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The Introduction of Contact Ecotoxicity Tests for the Assessment of Terrestrial Ecosystems
Modlitbová, Pavlína ; Kráčmar, Stanislav (referee) ; Beklová, Miroslava (referee) ; Demnerová, Kateřina (referee) ; Vávrová, Milada (advisor)
This dissertation thesis is focus on using toxicity tests with testing organism - terrestrial isopod Porcellio scaber. This organism is use for toxicity assessment of selected anorganic compunds which could contaminate terrestrial ecosystem. This organism was selected for several reasons - sophisticated test methodology; well known biology of these animals and their ecological relevance. Besides classical endpoints such as mortality, change in mass of test specimens and the effect of the presence and concentration of contaminants onto food consumption; we monitored changes in the morphometric characteristics at the level of tissues and cells, the cytotoxicity and possibly bioaccumulation in various parts of the body of the organism. Selected test substance in this thesis were nanoparticles and inorganic salts. Wherein for chosen gold nanoparticles was carried out a complete study presents several levels of organization of living matter (organism, tissue, cell). For selected salts compound, this study was carried out on the behavior of organisms and classical endpoints.
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Assessment of in vitro antioxidant properties of medicinal and edible plant extracts
Tauchen, Jan ; Kokoška, Ladislav (advisor) ; Jaromír , Jaromír (referee)
Identification and characterization of plant-based products with antioxidant and anti-proliferative effects has received much interest over the past few years as possible therapeutic mean for treatment of diseases likely to be associated to oxidative stress (such as atherosclerosis, diabetes, and cancer). This study provides characterization of in vitro antioxidant and/or anti-proliferative potential and phytochemical profile of (i) 39 wine samples of underutilized Georgian grapevine cultivars, (ii) extracts of 22 samples of medicinal plants from Ethiopia and (iii) 23 samples of edible and medicinal plants from Peruvian Amazon. For this purpose, 2,2-diphenyl-1-picrylhydrazyl radical assay (DPPH), oxygen radical absorbance capacity (ORAC) assay, total phenolic content (TPC), and cell viability assay based on metabolization of tetrazolium bromide (MTT) to formazan, together with methods based on high performance liquid chromatography (HPLC-ultra violet/visible spectrometry and HPLC-tandem mass spectrometry) were used. Georgian red wines (particularly Saperavi cultivars) exhibited higher antioxidant capacity (average DPPH and ORAC values at 5.1 and = 10.6 g TE/L wine, respectively) in comparison to Central and West European cultivars [Pinot Noir (DPPH = 3.1 and ORAC = 9.4 g TE/L wine), Cabernet Sauvignon (DPPH = 3.0 and ORAC = 7.3 g TE/L wine) and Cabernet Moravia (DPPH = 2.0 and ORAC = 8.5 g TE/L wine)]. Georgian wines contained significantly greater concentrations of quercetin (between 14.44 and 1.07 ug/mL), kaempferol (between 1.68 and 0.03 ug/mL) and syringic acid (between 12.59 and 4.72 ug/mL), whereas possessed lower quantities of resveratrol (between 5.11 and 0.32 ug/mL) in comparison to Central and West European wines. Amongst edible and medicinal plants from Ethiopia and Peruvian Amazon, only Dodonaea angustifolia (IC50 for DPPH = 22.2 ug/mL, ORAC = 767.6 ug TE/mg extract; IC50 for Hep-G2 = 120.0 ug/mL), Rumex nepalensis (IC50 for DPPH = 5.7 ug/mL, ORAC = 1061.4 ug TE/mg extract; IC50 for Hep-G2 = 50.5 ug/mL), Inga edulis (DPPH and ORAC = 337.0 and 795.7 ug TE/mg extract; IC50 for Hep-G2 and HT-29 = 36.3 and 57.9 ug/mL) and Oenocarpus bataua (DPPH and ORAC = 903.8 and 1024.4 ug TE/mg extract; IC50 for Hep-G2 and HT-29 = 102.6 and 38.8 ug/mL) have demonstrated combinatory antioxidant/anti-proliferative efficacy. Selective anti-proliferative activity was observed for Verbascum sinaiticum (IC50 for Hep-G2 = 80.6 ug/mL) and Annona montana (IC50 for Hep-G2 and HT-29 = 2.7 and 9.0 ug/mL, respectively). Above-mentioned plant material showed only weak or non-toxic effects towards normal cell line. Despite the fact that extracts of Jasminum abyssinicum (IC50 for DPPH = 26.3 ug/mL, ORAC = 1023.7 ug TE/mg extract), Rumex nepalensis (IC50 for DPPH = 5.7 ug/mL, ORAC = 1061.4 ug TE/mg extract), Mauritia flexuosa (DPPH and ORAC = 1062.9 and 645.9 ug TE/mg extract), Myrciaria dubia (DPPH and ORAC = 641.9 and 642.6 ug TE/mg extract) and Theobroma grandiflorum (DPPH and ORAC = 714.8 and 821.9 ug TE/mg extract) have exhibited considerable antioxidant effect, these species were found to possess moderate to low anti-proliferative potential or have shown to be toxic to normal cells line. In all cases it was detected that phenolic compounds content correlated strongly with antioxidant activity, however weakly with anti-proliferative effect. Results suggest above-mentioned species as prospective materials for further development of novel plant-based agents effective against oxidative stress related diseases. However, it is necessary to perform further research which would be focused on detailed characterization of their chemical composition, pharmacological effects and toxicological safety, in order to verify their possible practical use.
