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Name of the thesis Effect of nitric oxide on cerebral blood flow during neuronal activity Aim of the thesis The aim of this thesis is to determine whether the application of 7-nitroindazole, relatively specific inhibitor of neuronal nitric oxide synthase, affects the baseline blood pressure. Furthermore, to determine whether the application of the substance affects the baseline cerebral blood flow and whether it influences blood flow in brain during transcallosal stimulation with increasing frequency. Research method The research took place at the premises of the Institute of Physiology, Academy of Sciences of the Czech Republic. Experiments were carried out on laboratory albino Wistar rats. The group contained both experimental and control sample. General anesthesia was performed to rats, stimulating and sensing electrodes were implanted in epidural area of sensorimotor cortex and Laser Doppler flow probe was implanted into the contralateral hemisphere. A plastic catheter was applied in the carotid artery for measuring systemic blood pressure. In the first part of the experiment, we tested the effects of 7-nitroindazole on the systemic blood pressure. In the second part of the experiment, we investigated the effects of 7-nitroindazole on baseline cerebral blood flow. The third part of the...

Title: The role of nitric oxide in the pathophysiology of neurodegenerative diseases Objectives: The main objective of this thesis is to evaluate the effect of nitric oxide on the formation and development of neurodegenerative diseases. Another objective was to determinate, whether NO affects by its impact processes involved in apoptosis in the CNS. Methods: The thesis is prepared in the form of research, drawing from available relevant resources. Results: Nitric oxide is widely applied in the pathophysiology of selected neurodegenerative diseases, either directly or through other reactive nitrogen and oxygen. It also affects other factors that are involved in apoptosis in the CNS. Keywords: Nitric oxide, NMDA receptors, neurodegenerative diseases, excitotoxicity, apoptosis

The aim of this thesis was to contribute to the standardization and validation of expiratory flow parameters independent of NO testing, particularly alveolar concentration (CANO) and bronchial NO flux (JawNO) in children and adolescents with allergic respiratory diseases. The thesis was created in the time of increased interest of medicine professionals in non-invasive examination methods and targeted therapeutic influence of inflammation in small airways in asthmatics. After the standardization and validation of clinical benefits, the measurement of concentrations of nitric oxide (FENO) in the air exhaled through the mouth with the speed of 50 ml/s was validated for the evaluation of eosinophilic airway inflammation in patients with asthma and started to be widely used in clinical practice. Currently, there is an ongoing research focused on a new method for FENO measurement at several flow rates, which uses physiological models of NO transfer in exhaled air, allowing to quantify the amount of NO in an indirect way and to monitor the intensity of the inflammatory processes in distal and proximal airways non- invasively. In groups of children and adolescents with allergic rhinitis and / or bronchial asthma and their healthy peers (aged 6 -19 years), the concentration of FENO was examined using online...

Nitric oxide (NO) is an interesting molecule, which is involved in many important biological processes such as vasodilatation, neurotransmission, immune response and cell proliferation. This work presents the crucial role of NO during the Xenopus laevis embryonic epidermis development. The outer layer of the embryonic epidermis is composed of 4 cell types (small secretory cells, multi-ciliated cells, ionocytes and goblet cells). Embryonic epidermis composition reflects specialized epithelia such as a respiratory epithelium of mammals. Therefore, Xenopus embryonic epidermis has become a suitable model for the study of human mucosal and mucociliary epithelium and their defects. I found that NO is mainly produced in ionocytes and multi-ciliated cells by using molecular and immunohistochemical approaches. The study of molecular and cellular phenotype changes in embryos with inhibited NO production revealed the necessity of this molecule for correct formation and function of the mucociliary epithelium. NO inhibition caused reduction of specialized epidermal cell types (small secretory cells, ionocytes and multi-ciliated cells) and structural changes in multi-ciliated cells. I also showed that NO affects the development of the embryonic epidermis through the sGC-cGMP-PKG signaling pathway, probably by...

Sand flies (order Diptera) are vectors of Leishmania parasites (Trypanosomatida), which are inoculated into the host skin together with the vector saliva. Sand fly saliva plays the important role in the Leishmania transmission; in naive host it supresses the host immune response assisting Leishmania to establish the infection, while in repeatedly bitten host it elicits a protective immune response. The submitted thesis focuses on the effect of sand fly saliva on macrophages, the key cells in the infection control. In the first part of the thesis we established a laboratory model L. major - P. papatasi - Balb/c to describe the protective effect of saliva immunization on Leishmania infection development. Immunized mice were protected against Leishmania infection which was reflected in the ear lesion size, parasite load in the ear dermis and draining lymph nodes but also in cytokine production. On the contrary, produced lower amount of nitric oxide, while arginase activity was comparable with nonimmunized group. The IgG antibodies against saliva served as a marker of exposure to sandflies while IgG antibodies against Leishmania antigens served as a marker of infection severity. The experiments were aimed on the possibility of cross-protectivity in Balb/c mice against L. major between closely related...

