Tato práce se zabývá automatickou detekcí vysokofrekvenčních oscilací jakožto moderního elektrofyziologického biomarkru epileptogenní tkáně v intrakraniálním EEG, jehož vizuální detekce je zdlouhavý proces, který je ovlivněn subjektivitou hodnotitele. Epilepsie je jedním z nejčastějších neurologických onemocnění postihující 1 % obyvatelstva. Přestože jsou přibližně dvě třetiny případů léčitelné farmakologicky, zbylá třetina pacientů je odkázána zejména na léčbu chirurgickým zákrokem, pro nějž je zapotřebí přesně lokalizovat ložisko patologické tkáně. Vysokofrekvenční oscilace jsou v posledním desetiletí studovány pro jejich potenciál lokalizace patologické tkáně. Součástí této práce je shrnutí dosavadního výzkumu vysokofrekvenčních oscilací a výčet detektorů používaných ve výzkumu. V rámci práce byly vyvinuty či vylepšeny tři detektory vysokofrekvenčních oscilací, na jejichž popis navazuje evaluace z hlediska shody s manuální detekcí, přesnosti výpočtu příznaků oscilací a schopnosti lokalizace patologické tkáně. V závěru práce jsou představeny vyvinuté metody vizualizace vysokofrekvenčních výskytu oscilací a stručně uvedeny dosažené vědecké výsledky.

The Human brain consists of mutually connected neuronal populations that build anatomically and functionally separated structures. To understand human brain activity and connectivity, it is crucial to describe how these structures are connected and how information is spread. Commonly used methods often work with data from scalp EEG, with a limited number of contacts, and are incapable of observing dynamic changes during cognitive processes or different behavioural states. In addition, connectivity studies almost never analyse pathological parts of the brain, which can have a crucial impact on pathology research and treatment. The aim of this work is connectivity analysis and its evolution in time during cognitive tasks using data from intracranial EEG. Physiological processes in cognitive stimulation and the local connectivity of pathology in the epileptic brain during wake and sleep were analysed. The results provide new insight into human brain physiology research. This was achieved by an innovative approach which combines connectivity methods with EEG spectral power calculation. The second part of this work focuses on seizure onset zone (SOZ) connectivity in the epileptic brain. The results describe the functional isolation of the SOZ from the surrounding tissue, which may contribute to clinical research and epilepsy treatment.

The ATP-gated P2X7 receptor (P2X7R) is a non-selective cation channel which senses high extracellular ATP concentrations and has been suggested as a target for the treatment of neuroinflammation and neurodegenerative diseases. The use of P2X7R antagonists may therefore be a viable approach for treating CNS pathologies, including epileptic disorders. Recent studies showed anticonvulsant potential of P2X7R antagonists in certain animal models. To extend this work, we tested three CNS-permeable P2X7R blocker (Brilliant Blue G, AFC-5128, JNJ-47965567) and a natural compound derivative (tanshinone IIA sulfonate) in four well-characterized animal seizure models. In the maximal electroshock seizure threshold test and the pentylenetetrazol (PTZ) seizure threshold test in mice, none of the four compounds demonstrated anticonvulsant effects when given alone. Notably, in combination with carbamazepine, both AFC-5128 and JNJ-47965567 increased the threshold in the maximal electroshock seizure test. In the PTZ-kindling model in rats, useful for testing antiepileptogenic activities, Brilliant Blue G and tanshinone exhibited a moderate retarding effect, whereas the potent P2X7R blocker AFC-5128 and JNJ-47965567 showed a significant and long-lasting delay in kindling development. In fully kindled rats, the investigated compounds revealed modest effects to reduce the mean seizure stage. Furthermore, AFC-5128- and JNJ-47965567-treated animals displayed strongly reduced Iba 1 and GFAP immunoreactivity in the hippocampal CA3 region. In summary, our results show that P2X7R antagonists possess no remarkable anticonvulsant effects in the used acute screening tests, but can attenuate chemically-induced kindling. Further studies would be of interest to support the concept that P2X7R signalling plays a crucial role in the pathogenesis of epileptic disorders.