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New ligands for applications in medicine
Holá, Kateřina ; Holzhauser, Petr (referee) ; Kotek, Jan (advisor)
The aim of this Thesis was the preparation of macrocyclic ligands (Figure 1) for the complexation of lantanoides 3+ . Their complexes can be used in nuclear medicine (in case of ligand L1) or as MRI contrast agents (in case of ligand L2). Ligand L1 was designed to increase the rate of complexation due to the weakly binding pendant group based on iminodiacetic acid (IDA). The complexation mechanism was spectrophotometrically studied with Ce3+ . Spectrophotometric kinetic measurements proved that ligand L1 not only shown no improvement in the complexation rate but its complexation is rather noticeable slowed. Ligand L2 contains amino group whose protonization/coordination could be influenced by pH or temperature. It can be utilized in determination of physical status of different tissues. However, it is difficult to obtain the ligand L2 in sufficient purity for further studies N N NN COOHHOOC HOOC N COOH COOH P OH O L1 N N NN COOHHOOC HOOC NH2 L2 Figure 1: Ligands prepared in course of this Bc Thesis
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Methods of monitoring the profile of carnitine metabolites in urine and use in clinical diagnosis
NOVÁKOVÁ, Martina
This study deals with the profiling of carnitine metabolites in the urine of the patients with breast cancer. L-carnitine is a chemical compound that is very important for metabolism of lipids, and therefore some current research suggests that it could serve as a means for the treatment of cancer affecting various organs or tissues. The theoretical part deals with the general characteristics and a description of the L-carnitine and its effect on the body. This part is divided into chapters that summarize various information about L-carnitine and acylcarnitines, for example the metabolism, different effects in the body including a description of responses to the lack of these substances, or methods that can be used for the detection of L-carnitine levels. Methods of monitoring L-carnitine were described in detail and were compared with previously published scientific studies. The authors of scientific studies show procedures, advantages and disadvantages of the proposed methods for determining L-carnitine and acylcarnitines, that were examined in this work in detail and the method for the determination of carnitine metabolites in urine has been proposed. A brief description of the most using methods for the determination of L-carnitine metabolites by liquid chromatography mass spectroscopy (LC / MS) is done the end of this part. I have been motivated with a positive results of studies and I have used this method for analysis in my work. In the research part of my study, the sample preparation protocol and conditions for the application of the method are described. The samples from the hospital of České Budějovice have been obtained from patients with fresh breast cancer diagnosis. In this part, the treatment of samples prior to detection, the used materials, equipment, chemicals, process and device settings for analysis were described. The method provided chromatograms and mass spectra of the individual substances. Samples of patients were compared with samples of control group of healthy women, which enabled us to observe possible differences. For clarity, the graph was drawn up and it was possible to monitor only slight differences between the levels of carnitine metabolites in women with breast cancer versus the control samples. This difference is not statistically significant, especially because the variability between individuals is higher of an order of magnitude than the actual differences between the two groups. This result is due to the fact that the study has not included additional information on patients such as information about their medications or health style, the overall condition of the body, etc. A detailed study requires a systematic and very time consuming work, which was not the aim of my thesis. Liquid chromatography with hydrophilic interactions method associated with mass detector used for monitoring of L-carnitine and its metabolites profiles gives positive results, is not time consuming, affordable and above all allow suitable for the detection of substances that have been selected for my work.