This project is concerned with the preparation of electrospun polystyrene (PS) nanofiber materials with covalently bonded NO-photodonor and electrostatically attached tetracationic porphyrinoid photosensitizers. These photofunctional nanofiber materials exhibit effective simultaneous photogeneration of small antibacterial NO and O2(1 ∆g) species under irradiation with daylight creating an antibacterial surface and near surrounding. NO species can be also generated just by gentle heating. Nanofiber materials were analyzed with SEM, FTIR, emission and UV/vis spectroscopy and time-resolved emission and absorption spetroscopy. The antibacterial effect was tested on Escherichia coli. The dual antibacterial action, in combination with the nanoporous character of the material that detains pathogens like bacteria on its surface, is ideal for any application where a sterile environment is neces- sary. The known bimetallic cluster system [(PMe2Ph)4Pt2B10H10] that possesses the propen- sity to reversibly bind small gaseous molecules (O2, SO2, CO) was synthesized in good yields for NO reversible binding investigation. Seven new monometallic precursors (Pt, Pd and Ni) to new bimetallic species were succesfully synthesized with the aim of future study of NO reversible binding. All new compounds were purified by...

Nowadays, there is an increasing risk of bacterial infections from bacteria strains resistant towards antibiotics. Thus, it is of utmost importance to research novel therapies which can overcome this difficulty. The presented thesis focuses on the preparation, characterization and antibacterial evaluation of polystyrene polymer nanomaterials (nanofiber membranes and nanoparticles) modified with compounds that can efficiently inhibit bacterial growth either by their nature (polyethyleneimine) or by photoactivation upon visible light excitation (NO- photodonors, photosensitizers) and consequent production of highly reactive inorganic bactericidal species, nitric oxide (NO) and singlet oxygen (O2(1 g)). All materials were fully characterized by several independent methods. The concentrations of NO and O2(1 g) were measured by amperometric and time-resolved spectroscopic techniques and by variety of chemical analytic procedures. Due to the presence of bactericidal species and the efficient photogeneration of NO and O2(1 g) at physiological conditions, all materials exhibit strong antibacterial action tested on a Gram-negative bacterial strain Escherichia coli. Hence, these functionalized polymer nanomaterials may be intriguing systems for medical-, biological-, or environmental- application where a...

Nitric oxide (NO) has been proved to reduce parasite burden in vertebrates infected with Schistosoma, Fasciola, Brugia or Taenia. NO negatively influences parasite growth and development, which then leads to smaller parasite-caused damage to the liver during schistosomosis and stimulates healing processes in muscles infected with Toxocara canis. Peroxynitrite, formed from NO and superoxide, significantly reduces the viability of F. hepatica adults. In case of T. regenti, the neuropathogenic schistosome, the cells capable of NO production (macrophages, neutrophils, eosinophils, microglia and astrocytes) migrate to the site of the infection suggesting that NO might affect T. regenti infection as well. Therefore, the production of NO and its effect on the course of the infection was examined in vivo and the effect of peroxynitrite on T. regenti schistosomula was examined in vitro to assess the role of reactive nitrogen species during the infection. Our results from in vivo experiments demonstrate that although the infection did not significantly elevate nitrite/nitrate results in the sera, NO is locally produced in the early stages of the infection in both the skin and the spinal cord as shown by immunohistochemical detection of inducible NO synthase. Diminishing NO production by aminoguanidine...

The aim of present project was to uncover the effect of pharmacological increase in acute and chronic nitric oxide (NO) production on cardioprotective effect of chronic hypoxia. We studied the effect of NO donor molsidomine on hemodynamic conditions and ischemia - induced myocardium injury. Male Wistar rats were exposed to continual hypoxia in a normobaric chamber (10 % O2, 4 weeks). Rats received molsidomine either chronically (15 mg/kg/day) in drinking water or acutely (10 mg/kg) in saline infused 30 min before ischemia. Control rats were kept under normoxia and treated in a corresponding manner. Adaptation to chronic hypoxia resulted in development of pulmonary hypertension. Chronic treatment with molsidomine slightly reduced these consequences of chronic hypoxia but it had no effect on increased cardiac ischemic tolerance in chronically hypoxic rats. On the other hand acute treatment with molsidomine significantly reduced infarct size and increased the number of arrhythmias in both normoxic and chronically hypoxic animals. In conclusion, our data suggests that acute increase in availability of NO is cardioprotective in both normoxic and chronically hypoxic rats contrary to its chronic increase which seems to have no protective contribution.