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Spruce forest is an important production ecosystem for our civilization. Its development can be affected by three main types of disturbances: forest fire, windstorms and spruce bark beetle attack. Experiences with a massive and long-term attack of spruce bark beetle in the Šumava mountains and a strong windstorm in 2004 in the spruce forest in the High Tatra provoke questions, how natural are such severe disturbances. To understand the current development of forest ecosystems, it is necessary to study past structure of these forests and frequency of forest disturbances. This work is about locality Tatranská Lomnice located in the High Tatra mountains, in the belt of mountain spruce forest. In my work, I used mainly pollen and plant macro-fossils analyses. I compared my data with disturbance information originating from dendroecology for windstorms and from charcoal analysis for fires. My research found that according to the pollen assemblages, most of the disturbances did not have strong influence on the forest composition, and most likely they only caused the forest thinning. The most significant fire occurred around 1420. The most significant wind disturbance probably occurred in 1890. The was not found any effect of spruce bark beetle. In the past millennium disturbances affected spruce forest...

Intraneuronal filamentous lesions which are characteristic for Alzheimer´s disease, the so called neurofibrillary tangles (NFT´s), are misfolded beta-sheet like aggregates of tau protein. Their presence is closely linked to neurodegeneration and progression of the disease. Tau protein has been investigated as a promising target for therapeutic intervention of AD. Several monoclonal antibodies specific to tau protein were previously identified and recovered from human blood serum. As potential candidates for development of biopharmaceuticals, the anti-tau antibodies and their interactions with tau epitopes are to be investigated in detail. This work aims to develop novel methods for linear epitope mapping and to characterize binding sites of identified anti-tau antibodies on tau protein.

The objective of diploma thesis is to define specific groups of population, defined by sex, age or education, which are represented disproportionately in comparison of whole population of Czech Republic. In the thesis is described the population structure in terms of demography and economy, as well as factors which influence the labor market from short-term and long-term point of view. To achieve all marked out objectives is made explanatory statistics for unemployment of specified groups of population and the index analysis that allows to examine the inter-annual development of unemployment rate in terms of age, sex and education. Based on the index analysis are defined critical groups, which are evaluated by proportionality among unemployed persons. The analysis showed that critical groups are considered young people with lower level of education and women with high school education or women 30-39 years old.

Bachelor thesis deals with comfort of a mother-to-be during spontaneous and induced labour. Spontaneous labour has assumed on the basis of natural mechanisms of a woman´s organism and was going on without obstetrician´s interventions. In the opposite we understand with the induced labour an artificial evocation of the womb activity in order to end pregnancy with a vaginal delivery. Comfort is closely connected with the delivery; it is defined as situation of physical, psychic and social well-being or peace. Bibliography distinguishes between a physic and social comfort and an environmental comfort. In the theoretical part of this work is this closer described based on quoted bibliography and newest findings in this field. Further is in detail elaborated the nursing care of women in induced and spontaneous labour and the role of midwives during all delivery phases. The practical part of the thesis is aimed to perception of comfort differences at spontaneous and induced delivery by women during the labour. For this thesis was set only one goal, just to find out exactly the difference in perception of spontaneous and induced delivery by mothers-to be during the delivery. Two research questions were determined. Wording of the first question: "How do women evaluate the difference of pain perception of the womb activity at spontaneous and induced labour?" Second questions pursued differences in the medical procedures: "How do women evaluate the difference in medical procedures in case of a spontaneous and induced delivery?" For processing of this problematic and finding out of the given goal was chosen a quality research. For the data collection was used the technique of half structured interviews. These interviews were later processed with the method of content analysis. The research array were women after labour at the puerperium department of hospital Nemocnice Strakonice, a. s. For the quality research were eight women selected, who have the experience with both types of the labour (spontaneous and induced). Based on these quality research results were reached further described conclusions and answers to research questions. The majority of respondents handled better the spontaneous labour. Only one respondent was more satisfied with the progress of inducted delivery. All approached respondents realized that inconveniencies and pain, which accompany the labour, are inseparable part of the labour action, however there were detected significant differences primarily in experiencing of womb contractions. During the spontaneous delivery expectant mothers reported gradual entrance of contractions, so they could have used to the pain and get ready for it. During the induced delivery was the ache sudden, sharper and permanently intense. The biggest difference in the delivery leading experienced all respondents (without one) in the possibility of free movement, which was possible only during the spontaneous labour. During the induced labour was the motion supposedly restricted by frequent CTG-checks and often repeated vaginal screening. Further they could have drunk and eaten during the spontaneous delivery. They have used more non-pharmacologic relief methods by the spontaneous delivery too - like relief positions, aromatherapy, gymnastic ball, massages, shower etc. During the induced delivery they have already asked for pharmacologic methods. An effort of the expert public, mostly of the midwife, is to create the best possible level of comfort to women in labour, so they are satisfied with the labour progress and manage the own delivery best possibly. This bachelor thesis can serve as information material for pregnant women, who are actively getting ready for coming labour. Even future midwives, who prepare their selves for this job can use it, alternatively is it aimed for special midwives ("duly") and other persons attending the delivery.

