|
Interactions of new compounds with constitutive androstane receptor
Lágnerová, Jitka ; Pávek, Petr (advisor) ; Trejtnar, František (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Jitka Lágnerová Supervisor: Doc. PharmDr. Petr Pávek, Ph.D. Title of diploma thesis: Interactions of new compounds with constitutive androstane receptor Nuclear receptors (NRs) are ligand-activated transcription factors that regulate the expression of genes involved in a broad range of biological processes. Pregnan X receptor (PXR) and constitutive androstane receptor (CAR) are members of the orphan NR subfamily, that were originally defined as xenobiotic sensors regulating the expression of drug-metabolizing enzymes in order to protect the body from toxic chemicals. Alteration in CAR function may impact not only pharmacokinetics, efficacy, and toxicity of drugs but also endocrine homeostasis, energy metabolism, and cell proliferation. The aim of this study was to verified interactions of newly synthesized compounds with human CAR. Using the methods of gene-reporter assay and two- hybrid assay we tested 40 agents in various concentrations in HepG2 and LS174T cell lines. As the direct ligands of hCAR with significant transcriptional activity we have identified two substances, namely 2 - (3-methoxyphenyl)quinazoline-4-thiol, resp. -ol used in concentration of 10 µM. In the future, the...
|
|
Identification of antimicrobila peptides in spider venom
Benýšek, Jakub ; Liberda, Jiří (referee) ; Tichá, Marie (advisor)
Still increasing resistance to antibiotics leads to the need to find new active compounds with antimicrobial properties. This work is focused on the occurrence, chemical and physical description, mechanism of action and biological activity of such substances, found in spider venom. The second part is focused on isolation and identification of compounds with these properties from the venom of wild bees and a one spider. A novel peptide was isolated and identified from venom of bee Trachusa byssina. This novel peptide possess antimicrobial properties and low hemolytic activity. Molecular weight was estimated to 1749,9 ? 0,1contains 16 amino acids and is amidated on its C-terminus. Its primary structure GILSVLKNLLKKHMAS-NH2 was determined by using Edman degradation and ESI-QTOF mass spektrometry.
|
|
Functional and structural study of thermally activated TRP ion channels: The role evolutionarily conserved motifs in the TRPA1 modulation
Kádková, Anna
Ankyrin receptor TRPA1 is an ion channel widely expressed on primary afferent sensory neurons, where it acts as a polymodal sensor of nociceptive stimuli. Apart from pungent chemicals (e. g. isothiocyanates, cinnamaldehyde and its derivatives, acrolein, menthol), it could be activated by cold temperatures, depolarizing voltages or intracellular calcium ions. TRPA1 channel is a homotetramer in which each subunit consists of cytoplasmic N and C termini and a transmembrane region. The transmembrane part is organized into six alpha- helices connected by intra- and extracellular loops. The N terminus comprises a tandem set of 16 to 17 ankyrin repeats (AR), while the C terminus has a substantially shorter, dominantly helical structure. In 2015, a partial cryo-EM structure of TRPA1 was resolved; however, the functional roles of the individual regions of the receptor have not yet been fully understood. This doctoral thesis is concerned to elucidate the role of highly conserved sequence and structural motifs within the cytoplasmic termini and the S4-S5 region of TRPA1 in voltage- and chemical sensitivity of the receptor. The probable binding site for calcium ions that are the most important physiological modulators of TRPA1 was described by using homology modeling, molecular-dynamics simulations,...
|
|
Study of photosynthetic response of green algae on presence of selected heavy metals
NOVÁK, Jan
With the world population growth, more and more agricultural areas fall into regions affected by waste products. Therefore it is important to determine their impact on environment toxicity. The aim of this thesis is to evaluate the effect of selected ions of heavy metals (CoSO4, CuSO4, CdSO4, ZnSO4 and K2Cr2O7) on the process of photosynthesis in a selected green algae species (Scenedesmus quadricauda) by means of monitoring of dynamics of oxygen release into solution of nutrient medium. The experiments with individual compounds proceeded by pH 7, temperature 25°C and light 450 ?mol m-2 s-1. Algareact device developed by the company BVT Technologies was used for measuring. Comparison with growth curves Scenedesmus quadricauda by different pH was used for the validation of the method. The method of the measurement of dissolved oxygen in medium is suitable for the measurement of immediate impact of chemical compounds on photosynthetic activity of examined organism. The method is restricted by the fluidity of measured solution. The lowest quantity of oxygen disolved inmedium was measured by CdSO4 in concentration 0,1 M.
