|
|
Intravital diagnostics of neurodegenerative diseases
Baranová, Soňa ; Holada, Karel (advisor) ; Kaňovský, Petr (referee) ; Žáková, Dana (referee)
Prionopathies, also called transmissible spongiform encephalopathies (TSE) and synucleinopathies are neurodegenerative diseases that are associated with the accumulation of misfolded proteins (prion and α-synuclein) mostly in the central nervous system. To this day, early and definite diagnosis remains unavailable during the patient's lifetime, mainly due to the absence of reliable biomarker which makes clinical diagnosis more challenging. Therefore, the gold standard in diagnostics remains direct post-mortem evaluation of misfolded proteins within brain tissue by western blot and immunohistochemistry. In the recent years, seeding amplification assays (SAAs) like Real-Time Quacking-Induced Conversion (RT-QuIC) emerged for ultra-sensitive ante-mortem diagnosis of neurodegenerative diseases. SAAs exploit ability of pathological misfolded proteins present in patient's samples to change the conformation and initiate aggregation of native recombinant protein substrate by prion-like seeding mechanism. In the presented dissertation thesis, we exploited second-generation RT-QuIC assay (55řC, 700 rpm, cycles of 1 min double-orbital shaking and 1 min incubation) utilizing recombinant hamster shortened prion protein (rHAPrP90-231) to evaluate prion seeding activity in post-mortem TSE (n=38) and non-TSE (n=30)...
|
guest ::