Bacillus subtilis, a model Gram-positive soil bacterium, employs two distinct mechanisms in its membrane adaptation to low temperature: 1) Long-term adaptation to suboptimal temperature is accomplished by increasing the ratio of anteiso- to iso-branched fatty acids in the membrane lipids. 2) After a sudden temperature decrease, the oxygen-dependent fatty acid desaturase (Des) is induced which desaturates fatty-acyl chains incorporated in membrane lipids. The transcription of the gene encoding desaturase, des, is activated by the decrease of the membrane order, via two- component system DesK-DesR. In this work, I studied the influence of cultivation conditions on the mechanisms of B. subtilis membrane adjustments for a low temperature employing fatty acid analysis, fluorescence spectroscopy, differential scanning calorimetry and methods of molecular biology. In the first part of this work, I examined the impact of the cultivation medium on the composition and biophysical features of the B. subtilis cytoplasmic membrane during growth under the optimal (40 řC) and suboptimal (20 řC) cultivation temperature. I compared the nutrient-rich complex medium containing glucose and the mineral medium supplemented with either glucose or glycerol. The results obtained showed the crucial importance of medium...

In my Bachelor´s Report I try to introduce the area of ``Public relations{\crqq} (PR) and relate disciplines which are closely connected with this area/discipline. The different questions related to PR are described in every single part of this report. Closer view is devoted to one of PR tool which is an audiovisual presentation. In practical part of this report I concentrate on concrete function of kindergartens and on the forms of presentations which are used by kindergartens. The screenplay for audiovisual presentation of kindergarten is the output of this part. It includes the creating of DVD presentation of concrete kindergarten.

This thesis deals with metal nanoparticles and their antibacterial effects. Nanotechnologies have been paid intensive attention both in scientific and public area. Some metal nanoparticles possess provable antibacterial potential. Nowadays, when the growing resistance of pathogenic bacteria represents an important issue, metal nanoparticles could be one of the potential solutions to the problem. Interactions of metal nanoparticles with bacteria have however not yet been fully understood. It is also evident that their effects depend on various physical and biological factors. The cytotoxicity of nanoparticles to eukaryotic organisms has not been explored in depth either. These all are the reasons why the practical use of nanoparticles has still many limitations. This thesis summarizes information available on the topic of bactericidal effects of metal nanoparticles.

