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Charles University Faculty of Pharmacy in Hradec Králové Department of pharmaceutical botany and ecology Candidate: Bc. Andrea Gunčagová Supervisor: prof. RNDr. Luděk Jahodář, CSc. Title of diploma thesis: The analysis of Psilocybe constituents II. The aim of this thesis was to analyse Psilocybe constituents, to plot the places of occurence of chosen Psilocybe brands and also to determinate their water content. The thesis deals with 22 samples that originate in 10 habitats in Slovakia and Moravian- Silesian region. Accuracy of brand specification was supervised by a member of mycological society. Psilocybe serbica var. bohemica was identified in 2, whereas Psilocybe semilanceata in 8 habitats. The average content of H20 in samples was 87,07 % ± 10,91 %. All of the samples were analysed by LC-MS/MS (LIT) method. The qualitative analysis of determinated substances showed the presence of psilocine (PSC) and psilocybine (PSB) in all samples. The results of quantitative analysis (expressed in percentage of dry matter) are similar in particular habitats, but there is a significant difference between two specific brands of Psilocybe mushrooms. The content of determinated alkaloids in P. bohemica was: PSC 0,001-0,011 % a PSB 0,01-0,07%. P. semilanceata contained 0,0005 - 0,011 % PSC a 0,074 - 0,763% PSB....

The work is devoted to the influence of dust particles on the operation of rotating machines with sliding contact. In the work are described and characterized dust particles from environment of selected machines. Their influence on sliding contact and influence on the insulation system of these machines is investigated.

The aim of work was an optimization of separation deoxyribonucleic acid (DNA) with the use of nucleic acid reversible adsorption to the surface of magnetic particles coated by functional groups. Six carriers were verificated for DNA isolation: P (HEMA-co-GMA) ox, F-kol B 30 ox, F-kol 77 ox, F-kol B100 ox, F-kol 135 ox, coated with carboxyl groups and Perovskit 439 (coated by silicone). Bacterial DNA was isolated by phenol extraction procedure, first. DNA was reversibly bond to magnetis carrier in the presence of high concentration of NaCl ( 5 M) and poly (ethylene glycol) (PEG 6000). The final PEG and NaCl concentrations of 16.0 % (w/v) and 2.0 M, respectively, were used.DNA was eluted into TE buffer. The quality of extracted DNA was checked by PCR amplification. It was found out that although different quantities of DNA were isolated, the quality of isolated DNA was always compatible with PCR. Nanoparticles Perovskit 439 had the best separative characteristics in comparison to the other magnetic carriers because highest amounts of DNA was isolated. However, next optimisation of DNA separation procedure is required for the use of studied microspheres in real samples.

The aim of the proposed diploma thesis is to optimize the procedure of collagen isolation from animal tissues and methods for molecular characterization of isolates. In experimental part, isolations of collagens from animal tissues (chicken, hen, carp, and pork skin) were performed. Total protein concentration was determined by Biuret method and by TNBSA method. Both methods were used also in order to study physiological and thermal stability of the isolates under model conditions. Isolated collagens were characterized at molecular level using PAGE–SDS and microfluidic electrophoresis. Further, amino acids composition and microelement content were analyzed. Finally, the stability of isolated collagens in several types of simulated physiological and pathological conditions was tested too. During isolation relatively sufficient collagen yields were obtained (about 15 % of purified collagen per original mass of biological material). In all analyzed tissues Type I collagen was found. Thermal stability of individual samples differed according to biological source type and tissue age. Viscosimetry measurements confirmed higher stability in collagen samples of older animals. Collagenase exhibited the lowest degradation effect to bovine collagen, while selected mixed microbial hydrolases differed according to enzyme preparative type. Incubation of collagen isolates with selected human pathogens confirmed higher resistance of bovine collagen to biofilm formation when compared with the chicken one.

