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Synthesis of unsymmetrical derivatives of azaphthalocyanines V.
Nejedlá, Marcela ; Zimčík, Petr (advisor) ; Miletín, Miroslav (referee)
Title of diploma thesis: Synthesis of usymmetrical derivates of azaphthalocyanines V. Author: Marcela Nejedlá The aim of diploma thesis was synthesis of unsymmetrical derivates of azaphthalocyanines (Aza- Pc) with carboxy or hydroxy group, and the preparation of suitable precursors of Aza-Pc. Required zinc and magnesium complexes were synthesized using statistical condensation of two different precursors. The desired complex was isolated from the mixture, purified and characterized. The preparation of the precursor 6-(3-tert-butylsulfanyl-5,6-dicyanopyrazine-2- ylamino)hexanoic acid was successful. The cyclization of two precursors yielded the following Aza-Pc: 3-carboxy[2',3'-b]quinoxalino-11,12,18,19,25,26-hexakis(tert- butylsulfanyl)tripyrazino[g,l,q]porphyrazinato zinc (II); 3-carboxy[2',3'-b]quinoxalino- 11,12,18,19,25,26-hexakis(tert-butylsulfanyl)tripyrazino[g,l,q]porphyrazinato magnesium (II). The modification of the preparation of precursor 5,6-dioxo-1,4,5,6-tetrahydropyrazine-2,3- dicarbonitrile by decreasing the reaction temperature has shown to be unsuitable
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Intracellular trafficking of an anti - Amyloid Protein Precursor antibody.
Zitko, Jan ; Doležal, Martin (advisor) ; Trejtnar, František (referee)
Intracellular trafficking of an anti-Amyloid Protein Precursor antibody Alzheimer's disease is characterized by over-accumulation of beta-amyloid peptide (Aβ) in the brain. Aβ is produced by proteolytic cleavage of beta-amyloid precursor protein (APP) by β- and γ-secretases. Novel monoclonal antibody, 2B12, has been shown to bind to β-secretase cleavage site of APP, reducing the production of APP, presumably by preventing the cleavage by steric hindrance. 2B12 is hypothesized to bind to APP molecules exposed on the cell surface and to be internalized in the form of complex with APP via natural endocytic pathway. This hypothesis was confirmed by San Pei Ho's (2007), who followed the internalization of 2B12 in living astrocytoma MOG-G-UVW, cells in time-course experiment. This project is focused on intracellular trafficking of 2B12 and its localization within specific cellular compartments. Experiments were performed with fixed astrocytoma MOG- G-UVW cells (constitutively expressing APP). Originally planned experiments with live cells could not be performed due to decreased stability of 2B12 (causes remain unknown). 2B12 was tested for colocalization with polyclonal affinity purified antibodies labelling subcellular markers (proteins) associated with compartments known to participate in APP...
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Endoparasitic infections in farmed ring-necked pheasant (Phasianus colchicus) and red-legged partridge (Alectoris rufa) (Galliformes: Phasianidae)
Máca, Ondřej ; Hejcmanová, Pavla (advisor)
The entire infection process was monitored over four seasons (2012-2015) by a targeted and systematic study of ring-necked pheasants and red-legged partridges from an intensive artificial breeding programme in the Czech Republic, with focus on studying the transmission dynamics throughout the breeding process, from hatching up to release of birds to open area, in connection with timing of appropriate and effective application of drugs. 1752 pooled excrement samples were examined (745 from pheasants and 1007 from partridges). Post-mortem examinations were performed on 625 dead birds (514 pheasants and 111 partridges). Main infections of A. rufa and P. colchicus comprised representatives of the protozoa genera Eimeria and Cryptosporidium; as well as nematodes Capillaria
s.l., Heterakis gallinarum and Syngamus trachea.
Identification of oocyst isolates of Cryptosporidium baileyi, C. meleagridis anf C. galli were supported
and characterized by nested PCR of heat shock protein 70
(HSP70), actin and 60 kDa glycoprotein (GP60) genes.
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Biotransfer of selected risk metals into plants and their accumulation and distribution in plant organs
Le Minh, Phuong ; Lachman, Jaromír (advisor)
Contamination of soils with heavy metals is one of the serious environmental problems threatening human being. Heavy metals are considered as the special hazard of soil pollutants because of the adverse effects on the plant growth, the amount, activity of useful microorganisms in soils and the quality of food. Regard to the persistent and toxicity, the heavy metals are toxic when we consider different kinds of pollutants in soils. In the soil, zinc (Zn), cadmium (Cd), lead (Pb) and mercury (Hg) toxicities frequently occur than the other metals because of their precipitation and sorption by the soil. It is a very dangerous situation because when these metals are taken up by plants, they can be transported to the food web and food chains.
