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Postranslational modifications in DNA damage response
Moudrý, Pavel ; Hodný, Zdeněk (advisor) ; Malcová, Ivana (referee) ; Mistrík, Martin (referee)
Our genetic material is continually challenged by varieties of factors and processes that represent enormous threat to our ability to transmit genetic information to our offspring and to our survival as well. Cells possess many mechanisms capable of detecting and repairing damaged DNA, collectively known as DNA damage response (DDR). DDR activities must be tightly regulated in a spatiotemporal and DNA lesion-specific manner to optimize repair and prevent alterations in DNA during normal cellular processes. Defects in DNA repair drive genomic instability and may ultimately lead to cancer and neurodegenerative diseases. This work is a compilation of 4 projects focused on molecular mechanisms of DNA damage response and assembly of multiprotein signalling and repair complexes at sites of DNA damage. We identified nuclear pore component NUP153 as a novel factor specifically required for 53BP1 nuclear import and DDR. We showed that NUP153 knockdown prevents 53BP1 from entering the nuclei in newly forming daughter cells, which translates to decreased IR-induced 53BP1 foci formation, delayed DNA repair, impaired survival after IR and exacerbates replication stress-induced DNA damage. Furthermore, we identified ubiquitin-activating enzyme UBA1 as the apical E1 enzyme required for response to IR and replication...
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Macrophages and nitric oxide in leishmania - sandfly - host interactions
Kratochvílová, Tereza ; Fialová, Anna (referee) ; Kolářová, Iva (advisor)
Leishmania reside fagolysosome of macrophages immediately after their entry to host where they multiply and consequently infect other macrophages or eventually other cells. A synthesis of a reactive reactant of oxygen and nitrogen is one of the mechanisms that some mammal cells are equipped with and that also contributes to eradication of leishmania. Nitric oxide rising during a metabolic change of L-arginine under the catalysis of NO synthase is of a large importance. Beyond cytotoxic function, nitric oxide is involved in signalling pathways for a neurotransmission (nNOS) and vasorelaxation (eNOS). Not all types of macrophages have ability to produce NO (iNOS). It is a heterogeneous group differing in immunological function and also in physiology. A group of classical activated macrophages represents an effective APC capable of efficient killing of intracellular pathogens. In addition to NO, they also secrete an inflammatory cytokines, which evolve an immune reaction towards to Th1. Contrary to this, a group of alternative activated macrophages is not capable of any efficient antigen presentation and nitric oxide production but produces L-ornithine, which is a precursor of polyamines, which leishmania utilizes for its own intracellular growth. For the mouse model, status of resistance and/or...
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The efficiency testing of production commercial feeds for african catfish (Clarias gariepinus)
PETR, Miloš
In my diploma thesis I was tested six types of feed. I tested in 3 repetitions in the recirculating system with biological treatment of water. Tested fishes african catfish (Clarias gariepinus) weighted between 200 ? 800 g per piece, were deployed into 18 aquariums with volume of 190 litres per aquarium. The whole experiment took place in five partial 3-week periods in the intensive breeding of the laboratory - controlled reproduction of fishes FROV JCU. The average water temperature during the experiment was 27 °C. The main objective of the thesis was to observe production efficiency of tested types of feed, speed of the growth of fishes, feed rates and financial costs. Afterwards, determinativ of yield and organoleptic evaluation was determined, followed by laboratory examination of composition of meat. The results of production indicators (Specific Growth Rate, Relative Growth Rate, Food Conversation Ratio) were most favorable for 3 types of feed ? Haltáp (SGR=0,81 %; RGR=126 %; FCR=1,45), Coppens TROCO SUPREME-22 (SGR=0,73 %; RGR=105 %; FCR=1,19), Coppens TROCO PRIME-18 (SGR=0,68 %; RGR=97 %; FCR=1,26). The lowest costs for 1 kg of fish increment were achieved for these specific types of feed Haltáp and Aller Aqua Focus (45 Kč a 46 Kč). In the organoleptic analysis was found positive evaluation of meat of all tested fishes. Chemical composition of meat was (in term of proteins and fats content) most favorably documented for the hungarian feed called Haltáp (proteins=19,22 % and fats=4,85 %).
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Potential of cholesterol-loaded cyclodextrins and low density lipoprotein for increasing the efficiency of cryopreservation of epididymal sperms in stallions
Janošíková, Martina ; Rajmon, Radko (advisor)
In the case of sudden death or injury stallion reproductive system or necessary castration preventing the collection of semen can to preserve the genetic potential of exceptional individuals take advantage of epididymal sperm. Virtually the only way, how to preserve fertilizing sperm POTENTIAL for long time IS cryopreservation. However, this process induces irreversible changes that are the cause of the reduced number of sperm capable of fertilization after thawing. Epididymal sperm cells have properties different from the ejaculated, harder to keep. Therefore, they are constantly looking for ways to streamline their cryopreservation.
