Žabovřeské and Komínské meadows are divided acording to their names, but in the real landscape it is just only fluent space of large river valley. The space was created by river fo ages, so I let the space to float with the river. All important division of the space are done in parallel with the river, that itself classife the calm, narrower and more familiar right bank and more open, larger left bank of the river. The river shaped the linear charakter of the park and in the same time identify the circle – the space of stop in. The superficies of the area: 141,1 ha.

Alba family proteins are highly conserved in all domains of life. They are involved in RNA metabolism in Archae and Eucarya, while they are involved in the chromatin organisation in Crenarchaea. In animals, ALBA proteins were identified to associate with RNase P/MRP subunits. The objective of my thesis was the characterization of ALBA family proteins in a model plant Arabidopsis thaliana. The Arabidopsis genome contains six genes with close homology, three from Rpp20-like subfamily and three of Rpp25-like subfamily. Here I present the localization of GFP-fused proteins in Arabidopsis stable lines harbouring constructs cloned by Gateway® Technology. ALBA proteins were localized in the cytoplasm and undefined particles in root differentiation zone and in mature pollen. The characterization of the respective T-DNA insertion lines did not reveal significant phenotype defects in growth and development of sporophyte and gametophyte in comparison to Columbia-0 plants, probably because of likely functional redundancy od the paralogs. Expression profiles and localization of ALBA proteins suggest their possible role in differentiation and dehydration stress response in Oryza. They were also observed to associate with repressed mRNA transcripts in storage EPP particles. Collectively, I propose the likely role...

0 Abstract Cystic fibrosis is an autosomal recessive disease caused by mutation in CFTR gene coding for a chloride channel in apical membrane of epithelial cells. This disorder leads to the change in ion transport causing the increase in mucus viscosity in airways as well as changes in glycosylation of saccharide structures on the cells. Because of that these cells are the target for bacterial adhesion. Chronic bacterial infections, which lead to gradual decline of lung function and damage of lung tissue, are the major cause of death of patients suffering with cystic fibrosis. Pseudomonas aeruginosa is the main pathogen causing chronic infections in cystic fibrosis patients. This bacterium produces a biofilm protecting them from host immune system and antibiotics. Once the colonization with PA occurs, it is difficult to get rid of this pathogen. The prophylactic treatment with orally administered hen antibodies against the PA virulence structures could be a prevention of chronic PA infections. In this work we tested the antibody against the bacterial lectin PA-IIL, which is suggested to be involved in the adhesion of the pathogen on epithelial cells. First, it was verified that the prepared antibody from egg yolks of a hen immunized with the bacterial lectin PA-IIL recognizes this antigen expressed...

Maginot line is one of the essential issues of military history between two world wars. This graduation thesis situates this phenomenon in wider political and military context. The period of interest is between the First and the Second world war, but for explanation of full coherence, sometimes it was necessary abandon this frame. The main object of interest is not the history of Maginot line itself, but everything that shaped the points of view of generals and politicians about its construction. The answered questions are, why did they start to build this bulwark, what were they waiting for and what advantages and disadvantages they assigned to this line. The negativ opinions of political left were also analysed, just as the french interwar fortification school. There is an attention on reflections of Maginot line in civil and military periodicals that together with analysis of opinions of important political and civil persons enabled to answer the question, whether it is possible to speak about myth of Maginot line. This graduation thesis shows clearly that it is very difficult to think so, because construction of Maginot line didn not influence the life in France so much and it wasn not the cause of fallacious imaginations either.

Horizon as a line that can contain a mystery. Without any futilities, just because it goes from one point to another. It doesn`t matter what scale we find it in. The Possible Borderline will consist mostly from photographs, but also from drawings and watercolours. Another part will be a subtle wire installation dealing with the horizon topic in a metaconcrete level – at someone`s home. The entire artwork should create a gentle story, a personal, imaginary horizon.