In the present study, the accumulation of four heavy metals (mercury, zinc, lead and cadmium) in the whole grain of spring accessions of emmer, einkorn and common spring wheat cultivars and potato (Solanum tuberosum) is reported. Heavy and essential elements were monitored in potato cultivars in the exact field experiments and in hydroponically grown plants. The elements were determined by methods FAAS, ET AAS, and AMA (Advance Mercury Analysis). Statistical analyses were performed using SPSS 9.0 with the Tukey HSD (Honestly Significant Difference) test (alpha equal to 0.05).
In our study, the concentration of heavy metals decreased in the order zinc (Zn) > lead (Pb) > cadmium (Cd) > mercury (Hg) in the wheat grain. The comparison between three varieties of investigated wheat revealed that the emmer wheat was rich in zinc content (62.12 mg kg-1 dry matter), while the spring wheat had the lowest average concentration of zinc in the grain (40.99 mg kg-1 dry matter). Generally, the values of lead concentration in grain wheat varieties were low (ranging from 0.1268 mg kg-1 dry matter to 0.2950 mg kg-1 dry matter).
The concentrations of mercury in four typical growth stages of wheat (boot stage 10, heading stage 10.2 1/4 of head emerged, leaf-stage 10.2 and stage ripening 11 according to Feekes) were also determined. It has been shown that the concentrations of mercury in different wheat varieties were absorbed differently at different growth stages of plant. Stage 10.2 and leaf stage 10.2 showed the high mercury content (0.0152 mg kg-1 dry matter and 0.0214 mg kg-1 dry matter, respectively). Among individual varieties significant differences were determined.
Amounts of toxic and potentially toxic elements detected in investigated potato tubers are characterized by a large variability within investigated groups. Performing statistical analysis (one way ANOVA) showed that there were no significant differences between two investigated groups of samples (samples from Uhříněves and Valečov in the year 2013 and 2014) considering either one of investigated metals. Measurable levels of mercury were found in smallest amounts in all investigated potato samples comparing to other metals (Cd, Pb).
Plant cells compared to animal cells are characterized by the formation of cell walls. Plasma membrane or cell membrane is a biological active membrane separating the interior of cell from the outside environment. An adjusted method for isolation of protoplasts was developed and adapted for isolation of protoplasts from plant material (potatoes). In our experiment, the plants were grown hydroponically in the Research Institute of Plant Crops Prague-Ruzyně. If we examine the plant membrane, one option is to remove the cell wall by means of special mixture enzymes. Protoplasts were released in the dark at 25 degrees of Celsius for 18 hours. The 70 and 90 microns sieve was used to filter and then centrifugation for 5 minutes at 100 x g. All the steps were carefully carried out to prevent the damage or breakage of protoplasts.
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The role of ABC transporter in clinical practise
Tornová, Petra ; Pávek, Petr (advisor) ; Čečková, Martina (referee)
The role of ABC transporters in clinical practice ATP-binding cassette (ABC) transporters are a family of transporter proteins that contribute to drug resistance via ATP-dependent drug efflux pumps. There are seven subfamilies classified as ABC transporters (ABCA through ABCG) that are expressed in both normal and malignant cells. They are involved in the transport of many substances, including the excretion of toxins from the liver, kidneys, and gastrointestinal tract, and they limit permeation of toxins to vital structures, such as the brain, placenta, and testis. The best-characterized transporter protein is MDR1/P-glycoprotein, and a number of clinical investigations have suggested that its intrinsic or acquired overexpression resulted in a poor clinical outcome of chemotherapy. Conventional cancer chemotherapy is seriously limited by the multidrug resistance (MDR) commonly exhibited by tumour cells. One mechanism by which a living cell can achieve multiple resistances is via the active efflux of a broad range of anticancer drugs through the cellular membrane. Various types of compounds and techniques for the reversal of ABC transporters mediated MDR have been developed, and efforts have concentrated on the inhibition of function and suppression of expression. Increased drug accumulation and...