A natural arsenic anomaly at Mokrsko is a site of biogenic precipitation of realgar (As4S4) in stream sediment. The extent of the phenomenon in Mokrsko is globally unique. A previous study showed the ability of microorganisms to solubilize arsenic from its secondary minerals and to produce realgar precursors, i.e. sulfides and arsenites, by anaerobic respiration. The study also raised questions about the mechanism of realgar precipitation since physicochemical conditions favoring this reaction were never detected despite significant sampling efforts. We chemically and microbiologically analyzed sedimentary profiles to the depth of ca. 120 cm in order to understand the functioning of the biogeochemical system. The profiles comprised both the unsaturated and the saturated zone. We distinguished six different domains representing environments from the surface soil to anoxic sedimnent containing realgar-encrusted wood. An analysis of phylogenetic dissimilarity revealed that microbial communities from the various domains form distinct clusters. This suggests that different conditions prevail in the various domains, and that different biogeochemical processes take place there. Incubation (microcosm) experiments showed that bioprecipitation of realgar can be conducted in vitro. It requires a suppression...

Early DNA-based therapies were tested for therapeutic applications, but they sooner or later revealed multiple hurdles and risks preventing their use in further clinical trials. Recently, they have been replaced by rapidly evolving gene editing using programmed nucleases capable of precise genome modifications by cleaving specific DNA sequences. Zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and CRISPR/Cas9 system are currently under investigation as potential therapeutics. However, their off-target effects must be controlled. Targeted delivery of nucleases in a form of mRNA seems as the most promising method. Various types of nanoparticles enable mRNA transfer and could be used to facilitate the nuclease application. Some of these nanoparticles together with characterization of the programmed nucleases are described in this thesis.

Bacterial adhesion is a process of attachment of a planktonic cell to the surface and the necessary step for further biofilm formation. Most bacterial species are capable of biofilm formation. Bacteria within biofilm exhibit increased resistance to antibiotics and other external factors, which makes the eradication of adhered bacteria rather difficult. Bacterial adhesion is very complex process affected by many physical and chemical factors as well as by the adhesive molecules present on the substrate and cell surface. Accurate description of these factors enables to design appropriate modifications of both the substrate and the environment with the aim to fabricate the antiadhesive and antibacterial surfaces or to increase the efficiency in bacteria-based biotechnological processes. Whereas in medical applications the main goal is to decrease bacterial adhesion, the increased adhesion is desirable in certain processes employing bacteria, as is for example the wastewater treatment. This thesis reviews main theories used for prediction of bacterial adhesion, describes the process of adhesion and physical, chemical and biological factors that influence it. It also contains an overview of antiadhesive and antibacterial surfaces. Key words: Bacteria, adhesion, pili, flagellum, antiadhesive and...

Nanocrystalline diamond (NCD) films possess great mechanical properties (low friction coefficient, high hardness etc.), chemical properties (e.g. low corrosivity or chemical inertness) and good biocompatibility. This makes them perspective materials for protective coatings of medical implants and devices. As bacteria biofilms are often very resistant to antibacterial treatment, materials with anti-bacterial or at least anti-adhesive properties are needed. The interaction of NCD films with bacteria has not been properly examined yet. The aim of this thesis was to introduce and optimize the methods for routine bacterial biofilm cultivation and analysis, use them to investigate the ability of NCD films to inhibit the attachment and biofilm formation of Escherichia coli and correlate it with the NCD surface hydrophobicity. The materials used for the study were hydrogenated NCD (hydrophobic), oxidized NCD (hydrophilic) and uncoated glass. For bacterial biofilm growth, cultivation in six-well plates and continuous cultivation in CDC Bioreactor was used. Several methods were tested for quantitative biofilm detachment and analysis. The putative anti-bacterial properties of NCD material were not confirmed in this work. Higher bacterial attachment to NCD films in comparison to the uncoated glass was...

The genus Streptomyces produces more than a half of the known bioactive substances, ranking it among the most important bacterial taxons. Streptomyces lincolnensis ATCC 25466 encodes a biosynthetic gene cluster for lincomycin biosynthesis in its genome. Apart from the biosynthetic and regulatory genes, the cluster also contains three resistance genes, lmr(A), lmr(B) a lmr(C), which could protect of the host from the toxicity of a synthesized antibiotic. The Lmr(C) protein belongs to ARE proteins which generaly confer resistance to clinically important classes of antibiotics: macrolides, streptogramins, lincosamides and pleuromutilins. In addition to antibiotic producers, ARE proteins are also present in pathogenic microorganisms. However, the resistance mechanism conferred by these protins which belong to ABC transporters, even though they lack the transmembrane domain, have not been characterized yet. This makes the ARE proteins an interesting subject of the research. Using deletion mutants in resistance genes lmr(A), lmr(B) a lmr(C) we studied their effect on the lincomycin production and resistance to lincosamides, lincomycin and clindamycin with special focus on the function of the lmr(C). We have found that deletion of lmr(C) does not significantly influence lincomycin production and...

