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Molecular Pathology of Acute Intermittent Porphyria
Hrdinka, Matouš
Molecular Pathology of Acute Intermittent Porphyria Author: Matouš Hrdinka Acute intermittent porphyria (AIP) is autosomal dominant disorder caused by the partial deficiency of porphobilinogen deaminase (PBGD), the third enzyme in the heme biosynthetic pathway. AIP is manifested by life-threatening acute neurological attacks that can be provoked by various factors such as drugs or alcohol. Objective of this study was to identify and characterize the molecular lesions in Czech and Slovak AIP patients. To identify disease-causing mutations screening was performed by PCR, denaturing gradient gel electrophoresis (DGGE), automated DNA sequencing and restriction fragment lenght polymorphism (RFLP) assays. Total of 35 individuals from 8 families were analyzed to detect asymptomatic carriers. Seven mutations were identified, including 3 novel mutations. Of particular interest, one patient had two mutations R173Q and Q204K, both located in exon 10 on the same allele. To further characterize these mutations, human PBGD was cloned into the pGEX-4T-1 vector and mutations were generated by site-directed mutagenesis. The wild-type and mutated enzyme species were expressed in E. coli as GST fusions and purified by affinity chromatography. Conditions of the PBGD enzyme assay were determined and specific...
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The molecular mechanism of CSL protein participation in oxidative stress response in Schizosaccharomyces pombe
Daněk, Petr ; Převorovský, Martin (advisor) ; Sipiczki, Matthias (referee)
Redox homeostasis maintenance is important for proper organism and cell function, for while relatively low amount of reactive oxygen (and nitrogen) species contributes to the fine tuning of signal transduction, excessive concentration of ROS (oxidative stress) has demonstrably harmful effects and is tightly connected to many pathological states. Cells therefore evolved broad palette of antioxidant mechanisms that express striking level of conservation among different species. Large, intricate stress response signaling networks have been already described; nonetheless, novel molecules employed in stress-related signaling are still being discovered. Several studies recently suggested transcription factors CSL, proteins essential for regulation of metazoan development as effectors of Notch signaling, are also involved in response to oxidative stress. The fission yeast Schizosaccharomyces pombe, well established model of response to various stresses, comprises two paralogs of CSL proteins - Cbf11 and Cbf12. We have found cells depleted of cbf11 are highly resistant to hydrogen peroxide. This resistance appears to be caused by upregulation of important stress responsive genes including ctt1, gst2, pyp2, and atf1. Cbf11 is therefore negative regulator of these genes, which suppresses their expression...
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Evaluation of system of fenced and unfenced plots in the Horovice Division, Military Forests and Farms, state enterprise
KOHOUTOVÁ, Monika
The aim of the bachelor thesis Evaluation of system of fenced and control plots in the Horovice Division, Military Forests and Farms, state enterprise, was to introduce the complex issue of game damage in forests and the methods of assessing it, and to conduct a specific analysis of the functionality of fenced and control plots. This was done in the area of Horovice Division and its five administrations, all located in Brdy forests, Central Bohemia. Focusing on species representation, abundance of regeneration trees and browsing of terminal shoot, the analysis indicated some problems with the system of fenced and control plots in the study area, later also verified in the field. The major flaw was inadequate location and technical state of the plots, which limited their functionality and hence also usability of data for interpretation on game damage in the area.
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The influence of oxidative stress on mammalian sperm quality and fertilization ability.
Dudková, Barbora ; Komrsková, Kateřina (advisor) ; Svoboda, Petr (referee)
This bachelor's thesis deals with the oxidative stress and its influence on structures and physiologic functions of mammalian sperm. Oxidative stress is a consequence of imbalance between pro-oxidants and anti-oxidants in the body. It is associated with impaired quality of sperm and it is considered to be one of the major reasons of male infertility, including humans. This work is presented as a research review targeting how and where reactive oxygen species are generated in the male reproductive tract. It summarizes specific structures and physiologic functions that are influenced by reactive oxygen species either positively or negatively and it also covers a protective role of antioxidants in seminal fluid.
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Přirozená obnova listnatého lesa na exponovaných stanovištích VLS Lipník nad Bečvou
Pálka, Ondřej
The bachelor thesis deals with natural regeneration of deciduous forest at exposed sites in Czech military forests (division Lipník nad Bečvou), forest district Velký Újezd. The main goals of this study were to evaluate success of different-aged natural regeneration stands at different exposures (north, south, east, and west). In case of all analysed naturally regenerated stands, the Fagus sylvatica species is dominated, the Fraxinus excelsior, Tilia cordata, Acer pseudoplatanus, Acer platanoides are admixed species. Under parent stands, the highest natural regeneration density was detected at north exposure sites (222,500 piece/ha with the highest proportion of individuals in the height class of 1-20 cm). In two years naturally regenerated growths (in the open sites), the highest density of regenerated young individuals was detected at west exposure sites (115,000 piece/ha), with the highest proportion of regenerated individuals in 2 -- 3 meter height class. The characteristics of all investigated stands of natural regeneration fulfil the criteria for reforestation according to norm no. 139/2004 Sb. § 2. The results including analysis of different forest regeneration ways confirm that regeneration by border felling (width of 20 m), oriented along the slope (perpendicular to the contour) with regeneration period of 40 years is suitable silvicultural way for successful natural regeneration.