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Identification and molecular characterization of the putative immunophilins (IMMs) in the oilseed rape pathogens Leptosphaeria maculans, Leptosphaeria biglobosa, and Plasmodiophora brassicae
Sandhu, Khushwant Singh ; Ryšánek, Pavel (advisor) ; Radovan, Radovan (referee)
Oilseed rape is largely infected by several phytopathogens and two most economical important diseases are blackleg caused by fungus species complex Leptosphaeria maculans and L. biglobosa and clubroot caused by protist P. brassicae. The sequenced genomes of these phytopathogens provide opportunity to uncover various aspects related to disease infection, host pathogen interactions, plant disease resistance, and evolution of pathogens. Considering these we focused on one of the most conserved family called immunophilins (IMMs) in these genomes. IMMs are comprised of three structurally unrelated sub-families including cyclophilins (CYPs), FK506-binding proteins (FKBPs), and parvulin-like proteins (PARs). We identified putative members of IMMs in each phytopathogen using bioinformatics approaches. We further characterized the IMMs based on domain architecture, subcellular localization, exon-intron organization, transcriptomic expression patterns, gene ontology terms, conserved motifs presents and evolutionary analyses. IMMs are performing several vital roles in plants, animals and fungi. However, in phytopathogens their roles are not well established except for cyclophilin that implicates in pathogenicity in some phytopathogens. Therefore, we exploited the role of cyclophilin in L. maculans and L. biglobosa using expression profiles and in P. brassicae using Magnaporthe oryzae cyclophilin deletion mutant. Overall, we concluded that the cyclophilin acts as a virulence determinant in our studied phytopathogens. However, delineating the precise role of other IMMs would also be imperative. Taken together, our findings for the first time shed light on the highly conserved IMM family in the oilseed rape pathogens.
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Mapping project dependencies
Šárová, Irena ; Chocholatý, Drahomír (advisor) ; Roháčová, Barbora (referee)
This thesis describes the issue of project dependencies. In managing projects we meet almost every time some kind of dependency. Currently, the project is almost never an isolated group of activity that is able to independently bring same value or result. Therefore, this issue occurs in many areas of managing project. The first theoretical part is focused on some of these areas. It describes disciplines such as Project, Program and Portfolio management. These areas are characterized and then described how they can solve project dependencies. The second part is focused on matrices DSM and DMM, which can serve as one of the methods to effectively manage all connections and dependencies in projects. Area DSM and DMM matrix is first explained and then are expressed its possibilities. In the last part are the information gained in the previous phase applied to a practical example in the banking environment. The method is applied to 25 projects that are guided in an unnamed banking institution. For these projects are set dimensions and are subsequently compiled specific DSM (Dependency Structure Matrix) and DMM (Domain Mapping Matrix) matrices. To certain dimensions is then used clustering method and on the resulting matrix is used simple matrix notation, such as transposition and matrix multiplication. It will create basically a tool, which is a network of all the dependencies between dimensions and projects that can simulate pre-defined scenarios.
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RNA elements in the genus Armillaria
Dvořák, Jiří ; Gabriel, Jiří (advisor) ; Tomšovský, Michal (referee)
Mycoviruses are widespread among the fungi. Most mycoviruses reported have dsRNA genomes. The first mycovirus was found in the edible mushroom Agaricus bisporus. Mycoviruses do not have an extracellular phase of their life cycle and are transmitted only by intracellular routes. Many mycoviruses reveal no apparent effect on their hosts. The genus Armillaria represents economically important edible fungal pathogens causing root rot in conifers and broadleaves trees. From this viewpoint, genus Armillaria presents an important subject of research studies. In this study we tried to find evidence for the presence of RNA elements in the respective genus. Total amount of collected strains was 52. CF-11 method for nuclei acids extraction was applied to 40 strains. The method is based on the affinity of cellulose powder for nuclei acids and specifically, the adsorption of dsRNA at ethanol concentrations of 15%. According to our results, there is no evidence for dsRNA elements occurence in the tested strains. The study was also focused on basic characterization of physiology of isolates, esp. to optimum growth conditions on agar plates and milled straw as well as on rhizomorph formation and fructification. No effect of pH changes and addition of various alcohols on rhizomorph formation was found, while primordia...
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Characterization of hydrogenosomal iron-sulfur flavoproteins from Trichomonas vaginalis
Pilařová, Kateřina ; Hrdý, Ivan (advisor) ; Kašný, Martin (referee)
Trichomonas vaginalis is flagelated microaerophilic protozoan parasite from Excavata group, which causes trichomoniasis, the most common nonviral sexually transmitted disease in the world. It causes vaginitis in women and uretritis in man and it can also cause problems for example during pregnancy. This thesis is aimed on the characterisation of hydrogenosomal iron-sulfur flavoproteins (ISF) from Trichomonas vaginalis, proteins, which were only recently discovered in the proteome of hydrogenosome of T. vaginalis. Specifically, we have focused on characterisation of ISF3 which is, according to our data, active homodimer and binds flavin mononucleotide (FMN) and iron-sulphur centre in its active site. The iron- sulphur centre is not characterised yet. ISF3 is able to reduce oxygen, hydrogen peroxide, sodium nitrate and metronidazole also in the enzymatic system with PFO and ferredoxin. Next, I tried to reduce ammonium sulphate with ISF3, but unsuccessfully. These results correspond with the activities obtained for ISF from Methanosarcina thermophila, where ISF reduces oxygen and hydrogen peroxide to water. In addition, ISF3 is able to reduce nitrogen compounds. It is important according to the fact, that metronidazole is a drug from the group of 5−nitroimidazoles. The other results show the decrease...
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