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The introduction of diagnosis of new bacterial pathogens Ralstonia a Achromobacter isolated from patients with cystic fibrosis and determining their sensitivity to antibiotics
Michálková, Alice ; Melter, Oto (advisor) ; Bébrová, Eliška (referee)
Cystic fibrosis (mucoviscidosis) is an incurable genetic disease caused by mutations in the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator Gene). The most common causes of increased mortality and morbidity of patients include bacterial respiratory infections which may occur even due to less frequent pathogens. Bacterial species of the Ralstonia and Achromobacter genera are not considered pathogenic for healthy people, but they have been established as pathogens in the sputum of patients with CF. However, due to their phenotypic similarity to other bacterial pathogens encountered in patients with CF, microbiologists often do not pay attention to them. The aim of this thesis was to propose some methods of identification of the genus Ralstonia, to determine both quantitative and qualitative susceptibility towards antibiotics, and to make a bibliographical search focused on the issue of the Achromobacter genus. Strains of Ralstonia spp. were identified using phenotypic and genotypic methods and were tested for the susceptibility towards antibiotics. The thesis also proposes a new method of genotypic indentification of R. respiraculi.
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Isolation of bacterial flagellin as an immunogen for the preparation of antibodies for cystic fibrosis patients
Supová, Veronika ; Hodek, Petr (advisor) ; Kubíčková, Božena (referee)
Cystic fibrosis (CF) is an autosomal recessive disease caused by CFTR gene mutation. The consequence of this mutation is an improper function of the chloride channel consisting of CFTR protein. Due to this abnormality, the transport of chloride ions is either reduced or inhibited completely, which leads to a mucus secretion. Mucus is mostly created in the lungs and it is the ideal environment for pathogenic bacteria like Pseudomonas aeruginosa (PA). Pseudomonas aeruginosa is an aerobic, gram-negative conditioned pathogen occurring in patients with weakened immunity such as patients with CF, who are often hospitalized. PA has one polar flagellum which contains filament composed of a protein called flagellin. The flagellum is one of the most important virulence factors of PA bacteria. This thesis focused on the isolation of flagellin from Pseudomonas aeruginosa flagella. The isolated flagellin will serve as an antigen for the preparation of prophylactic antibodies for CF patients. The isolation of flagellin was carried out by four methods using combinations of precipitation and centrifugation. The published methods were optimized to achieve the isolation of flagellin. Final samples and intermediates were analyzed by SDS-electrophoresis on polyacrylamide gel and the presence of flagellin was verified...
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New approaches to determination pathophysiological changes in patients with cystic fibrosis
Doušová, Tereza ; Dřevínek, Pavel (advisor) ; Kreslová, Marcela (referee) ; Gayillyová, Renata (referee)
New approaches to determination pathophysiological changes in patients with cystic fibrosis Cystic fibrosis (CF) is a life-limiting disease caused by mutation in the cystic fibrosis transmembrane regulator (CFTR) gene. To date, more than 2,000 mutations in the CFTR gene have been described, of which only 360 are directly related to CF. In a group of patients carrying mutations of unknown or variable clinical significance, it may be difficult not only to diagnose CF but also to facilitate clinical studies to determine the efficacy of new low - molecular weight compounds targeting disrupted CFTR protein. These so-called CFTR modulators have opened a new era in causal treatment of CF. To maximize the effect of these new therapies, not only the patient's genotype, but also the individual rate of response is crucial. In recent years, intestinal organoids have been used as an ex vivo model to determine the degree of CFTR function and at the same time to predict the therapeutic response to available therapeutic molecules. In our project, using the patient's native tissue and cultures of intestinal organoids derived from this tissue, we demonstrated varying degrees of CFTR residual function in a total of 14 patients with CF (0-39.7% of healthy control function). We characterized de novo mutation of the CFTR gene in...
