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...

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Student: Bc. Lucie Polenová Supervisor: MUDr. Pavla Paterová Title of diploma thesis: Detection of methicillin - resistant Staphylococcus aureus (MRSA) in Hospital České Budějovice, a. s. and in University Hospital Hradec Králové The resistance of microbes to antibiotics belongs to worldwide health problems. Infections, which are caused by methicillin-resistant Staphylococcus aureus (MRSA), occur mainly in hospitals. It is because of incorrect and excesive consumption of antibiotics. Insufficient observance of hygienic-epidemiological measures helps to spread resistant strains. Background: The aim of the study was to determine the incidence of MRSA strains in two Czech hospitals - in Hospital České Budějovice, a.s. and in University Hospital Hradec Králové. To compare results with figures in the previous years and sort isolated strains by different characteristics in both hospitals. Methods: Results of the study are based on retrospective data analysis from electronic database. All data from hospitalized patients or outpatients during 1.1.2015 - 31.12.2015 with isolation of MRSA were counted. Processing and cultivation of isolates were made by standard microbiology measures. Strains,...

The latest model of tertiary structure of capsid protein of potato virus X (PVX CP) was used as a template to design new insertion sites suitable for the preparation of PVX-based antigen presentation system. Based on this model, seven insertion sites (A-G) located in putative surface loops were tested. As an antigen inserted into these sites was used 17 amino acids long epitope derived from human papillomavirus type 16 E7 oncoprotein (E7 epitope) fused with either 6xHis tag or StrepII tag in both possible orientations (6xHis-E7 and E7-6xHis, StrepII-E7 and E7-StrepII). Prior to plant expression, modified PVX CPs were expressed in Escherichia coli MC1061. The results showed that only PVX CP carrying StrepII-E7 or E7-StrepII in the insertion site A formed virus particles. The results from transient expression experiments with modified PVX CPs in Nicotiana benthamiana showed that only the insertion site A (located between 24th and 25th amino acid in the PVX CP) could tolerate all tested inserts. Importantly, viral particles were detected only in the presence of StrepII tag and their stability was affected by the insert orientation (StrepII-E7 vs. E7-StrepII) as only the viral particles presenting E7-StrepII could be purified. Besides the preparation of PVX-based antigen presentation system, an...

The main aim of this thesis is to investigate the role of Impl2 gene in the regulation of immune response to bacterial infection in Drosophila melanogaster. The theoretical part provides a conceptual background concerning immunity in drosophila and also it summarizes what is known about the Impl2 gene. In the practical part, the function of Impl2 gene during infection is examined, starting with the location of its production during immune response, the changes of Impl2 gene amount during infection and its essentiality for a proper immune response. Furthermore, it examines the metabolic changes underlying an immune activation in connection to Impl2 gene. Last but not least, it tests how Impl2 gene is connected with the mobilization of energy during Chill Coma Recovery assay.

The paper analyzes the factors affecting the quality and nutritive value of wilted grasssilage. Grass silage sample were collected in the operating conditions for the firstmowing. Three groups were formed after 12 samples. The first group was the controlwithout using preservatives. The sekond group were included in the silage treated withbacterial agents. The third group included the bacterial-enzymetreated silage additives. There were no statistically signifiant differences in lactic acid kontent (P < 0.05) andvalues of thedegree of proteolysis (P < 0.05) between the kontrol group without the useof silage additives and the group, which was used for bacterial and bacterial-enzyme additives.

Total hip and knee arthroplasty surgeries form an integral part of orthopaedic practice. With an increase of the primary arthroplasty surgeries performed, comes also an increase in the number of complications. The most common complications of these otherwise very successful procedures is periprosthetic joint infections which are also one of the most difficult to treat. In the first part of the study, we evaluated the primary bacterial resistance of the 14 most commonly used materials in the construction of joint prostheses. More specifically, we concentrated on how their surface treatment resists colonisation by specific bacterial species (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Enterococcus faecalis and Escherichia coli). The studied materials included metal alloys that are commonly used in the weight bearing parts of implants - CoCrNo, FeNiCr and Ti6A14V - but also polymeric and ceramic materials used in the bearing materials, represented by ultra-high- molecular-weight polyethylene (UHMWPE) and aluminium oxide (Al2O3). Our aim was to assess the relationship between material surface roughness and the sensitivity to colonisation by specific bacterial strains and to evaluate their affinity to different materials. Previous studies have proven that the roughness of...