1 Abstract: Secondary metabolism of Gram-positive soil bacteria from the genus Streptomyces is a inestimable source of natural products including manumycins, which belong to a polyketide group. These products possess weak antimicrobial, but important antiinflammatory, and antitumor activities. Streptomyces sp. offers broad amounts of yet undiscovered antibiotics, potentially utilizable in clinical medicine. This fact makes out of these organisms a promising solution to our present problem with rising antibiotic resistance among microorganisms. Two main ways are applied in this research: There are efforts of prepairing new derivates based on known products and creating various modifications in their structure. Next, new producers are discovered by "genome mining" methods, activation of silent gene clusters, followed by improvements of antibiotic production. One of those silent clusters was found in the Saccharothrix espanaensis DSM44229 strain. The genetic information has been transferred to a heterologous host in order to characterize its product. Cluster activation and production of novel manumycin-type metabolites occurred in the host after the transfer.

This work compiles studies that lead to new applications of viral nanoparticles with an emphasis on utilizations in biomedicine. Firstly the work follows the characteristics of viral nanoparticles and some general aspects of nanoparticles. Further described herein are exemplary studies that are using viral nanoparticles as a tool for the treatment of cancer, viral, bacterial or other diseases. This review also includes examples of researches leading to the usage of viral nanoparticles for diagnostic purposes. Lastly, studies in which viral nanoparticles served as nanoextractors, nanoreactors or nanotemplates for the synthesis of metal nanoparticles and galvanic cells are mentioned here too.

Defensins are peptides with antimicrobial, antifungal and antiviral activity. Defensins are likely to occur in all vertebrate species. There are three subfamilies of defensins - α, β and θ-defensins. The α-defensins exists only in mammals and θ-defensins were described only in macaques and baboons but their genes occur in genomes of all old world monkeys except Homininae subfamily. In humans no θ-defensins are produces due to premature stop codon in their coding sequence. However, they were produced synthetically in vitro and named retrocyclins. Retrocyclins not only maintain antimicrobial activity - but also exhibit unusual antiviral activity. Retrocyclins successfully protect cells from HIV, HSV, influenza and dengue fever virus infections with no detectable cytotoxicity to host cells. Retrocyclin RC- 101 is close to the clinical testing as microbicide to prevent heterosexual transmission of HIV-1.

Two component systems were traditionally considered as main phosphorylation systems of bacteria involved in cell signalling. Recently, attention focuses increasingly on bacterial eukaryote-like Ser/Thr protein kinases (eSTKs). These protein kinases are structurally similar to their eukaryotic counterparts. Some eSTKs possess additional domains such as extracellular PASTA domains that were discovered in a variety of gram-positive bacteria. It has been proved that these domains can act as sensors for unlinked peptidoglycan fragments. However, majority of environmental signal molecules still remains unknown. eSTKs phosphorylate a broad spectrum of substrates including proteins involved in various cell processes such as virulence, cell wall biosynthesis, cell division, and central and secondary metabolism. Cross talk between eSTKs and two component systems also occurs. In this thesis, the current knowledge about eSTKs and their significant substrates in different bacterial species is discussed.

