 Keywords: Lung Tx, EVLP, NHBD, IR injury, ROS, hypercapnia, gender differences This dissertation thesis deals with a very topical issue of the lack of donor organs for lung transplants. As with other organs, the number of patients on waiting lists in terminal stages of their diseases is also constantly rising but there is not an adequate increase in donor organs. We focused our experimental work on the development of research concerning the solution which is most successful in a long-term perspective, i.e. organ transplants from non- heart-beating donors (NHBDs), in an ex vivo lung transplant protocol (EVLP) on an animal model (Wistar rats). This is a method which is clinically established worldwide (in the Czech Republic only experimentally) and is constantly subject to further research. Based on earlier studies, we focused the first experimental part of this work on the potential protective effect of hypercapnic ventilation on ischemia-reperfusion (IR) lung injury in EVLP. The study proved that the hypercapnic ventilation has a protective effect on the generation of reactive oxygen species (ROS) in IR lung injury, but only when used in the period of reperfusion. In the second experimental study, we followed up a very topical issue of the effect of gender on IR lung injury in EVLP in...

This thesis deals with the segmentation of lung parenchyma and extraction of airways tree from three dimensional CT scans. The external mask of the lungs is created and subsequently used to ease the process of airway segmentation. In this work, some published methods for airways segmentation are described with focus on one, which is described more in detail and also implemented in MATLAB. The proposed approach is based on morphological grayscale reconstruction. Method is tested on the real patient CT scans and finally, the results are discussed.

The thesis is focused on the comparison of functional examination of the patients lungs, before and after coronavirus disease 2019 (COVID-19), as well as on the comparison of physiological data for a individual person with measured values after the disease. The relationship between lung function and smoking, sex, age and time gap after COVID undergone was monitored. The screening was accomplished by spirometry in a pneumology surgery and the obtained data were processed in practical part of my thesis. The theoretical part of the diploma thesis describes the respiratory system and lungs functional testing. In addition, infectious disease COVID-19 is described herein. The aim of the thesis was to select suitable patients, who have suffered from the COVID-19 disease, but without other serious respiratory problems, make a measurements and statistically evaluate the correlation between the measured results. The experiment involved 66 people aged between 21-94, of which 43 were men and 23 women. It has been shown a predominantly negative effect of COVID-19 in the values of the examined indicators in spirometric measurements, similarly, the impairment was seen more in men than in women. The role of younger age has been proven in the better results of spirometric measurements. However, the differences in...

Human health is significantly influenced by inhaled aerosols. Insight to the aerosol transport and deposition mechanisms is a prerequisite for both, toxicological protection against harmful particles and efficient application of inhaled therapeutic aerosols. The purpose of this doctoral thesis was to gain new knowledge of this topic on the basis of in vitro measurements. Phase-Doppler Anemometry was chosen for aerosol transport measurement, for it allows simultaneous measurement of particle size and velocity. Results were processed by means of statistical methods and frequency analysis. Deposition of spherical aerosol particles was measured by Positron Emission Tomography, while deposition of fibrous aerosol was measured by Phase-Contrast Microscopy combined with automated image analysis. All experiments were performed on physical models created on the basis of the real lung geometry. New knowledge of flow characteristics, transition from laminar to turbulent flow, effect of breathing pattern or particle size on aerosol transport and deposition in human lungs are outcomes of this work. Significant effect of the oral cavity was ascertained due to comparison of aerosol deposition in realistic and semi-realistic model with cylindrical smooth walls. Acquired data not merely extended our knowledge of aerosol behavior in lungs but it can also be used for validation of numerical simulations.

This diploma thesis deals with chronic obstructive pulmonary disease (COPD) and its influence on the musculoskeletal system, lung transplantation and pulmonary rehabilitation. It discusses the role of POWERbreathe, a breathing training device, which is primarily used to strengthen the inspiratory muscles. The practical part was performed as a randomized control pilot study to assess the effect of POWERbreathe on the strength of the inspiratory muscles, the thorax dynamics and selected pulmonary function. The aim of the study is to determine the benefits of using the POWERbreathe for COPD patients during the pre-transplant period. In total, 18 patients with COPD IV. grade participated in the study (6 women and 12 men) with an average age of 59.8 ± 5.53 years. Probands were divided into two groups, only one of which used a POWERbreathe. Pulmonary functions (FEV1, VCin, FVC, PIF), muscle strength (SIndex), and chest dynamics (chest disturbances in the mezosternal and xiphosternal region with maximum inspiration and exhalation and respiratory amplitudes in these areas) were evaluated. The results did not show a significant inter-group difference in the strength of the inspiratory muscles. Within pulmonary function, this difference was evident in the FEV1 parameter and the chest dynamics increased...

