1 ABSTRACT Kučerová J.: Biological activity of secondary plants metabolites VI. Alkaloids of Vinca minor L. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of pharmaceutical botany and ecology, Hradec Králové 2016, 67 pages. Isolation of alkaloids was done from extract of Vinca minor, family Apocynaceae. Individual substances obtained were examined for their ability to inhibit human acetylcholinesterase and butyrylcholinesterase. The basic extract was after an initial extraction with 95% ethanol pre-purified by extraction with chloroform. This chloroform extract was divided further by column chromatography on a total of 531 fractions. The fractions obtained were, based on the similarities identified by thin layer chromatography, interconnected in a final number of 17 fractions. The mobile phase for column chromatography were mixtures of chloroform and medical gasoline, chloroform and ethanol, and ethanol itself. For further testing were used fractions number 2 and 5. From the fraction 2 was by preparative thin layer chromatography and subsequent crystallization isolated substance number 1. After analysis of this substance by GC/MS was found that it is a (+) - vincaminorein. Further was examined it's biological activity towards cholinesterases, which found a...

Work of groups in the extreme, isolated and confined environment relates to the high risks and demands as well. One of the groups working in that specific conditions are the space crews. Attention has been focused recently on preventing the risk of their failure. For a successful fulfilment of the space missions, it seems that the relations between the crew members are crucial. They manifest in various areas: communication, cooperation, group cohesion, mutual trust, support and others. The theoretical part of this thesis describes selected factors related to interpersonal relationships. Two groups of factors are discussed - factors connected with a diversity of the crew and the and the situational factors. There is a discussion of the methods used for measuring and assessing the interpersonal relations, their advantages and limitations. As a follow-up to this part, socio-diagnostic method called "sociomapping" is described in detail because of its use in the empirical part of the thesis. The goal of the empirical part is to analyse two areas: the development of the group dynamics during a mission and the connection between the gender-mixed crew and the tendency to creating the subgroups. Previous research show ambiguity in the research findings. For this purpose, data from 17-day simulation of the...

Šípková, P.: Biological activity of secondary plants metabolites X. Alkaloids of Vinca minor L. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové, 2019, 66 s. Key words: Apocynaceae, Vinca minor, alkaloids, isolation, biological activity, screening Alzheimer's disease is a progressive neurodegenerative disease. The number of affected patients is constantly increasing. This disease cannot be treated casually, therefore discovering and testing new substances that could potentially be used in a treatment is very important. The Vinca minor L. fraction after column chromatography was separated by flash chromatography. Isolation of the individual alkaloids was performed by preparative TLC. Based on NMR and MS analyses and comparison with literature, alkaloids were identified as vincarubine and (-)-vinoxine. Modified Ellman's method was used to test cholinesterase inhibitory activity of isolated alkaloids. Acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) are enzymes, that play a very important role in the pathofysiology of Alzheimer's diasease. (-)-Vinoxine showed relatively high activity against BuChE (IC50 = 24,61 ± 1,71 µM), inhibitory activity against AChE was insignificant (IC50 > 1000µM). Vincarubin did not...

The theoretical part of the thesis describes the use, working process and application of natural materials in family house construction, as well as their impact on the health aspects of building users. The thesis defines the environmental footprint of each particular material. The practical part of the thesis is primary focused on the building design which combines different structural systems from solid wood. The visual concept of the construction in particular was designed with the emphasis and respect to the surrounding countryside which belongs to Protected Landscape Area and is liable to special requirements. Requirements for permanent residence under the applicable legislation in the Czech Republic were respected with a maximum effort to use natural materials in the construction itself.

The hereditary cataract is one of the most common eye disease in dogs. The expansion of this disease in the Staffordshire bullterrier breed has been so massive that in the Czech Republic was introduced the rule of mandatory testing of at least one of a breeding pair. This is a degenerative disease of the lens causing total blindness of the affected animal within three years. Since some time ago there are no more dogs affected by the disease in the Czech Republic, there are however still hidden carriers which need to be discovered to the complete extinction of the disease in the genome. The goal of this study was to test simple ways of collecting biological samples, try them in practice and to verify whether they are suitable for the DNA isolation and also to test an alternative method of molecular detection of this disease. In total there have been 23 buccal swabs collected from male and female Staffordshire Bullterrier examples. The detection of the hidden carriers of the hereditary cataract was carried out by PCR analysis with specific primers. The obtained amplicons were detected by both gel and chip electrophoresis and by using fragment analysis. This detection of the carriers was based on the presence of two amplicons (heterozygotes). I came to conclusion that to detect hidden carriers it is neccessary to use the fragment analysis because of the difference of only one base in the reference section of DNA. Neither gel nor chip electrophoresis does provide sufficiently high resolution and it is not possible to detect two fragments that differ only by one bp. As the most appropriate sampling method I have chosen the buccal smear by cytological brush followed by isolating the DNA by Chelex with purification of the sample subsequently.

