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Determination of the concentration of alpha-1-antitrypsin in the stool by immunoassay method
Plačková, Marie ; Kocna, Petr (advisor) ; Průša, Richard (referee)
Determination of the concentration of α1 - antitrypsin in the stool is a diagnostic indicator of inflammatory diseases of the small and the large intestine, especially malabsorption syndrome. α1 - antitrypsin belongs to the family of plasma proteins with antiproteinase effect. α1 - antitrypsin is synthesized in liver, in small amount in macrophage and is a protease inhibitor of serine proteases sercected from neutrophils. α1 - antitrypsin is acute phase protein. Higher α1 - antitrypsin values are in early phase of inflammation associated with raised CRP and other pozitive acute phase proteins. Fecal α1 - antitrypsin clearance is a sensitive and specific marker of protein loss. For α1 - antitrypsin determination in stool samples ELISA method can be used. ELISA is noncompetetive immunoassay used to detect presence of antibody or an antigen in a sample. The aim of this work was to compare two ELISA sets (Immundiagnostik and Ridascreen) used for determination α1 - antitrypsin in the stool. Then examine stability of α1 - antitrypsin in the stool and in extract prepared from stool in various storing conditions temperature and time. After this establish this method as routine in laboratory. 20 patient stool samples were examined to compare ELISA sets. Samples were suggested to be α1 - antitrypsin...
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Effect of selected inflammatory agents on the osteoclastogenesis
Škubica, Patrik ; Daňková, Pavlína (advisor) ; Hušáková, Markéta (referee)
Introduction: Bone is a highly active tissue throughout life and is a subject to constant remodelling. Main cells responsible for continuous resorption and de novo synthesis of bone matrix are osteoclast, osteoblasts and osteocytes. Osteoclasts are the only known type of cells able to resorb bone. These cells are formed by fusion of precursor cells in bone marrow or peripheral blood in a process called osteoclastogenesis. Formation of osteoclasts may be of importance concerning chronic inflammatory diseases that are linked with higher risk of developing osteoporosis during lifespan. Celiac disease is one of those diseases, which is characterized by destruction of intestinal mucosa after ingestion of gluten by susceptible individuals followed by induction of chronic inflammation. In this work, we focused on the potential role of osteoclastogenesis in the development of osteoporosis in patients with celiac disease and we studied roles of selected inflammatory agents (TNF-α, IL-6, IFN-γ a cfDNA) with supposed or hypothesised effects on osteoclastogenesis. Material & Methods: We obtained plasma and serum samples from newly diagnosed patients with celiac disease, patients on gluten free diet and healthy controls and analysed concentrations of cfDNA and inflammatory cytokines TNF-α, IL-6 and IFN-γ in...
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Microvesicle and exosome detection in immune-related diseases
Šťastná, Evelína ; Drbal, Karel (advisor) ; Fabišik, Matej (referee)
Exosomes (ES) and microvesicles (MV), collectively called extracellular vesicles (EV), are submicroscopic vesicles encapsulated by a phospholipid bilayer. Smaller ES (40 - 100 nm) originate in endosomal compartment, while larger MV (50 - 1000 nm) shed from cell plasma membrane. EV are secreted by all types of cells. They consist of lipids and proteins, but their composition varies according to the cell they originate from. In addition, they differ in the cargo they transport (DNA, RNA and proteins). They occur in every bodily fluid in much higher amounts compared to the original cells themselves, what makes them an attractive and accessible biomarker of autoimmunity diseases, cardiovascular diseases or tumours. For detection of EV, sensitive flow cytometry (FCM) is used, which I am going to compare to alternative methodologies. Part of this work will be description of EV biogenesis and then I will focus on the role of EV in coagulation and inflammation related to autoimmune diseases, more specifically in rheumatoid arthritis (RA).
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Determination of the concentration of alpha-1-antitrypsin in the stool by immunoassay method
Plačková, Marie ; Kocna, Petr (advisor) ; Průša, Richard (referee)
Determination of the concentration of α1 - antitrypsin in the stool is a diagnostic indicator of inflammatory diseases of the small and the large intestine, especially malabsorption syndrome. α1 - antitrypsin belongs to the family of plasma proteins with antiproteinase effect. α1 - antitrypsin is synthesized in liver, in small amount in macrophage and is a protease inhibitor of serine proteases sercected from neutrophils. α1 - antitrypsin is acute phase protein. Higher α1 - antitrypsin values are in early phase of inflammation associated with raised CRP and other pozitive acute phase proteins. Fecal α1 - antitrypsin clearance is a sensitive and specific marker of protein loss. For α1 - antitrypsin determination in stool samples ELISA method can be used. ELISA is noncompetetive immunoassay used to detect presence of antibody or an antigen in a sample. The aim of this work was to compare two ELISA sets (Immundiagnostik and Ridascreen) used for determination α1 - antitrypsin in the stool. Then examine stability of α1 - antitrypsin in the stool and in extract prepared from stool in various storing conditions temperature and time. After this establish this method as routine in laboratory. 20 patient stool samples were examined to compare ELISA sets. Samples were suggested to be α1 - antitrypsin...
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