GENERAL most commonly used cryoprotectants include glycerol and Egg yolk. Despite the positive effects is glycerol toxic to cells and Egg yolk also contains ingredients that act on sperm metabolism negatively. Increasing the efficiency of cryopreservation protocols themselves likely to be achieved by replacing toxic components while leveraging industry-produced active yolk fractions.
Submitted dissertation topic itself will deal with the use of cholesterol-loaded cyclodextrins Along with low density lipoproteins, the cryopreservation of epididymal sperm.
Main method: HPLC, Western blot, electrophoresis, fluorescent labels, computer-assisted sperm analysis (CASA).
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Phylogeography and conservation genetics of endangered saproxylic beetles in Europe
DRAG, Lukáš
This thesis introduces the use of molecular methods for the conservation of several species of endangered saproxylic beetles in Europe. It focuses on the questions related to the DNA preservation and microsatellites development, as well as the evolutionary history and conservation of threatened species. Using the combination of mitochondrial and nuclear markers, the genetic diversity and reintroduction history of Cerambyx cerdo was assessed and the phylogeography of Rosalia alpina from the whole range of its distribution was studied. This information is valuable for designing more efficient conservation strategies.
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Estimating the flow capacity of river
Lorenc, Václav ; Dráb, Aleš (referee) ; Duchan, David (advisor)
The bachelor thesis is dealing with the watercourse`s capacity assessment of the river Jihlava on the kilometers 136,5 to 139,5 in the village Maly Beranov. In the chosen locality was made a model of the surface water flow during the flood flow rate Q1, Q5, Q20 and Q100. Based on the gathered information about the area`s utilization, errand and following calculation of the flow, the areas for a potential flood protection were identified. In the problematic locations was suggested to dredge the bottom. In selected sections was made a draft of protective earth dam and concrete wall. According to the draft of flood protection, the model of surface water flow was adapted and the efficiency of flood protection was verified by calculations. All the results are published in clear maps.
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Analysis of micloflora in cheeses using DGGE
Čakajdová, Martina ; Rittich, Bohuslav (referee) ; Trachtová, Štěpánka (advisor)
Molecular biological methods are fast and efficient tool for the identification of microorganisms in real samples. The aim of this diploma thesis was analysis of microflora in control and contaminated brined cheeses. DNA isolated from analyzed samples was used to optimize the PCR course using primers with GC clamp on the distribution of amplicons using DGGE. DGGE products were reamplified after optimization and prepared for DNA sequencing. DNA isolated from analyzed samples was used in real-time PCR with high resolution melt analysis of the amplicons (HRMA). Samples of cheese and bacterial cultures isolated from cheeses were compared by DGGE and HRMA. Comparing the position of the amplicons was found that contaminants may be Bacillus licheniformis, Staphylococcus epidermidis, and Acinetobacter baumanii/ calcoaceticus. Sequence analysis of cheese and pickles amplicons, the presence of Bacillus sp. and other microorganisms spree in five genera were detected. Representatives of the tree genera were cultured. It is considered contamination Bacillus sp., or microorganisms which are not culturable methods used. The method is suitable for the analysis of complex microflora in cheese and pickles after further optimization.
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Porovnání různých metod izolace genomové DNA hmyzu
Novák, Vojtěch
This thesis deals with description and verification of different methods of DNA isolation from insect tissue of order Lepidoptera -- butterflies with an emphasis on family Sphingidae -- hawk moths and subsequent amplification of COI gene by PCR method. All tested methods were compared and evaluated in terms of efficiency, yield and purity of the DNA isolation, duration and financial costs. We selected to establish a methodology which is sufficient for obtaining COI gene sequences using genetic analyzer, its application in the system BoldSystems and subsequent determination of the sample. This is important in effort to map the species diversity.
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Metody izolace a archivace DNA z živočišných tkání
Dratvová, Lenka
n bachelor thesis methods of isolation and archiving of DNA from animal tissues I deal with different samples of animal tissues suitable for efficient isolation of DNA. It selected simple and modern methods of DNA isolation. The work also deals with the possibility of short-term and long-term storage of DNA with a description of the selected archival methods. The method of DNA compared in terms of quality, quantity, and labour, time and monetary perspectives. The individual methods of archiving are also evaluated at the end of the work. The work also includes a practical part, which deals with the isolation of DNA mainly from animal products (milk and milk products, blood, meat and meat products), but also other biological tissues. The procedure and results are presented in chapters "Material and Methods", and "Results and Discussion".
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