NK buňky (z angl. natural killer cells, přirozeně zabíječské buňky) hrají klíčovou roli při rozpoznávání a ničení nádorových, infikovaných nebo jinak pozměněných buněk. Na svém povrchu nemají antigenně specifické receptory, proto je řadíme mezi složky přirozené imunity. K rozpoznání cílových buněk slouží řada jiných povrchových receptorů. Inhibiční receptory zajišťují buněčnou toleranci, naopak aktivační receptory spouští cytotoxické mechanismy vedoucí k apoptóze a tedy lýzi buňky. Díky této vlastnosti jsou NK buňky intenzivně studovány v souvislosti s imunoterapií nádorových onemocnění. Jedním z aktivačních receptorů je NKp80 rozpoznávající svůj ligand AICL. Oba proteiny patří do rodiny receptorů podobných lektinům C-typu. Tento komplex se účastní nejenom přímé lýze maligních buněk myeloidního charakteru, ale má také důležitou roli v imunomodulaci zánětu. Předmětem této diplomové práce je příprava receptoru NKp80 a jeho ligandu AICL. Receptor NKp80 byl připraven v linii lidských embryonálních ledvinných buněk (HEK 293S GnTI- ). Byly připraveny stabilně transfekované linie produkující protein NKp80 konstitutivně nebo indukovatelně. Zapojení disulfidických můstků a obsazení N-glykosylačních míst proteinu NKp80 bylo ověřeno hmotnostní spektrometrií. Dále byl optimalizován postup Mgr. Jiřího Nového na...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Jesika Štorkánová Supervisor: RNDr. Eva Novotná, Ph.D. Title of diploma thesis: Study of the cytotoxicity of selected chemotherapeutics for the treatment of leukemia in human tumor cell lines Leukemia represents a diverse group of malignant diseases with a hematopoietic disorder with different prognoses. As the incidence of patients with leukemia is increasing, is an effort to establish the treatment that will lead to successful therapy. One of the basic approaches to the treatment of leukemias is chemotherapy. Today it is known that the effectiveness of chemotherapy is influenced by a number of factors which can significantly affect the treatment strategy and thus decide on the outcome of the treatment itself. An important approach in chemotherapy is the selection of cytostatics with maximum efficacy for oncological disease and elimination cytostatics to which the cells are resistant based on the findings in in vitro conditions. The aim of this diploma thesis was to determine the inhibitory effects of in vitro selected chemotherapeutics in cell tumor lines. For determine the inhibitory effect, HCT116, HepG2 and HL-60 cell lines were selected using a colorimetric method based on the...

Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Helena Zálešáková Supervisor: PharmDr. Marie Vopršálová, CSc. Title of diploma thesis: In vitro Methods for the Prediction of Blood Brain Barrier Penetration This thesis deals with the correlation between two in vitro models simulating the blood- brain barrier (HEB, hematoencephalic barrier) and their comparison in terms of practical use. These are the PAMPA (Parallel Artificial Membrane Permeability Assay) method and the MDCK (Madin-Darby Canine Kidney) cell line, which are models for potential central nervous system (CNS) penetration screening. Within this work, a set of sixteen standard drugs were measured. The procedure was similar in both methods in order to obtain information on the amount of test substances passing through the membrane from the donor portion of the plate to the acceptor. The concentration in the donor portion was measured by UV-VIS spectrophotometry. The main difference between these methods is the membrane through which the substances penetrate. In the case of PAMPA, a lipid solution that has been isolated from pig brain (PBL, polar brain lipid) is used. This lipid simulates the phospholipid membrane of the brain capillary endothelium. In the MDCK model, the membrane...