Biofilm formation is one of the most common bacterial survival strategies. Majority of bacterial species are able to form these three-dimensional structures, including pathogens like Mycobacterium tuberculosis. Representatives of Mycobacterium genus widely occur in the nature, although they can cause serious problems when they appear in medical equipment and artificial replacements of the human body. Non-pathogenic Mycobacterium smegmatis mc2 155 was used as a model organism in our experiments. We investigated morphology of the three- and six-day-old colonies (in fact biofilms) on agar and agar covered with cellophane using Stereo microscope and Scanning Electron Microscope. We found that a type of surface as well as a carbon source has a great influence on the morphology of the M. smegmatis colonies. We isolated proteomes from the agar and cellophane cultures and from planktonic culture. Two-dimensional electrophoresis was used as the main proteomic method. Proteomic data were analyzed using PDQuest software. Then the sets of proteins detected by qualitative and quantitative analyses were compared using Venn diagrams. As a result, we recognized 7 unique proteins that might be specific for recognition and adhesion of bacteria to the cellophane, no unique protein in agar proteome and 46 unique...

Pathogenic yeasts are nowadays a serious threat for mammalian host. They can cause dangerous diseases, which in many cases even result in death. Pathogens increase the chances of systemic infections by many virulence mechanisms. Experiments addressing the pathogenhost interactions are crucial for defeating these types of infections within the human body. Host-pathogen interactions are very complex and include all components of multicellular host organism. Recently, scientists have focused on the interaction of the mammalian immune system and pathogenic yeasts in more detail. This work summarises interactions of pathogen with selected host cells, especially with macrophages. Yeast pathogens, especially Candida albicans, are capable of influencing the gene expression in interacting cells. These pathogens are capable of modulating the expression while engulfed inside macrophages and other cells of the immune system. Pathogenic yeasts can also change the overall characteristics of their surrounding environment. C. albicans can sense pH and influence it. Therefore, it can increase its virulence by the changes of pH leading to autoinduction of morphological transitions. This work briefly reviews how selected yeast pathogens influence their surroundings while interacting within the host organism. Deeper...

Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are causative agents of Lyme disease and human granulocytic anaplasmosis. Their common vector in Europe are the ticks from the genus Ixodes. In our work, we focused on interaction of innate immune cells with the causative agent of Lyme diseases, that are insubstitutable in their function in the early phase of the disease. Anaplasma phagocytophilum is hard to cultivate, the only possibility is to cultivate it in cell cultures. Successful cultivation of Anaplasma phagocytophilum acquired from patients in our geographic area is crucial for following experiments and for diagnostics too. In our experiments, we used validated cell cultures of HL-60 cells, canine monocytes DH82 and murine monocytes P388D1. During our studies of interaction of the causative agent of Lyme diseases with cells, we used two strains of different species Borrelia. Borrelia garinii M192 and Borrelia burgdorferi sensu stricto B31. These strains vary in virulence. The strain M192 is virulent, but the strain B31 lost its virulence by passages. We specialised in study of morphological changes using light microscopy (observation of dyed and fixed preparates and observation in dark field), eventually by transmision electron microscopy. During our experiments, we concluded that HL-60...

1 Abstrakt Aby bakteriální buňka přežila neustále se měnící podmínky, musí se na ně adaptovat. Tato adaptace je podmíněna změnou genové exprese. Klíčovým krokem genové exprese je transkripce. Hlavním enzymem bakteriální transkripce je RNA polymerasa (RNAP), což je esenciální vícepodjednotkový enzym. RNAP je nejvíce prostudována u Escherichia coli, modelového organismu gram negativních bakterií. Porovnala jsem E. coli a Bacillus subtilis (zástupce gram pozitivních bakterií) a shrnula jsem rozdíly v RNAP a transkripci. Jejich RNA polymerasy se liší přítomností podjednotky δ u gram pozitivních bakterií. Tato podjednotka zvyšuje promotorovou selektivitu, recykluje jádro RNAP a celkově stimuluje syntézu RNA. Podjednotka δ ovlivňuje sporulaci a virulenci některých bakterií. V této práci jsem shromáždila současné poznatky o jednotlivých částech genové exprese, zejména o regulaci iniciace transkripce a o podjednotce δ RNAP.