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The role of oxygen radicals in the early phase of exposure to hypoxia in the development of hypoxic pulmonary hypertension
Lachmanová, Věra ; Herget, Jan (advisor) ; Geršl, Vladimír (referee) ; Neckář, Jan (referee)
A pulmonary vascular bed is low-pressure system at adult subjects. Pulmonary vessels react to hypoxia by two different processes. These are hypoxic pulmonary vasoconstriction (HPV) and hypoxic pulmonary hypertension (HPH). They differ in mechanism of origin, but there seems to be important role of reactive oxygen species and nitric oxide. It was assumed in the past that HPV is isolated reaction of small pulmonary arteries to acute hypoxia and HPH to chronic hypoxia. Recently we believe that HPH is developed on the basis HPV (Crossno, Garat et al. 2007) and remodelation of peripheral pulmonary vessels (Reid 1986). Our main task was to learn, whether antioxidants given in the early phase of exposure to hypoxia influence pulmonary hypertension more than its late administration, in the period of already developed damage of pulmonary vessels. We have used N-acetyl-L-cysteine (NAC) as an antioxidant substance. We measured changes in resistance of pulmonary vascular bed, changes of reactivity of pulmonary vessels in dependence on concentration of oxygen in the inhalated air. Measurements were performed on the model of rat isolated perfused lungs. In addition we have observed influence of the early and late treatment of NAC on the pulmonary artery pressure at rats kept in hypoxic conditions. Our results show that...
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Rostliny tradiční čínské medicíny
Barcalová, Šárka
The bachelor's work concerns with flowers used by traditional Chinese medicine, which can be also grown in Czech republic. There are botanically described 25 species of plants. For all species, except Adenophora tetraphylla and Rhododendron molle, were proved antioxidant effects of drug. Availability of the plants in the form of seeds and seedlings and their location in botanical gardens of Czech republic is also added. Hereafter are mentioned 28 species of flowers, that belong to traditional Chinese medicine that can be found in Czech republic. A list of species, that belong to traditional Chinese medicine and are in experimental garden ZF MENDELU and a list of other flower species, which could be located here in the future, are also included.
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Horticulture production in conditions of Sub-Sahara Africa
Milanda, Johannes
This paper was written to provide the literature review on the topic of horticultural production in Namibia or in neighboring countries, with a focus on vegetable species. To suggest a prospective representative vegetable species from the study area, which could have production potential in the Czech Republic. Four chosen cultivars (Metro, Nakare, Bira and Shidimba) from the Cowpea (Vigna uinguiculata L Walp) species were experimentally tested in conditions of the Czech Republic. They were grown naturally extensively with no fertilizers, herbicides or pesticides used. They're morphological parameters were then recorded and laboratory analyzed using ANOVA and three valuables [flavonoids, total antioxidants capacity (TAC) and polyphenols] were determined using different methods with the spectrophotometry.
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Accumulation and biotransformation of nitramines in plant tissue
Šídlová, Petra ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Explosives are toxic, recalcitrant to degradation and contaminate large areas of land and ground water. Remediation of these xenobiotics is difficult and an enormous logistical task. Phytoremediation is a technique that offers an environment friendly, low-cost alternative to current remediation techniques. However this approach is hindered by the low metabolic abilities of plants towards these xenobiotic compounds and the phytotoxicity of these compounds. The fate of the explosives RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) and HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine) on exposure to plant was investigated in hydroponic system of sunflower (Heliantus annuus L.) and hemp (Cannabis sativa L.) approximately for more than 10 days. Exposure levels were: 10mg/l and 50mg/l RDX, respectively HMX for sunflower; 25mg/l and 50mg/l RDX , 5mg/l and 50mg/l HMX for hemp. Their ecotoxicites were characterized by the test based on the inhibition of elongation of primary roots 3days-old mustard and hemp seedlings. Extracts of plants were analyzed by HPLC (liquid chromatography). These study showed that RDX and HMX was taken up by both plant species. The accumulation of these nitramines was observed more in the roots than in the aerial parts of the plants, except for RDX in hemp, where the...
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Influence of genotype on rutin content in plants from genus \kur{(Fagopyrum)}
KRÁLOVÁ, Martina
The bachelor work is focused on evaluation of the influence of buckwheat species and variety on rutin containt in particular plant parts, on the total rutin yield per hectare production and on comparison varietal differences between both species, common buckwheat \kur{(Fagopyrum)}esculentum)} Moench) and tartary buckwheat \kur{(Fagopyrum tataricum)}. Rutin is a flavonol (quercetin-3-O- rutinosid) which is also called P vitamin. As well as it is known as the permeability factor or factor allowing permeability of the blood vessel. Rutin stimulates the C vitamin effect. Rutin has as well a high influence on hypertension, hypercholesterolemia and other cardiovascular diseases due to antioxidant properties. Buckwheat is the most important source of rutin. It is annual plant which belongs to Polygonaceae family and has a beneficial effect on human body. The common grown type is common buckwheat \kur{(Fagopyrum)}esculentum)} Moench) and less used type is buckwheat tartary \kur{(Fagopyrum tataricum)}. Tartary buckwheat \kur{(Fagopyrum tataricum)} contains several times higher amount of rutin than common buckwheat \kur{(Fagopyrum)}esculentum)} Moench). The highest amount of rutin is presented in the leaves and blossoms and the lowest one in seeds. Tartary buckwheat \kur{(Fagopyrum tataricum)} has much higher amount of rutin in seeds than common buckwheat \kur{(Fagopyrum)}esculentum)} Moench). The varietal differences in rutin content are considerable.
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