Marečková, L .: Biological activity of secondary plant metabolites VII. Alkaloids of Vinca minor L. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Kralove, Hradec Králové, Department of pharmaceutical botany and ecology, Hradec Králové 2018 There was prepared an extract from aerial parts of the plant Vinca minor L. This prepared extract was separated by the column chromatography into individual fractions. One chosen fraction was separated by TLC on silicagel. Two substances were isolated and they we identified by GC/MS and NMR as (-)-minovin and (+)-minovincin. These two substances were tested for their inhibition activity against human acetylcholinesterase (hemolysate of human erythrocytes) and butyrylcholinesterase (human plasma). Their inhibition activity was compared to standards which were Galanthamin, Huperzin A and Eserin. (+)-Minovincin did not show significant inhibitory activity against either enzyme (IC50 = AchE > 1000 µM, BuChE 655,05 ± 35,37 µM) and it is not suitable for further investigation. (-)-Minovin showed no significant activity against AChE (IC50 = 234,27 ± 63,40 µM), activity against BuChE (IC50 = 26,32 ± 2,52 µM) was higher than inhibitory akctivity of Galanthamin, but still relatively low for further investigation. The aim of this work was mainly the...

This thesis primarily focuses on one segment of freshwater Betaproteobacteria, the Limnohabitans genus (including the RBT lineage). As opposed to other recent research directions, the major aim was to recover the members of the previously uncultured RBT lineage through their isolation from various freshwater habitats. However, the results presented in this thesis have also ambitions to go far beyond the taxonomic descriptions only; the dissertation intends to contribute significantly to unveiling of important ecophysiological characteristics of the studied lineage in a set of both laboratory and field research. Therefore, understanding of growth characteristics, mortality, diversity and life strategies of aquatic microbes is of highest importance regarding profound human impact on water quality and increasing need of drinking water supplies.

The topic of this publication is a comparison of insulated sandwich panels and materials of organic origin - wood based materials. There were compared insulative properties, lifetime and finacial costs of these materials.

Extracellular vesicles (exosomes) are the subject of current nephrology proteomics research as they are considered as a promising source of potential biomarkers of kidney disease. This work is focused on discovery of the most appropriate procedure for the urinary exosomes isolation. We have compared already described methods, based on different physicochemical principles of isolation: hydrostatic filtration dialysis (HFD), differential ultracentrifugation, ultrafiltration through a 100 kDa filter, or sample precipitation with Total Exosome Isolation (from urine) kit. Characterization of individual isolated exosomal fractions was performed using SDS-PAGE method (presence of contaminating proteins), western blot analysis (detection of exosomal markers TSG101, alix), nanoparticle tracking analysis (NTA, vesicle size and concentration) or transmission electron microscopy (TEM, vesicles morphology). Due to the presence of contaminating proteins in urine samples, which could distort the results of subsequent proteomic assays, the conditions for the cleavage of undesirable proteins by proteinase K prior to their own isolation were optimized. It has been found that the best yield and purity of the isolated exosomal fractions were provided by a process combining HFD with differential ultracentrifugation...

Hodková, Gabriela - Metabolic Effects Of Hyperthermic Isolated Limb Perfusion (HILP) in Malignant Melanoma Patients First Faculty of Medicine, Charles University in Prague, Praha 2, Kateřinská 32 Head of the work: Doc. MUDr. Michal Semrád, CSc. Supervisor - consultant: MUDr. Miroslav Špaček, Ph. D. The aim of the study is to assess the metabolic consequences of mechanical isolation and hyperthermic cytostatic perfusion in a limb affected by malignant process. The theoretical part refers to a topic of malignant melanoma, its clinical evaluation and treatment. Methods based on conservative and surgical treatment are described. The isolated hyperthermic cytostatic limb perfusion is a consecutive local treatment indicated in cases of recurrent malignant lesions following surgical resection, when next surgery is impossible. In the practical part, the laboratory samples and clinical data were recorded in patients who had undergone hyperthermic cytostatic limb perfusion in the 2nd Surgical Department of The General Teaching Hospital and First Faculty of Medicine, Charles University Prague. The affected limb was flushed with a warm oxygenated blood containing cytostatic drugs using an extracorporeal circuit apparatus. Selected arterial blood gas, metabolic and hematologic parameters were studied intra and...