šima, Aleš ; Stoupalová, Michaela (referee) ; Vávrová, Milada (advisor)
The most important substances of the group of perfluorinated compounds are perfluorooctyl sulfonate (PFOS), perfluorooctanoic acid (PFOA) and perfluorooctane sulphonamide (FOSA). Perfluorinated compounds have special physical and chemical properties, which make them valuable for usage in different industrial branches. These properties, which make them so useful are also the reason, why they are so harmful for the environment. The target of theoretical part of the diploma thesis was processing of literature searche on the topic: Evaluation of environmental load by perfluorinated compounds. In the experimental part was developed and optimized a method that was used for the identification and quantification of selected pollutants contained in real soil samples.
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