Tumors are one of the leading causes of death worldwide. Generally, the prognosis is better if the treatment begins at an early stage. Nowadays, the conventional chemotherapy treatment of cancer, known for its limited efficacy and side effects, is being gradually replaced by targeted biological treatment, which is used when specific genetic mutations are found. A part of the treatment is a detection of a potential progression, which is mainly based on the tumor biomarkers monitoring. Currently, further investigation of a so-called liquid biopsy method are ongoing, on which this thesis is focused. The main aim of this work was the experimental development and validation of the method for detection of the ctDNA in the plasma samples based on the somatic mutations presence. For the development and optimization of the system on the principle of denaturation capillary electrophoresis, the samples of cancer patients with KRAS mutation were used. Subsequently, a clinical part of the research was performed on a pilot set of 21 plasma samples. Finally, the method was optimized for the detection of BRAF and EGFR markers. A partial objective was to improve the detection sensitivity and increase the capture of the ctDNA in patients with advanced stage of the disease. The results of this work suggest the...

Non-small cell lung cancer belongs to most frequent malignant tumours at all worldwide. Despite significant progress in knowledge about etiopathogenesis and targeted anticancer therapy, basic scientific research in this particular field and development of more effective treatment remains challenging. In case of its inadequate activation, the Hedgehog signaling pathway is involved in non-small cell cancer development. P53 is well known tumour suppressor gene, that serves as anticancer barrier. Its activity is mostly determined by the transcriptional activation of many pro-apoptotic genes, one of which is SIVA-1. Recently, it has been surprisingly shown, that SIVA-1 has also pro-oncogenic properties in a mouse model of non-small cell lung cancer. The aim of this study was to clarify the importance of Hedgehog signaling pathway and protein SIVA-1 and their potential relationship in development and progression of human non-small cell lung cancer. In selected cell lines of human non-small cell lung cancer, expression of each single component of Hedgehog signalign pathway was detected. In the tissue samples of tumour obtained from 39 patients that underwent surgery for non-small cell lung cancer and selected cell lines of the same tumour, expression of SIVA-1 protein was revealed. These findings indicate...

Several experiments were designed for utilization of fluorescence spectroscopy as a method of aerosol particle detection in a model of lungs. One of the experiments was to arranged use luminescent properties of DEHS (bis(2-ethylhexyl)decandioate) for calculating aerosol deposition. The outcome of this experiment was confirmation of clusters existence, which causes luminescence of DEHS. But the luminescence is not enough dependent on concentration and as such is not suitable for calculation of aerosol deposition. As the next experiment DEHS-fluorescein particles were generated by condensation monodisperse aerosol generator (CMAG), where water was used instead of isopropyl alcohol as a solvent. By this alteration the negative influence of DEHS was eliminated, which caused results refinements of aerosol deposition. Generation of fluorescein sodium salt particles by small-scale powder disperser (SSPD) was designed as a last experiment. The lower deposition efficiency measured by this method was caused by particles polydispersion. Photo records were used for documentation of Hot-spots. Outcomes of this study are new knowledge of fluorescence spectroscopy utilization for study of aerosol deposition and possibilities of fluorescent aerosol particles generation. Acquired data can serve for knowledge extension of possible detection methods for aerosol particles in the model of lung and can serve for validation of numerical simulations.

Human health is significantly influenced by inhaled aerosols. Insight to the aerosol transport and deposition mechanisms is a prerequisite for both, toxicological protection against harmful particles and efficient application of inhaled therapeutic aerosols. The purpose of this doctoral thesis was to gain new knowledge of this topic on the basis of in vitro measurements. Phase-Doppler Anemometry was chosen for aerosol transport measurement, for it allows simultaneous measurement of particle size and velocity. Results were processed by means of statistical methods and frequency analysis. Deposition of spherical aerosol particles was measured by Positron Emission Tomography, while deposition of fibrous aerosol was measured by Phase-Contrast Microscopy combined with automated image analysis. All experiments were performed on physical models created on the basis of the real lung geometry. New knowledge of flow characteristics, transition from laminar to turbulent flow, effect of breathing pattern or particle size on aerosol transport and deposition in human lungs are outcomes of this work. Significant effect of the oral cavity was ascertained due to comparison of aerosol deposition in realistic and semi-realistic model with cylindrical smooth walls. Acquired data not merely extended our knowledge of aerosol behavior in lungs but it can also be used for validation of numerical simulations.

Objective: The goal of the thesis "Monitoring of the Respiratory Functions in patients after the Rib Cage Surgery according to Nuss" was to confront the present knowledge about respiratory functions and the strength of respiratory muscles of patients with inherited pectoral deformity pectus excavatum, to compare it with my own measuring and to find out about their development after the surgical correction according to Nuss. Methodology: There were 15 patients being monitored (13 men and 3 women), who underwent spirometric and bodypletysmographic examination and the occlusal mouth pressures were determined before the surgery. The same pulmonary function tests were run in the course of 4 to 10 months (7.1 months on average) on all the patients, the control determination of mouth pressures is available for 13 of them. Results: In the entry tests, compared to adequate values the patients in this file had increased residual lung capacities (RV 142%, p=0.001; ITGV 116.2%, p=0.004; RV%TLC 133.8%, p=0.002; ITGV% 116.1%, p=0.001) at the expense of statistically lower vital lung capacity (VC 81.9%, p=0.0004) and inspiratory reserve capacity (IC 84.2%, p=0.0003), while the total lung capacity remained unaltered (TLC 99.7%, p=0.877). The obstructive parameters of these patients were significantly altered (FEV1 95.7%,...