|
|
Access to Justice in Environmental Matters
Bernard, Michal ; Damohorský, Milan (advisor) ; Smolek, Martin (referee) ; Sobotka, Michal (referee)
The aim of this work on Access to Justice in Environmental Matters is to describe possibilities of access of natural and legal persons to the judicial protection of the environment in the Czech Republic. Judicial protection is an important element of ensuring the right to favourable environment. To ensure effective environmental protection and the rights of individuals to the favourable environment, it is necessary to ensure the widest possible access of natural and legal persons to public authorities and courts claiming the illegality of decisions, acts and omissions of public authorities or third parties. Broad judicial protection of the right to favourable environment is necessary and indispensable part of democratic system and a prerequisite for the functioning of basic human rights in practice. This work must first address the question of substantive legal right to favourable environment -its content and range of entities to which this fundamental constitutional right belongs. Right to favourable environment means the right to an environment which is polluted below given limit. Thus, on environmental polluted and stressed as a consequences of human activities, including the introduction of physical, chemical or biological agents emissions and agents in extent to which the legal regulations on...
|
|
Development and applications of affinity carrier for isolation of human carbonyl-reducing enzymes
Andrýs, Rudolf ; Wsól, Vladimír (advisor) ; Šebela, Marek (referee) ; Šatínský, Dalibor (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Rudolf Andrýs Supervisor: Prof. Ing. Vladimír Wsól, Ph.D Title of dissertation thesis: DEVELOPMENT AND APPLICATION OF AFFINITY CARRIER FOR ISOLATION OF HUMAN CARBONYL-REDUCING ENZYMES For several millennia the human medicine is based on application of small bioactive molecules that are administered in the form of plant extracts or synthetic compounds. However, their use in modern medicine is not possible without a detailed understanding of their biochemical effects and identification of their molecular targets. Chemical proteomics based on the specific recognition between the bioactive molecule and the target molecule is currently the most widely used techniques for identification of molecular targets of small molecules. Compared to conventional biochemical methods (e.g. 2D electrophoresis), chemical proteomics represents particularly sensitive and very selective technique that enable successful identification of biomolecules from complex biological samples that are naturally presented in very small concentrations. Carbonyl- reducing enzymes, which play an important role in physiology due to their involvement in metabolism of various endogenous (e.g. prostaglandins, steroid...
|
|
Effect of substrate on the production of earthworms Dendrobena veneta
SCHMIDTMAJEROVÁ, Eva
The importance of biological processes for processing of organic wastes is widely acknowledged. My thesis focuses on one of the most efficient methods for conversion of organic wastes into precious products, which are at the same time environmentally friendly. Vermicomposting includes complex food webs and simultaneously modifies various chemical forms of basic nutrients into inorganic compounds, useful for plant nutrition. The thesis briefly describes biology, distribution and basic ecological categories of earthworms and shows their importance in decomposition organic wastes. I describe here the main earthworm species and then concentrate on Dendrobaena veneta. I then compare 5 substrates from the point of view of their suitability for production of this earthworm species and of the suitability of the resulting substrate as fertilizer. I have found that the best substrate for production of Dendrobaena veneta was the output of a sewarage plant and pure celulose.