Isogenic lines in fish represent a fundamental approach to control the genetic background of experimental animals. All individuals from a given isogenic line share the same genotype. So far, isogenic fish lines have been produced only by repeated uniparental inheritance - androgenesis and gynogenesis. Homozygous progeny is produced in the first generation of uniparental inheritance, and each homozygous individual produces a different isogenic line after second generation of uniparental inheritance. Despite optimized procedures for inducing uniparental inheritance, isogenic lines have been successfully produced in only a few species of fish. Doubled haploids after first uniparental inheritance have affected fitness as well as reproductive performance. Long-term maintenance is considerably problematic even when isogenic line is established already, due to low viability and poor reproductive characteristics. The situation is further complicated by the fact that isogenic lines are usually naturally monosex, thus uniparental inheritance must be re-used for further reproduction, or sex reversal needs to be applied in part of isogenic line. Several types of germ cell manipulation were performed in presented thesis. Protocols for cryopreservation of spermatogonia and oogonia have been developed and optimized to maximize post-thaw viability. The physiological activity of cryopreserved cells was confirmed by transplantation into a surrogate host. Cryopreserved and subsequently transplanted cells retained colonization activity comparable to non-frozen control germ cells. More importantly, male germ cells were able to transdifferentiate from oogonia. The success of transplantation was confirmed by detection of expression of genes associated with gametogenesis in carp by RT-PCR. In the next study, the results of cryopreservation experiments were followed, where sterile goldfish was identified as a suitable host for homozygous carp cells. Germ cells obtained from several homozygous individuals were individually transplanted into sterile goldfish. This procedure has a potential to increase the chance of producing a viable gamete for isogenic line production. Germ cells from homozygotes with affected gametogenesis can be transferred to fully viable recipients, thereby increasing the efficiency of isogenic line production overall. In addition, the use of a goldfish as a surrogate parent will ensure that part of the germline chimeras will be male and female, thus isogenic gametes of both sexes can be obtained and no further intervention for further reproduction of the isogenic line. The suitability of triploid zebrafish, which can potentially be used as recipients for cells from homozygotes to produce isogenic lines, has been confirmed for zebrafish. Spermatogonia and oogonia from diploid donors were transplanted into artificially induced triploid larvae. Donor-derived sperm was were obtained upon maturation of triploid recipients. Transplanted oogonia transdifferentiated into spermatogonia and spermatozoa with female sex chromosomes have been produced, which may be of interesting for further studies of sex determination in zebrafish. A new germline transfer technique has been developed using striped embryos. Donor cells were transplanted from the blastula stage to the swim-up larvae. With this approach, undifferentiated primordial germ cells were able to colonize the genital groove and initiate gametogenesis. After reaching sexual maturity, germline chimeras were obtained with gametes and viable progeny. Although the overall efficacy of this method was lower compared to other transplantation methods, this study may be of relevance for germline rescue in poorly viable embryos or lethal mutants.

The induction of germ-line chimerism is an expanding focus of fisheries research. This technique is having a potential to enhance the production of gametes of species that are commercially valuable, endangered, species with problematic reproduction, using a more common or easily available species or species adapted to artificial reproduction as a surrogate host. The main goal of this technology is to establish a small-bodied surrogate broodstock producing functional donor gametes based on germ cell transplantation. Extent preliminary experiments, including documentation of donor/host embryonic and larval development, characterization of germ cells enriched by documentation of their migratory activities, sterilization of the host, isolation and cryopreservation of donor germ cells, are key factors for launching this biotechnology. All these crucial points were the main objective of the present work. The whole thesis provided the focus on two different fish species. First, our commercially valuable fish, the tench, where we would like to apply our current knowledge to create a germ-line chimera within cyprinids by transplantation of tench germ cells to smaller and faster-reproducing fish species as white cloud mountain minnow. Secondly we focused on the endangered species (listed in IUCN Red List) of large body size with long reproductive cycle, the sturgeons. In this case, we have chosen sterlet as a host, providing an advantage of shorter generation interval and smaller body size, to produce gametes of donor, a critically endangered species of large body size with long reproductive cycle, such as beluga. This innovative technology could result in collection of sperm and eggs in shorter time from small-bodied host. In tench we firstly focused on embryonic and larval development documentation together with description of origin and migration pathways of primordial germ cells (PGCs). PGCs represent a powerful tool for creation a germ-line chimera within fish species because they transmit genetic information to the next generation (Linhartova et al., 2014a). Secondly, we reported a practical technique for isolation and cryopreservation of early stages of germ cells (GC), including spermatogonia (SG) and spermatocytes (Linhartova et al., 2014b). In case of sturgeons, Saito et al. (2014) firstly described the origin and migration patterns of sturgeon PGCs deposited at the vegetal pole of the egg similar to that in anurans. Secondly, Psenicka et al. (2015) reported isolation and cryopreservation of female and male GC, SG from testes and OG and pre-vitellogenic oocytes from ovary, of 2-4-year old Siberian sturgeon. Moreover the isolated GC were transplanted into host (sterlet) and process of transplantation resulted in successful colonization of sterlet genital ridge. The potential host for germ-cell tranplantation, sterlet, was sterilized by knock-down of germ cell specific gene, the dead end gene, by the morpholino antisense oligonucleotide (MO) agent (dnd-MO). These results reported the first known and functional method of sturgeon sterilization (Linhartova et al., manuscript). We provided important information on morphology and ultrastructure of beluga spermatozoa structure by scanning and transmission electron microscopy to increase knowledge of evolutionary and taxonomic relationships among sturgeons (Linhartova et al., 2013). Finally, this thesis presents several studies with differing focus of research but with one target goal to induce germ-line chimerism in fish. All these results are prerequisite of future application and development of surrogate production in these species.