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Soil microflora of spruce forests of the Bohemian Forest
Nováková, Alena ; Lukešová, Alena
Species composition and quantity of microalgae and micromycetes inhabiting soils of original spruce forest were studied in 3 localities of the Bohemian Forest (Trojmezná, Smrčina, Boubín) in years 2000-2003. Both intact plots, and those ones with selective tree cutting were compared in Trojmezná and Smrčina. In total, 46 species of algae and 72 taxa of micromycetes, represented by common soil species, were found in studied soils. Green algae (mainly Chlorophyceae and Charophyceae) prevailed both qualitatively and quantitatively among soil algae which was connected with low pH. All isolated micromycetes belonged to saprotrophic fungi. Total numbers of algal species ranged between 20-33 and total algal abundance between 48,1-4210,0 cells per g of dry soil in studied plots. Higher algal species richness was recorded in plots with selective tree cutting then in intact plots. Oposite trend of average total algal abundance was observed, despite big variations in particular sampling occasions. Numbers of taxa of saprophytic mycromycetes ranged between 32-40, lenght of mycelium between 6,1-84,5 m per g of dry soil, and numbers of CFU between 57,5-2413,4. No differences in species composition and quantity of saprotrophic micromycetes between intact plots and plots with tree cutting were found.
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Metabolic Effects Of Hyperthermic Isolated Limb Perfusion (HILP) in Malignant Melanoma Patients
Hodková, Gabriela ; Semrád, Michal (advisor) ; Štádler, Petr (referee)
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...
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Differentiation of semiochemicals among lineages of the bed bug (Cimex lectularius)
Pilařová, Kateřina ; Balvín, Ondřej (advisor) ; Kateřina, Kateřina (referee)
Lineages of the common bedbug Cimex lectularius associated with bats and humans separated from each other tens or hundreds of thousands years ago. Currently, they are genetically isolated and some degree of postcopulatory reproduction barrier has been shown. This thesis aims to test the possible mechanisms of precopulatory isolation. Possible differences of semiochemicals between the host lineages were examined by experiments in arenas (four shelters, two unconditioned, one conditioned by scent of specific lineage and one by scent of non-specific lineage, put together in one arena, to which one bedbug male was introduced every time). For the experiment, six breeds (three from bat lineage and three from human lineage) which originated from different parts of Czech Republic were used.
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Diversity of insect communities of non-indigenous forests of black pine (Pinus nigra) in the western part of Central Bohemia
Tůmová, Klára ; Kadlec, Tomáš (advisor) ; Jakubíková, Lada (referee)
Due to the rapid intensification of agriculture, there were a bigger and more often habitat loss and their fragmentation. These led to the decreasing of the size of suitable habitats and their isolation. The isolation of landscape fragments is a basic geographical characteristic, which plays an important role in the ecological invasions. The invasions generally cause major economic damages and they affect the loss of biological diversity in the stands.
In this work, we focused on the effects of black pine (Pinus nigra). This wood plant has been occurred in the Czech Republic since the Tertiary period and disappeared by the subsequent disintegration of continents. The aim of this study is to determine the influence of black pine (Pinus nigra) on the invertebrates compared with the native stands of scots pine (Pinus sylvetris).
The experiment was conducted at 35 plots in Central Bohemia (18 plots of Scots pine, 17 plots of black pine). The invertebrates were captured by light traps. The total abundance and biomass of invertebrates were determined from the captured samples. From the processed data was not proven negative impact on non-native black pine to overall abundance and biomass of invertebrates. This fact is most likely due to the phylogenetic relationship of both pine species.
There were not found any differences in abundance and biomass of most herbivorous groups of invertebrates. However, in the black pine stands was reported a higher incidence of carnivorous species of invertebrates.
The stands of the black pine have no negative impact on number of invertebrates in the Czech Republic, actually there was demonstrated even higher affinity of carnivorous species of invertebrates to the black pine trees.
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Specifická izolace microRNA pomocí magnetizovatelných mikročástic
Vlahová, Veronika
MicroRNAs are small non-coding RNA molecules with length of about 22 nt. These molecules participate on regulation of gene expression at the post-transcriptional level. They represent the largest group of regulators in the cell and therefore are also involved in all key processes such as proliferation, differentiation or apoptosis. Moreover, they participate in tumor transformation. These small molecules have a great potential to be diagnostic markers or assist in the treatment and prevention of diseases. This research was focused on the development of isolation method using magnetic particles with subsequent electrochemical detection of microRNA. Optimization steps were performed and then the entire method was successfully applied to real samples of HEK293 cells expressing increased levels of miR-124. The developed method proved to be sufficiently specific and applicable to the analysis of microRNA.
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DNA Extraction of Sweet Cherry (Prunus avium L.)
Hřebcová, Kateřina ; Sedlák, Petr (advisor) ; Korecký, Jiří (referee)
Isolation of high quality DNA in satisfactory yield and purity is a fundamental and essential step for all molecular-biological studies and analyses. The process of its extraction can be complicated by many of materials like are polyphenols, polysaccharides, proteins and other metabolites that can be co-isolated with nucleic acids and can act as inhibitors of PCR and cause deterioration of samples for further analyses.
In this thesis, mostly used methods of plant DNA isolation were mapped, and, in experimental part, results, regarded to the yield and purity, of selected plant DNA isolation methods were compared. DNA was obtained from various tissues of Prunus avium L. species, namely from fresh leaves, buds and from frozen embryos of several varieties. Comparison of the two commercial isolation kits (DNeasy Plant Mini Kit by Qiagen and GeneEluteTM Plant Genomic DNA Miniprep Kit, Sigma Aldrich) was the original intention. The first of the kits was replaced by simple and quick DEP-25 DNA Extraction Kit, Top-Bio and the experiment was extended with CTAB DNA isolation protocol, both with and without application of RNase into the protocol.
