Tellurium and bismuth are non-biogenous elements, which can be accumulated in human body. The aim of this work is focused on the determination of these elements by the electrochemical hydride generation with non-membrane electrolytic cell with quartz tube - atomic absorption spectrometry. The first step of the work was to optimize parameters for the continuous setup measurements and to obtain its figures of merit. After this, the arrangement with most suitable conditions for determination of these elements was converted to the flow injection setup (FIA) and all experimental parameters were again optimized and its figures of merit were obtained. The conditions of hydride generation were studied for platinum, lead and silver cathodes with hydrochloric and sulphuric acids and their sodium salts as electrolytes. The optimizations of the FIA setup were performed on platinum cathode in hydrochloric acid for bismuth and on lead cathode in sulphuric acid for tellurium. The limit of detection achieved for the generation of tellurium hydride was 1.1 ppm with a linear range up to 20 ppm; for the bismuth hydride the limit of detection was 9.5 ppm and limit of linearity 50 ppm. Keywords Bismuth, tellurium, electrochemical hydride generation, atomic absorption spectroscopy, flow injection analysis

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Complex Campanula rotundifolia agg. is rich in endemic species, especially in the high- elevated mountain ranges. Extreme climatic conditions in the subalpine habitats can lead to a convergence in plant's morphology between plants isolated in different mountain ranges. Because of the lack of morphological differentiation, it is very difficult to reconstruct the evolutionary history of bellflowers C. scheuchzeri from the Alps, C. bohemica from the Krkonoše Mountains and C. tatrae from the Western Carpathians. A similarity in morphology and the same ploidy level can suggest vicariance from a large area of an ancestor species. However, regarding the continuous morphological variation, the high-altitudinal endemics could have originated by local adaptation from originally low-land species. DNA-ploidy level and genome size were detected by flow cytometry. Taxonomically important characters were found using multivariate morphometric analysis. The phenotypic plasticity of the taxon C. tatrae was tested by a cultivation experiment. Genetic structure of the studied species was revealed using molecular marker - microsatellites (7 variable primers). Studied taxa were tetraploids, but some possessed different genome size. The morphological differentiation was mainly due to characters like calyx lobes length...

Cleft lip is one of the most common human birth deffects. Its etiopathogenesis is multifactorial and many aspects of its occurrence remain unknown in the fields of both genetics and teratology. One of the set of known negative external factors causing cleft lip is chemical hydrocortisone. Its effect on cell proliferation is highly heterogeneous and depends on attributes of a specific cell population. In this work we studied the cleft beak origin after the hydrocortisone treatment on the basis of Chick Embryotoxicity Screening Test (CHEST). Our main aim was to detect cell cycle changes in the chick frontonasal process after hydrocortisone injection via flow cytometry analysis. Hydrocortisone caused S phase arrest within a minor subpopulation of highly granular cells with specific cell cycle. This sensitive subpopulation was localized in the areas of previously defined proliferative centers within the frontonasal process using immunohistochemistry of frozen sections. Quantitative analysis of cells in these areas revealed significant decrease of M phase portion in the hydrocortisone treated samples in comparison with the control samples. The TUNEL staining of histological sections was used to determine the apoptotic rate in the frontonasal process. The comparison between the control and the...

Callitriche (water-starwort) belongs among difficult and insufficiently known genera of the Czech flora. The presented work provides the first critical taxonomic revision of the genus in the Czech Republic, with information relevant also to the broader region of Central Europe. Morphological and cytometric investigation resulted in identification of six Callitriche species in the Czech Republic. The genome size of all six Czech representatives of the genus was estimated using flow cytometry. The hybrid C. × vigens Martinsson (C. cophocarpa × C. platycarpa) was found for the first time in the Czech Republic. An individual related to C. hamulata was found in the Tichá Orlice river, which has aberrant genome size and aborted flowers; further study of this taxon is necessary. Multivariate morphometric analyses of fruits and cultivation were used to check and define reliable distinctive features and an impact of phenotypic plasticity. Key to the identification of species (including the first key for sterile plants), its detailed descriptions and ecological demands were provided. Each species differ significantly from the others in genome size, morphological features as well as in ecology. The reproductive strategy has the main importance for ecology and morphology of the studied species. The distribution of...

Nuclear DNA content (genome size) is one of the basic characteristics of living organisms. In the Angiosperms, the range of genome size is 2,300-fold, which raises questions about the causes and consequences of this tremendous variation. This thesis deals with genome size in plants from the level of intraspecific homoploid variation, through intraspecific ploidy variation, to interspecies comparisons. On various study systems we investigated the dynamics and ranges of genome size variation, tried to reveal possible associations between genome size and selected biological traits, and assessed the extent to which differences in genome size are manifested at the ecological and evolutionary level. As a means of estimating genome size we applied flow cytometry (FCM). In Taraxacum stenocephalum we conducted a detailed study of its enormous genome size variation. We carried out crossings of parents with various genome sizes and compared these parental genome sizes with those of F1 offspring. We also attempted to reveal the association of genome size with various growth traits. In Galium valdepilosum and Arabidopsis arenosa we carried out an extensive flow-cytometric ploidy level screening and compared the distribution and ecological preferences of detected cytotypes. We studied the Andean genus...