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Cardiovascular Risks in Chronic Airway Disease in Childhood
Kreslová, Marcela ; Sýkora, Josef (advisor) ; Fila, Libor (referee) ; Kopřiva, František (referee)
1 Cardiovascular risks in chronic airway disease in childhood The aim of this thesis was to evaluate cardiovascular risk by using a combined diagnostic approach by measuring RHI and specific biochemical markers in patients with chronic respiratory disease, where we could assume a possible risk of CVD. A total of 119 probands were examined, including 22 patients with cystic fibrosis (CF) and 52 asthma patients. We evaluated RHI using a new plethysmographic method that has a number of advantages over the ultrasonographic methods used in other studies, including non-invasiveness, high sensitivity, low biological variability and objectivity due to automatic processing. Of the biochemical parameters, we measured 4 biomarkers in relation to endothelial dysfunction (ED): hsCRP, ADMA, E-selectin, and VCAM-1. We compared RHI and biomarkers in CF and asthma patients with healthy controls and sought mutual correlations. We did not prove a statistically significant difference in RHI between the test groups with CF children but we confirmed the decreasing trend of RHI since adolescence and significantly lower RHI values in CF adults, confirming the progressive development of atherogenesis and worsening of ED with age. Biochemical parameters showed significantly higher levels of hsCRP, sVCAM-1 and E-selectin in CF...
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Use of chicken antibodies against PAIIL lectin for prevention of Pseudomonas aeruginosa infections in patients with cystic fibrosis
Kubíčková, Božena ; Hodek, Petr (advisor) ; Anzenbacherová, Eva (referee) ; Eckschlager, Tomáš (referee)
Cystic fibrosis (CF) is one of the relatively common inherited diseases caused by a mutation in the gene encoding for CFTR protein, which forms a chloride channel that significantly affects ion homeostasis and the associated fluid management of the cell. This disease mainly affects the respiratory and digestive systems, being the most life-threatening in the respiratory tract. Patients with CF suffer from frequent and recurrent respiratory infections that lead to the development of chronic inflammation and gradual destruction of lung tissue. These lung infections, which are caused mostly by the opportunistic pathogen Pseudomonas aeruginosa, are the most common cause of morbidity and mortality in these patients. At present, antibiotics are used in the treatment of Pseudomonas aeruginosa infections, but new methods of antibacterial therapy need to be found to overcome the development of resistance. In addition to active immunization of CF patients against Pseudomonas aeruginosa, their passive immunization with specific chicken antibodies directed against this pathogen offers promising possibilities. This dissertation thesis is aimed to verify the prophylactic potential of hen IgY antibodies against the virulence factor Pseudomonas aeruginosa - lectin PAIIL, and to further develop an experimental...
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Interaction of bacterial lectins with human lung epithelium
Vyhnalová, Kateřina ; Hodek, Petr (advisor) ; Nosková, Libuše (referee)
Recessive autosomal disease cystic fibrosis (CF) is caused by a mutation in the CFTR gene ("regulator of cystic fibrosis transmembrane conductance"), which encodes the same named chloride channel. This mutation leads to incorrect ion transport, which causes the formation of an excessively viscous mucus on the surface of the airways and subsequently to the susceptibility to bacterial diseases. This disease mainly affects the respiratory system, where infections are associated with various causes of death in patients with CF. The most common pathogen causing infections is Pseudomonas aeruginosa (PA), which uses many virulence factors, such as pili or adhesins. Lectin PA-IIL, from the group of PA adhesins, is characterized by a high affinity for L-fucose, so it contributes to the adhesion of PA to the low sialylated epithelium of CF patients. In this work the interactions between PA-IIL and lung epithelium were investigated. The cell lines CuFi-1 (CF patient) and NuLi-1 (healthy individual), which were examined ex vivo, were used. A part of these cell lines were exposed to neuraminidase. The PA-IIL lectin was isolated from the E. coli cell line pET25_PAIIL and subsequently fluorescently labeled with DyLight 488. The activity of mentioned lectin was verified by red blood cell agglutination. The...
