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Study of biomolecular interactions by the method of surface plasmon resonance
Bocková, Markéta ; Homola, Jiří (advisor)
Title: Study of biomolecular interactions by the method of surface plasmon resonance Author: Markéta Bocková Department: Department of Biochemistry, Faculty of Science Supervisor: Prof. Jiří Homola, Ph.D., DSc., Institute of Photonics and Electronics of the Czech Academy of Sciences Abstract: In this diploma work, a surface plasmon resonance (SPR) biosensor technology was exploited to study two different classes of molecular interactions: hybridization of partially complementary oligonucleotides (ONs) and interactions between enzymes and peptides involved in pathogenesis of Alzheimer disease (AD). In particular, the study of interactions between ONs was focused on determining kinetic parameters of the hybridization proces. It was found that the association equilibrium constant decreases with the increasing surface probe density. An SPR biosensor-based method for rapid and sensitive detection of the 17-HSD10 enzyme and its complex with amyloid β (Aβ) peptides was designed and developed. It was demonstrated that the developed method is capable of detecting 17β-HSD10/Aβs complex in real cerebrospinal fluid (CSF) samples and that CSF samples from AD patients exhibited elavated levels of 17β- HSD10/Aβs complex compared to the control groups. The research reported in this thesis resulted in two publications in...
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Retinal nerve fiber layer measurement in patients with Alzheimer's disease
Kasl, Zdeněk ; Jirásková, Naďa (advisor) ; Pašta, Jiří (referee) ; Skorkovská, Šárka (referee)
Retinal nerve fiber layer measurement in patients with Alzheimer's disease The current ophthalmologist's possibilities in diagnostics of Alzheimer's disease Alzheimer disease (AD) is the most common cause of dementia syndrome and mild cognitive impairment. To enroll the disease most securely there are used so called biomarkers using evidence of changed brain metabolism by pozitron emission tomography (PET) and in cerebrospinal fluid or the brain's structure magnetic resonance imaging (MRI). These methods are expensive, organisationally and temporally challenging and burdening for the patients. According to that reasons we are still seeking for alternative attitudes suitable for early diagnosis. The evaluation of thickness of retinal nerve fiber layer (RNFL) which is well accessible to examination through optical aparatus of the eye could be one of the options. The aim of our work was to present current knowledges about Alzheimer's disease targeting relations of Alzheimer's disease and an ophthalmological finding. In the next part of this paper we introduce the retinal nerve fiber layer measurement by optical coherence tomography (OCT) as a potential diagnostics method by screening of patients with Alzheimer's disease and to present our results measured in our cohort of patients. The studied cohort...
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Yeast as the model for for neurodegenerative diseases
Motyčková, Věra ; Schierová, Michaela (advisor) ; Zikánová, Blanka (referee)
Yeasts are traditional model organisms, which can be, surprisingly, used for studying neurodegenerative diseases such as Parkinson, Alzheimer and Huntington diseases. These diseases affect mostly elder population. It's because of decrease of cell control. Common denominator of these diseases are impaired vesicular transport, protein synthesis and degradation, increasing oxidative stress and ROS. The main cause of neurodegenerative diseases is protein misfolding, which provokes cytotoxic protein aggregation. Modified yeast strains can be used to identify genes and proteins that affect cytotoxicity of these key proteins. If these genes have human orthologues, they could be therapeutic targets. The aim of this work is to summarize methods that study causes of neurodegeneration in yeast and to describe the main results discovered in yeast models. Key words: yeast, model system, Parkinson disease, Alzheimer disease, Huntington disease
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Impact of antidiabetic substances to development of insulin resistance and neurodegenerative changes in mouse models of type 2 diabetes
Mikulášková, Barbora ; Čabala, Radomír (advisor) ; Kuneš, Jaroslav (referee)
Numerous epidemiological and experimental studies have shown that patients suffering from metabolic disorders such as type 2 diabetes mellitus (TDM2), insulin resistance or obesity are at a higher risk of cognitive functions impairment and developing Alzheimer's disease (AD). Impairment of insulin signalling in the brain could contribute to two pathological changes which leads to AD development that include insoluble senile plaques and neurofibrillary tangles, containing an abnormally hyperphosphorylated tau protein (Tau). This work is focused on investigating of insulin signaling in hippocampi in the brains of mice models of insulin resistence, impact of disturbed insulin signaling on hyperphosphorylation of Tau, and possible benefical efect of insulin sensitizing agents on insulin signaling and Tau phosphorylation in the hippocampi of diabetic mice. The first, we examined insulin signaling and phosphorylation of Tau in hippocampi in two mouse models of TDM2 - lipodystrofic A-ZIP F-1 mice and monosodium glutamate obese mice (MSG mice). We did not observe any changes in insulin signaling and Tau phosphorylation in hippocampi of A-ZIP F-1 mice compared to controls. In the hippocampi of MSG mice there was attenuated phosphorylation of kinases of insulin signalling including Ser9 of glycogen synthase...