This diploma thesis deals with the determination of creatinine using a combination of flow injection analysis (FIA) or high-performance liquid chromatography (HPLC) with pulse amperometry, an electrochemical technique based on the application of potential pulses on a gold working electrode. The determination was performed in a basic environment of borate buffer with creatinine concentration of 1∙10-4 mol∙l-1 . The lenght of the cleaning and activation pulse was optimized as well as the pH of the running buffer. A cleaning pulse of +1.8 V was first applied to the electrode for 100 ms, then an activation potential of -0.5 V was applied for 150 ms and then a measuring potential of +0.2 V for 300 ms. The optimal pH was selected as pH=9,4. Methanol and acetonitrile were added to the borate buffer to test whether creatinine could be determined in presence of these organic solvents and whether flow injection analysis could be transformed into HPLC. Methanol in the system caused peak deformation, acetonitrile did not cause the peak deformation in the system, at higher contents the baseline was destabilized. Furthermore, the calibration dependence in the range of concentrations from 2.5∙10-4 mol∙l-1 to 5∙10-6 mol ∙ l-1 was measured using PAD in combination with FIA. At higher concentrations, peaks splitted....

Presented diploma thesis is focused on testing encapsulation methods of enzymes and probiotic bacteria. In the theoretical part a summary of different encapsulation techniques used in food industry is given. Further, materials for encapsulation, above all polysaccharides are presented. Next, some procedures of encapsulation of biopolymers and microorganisms – mainly enzymes and probiotic cultures are discussed. In the experimental part methods for preparation of several types of particles based on polysaccharides and liposomes are introduced. Particles were used for encapsulation of selected hydrolytic enzymes and probiotic strains Bifidobacterium breve a Lactobacillus acidophilus. The encapsulation effectiveness was evaluated by analysis of total proteins and enzyme activities. Particles sizes and their stability in water, in selected model foods and model body fluids were observed, too. According to results obtained in this work it was found that encapsulation of enzymes into polysaccharide particles were succesfull in all types of particles (encapsulation effectivness was more than 50 %). Polysaccharide particles showed a very good stability in body fluids as well as in model foods. As the most suitable materials for enzymes encapsulation chitosan and liposomes were found. Polysaccharide particles were used also for the encapsulation of microorganisms. The stability of particles with lactic acid bacteria was similar to particles containig enzymes, very good stability was verified aslo in model foods and model body fluids. Encapsulation enables long-term stabilization of biologically active compounds as well as posibility of their transport and controlled releasing in gastrointestinal tract. Encapsulation of probiotic bacteria could preserve their viability and long-term survival until the product expiration date. Thus, encapsulation is one of the most promissing procedures for production of foods and food suplements of great quality and high additional value.

Ranunculus trichophyllus agg. (thread-leaved water crowfoot) represents a taxonomically challenging group of aquatic plants in which the presence of several significantly different genotypes and the genome size variation have been recently revealed. The results of previous studies suggest that cryptic taxa occur in this group, being so far overlooked due to considerable morphological reduction and extensivephenotypic plasticity. In this thesis, the variation and genetic relationships of four morphologically similar homophyllous water-crowfoot species was critically assessed in the area of Central Europe, using a combination of modern biosystematic methods (flow cytometry, direct DNA sequencing, morphometric analyses), specially focusing on the complex of R. trichophyllus.. The genome size analysis via flow cytometry was confirmed as a suitable method for determining the studied species; further, several hybrid combinations were revealed using this approach. However, recent interspecific hybridization is rather infrequent in the interest group. The results of DNA analyses indicate an importance of hybridization events in the evolution of sect. Batrachium: all the polyploid taxa studied are probably of allopolyploid origin. Two cryptic taxa within the traditionally recognized species R. trichophyllus have...

This diploma thesis is focused to the determination of procaine using flow injection and sequential injection analysis coupled with spectrophotometric detection. This determination is based on the reaction of procaine with a colouring agent, 1,2-naphthoquinone-4-sulfonic acid. An orange coloured product is formed, which is determined spectrophotometrically at the wavelength 484 nm. The high of the absorption signal of the product is directly proportional to the concentration of procaine. The aim of this work was to optimize the parameters of both methods of flow analysis so that the limit concentration of procaine can be as low as possible and at the same time, high sensibility is achieved. The next step was to apply these methods on the determination of procaine in real samples. It was found that in flow injection analysis, the absorbance of procaine hydrochloride obeys Beer's law for concentrations from 2.5 to 120 µg/ml. The linear regression equation of calibration graph was y = 0.0059x - 0.0051, with a linear regression correlation coefficient 0.9993. Limit of detection was 0.72 µg/ml. Effects of standing time (stopped-flow), flow rate, concentration of colouring agent, pH and the volume of the sample loop have been examined and optimized. It was also found that in sequential injection...