|
|
Use of Imunohistochemical Methods in Diagnostics of Neoplasms
VELKOVÁ, Michala
This thesis deals with the application of immunohistochemical methods in the diagnosis of neoplasms. Indications of immunohistochemical methods of particular departments in Hospital Písek, a.s. are outlined there. It describes the origins and development of this method in the Pathological Anatomy Department, Hospital Písek, a.s. In the theoretical part, I describe knowledge of single methods. As for immunohistochemistry, I focused on the determination of antigens and the use of suitable primary antibodies for diagnosing diseases, mostly neoplasms. There are 20 primary antibodies available at the Department of Pathological Anatomy. In the practical part, I compare conventional staining and immunohistochemical methods with conventional staining and special staining methods in histology for the diagnosis of tumours. Hematoxylin eosin serves for distinguishing of essential cellular components. Selective staining enables colour differentiation of various tissue components. Using immunohistochemistry, the presence of certain tissue antigens by antibody reaction with the immobilized chemical compounds is detected. This comparison I was applied at the differential diagnosis of malignant melanoma and myogenic tumours at the Pathological Anatomy Department, Hospital Písek, a.s. For the determination of malignant melanomas, initially the sample was histologically processed and stained with Hematoxylin eosin staining base. Secondarily, the argentaffin reaction was used to detect melanin pigment and immunohistochemical staining of antigen HMB 45 using a mouse monoclonal antibody against HMB 45 by Dako Company and S 100 protein using a polyclonal rabbit antibody against S 100 by Dako Company. 14 samples of patients with malignant melanoma were examined this way. HMB 45 was stained positively in 12 samples out of 14 in total. Monoclonal antibody against HMB 45 is for malignant melanoma quite specific. S 100 protein was detected in all tissue samples. Polyclonal antibody against S 100 shows higher sensitivity, however it is not very specific. It is more used to distinguish neuroectodermal and nonneuroektodermálních lesions. Melanin was detected in 11 tissue samples. Detection of melanin is also non-specific and complicated. Argentaffin reaction shows colouring of other substances contained in the cytoplasm, such as lipofuscin therefore it is necessary to use other methods for the melanoma diagnosis, e.g. immunohistochemistry. In the diagnosis of myogenic neoplasms, Hematoxylin eosin basic staining was used and subsequently immunohistochemical examination followed to detect smooth muscle actin and desmin using monoclonal mouse antibodies by Dako Company and transparent Hematoxylin van Gieson staining, which is used less than in the past. A total of 94 myogenic neoplasms (leiomyoms uterus) were examined. 72 neoplasms stained with Hematoxylin van Gieson showed a positive colour reaction of muscle mass and 22 neoplasms weak colour reaction. Smooth muscle actin and Desmin was detected by immunohistochemistry in 92 samples. The main problem of Hematoxylin van Gieson staining remains less specificity and sometimes ambiguous differentiability of colour tones in their own preparation impeding and preventing to set a reliable diagnosis in some cases. For this reason, immunohistochemical detection of Smooth muscle actin and Desmin (amongst others) is preferred as key parameters. Preferably, you can then use positive antibody reaction with Smooth muscle actinof myoepiteliels in the search for possible invasiveness mainly to differentiate in situ and microinvasive tumous in glandular organs. The differential diagnosis of myogenic origin tumours currently prefers immunohistochemical methods to special staining for high sensitivity and specificity, as well as in the differential diagnosis of malignant melanoma. When combined, both approaches complement each other well and provide relatively high degree of certainty in determining cells origin.
|
|
An Environmental Impact of Pyritic Bauxite from the Dajti Mine, Albania
Novák, Jiří Karel ; Minařík, Luděk ; Peza, Luftulla Hasan ; Melka, Karel
The accelerated chemical weathering of bauxite due to acid atmospheric deposition and breakdown of pyrite was simulated using short-termed laboratory acid leaching at 20 o C. Pyrite-bearing gibbsite-boehmitic bauxite giving rise to a number of questions concerning ecology was tested along with oxic boehmitic bauxite from the Dajti mine and that from Mali Korites (Mali Thate Mts., SE Albania). The leachte of pyrite-bearing and sulphate-producing bauxite is enriched in the following risk elements: Cd, Be, Zn, Ni, Co, and much less Cr, Cu, As, and Pb. Toxic soluble hydroxy-alumina ions may be also released into the aquatic environment, because of high concentration in output leachate (563 mg/l). In contrast, the leachates of oxic boehmitic bauxites commonly contain much lower concentrations of toxic elements with exception of Cu (Dajti mine) and Be (new occurence of Mali Korites).
|
|
Laboratory preparation of inorganic compounds
FRANTOVÁ, Jana
``Laboratory preparation of inorganic compounds{\crqq} is the fourth part of web-based data-base of chemical experiments. This compartment is suitable especially for secondary and high schools. Fifteen experiments are presented in total (on internet from December 2006). Didactic elaboration includs clips, instructions and principles of experiments. Teaching aplications are suggested by questions and tasks. http://home.zf.jcu.cz/public/departments/kch/didaktika/didaktika.htm
|
guest ::