In the food industry are requested speedy and accurate methods for identification of bacteria in microbiological testing of products. Molecular diagnostic methods are based on isolation of DNA from bacterial cells which is amplified in polymerase chain reaction (PCR). The result is fragment DNA about specific size, characteristic for genus or species of bacteria. The aim of the work was isolation of PCR-ready DNA. DNA has been isolated from 8 strains of genus Clostridium. Procedure of cell lysis was optimized in order to find the optimal concentration of EDTA and proteinase K in lysing buffer. DNA was isolated by phenol extraction and using magnetic microspheres. Concentrations 10 mM of EDTA and 10 l of proteinase K (100 g/ml) were the best for cell lysis for isolation of DNA by phenol extraction. Concentrations 10 mM of EDTA and 15 l of proteinase K (100 g/ml) were the best for cell lysis for isolation of DNA by magnetic microspheres. Isolated DNA was checked by gel electrophoresis, quantificated by spectrophotometry and tested in PCR. Individuals species were distinguished in denaturing gradient gel electrophoresis (DGGE).

The diploma thesis is consisting of three parts. The first part is a theoretical overview. It discusses the basic types of refrigeration systems and addresses used by air conditioners. The theoretical part contains describes the basic terms of the proposed refrigeration system, air filtration technology. In the second part of the project design air conditioning systems for offices three-story office building. Following heat balance the budget, the air conditioning system. Numerical solution is completed drawings and bill of material. The third part is experimental. Objective measurement is to determine the heat loss of the pipe without insulation, with normal insulation, new insulation and determine the applicability of a new type of insulation.

Autophagy is a nonspecific catabolic mechanism, important for maintaining the cellular homeostasis. Stress condition can increase the activity and support the cell survival. It is well known that normal and pathological aging are associated with a reduced autophagic potential and that experimental induction of autophagy leads to higher longevity of certain model organisms. This decreasing may be facilitated by other factors, such as chronic stress. Higher age and chronic stress are two factors contributing to the development of neurodegenerative diseases, in this context the mechanism of autophagy is now actively studied. We wanted to know if and how the combination of these two factors may influence the activity of autophagy. In our study, we investigated the role of aging and chronic stress, induced by social isolation in Wistar rats at the age of 6 (N= 16) and 12 (N= 16) months. The animals in experimental groups were separated in the individual cages for 4 weeks. Control groups animals were housed in the pair. We measured the markers of autophagy expression (Beclin-1, LC3-II, mTOR) in the hippocampus and striatum of socially isolated and control groups animals. We measured the expression of NMDA receptor subunits (NR1, NR2A, NR2B) in the hippocampus of the same animals. Our results showed that older...

This thesis deals with the design of the superstructure of the family house in the construction system of a wooden building. The main objective is to assess the original state of the house. According to this assessment the superstructure design with regard to applicable regulations and standards should be done. The static assessment of the truss roof and the assessment of two variants of the perimeter shell in terms of thermal engineering should be also considered. The superstructure design was done according to valid legislation and standards in AutoCAD. The static assessment of the roof was solved in the Pamite program by Mitek. The thermal engineering assessment was carried out in PHPP. The result of this work is the project documentation of the construction with a static assessment of the truss and the energy performance of the building.

Cytochromes P450 form a large family of hemoproteins. Some of them are responsible for the metabolism of endogenous substrates, but their major role is in detoxification of exogenous substrates (xenobiotics), some of them are activated to reactive species forming covalent adducts with DNA and increasing intracellular oxidative stress. Cytochrome P450 are considered by very promiscuous in terms of their substrate specificity, thus one enzyme can typically oxidize many substrates. Cytochrome P450 1A1 prefers a planar aromatic compounds (e.g. polycyclic aromatic hydrocarbons, azo dyes, etc.). Cytochrome P450 1A2 elicits similar substrate specificity, but prefers slightly basic aromatic derivatives, for example caffeine. This work focuses on (i) the preparation of expression vectors containing genes encoding human cytochromes P450 1A1 and 1A2, (ii) their consequent expression in heterologous system followed by (iii) isolation of corresponding proteins. The genes coding both proteins were modified and transferred from older vectors to the more efficient to expression plasmids pET-22b. However, the new constructs did not produce stable native proteins. The modified genes were therefore transferred to the original expression plasmids pCW. The problem with the incorporation of native human form of...