|
|
Preparation and characterization of chicken antibodies against pathogenic yeast Candida albicans
Vodecký, Matúš ; Hodek, Petr (advisor) ; Dračínská, Helena (referee)
The increase of infections caused by yeast pathogens is an alarming global public health problem. People with compromised immune systems, such as after infection with human immunodeficiency virus or diabetes mellitus, are most at risk. Diagnosis is often inadequate, and treatment relies on a limited selection of antifungal drugs, to which resistance is also increasing. These aspects represent a major problem, as second-choice drugs, which may have toxic side effects (e.g., nephrotoxicity in the case of amphotericin B), must then be applied to resistant pathogens. Given the severity of yeast infections, scientists are trying to find new ways of protection against these pathogens. One option is passive immunotherapy. In this bachelor thesis, antibodies from hen's eggs were chosen for this purpose, as they have the potential for wide therapeutic use. The preparation of hen's egg yolk IgY antibodies is both ethically and economically acceptable. Moreover, egg yolk antibodies show relatively high stability. Polyclonal antibodies were isolated from eggs of hens immunized with either cell wall or whole C. albicans cells in a medium simulating the vaginal environment (VSF). Antibody reactivity was tested by ELISA. The cell walls of C. albicans in VSF medium or YPD medium were used as antigen. The results...
|
|
|
Preparation and characterisation of antibodies against cytochrome P450 1A1
Vlasáková, Kateřina ; Hodek, Petr (advisor) ; Mácová, Iva (referee)
Cancer is the second most frequent cause of death in the Czech Republic. The malignant transformations of the tumour cells are thought to be caused by formation of DNA adducts or by reactive forms of oxygen. One of groups of enzymes, that catalyse the formation of these carcinogenic compounds, are cytochromes P450, for example the type CYP1A1 or CYP1A2. Recently, chemopreventive food supplements that should decrease the risk of cancer have become very popular. However, overusing of these supplements can induce CYP and increase the risk of cancer. Due to CYP proteins cognition could be the pharmacotherapy more efficient. Suitable tool for the study of CYP proteins is "immunoblotting", which utilizes specific bound between an antigen and an antibody. The chicken anti-peptide antibodies were used for this purpose. The goal of this experiment was to obtain chicken anti-peptide antibodies, using a peptide based on the sequence of rat CYP1A1. The chicken antibodies can be used in further research. (In Czech) Keywords: Yolk antibodies, peptide antigen, ELISA, Western blot
|
|
|
Development of a model system to study adhesion of Burkholderia cenocepacia to epithelial lung cells of patients with cystic fibrosis
Volejníková, Anna ; Bořek Dohalská, Lucie (advisor) ; Kubíčková, Božena (referee)
Cystic fibrosis (CF) is a common autosomal recessive disorder that primarily affects epithelial cells in respiratory system. As a consequence of lung damage the patients are chronically colonized by a number of pathogenic microorganisms such as Pseudomonas aeruginosa or Burkholderia cepacia komplex. Due to the seriousness of these bacterial infection and its primary resistance to commonly used antibiotics the suitable therapeutic alternatives are being sought. The passive immunization with yolk antibodies seems to be a suitable alternative. This thesis studied the bacterial adhesion of Burkholderia cepacia, the opportunistic human pathogen, to lung epithelial cells of healthy individuals and patients with CF, respectively. The aim of this study was to develop a model system suitable for this research. Two types of immortalised cell lines have been used for this purpose. CuFi-1 is the cell line derived from patient with CF caused by ∆F508 mutation , NuLi-1 is a healthy epithelial cell line. The strains of B. cenocepacia (ST28 a ST32) were fluorescently labeled with fluorescent cell linker PKH26. Furthermore, the way of fluorescent labeling of cell lines CuFi-1 and NuLi-1 using PKH67 dye was optimized. Using this model system, the adhesion of bacteria to cell line CuFi-1 was up to three times higher...
|
|
|
Preparation and characterisation of antibodies against cytochrome P450 1A1
Vlasáková, Kateřina ; Hodek, Petr (advisor) ; Mácová, Iva (referee)
Cancer is the second most frequent cause of death in the Czech Republic. The malignant transformations of the tumour cells are thought to be caused by formation of DNA adducts or by reactive forms of oxygen. One of groups of enzymes, that catalyse the formation of these carcinogenic compounds, are cytochromes P450, for example the type CYP1A1 or CYP1A2. Recently, chemopreventive food supplements that should decrease the risk of cancer have become very popular. However, overusing of these supplements can induce CYP and increase the risk of cancer. Due to CYP proteins cognition could be the pharmacotherapy more efficient. Suitable tool for the study of CYP proteins is "immunoblotting", which utilizes specific bound between an antigen and an antibody. The chicken anti-peptide antibodies were used for this purpose. The goal of this experiment was to obtain chicken anti-peptide antibodies, using a peptide based on the sequence of rat CYP1A1. The chicken antibodies can be used in further research. (In Czech) Keywords: Yolk antibodies, peptide antigen, ELISA, Western blot
|
guest ::