This master thesis deals with polyhydroxyalkanoates (PHA) and their role in bacterial biofilms. In the theoretical part the polyhydroxyalkanoates, bacterial biofilm and the relationship between them were reviewed. The experimental part focused on differences in PHA production by planktonic and biofilm cells. In order to study selected topic, bacterial strains of Burkholderia cepacia and Burkholderia sacchari were cultivated using a CDC biofilm reactor. The attention was paid to quantity and especially to the form in which PHA occurs in planktonic and biofilm cells. Results of Raman spectroscopy have shown that PHA exists exclusively in native amorphous form in planktonic bacterial cells. On the other hand, in biofilm PHA occurs also in a partially crystalline form. In addition, the resistance of planktonic and biofilm cells against various stress factors and the effect of osmotic stress on PHA production was tested too. According to the results of the experiment, when the bacteria were exposed to different stress factors (high temperature, low temperature, presence of detergent and so forth) biofilm cells showed a higher stress resistance than planktonic cells. Apart from slowing cell growth and reproduction, increased osmotic pressure in the culture medium also caused decrease of PHA production. In addition, planktonic cells responded to external stimuli more sensitively than biofilm ones.

The diploma thesis deals with droplet infections and finds out what knowledge, attitudes and behavior pupils of the lower secondary school have towards them. The theoretical part focuses on educational documents in the Czech Republic, including the occurrence of topics about droplet infections in RVP ZV, didactic game as a teaching method and a detail description of selected viral and bacterial droplet infections. In the practical part, the diploma thesis finds out what knowledge, attitudes and behavior students have about droplet infections. Subsequently, a didactic game is performed and the effectiveness of the didactic game is evaluated by comparing the pre-test and the post-test on the basis of changes in the knowledge, attitudes and declared behavior of the interviewed pupils. The results showed that due to the didactic game there was an improvement in the students knowledge and a desired change of attitudes. The declared behavior of students remained unchanged, as the entry level of students in this area was already at a high level.

Dynamic molecular interactions between the microbiota and the intestinal mucosa play an important role in the establishment and maintenance of mucosal homeostasis. Aberrant host- microbiota interaction could lead to many diseases such as inflammatory bowel disease. The aim of our study was to evaluate the commensal and probiotic bacteria activities and their ability to induce pathological or exert beneficial effects. The most important trigger for immune system development is an exposure to microbial components. Here, we show that there is a time window at about three weeks of age, which enables the artificial colonization of germ free mice by a single oral dose of cecal content. The delayed colonization by either inoculation or co-housing causes permanent changes in immune system reactivity, which may downgrade the results of experiments performed on first generation of colonized animals. In this thesis we report that even non-living commensal bacteria such as Parabacteroides distasonis (mPd) or well known probiotics such as L. casei DN-114 001 (Lc) possess anti-inflammatory effects in experimental model of colitis. The mechanisms that this effect is achieved by the lysate of L. casei DN-114 001 comprise: a) improvement in the gut barrier function, b) correction of the dysbiosis, and c) modulation of the...

5 Abstract Even though prophylactic vaccine against human papillomavirus (HPV) is currently licensed, infections by the virus continue to be the major health problem mainly in developing countries. Considerable effort is being devoted to preparation of therapeutic vaccine and to decrease of the production costs of current vaccine. Viral proteins such as the E7 oncoprotein and the L2 capsid protein from HPV type 16 are promising targets for the development of the experimental anti-HPV vaccine. The aim of our work was optimization of expression of mutagenized E7 oncoprotein (E7ggg) fused to the C-terminus of Tobacco mosaic virus (TMV) coat protein (CP) or Potato virus X (PVX) CP in viral vectors derived from these plant viruses. The impact of linkers connecting CP and E7ggg fusion partners on expression and stability of fusion proteins was examined. The fusion proteins were first expressed in Escherichia coli (E. coli) MC1061 to assess the characteristics of the recombinant protein prior to their transient expression in both non-transgenic or transgenic Nicotiana benthamiana (N. benthamiana). We have obtained the high level expression in E. coli, but most of the expressed proteins based on TMV CP remained in insoluble inclusion bodies. To increase the ratio of soluble protein various molecular...