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Quantification in descriptive and experimental study of functional
Tonar, Zbyněk ; Kočová, Jitka (advisor) ; Hach, Petr (referee) ; Míšek, Ivan (referee) ; Křen, Jiří (referee)
Dissertation abstract The dissertation thesis consists of seventeen papers dealing with quantitative histological assessment of vascular wall and with application of morphometry in atherosclerosis research as well as in biomechanics of elastic arteries. History of microscopic morphometry with an emphasis on stereology is reviewed in the Introduction. The principles of mathematical morphology and of stereological estimation of volume, surface, area, length, and numerical density of objects in tissue blocks are explained in the Methods. The key rules of unbiased sampling of histological series and designing morphometric studies are summarized. The distribution of the overall observed variance in biological experiments, its sources and management are discussed. Procedures necessary for three-dimensional reconstructions based on scanned histological series are described. We proved that inflammatory process was significant in patients with ruptured aneurysm of abdominal aorta (AAA). There were more pronounced inflammatory infiltrates in areas of neovascularization in ruptured AAA than in asymptomatic AAA patients. Histological findings were in agreement with higher cytosol cytokine levels (IL-6, IL-8, TNF-α), enhanced collagen III metabolism and degradation in ruptured AAA. We found lower volume fraction of the...
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Effect of inhaled metal oxide nanoparticles on organs of exposed mice
Mikuška, Pavel ; Večeřa, Zbyněk ; Dočekal, Bohumil ; Čapka, Lukáš ; Křůmal, Kamil ; Coufalík, Pavel ; Moravec, Pavel ; Buchtová, Marcela ; Míšek, Ivan ; Vrlíková, Lucie ; Smutná, Tereza ; Dumková, J. ; Hampl, A. ; Fictum, P.
Adult mice were continuously exposed to metal oxide nanoparticles for different time intervals. At the end of experiment, mice organs (lung, brain, kidney, liver and spleen) were collected for chemical, biochemical, histological and electron microscopical analyses.
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Evolutionary and developmental aspects of dentition of squamate reptiles
Zahradníček, Oldřich ; Horáček, Ivan (advisor) ; Míšek, Ivan (referee) ; Peterková, Renata (referee)
EVOLUTIONARY AND DEVELOPMENTAL ASPECTS OF DENTITION OF SQUAMATE REPTILES Summary of the PhD. thesis Dentition and its evolutionary modifications played important roles during the radiation of reptiles. It is generally accepted that a plesiomorphic state for reptiles is represented by homodont, polyphyodont dentition with conical or cylindroconical teeth. However, in contrast to mammalian dentition which is the object of intensive research, reptilian dentition is studied only rarely. I therefore focused on developmental and evolutionary aspects of squamate dentition and used histological, immunohistochemical and in situ hybridization methods, computerised tomography, rentgenography and scanning electron microskopy. The goals of this work were as following: (a) to choose the model taxa representing squamate lineages relevant from a phylogenetic position as well as possessing dissimilar types of dental adaptations; (b) to evaluate using of these taxa as model species; (c) in these taxa describe basic odontogenic features. As a species with generalized dentition gecko Paroedura picta was chosen, the up coming reptilian model species with features also present in a wide spectrum of other lizards. I focused mainly on the developmental differences between teeth of the null and functional generations, development...
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Quantification in descriptive and experimental study of functional
Tonar, Zbyněk ; Kočová, Jitka (advisor) ; Hach, Petr (referee) ; Míšek, Ivan (referee) ; Křen, Jiří (referee)
Dissertation abstract The dissertation thesis consists of seventeen papers dealing with quantitative histological assessment of vascular wall and with application of morphometry in atherosclerosis research as well as in biomechanics of elastic arteries. History of microscopic morphometry with an emphasis on stereology is reviewed in the Introduction. The principles of mathematical morphology and of stereological estimation of volume, surface, area, length, and numerical density of objects in tissue blocks are explained in the Methods. The key rules of unbiased sampling of histological series and designing morphometric studies are summarized. The distribution of the overall observed variance in biological experiments, its sources and management are discussed. Procedures necessary for three-dimensional reconstructions based on scanned histological series are described. We proved that inflammatory process was significant in patients with ruptured aneurysm of abdominal aorta (AAA). There were more pronounced inflammatory infiltrates in areas of neovascularization in ruptured AAA than in asymptomatic AAA patients. Histological findings were in agreement with higher cytosol cytokine levels (IL-6, IL-8, TNF-α), enhanced collagen III metabolism and degradation in ruptured AAA. We found lower volume fraction of the...
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Ontogenetický vývoj nasální oblasti u Discoglossus pictus (Anura: Discoglossidae)
Královec, Karel ; Roček, Zbyněk (advisor) ; Míšek, Ivan (referee) ; Slípka, Jaroslav (referee)
Using histological techniques and computer-aided three-dimensional reconstructions made from serial histological sections, I studied the ontogenetic development of the ethmoidal endocranium and nasal chemosensory organs in discoglossid frog Discoglossus pictus. The nasal cartilages, including the inferior prenasal cartilage, are de novo adult structures. The only larva-derived structures of the adult nasal capsules are the posterior aspects of the solum nasi and septum nasi. I identify a pattern of development for the suprarostral cartilage that differs from previous findings, and probably represents the ancestral anuran pattern. I also identify patterns of development for the ethmoid plate and postnasal wall that occur during early in ontogenesis. The pattern of timing of chondrification of the anterior nasal cartilages more closely coincides with that of the neobatrachian species than that recorded for the pelobatid frog Spea. The olfactory epithelium in larval D. pictus represents one continuous unit of tissue not divided into two separate portions. However, it forms distinct ventromedial diverticulum embeded into roof of the buccal cavity. This particular arrangement may enhance chemosensory sampling during larval period and this diverticulum might be homologous with the ventral olfactory...