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Central insulin signaling and phosphorylation of Tau protein in rat model of type 2 diabetes
Kasperová, Barbora Judita ; Čabala, Radomír (advisor) ; Mládková, Jana (referee)
Diabetes mellitus 2. typu (DM2) a Alzheimerova nemoc (AN) jsou jedny z nejzávažnějších nemocí naší doby. Právě DM2, spolu s obezitou a nedostatkem pohybu jsou jedny z nejrizikovějších faktorů vedoucí k poruchám kognitivních funkcí a rozvoji demence. Při rozvoji AN dochází k patologickým změnám v mozku. Dochází ke vzniku nerozpustných extracelulárních plaků amyloidního beta peptidu a hyperfosforylaci proteinu Tau. Tato bakalářská práce studuje inzulínovou signalizační kaskádu a fosforylaci proteinu Tau v hipokampech 12ti týdenních ZDF (Zucker Diabetic Fatty) potkanů, zvířecím modelu DM2, v brzkém věku života. Z inzulínové signalizační kaskády bylo detekováno signifikantně nižší množství inzulínového receptoru v hipokampech ZDF potkanů ve srovnání s kontrolami. U ostatních kinas této kaskády (PDK-1, Akt) nebyl nalezen rozdíl jak v celkovém, tak fosforylovaném/aktivovaném proteinu v hipokampech ZDF potkanů a kontrol. Rovněž u fosforylace/aktivace kinas proteinu Tau (GSK-3β, ERK, JNK) a fosforylace/aktivace proteinu Tau na několika epitopech (Ser396, Thr212, Thr231) nebyly pozorovány žádné signifikantní rozdíly mezi ZDF potkany a kontrolami. V hipokampech 12ti týdenních ZDF potkanů, ačkoliv silně hyperglykemických a obézních, nebyly zaznamenány změny v efektivitě inzulínové signalizační kaskády ani...
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Comparison of the Montreal Cognitive Assessment and the Mini-Mental State Examination in screening diagnostics of Alzheimer's disease
Orlíková, Hana ; Kulišťák, Petr (advisor) ; Hrachovinová, Tamara (referee)
This thesis deals with the neuropsychological diagnosis of Alzheimer's disease. The aim is to evaluate the psychometric characteristics ofthe new Czech translation of the Montreal Cognitive Assessment (MoCA) by comparison with the Mini-Mental State Examination (MMSE), a method widely used by doctors inscreening diagnostics of Alzheimer's disease. The theoretical part deals with the diagnostics of Alzheimer's disease and mild cognitive impairment. We describe international diagnostic criteria of cognitive disorders and provide an overview of the screening neuropsychological methods most commonly used by Czech specialists. We summarize the current psychometric and psychodiagnostic findings on these methods and focuse on description of MMSE and MoCA. In the empirical part we compare Czech version of MMSE and MoCA-CZ (the new Czech translation of the test). We examined 38 patients with Alzheimer's disease and 70 cognitively healthy seniors. The results show that MoCA-CZ is sufficiently valid and reliable screening method that accurately distinguishbetween healthy subjects and patiens with Alzheimer's disease. We believe that it can enrich screening tools that are available to Czech experts. Key words: Montreal Cognitive Assessment, Mini-Mental State Examination, Alzheimer's disease, psychodiagnostics,...
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Diagnostics of neurodegenerative diseases by means of Raman spectroscopy
Klener, Jakub ; Kopecký, Vladimír (advisor) ; Matějka, Pavel (referee)
Therapies of neurodegenerative diseases are often very difficult and their success depend on an early diagnose. From that reason we have been developing new diagnostic method for multiple sclerosis and Alzheimer disease by drop coating deposition Raman (DCDR) spectroscopy of cerebrospinal fluid (CSF) in this work. We found out conditions of measurements, where spectra were reproducible and accepted for standard diagnostic practices. We discovered that CSF has fast degradation at a room temperature, which was detectable in spectra after 5 hours, and degradation due to refreezing. DCDR spectra of CSF from individual patients were analyzed by factor and cluster analysis. Multiple sclerosis was manifested by lower intensity of a Raman band at 1080 cm−1 , which is probably connected with more general pathologic state. Spectral changes caused by Alzeheimer disease were more complex and beside changes mentioned above also changes connected with composition and conformation of proteins were identified in regions 1200-1800 cm−1 and 2870-2950 cm−1 . Additionally, we succeeded in distinguishing of young healthy patients from older patients in DCDR spectra. In this work were checked up, that DCDR is good diagnostic method for clinical practices for determining neurodegenerative diseases through the complex...
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