The results obtained proved quite significant differences between the methods used, both in yield and purity. The original assumption, supported by several studies, that commercial kits not always gain relevant results, regarded to ability to provide pure DNA, was not accurately proven, the assumption that the CTAB protocol can gain satisfactory results according to the DNA yield and purity was proved only with some tissues. The results of the spectrophotometry were supported with polymerase chain reaction (PCR) analyses conducted with the isolated DNA samples and after statistical evaluation were discussed.
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Characterization of bifidobacteria and lactobacilli isolated from intestinal tract of wild pigs
Šimurka, Julius ; Rada, Vojtěch (advisor) ; Švejstil, Roman (referee)
The microbiota of the gastrointestinal tract is considered as "acuqrated organ" located in a host organism. There are many performs of specific functions. The microbiota composition changes during the life of the individual, depending on his environment and nutrition. The process of identification and characterization of the bacteria from the intestinal microbiota is an important prerequisite for understanding the function of this "acuqrated organ".
This work deals with the anatomy and physiology of the digestive tract (DT) of pigs and evolutionary and eating differences of wild and domesticated pig, for understanding the natural environment of bacteria from DT, which in turn affects the bacterial diversity of the intestinal microbiota. Also mentioned is the process of domestication, which influenced the present form of domestic pig. The work includes the characteristics of lactobacilli and bifidobacteria, which contains the descriptions, taxonomy, metabolism and habitats. The work also shows the insulating media used in practice for the isolation of bifidobacteria and lactobacilli from DT of pigs and methods of their identification and related methods of characterization.
The available literature confirms the differences in the composition of microbiota DT between wild and domesticated pig, that correlate with other studies which study microbiota diversity of other domesticated and wild animals. The diversity studies of humans microbiota, also confirms the broad spectrum of bacterial species in primitive tribes of humans compared with urbanized man.
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Mitochondrial subpopulations in rat myocardium - effect of chronic hypoxia
Kovalčíková, Jana ; Nováková, Olga (advisor) ; Žurmanová, Jitka (referee)
Adaptation to chronic hypoxia induces endogenous cardioprotection and increases the heart resistance to ischemia/reperfusion injury. The heart mitochondria, which produce reactive oxygen species (ROS) in addition to ATP, play an important role in these processes. During ischemia/reperfusion, ROS are produced in excessive amounts and damage the cells. However, in lower concentrations, ROS are involved in the signalling pathway of cardioprotection induced by adaptation to chronic hypoxia. In the heart, two mitochondrial subpopulations have been observed, subsarcolemmal mitochondria (SSM) and intermyofibrillar mitochondria (IMFM), which differ in cell localization as well as in morphological and biochemical properties. The aim of this work was to introduce the method of SSM and IMFM isolation in our laboratory and to analyse their antioxidative capacity after adaptation to chronic hypoxia. Adult male Wistar rats were kept either under normoxic conditions or exposed to intermittent high-altitude hypoxia (IHA; 7000 m, 5 days a week/8 hours a day, totally 25 exposures). Mitochondrial subpopulations were isolated from heart left ventricle and their functionality was verified by measuring oxygen consumption and enzyme activities. The IMFM had higher oxygen consumption in comparison with SSM and activities...
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Degradation of complex carbohydrates by bifidobacteria
Makovská, Marie ; Bunešová, Věra (advisor) ; Švejstil, Roman (referee)
In my bachelor thesis I discuss different strains of bifidobacteria and their ability to utilize different complex carbohydrates as an energy source. For this bachelor thesis were used 112 strains of Bifidobacterium genus which came from different ecological niches. Official collection type and untyped strains were used, along with wild strains, some of which have been isolated by workers and students of the Faculty of Microbiology, Nutrition and Dietetics at the Czech University of Life Sciences. Pure cultures were inoculated into media enriched with different substrates (3 types of starch and Nutriose - resistant dextrin), while tested substrates were always sole carbon sources. Media contained bromcresol red as a pH indicator. A positive sample of the culture, which was able to utilize the substrate, exerted a color change, because of the formation of metabolites (especially acetic acid and lactic acid), which was resulted in lowering the pH.
Further, cultures were tested for ability to degrade starch with so-called plate method. Petri plates contained an agar enriched with starch and tested Bifidobacterium strains were inoculated in preformed holes. After an anaerobic cultivation, coloration by Congo red agent and rinse by NaCl, bright zones around the samples, which prove ability to degrade starch by the bifidobacteria culture, were evaluated.
By the evaluation of the tests has been found that degradation of starch is more frequent for animal strains (which are more species-diverse). Positively respond 59,7 % of total 61 animal strains (20 different species of total 35). Strains isolated from human sources have less capacity of starch degradation. There 26 % of total 42 strains were able to degrade starch. However these positive strains include 8 different species of total 14 tested. From 6 dietary sources of strains just 2 strains (2 different species of 4 tested) were able to degrade starch. One of them was isolated from probiotic product (drops) and the second one was isolated from airag (Mongolian traditional fermented drink, which is made from the milk of mares). All three strains, which were isolated from sewage, can degrade starch (each of them was different species).
The results suggest that some substrate preference of Bifidobacterium strains depends of their habitat.
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