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Influence of inhaled manganese oxides nanoparticles on mass of internal organs in mice
Zeman, T. ; Dočekal, Bohumil ; Mikuška, Pavel ; Buchtová, Marcela ; Míšek, Ivan ; Šerý, Omar ; Večeřa, Zbyněk
Results of the statistical analysis of a long-term inhalation experiment on laboratory mice are presented. During 17 weeks of the experiment, the experimental group was exposed to inhaled manganese oxides nanoparticles. Manganese oxides (MnO center dot Mn2O3) nanoparticles (MnONPs) were synthesized continuously via aerosol route in a hot wall tube flow reactor using a thermal decomposition of metal organic precursor manganese(II) acetylacetonate in the flow tube reactor (in vertical position) at temperature 750 degrees C in the presence of 30 vol% of oxygen. The concentration of produced MnONPs at the reactor output was in the range 1-3 x 10(7) particles/cm(3) and the size of generated nanoparticles MnO center dot Mn2O3 was in the range 7-50 nm. Before entering the inhalation chamber, MnONPs in the mixture of N-2, O-2 and air (at total flow rate of 3 L/min) were further diluted using filtrated humidified air (20 L/min) at temperature 21 degrees C resulting in the MnONPs concentration 2 x 10(6) particles/cm(3). At regular time intervals the mass of selected internal organs of mice from both experimental and control group was assessed. It has been proven that inhaled nanoparticles are able to influence the mass of internal organs of mice. Statistically significantly lower mass of kidneys, liver and spleen and higher mass of pancreas have been found in the experimental group compared to the control group.
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Influence of inhaled manganese oxides nanoparticles on mass of internal organs in mice
Zeman, T. ; Dočekal, Bohumil ; Mikuška, Pavel ; Buchtová, Marcela ; Míšek, Ivan ; Šerý, Omar ; Večeřa, Zbyněk
Results of the statistical analysis of a long-term inhalation experiment on laboratory mice are presented. During 17 weeks of the experiment, the experimental group was exposed to inhaled manganese oxides nanoparticles. Manganese oxides (MnO.Mn2O3) nanoparticles (MnONPs) were synthesized continuously via aerosol route in a hot wall tube flow reactor using a thermal decomposition of metal organic precursor manganese(II)acetylacetonate in the flow tube reactor (in vertical position) at temperature 750 °C in the presence of 30 vol% of oxygen. The concentration of produced MnONPs at the reactor output was in the range 1-3 × 107 particles/cm3 and the size of generated nanoparticles MnO.Mn2O3 was in the range 7-50 nm. Before entering the inhalation chamber, MnONPs in the mixture of N2, O2 and air (at total flow rate of 3 L/min) were further diluted using filtrated humidified air (20 L/min) at temperature 21 °C resulting in the MnONPs concentration 2 × 106 particles/cm3. At regular time intervals the mass of selected internal organs of mice from both experimental and control group was assessed. It has been proven that inhaled nanoparticles are able to influence the mass of internal organs of mice. Statistically significantly lower mass of kidneys, liver and spleen and higher mass of pancreas have been found in the experimental group compared to the control group.
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Unique exposure system for the whole body inhalation experiments with small animals
Večeřa, Zbyněk ; Mikuška, Pavel ; Dočekal, Bohumil ; Vojta, Šimon ; Míšek, Ivan ; Moravec, Pavel ; Smolík, Jiří
The especially inhalation chamber is constructed for chronic exposure studies of small animals to nanoparticles under strictly controlled conditions in frame of project „Study of transport of inhalated nano-sized particles (Pb, Cd) and their allocation in organs“. The inhalation chamber builds from a glass and stainless steel and makes possible long-term inhalation procedures (up to 60 days, 24 hours/day) with up to four discrete groups of mice population (up to 60 individuals each) or of rats population (up to 20 individuals each) under controlled illumination, temperature, relative humidity, doses (0.5 x 106 - 5 x 106 particles/cm3) and size (10-60 nm) of nano-sized particles which are synthesized via aerosol route in a hot wall tube flow reactor using metal organic precursors.
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Modulace intracelulární kaspázové mašinérie s využitím explantátových kultur
Matalová, Eva ; Fleischmannová, Jana ; Norek, Adam ; Míšek, Ivan
Organ explant cultures eliminate animal suffering during experiments but simultaneously allow investigations of intact organ, tissue and cell systems with preserved interactions corresponding to the situation in vivo. Moreover, the explants can be manipulated in several ways using modern micromanipulation techniques (e.g. electroporation, inhibition). We show possible usage of explant culture approaches in embryonal apoptosis research in three models – mouse limb digitalization, tooth germ morphogenesis, separation of middle ear ossicles. Data from specific pharmaceutical inhibitions focused on key apoptotic molecules – caspases - are demonstrated.
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