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Methylene-bis(phosphonate) derivatives for modification of biomolecules
David, Tomáš ; Lorenc, Miroslav (referee) ; Kubíček, Vojtěch (advisor)
5 Title: Methylene-bis(phosphonate) Derivatives for Modification of Biomolecules Author: Tomáš David Department: Department of Inorganic Chemistry, Faculty of Science Supervisor: RNDr. Vojtěch Kubíček, Ph.D. Supervisor's e-mail address: vvvojta@seznam.cz Abstract The aim of this work was to prepare derivatives of methylen-bis(phosphonic) acid. Thirty compounds (from which 29 are new) were prepared with moderate or good yields. All compounds were fully characterized by NMR and MS. Four new crystal structures were obtained by RTG monocrystal analysis. Derivative containing 4-nitrobenzyl group in the side chain was used for study of bis(phosphonates) adsorption parametres to hydroxyapatite (with quantification by UV-Vis spectroscopy). Several derivatives containing characteristic functi- onal groups (-NH2, -COOH, -NCS) were prepared. These compound are suitable for further derivatization, mainly for conjugation with more complex structures through amide bond or thiourea bridge. Conjugates of building blocks with fluorescent probe (fluorescein and dansyl) were prepared for potencial optical imaging. Further modification of one of the dansyl derivative afforded trifunctional molecule - contating bis(phosphonate) as a vector for a bone tissue, a fluorescent probe for a possible fluorescent imaging and a free...
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Cryopreserved semilunar heart allografts: Leaflet surfase damage in scanning electron microscopy
Burkert, Jan ; Špatenka, Jaroslav (advisor) ; Černý, Štěpán (referee) ; Černý, Jan (referee) ; Třeška, Vladislav (referee)
7. Anglický souhrn: Cryopreserved semilunar heart valve allografts: leaflet surface damage in scanning elect.-on microscopy. Objective: Allograft heart va1ves (AHV), biologica1 va1ves ofhuman origin, offer potential advantages over conventiona1 xenografts in terms of superior hemodynamics and, perhaps, better durability. The most important factors for long-term AHV c1inical peďormance are the processing and cryopreservation methods. The aim ofthis study was to eva1uate the impact of current processing protocol on va1ve tissue morphology, maínly to address the effect of successive processing steps on the leaflet suďace structure. For the detection offine changes in endothelia1 covering and underlying layers, our own modification ofthe scanning electron microscopy (SEM) technique was utilized. Material and methods: The study was based on an investigation of20 AHV (40 specimens). Fourteen valves came from heart-beating donors (multiorgan harvesting) when the heart could not be transplanted for any reason (donor criteria, availability of recipient and/or logistics). Six were obtained at the time of routine postmortems - non heart-beating donors (NHBD). AU specimens were initially flxed in Bakeťs solution. Tissue samples were dissected, dried with hexamethyldisilazane (HMDS), gold-coated, studied and...
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název v anglickém jazyce není uveden
Šenolt, Ladislav ; Pavelka, Karel (advisor) ; Rovenský, Jozef (referee) ; Pokorný, Jaroslav (referee)
Rheumatoid arthritis (RA) and osteoarthritis (OA) represent the most common forms of musculosceletal disorders that affect diarthrodial joints, lead to joint damage and disability. Extra-articular manifestations accompanied the joint disease only in RA. Diagnosis of both conditions most commonly bears on the conventional radiography. Mostly in OA, radiographic changes often occur late in the disease and are largely irreversible. Molecular markers could reflect joint damage, inflammation, or immune response. Current investigation revealed potential uses of molecular markers, ranging from understanding pathogenesis of the diseases to predicting and monitoring the outcome of the treatment. The aim of the thesis was to analyze several biochemical markers in serum, synovial fluid and synovial tissue samples from patients with RA and OA. and to evaluate their diagnostic and predictive values as well as their contribution to the pathogenesis of the diseases. We found increased serum pentosidine concentrations in OA patients that were ol a predictive value of the joint space narrowing in OA of the knee joint and correlation between pentosidine and cartilage oligomeric matrix protein (COMP) in synovial fluid that make pentosidine one of the new potential biomarkers of the OA. Scrum level of COMP was similar among...
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Role of Smarca5 (Snf2h) during transcription of transfected DNA template.
Zikmund, Tomáš ; Stopka, Tomáš (advisor) ; Smetana, Karel (referee)
Cellular and tissue characteristics are results of dynamic regulation of gene expression. DNA wrapped into proteins, referred to as chromatin, requires involvement of mechanisms guiding accessibility of specific sequences. In higher organisms, chromatin remodeling proteins are indispensable in regulating chromatin structure including ISWI ATPase SMARCA5. SMARCA5 is involved in almost any transaction on DNA including transcription, however precise in vivo role of SMARCA5 in these processes remains unknown. To advance understanding of specific role of SMARCA5 in the development of chromatin structure during transcription we devised cellular model in which SMARAC5 level is manipulated while chromatin structure development and transcriptional response are monitored. Our data indicate that the transfected DNA template that is transcribed is enriched with histone H3 and its specific methylation of Histone H3 lysine (K) 4, a mark of active chromatin structure. Overexpression of SMARCA5 results within the reporter gene coding sequence in ~2,5-3 fold increase of both H3 occupancy an its modification H3K4Me3. Increased DNA template commitment into chromatinization is associated with repression of reporter gene expression. These results are supported by studies indicating dynamic development of nucleosomal...
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