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Quality of life in patiens with cystic fibrosis
Šebková, Iveta ; Mádlová, Ivana (advisor) ; Raudenská, Jaroslava (referee)
Bachelor thesis "Quality of life in patients with cystic fibrosis" deals with difficulty about living with this diagnosis. The work is divided into theoretical and empirical part. The theoretical part describe disease cystic fibrosis and deals with the clarification of basic terms related to this disease, symptoms and treatment. Empirical part is processed on the basis of quantitative research using of short version WHOQOL - BREF questionnaire, which we have changed for needs of our thesis and filled in with question of our construction. Goal was to find out impact of this desease on profesional and personal live of our respondent, evaluate the level of quality of their life and satistfaction with care. Findings and evaluation defined goals are at the end of thesis.
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Attitudes of parents and teachers in the education of a pupil with a chronic disease at the 1 st stage of a regular primary school
Vopatová, Kristina ; Zulić, Marija (advisor) ; Hájková, Vanda (referee)
The diploma thesis deals with the attitude of parents and teachers of pupils with chronic diseases in the attenuation of compulsory school attendance at the 1st stage of ordinary primary school. The objective of the work is to identify the problems that parents and teachers of chronically ill students encounter during the educational process. From this basis, the purpose is also to find the basis for the best possible cooperation between parents and teachers. The theoretical idea is represented by individual chronic diseases, namely asthma, cystic fibrosis, pediatric oncological diseases, type 1 diabetes mellitus and epilepsy. In addition, the legislative framework for the education of pupils with these diseases, the methodology of working with pupils with chronic diseases and a search of domestic and foreign research conducted on a similar topic are addressed. The methodological part is devoted to qualitative research, the main goal of which is to find out the attitudes of teachers and parents to the educational process of pupils suffering from the above diseases. Using a questionnaire survey and a semi-structured interview, it is ascertained what are the attitudes of parents and teachers towards the education of pupils with the above-mentioned types of chronic diseases in ordinary primary school....
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Inclusion of pupil with cystic fibrosis in physical education
Kyndlová, Tereza ; Daďová, Klára (advisor) ; Majorová, Simona (referee)
Title: Inclusion of a pupil with cystic fibrosis in physical education Objectives: The aim of this thesis was to point out through case studies the possibilities of inclusion of a pupil with cystic fibrosis in physical education and to investigate the influence of physical applied activities on pupil's physical abilities. Part of the research was also working with a girl in physical education classes. Methods: Due to the uniqueness of the disease, was chosen for this thesis the method of a case study of schoolgirl aged 12 with CF. Methods of data collection included interviews with pupil's mother, teacher and teaching assistant, questionnaires, observations and MABC-2 test set. Results: We found out, that the inclusion of a pupil with CF is possible, but it is necessary to have prepared alternative methods and modification of physical activities in order not to damage the health of the individual. The individual approach to the student in class also proved to be effective. The research shows that physical activities as a part of physical education have a positive effect on some motor skills of a pupil with CF. Keywords: integration, cystic fibrosis, joint education, primary school, physical activities
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Biochemistry and pathobiochemistry of phylloquinone and menaquinones
Dunovská, Kateřina ; Klapková, Eva (advisor) ; Uřinovská, Romana (referee) ; Skoumalová, Alice (referee)
Vitamin K belongs to the family of fat-soluble vitamins, which is not determinated in clinical laboratories. It is a cofactor necessary for posttranslational γ-carboxylation of glutamyl residues in selected proteins such as the osteocalcin, and procoagulation factors II, VII, IX, X. Vitamin K deficient individuals appear to have more undercarboxylated proteins, which are functionally defective. Lack of this vitamin has been associated with risk of developing osteoporosis and cardiovascular diseases. The aim of this work was to develop and validate the HPLC method and the LC-MS/MS method for determination of three vitamin K's forms - vitamin K1, MK-4 and MK-7 in serum. After successful validation of both methods, patient samples and healthy population samples were measured. There were measured 350 patient samples by HPLC method. These samples were divided into two groups - patients with diagnostic of osteoporosis and patients without osteoporosis. We measured 946 samples by LC-MS/MS method. Samples were divided into groups: patients with osteoporosis, patients without osteoporosis, healthy population, patients with osteopenia and patients with cystic fibrosis. The reference range of vitamin K in healthy population was obtained by LC-MS/MS method. The next aim was to compare the effectiveness of...
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