Zusammenfassung der Arbeit Beteiligung purinerger Rezeptoren am Schmerzgeschehen und an der neuronalen Entwicklung Patrizia Rubini Illes Rudolf-Boehm-Institut für Pharmakologie und Toxikologie, Universität Leipzig Diese kumulative Arbeit umfasst neun Publikationen, welche die Beteiligung purinerger P2-Rezeptoren (P2R) am Schmerzgeschehen sowie der neuronalen Entwicklung belegen. Im Fokus stehen zwei ATP-aktivierte Rezeptor-Kanäle (P2X3R, P2X7R) und ein G-Protein-gekoppelter, ATP-empfindlicher Rezeptor (P2Y1R). (1) Da sich humane P2X3R an peripheren und zentralen Endigungen primärer-sensorischer Afferenzen befinden, ist die Entschlüsselung der Agonisten-Bindungsstelle für die Entwicklung potenziell analgetisch wirkender, kompetitiver Antagonisten von großer Bedeutung. Wir identifizierten Gruppen konservierter und nicht-konservierter Aminosäure-Reste, die an der Schnittstelle zweier benachbarter Untereinheiten, die Agonisten-Bindungstasche auskleiden. Abhängig vom Mengenverhältnis der zur Transfektion verwendeten cDNA, bilden P2X2-, P2X3- und P2X6-Untereinheiten heteromere Rezeptoren (P2X2/3R, P2X2/6R) in einer variablen Stöchiometrie. (2) Adulte neurale Progenitorzellen (NPZ) der Subventrikulärzone der Maus besitzen Apoptose- bzw. Nekrose-vermittelnde P2X7R. Diese kommen in Assoziation mit P2X4R vor, bilden jedoch wahrscheinlich keinen heteromeren P2X4/7R. P2X7R könnten zum Absterben der, nach metabolischen Schäden im Überschuss gebildeten, NPZ beitragen. Im Laufe der neuronalen Entwicklung verlieren die NPZ ihre P2X7R. Im Gegensatz hierzu bleiben diese bei ihren astrozytären Nachkommen erhalten. (3) Humane mesencephale embryonale NPZ sind mit P2Y1R ausgestattet, die über eine Ca2+-Wellenaktivität die Proliferation der NPZ beschleunigen und in Differenzierungsvorgänge eingreifen. De-differenzierte, kultivierte Neurone des Ratten-Striatum enthalten ebenfalls P2Y1R. Diese setzen Ca2+ aus seinen intrazellulären Speichern frei und öffnen somit Ca2+-selektive „store-operated channels“ in der Zellmembran. Wir konnten zeigen, dass der P2Y1R-stimulierte Anstieg der intrazellulären Ca2+-Konzentration in den striatalen Neuronen durch die Aktivierung von Dopamin D1- und D2R inhibiert wurde. Die vorliegende Arbeit belegt die Bedeutung einiger P2R für zahlreiche, auch potentiell therapeutisch bedeutsame Funktionen des menschlichen und tierischen Organismus.:Inhaltsverzeichnis Kapitel Seite Abkürzungen …………………………………………………………………………………. 5 1. Einführung in die Thematik.......................................................................... 7 1.1. Freisetzung von Nukleotiden und ihre Metabolisierung im Extrazellulärraum …7………………………………… 1.2. P2X-Rezeptoren…………………………………………………………………….. 9 1.3. P2Y-Rezeptoren……………………………………………………………………. 12 1.4. P2-Rezeptoren und Schmerz……………………………………………………... 14 1.5. P2-Rezeptoren und neuronale Entwicklung; embryonale undadulte neurale Vorläuferzellen……………………………………………………. 17 1.6. Referenzen………………………………………………………………………….. 19 2. Wissenschaftliche Ergebnisse……………………………………………………… 23 2.1. Komplex I. Struktur-Wirkungs-Zusammenhänge bei rekombinanten, humanen P2X3- und P2X2/3-Rezeptoren in Expressionssystemen…………. 24 2.1.1. Amino acid residues constituting the agonist binding site of the human P2X3 receptor……………………………………………………………… 24 2.1.2. ATP binding site mutagenesis reveals different subunit stoichiometry of functional P2X2/3 and P2X2/6 receptors……………………………………... 26 2.1.3 Flexible subunit stoichiometry of functional human P2X2/3 heteromeric receptors……………………………………………………………... 28 2.2. Komplex II. Vorhandensein und Wirkung von P2X7-Rezeptoren an kultivierten adulten, neuralen Vorläuferzellen und Astrozyten………….. 29 2.2.1. P2X7 receptors at adult neural progenitor cells of the mouse subventricular zone………………………………………………………………… 29 2.2.2. Co-expression of functional P2X4 and P2X7 receptors at adult neural precursor cells of the mouse subventricular zone……………………… 31 2.2.3. Functional P2X7 receptors at cultured hippocampal astrocytes but not neurons…………………………………………………………………….. 32 2.3. Komplex III. Vorhandensein und Wirkung von P2Y-Rezeptoren an embryonalen neuralen Vorläuferzellen und dedifferenzierten kultivierten striatalenNeuronen... 33 2.3.1. Increase of intracellular Ca2+ by adenine and uracil nucleotides in human midbrain-derived neuronal precursor cells…………... 33 2.3.2. Regulation of intracellular Ca2+ by P2Y1 receptors may depend on the developmental stage of cultured rat striatal neurons…………………... 35 2.3.3. Modulation by D1 and D2 dopamine receptors of ATP-induced release of intracellular Ca2+ in cultured rat striatal neurons 36 3. Zusammenfassung und Ausblick………………………………………… ………... 37 4. Erklärungen, Danksagung…………………………………………… 38 Erklärung über die eigenständige Anfertigung der Arbeit………… 38 Erklärung über den eigenen Anteil an den einzelnen Arbeiten…………….. 39 Danksagung………………………………………………………………… 42

Jakub Žalud PhDr. Alena Javůrková Ph.D. adverse effects of antiepileptics, their classification and expression from the patient's subjective assumption was that the patient's experience of the adverse effects of antiepileptic drugs will significant overall (p˂0.05; R2 = 0.20). ts was not statistically significant (p˃0.05; R2 = 0.21). (p˃0.05; R2 = 0.16).

The Diploma thesis is focused on attitudes of helping profession to epilepsy. The aim of theoretical part is to introduce life with epilepsy and to increase public awareness of the illness. As a part of the thesis there is refuting myths and prejudices that are often cause of negative attitudes. Helping professions are a main part of life both of epileptics and their families as well their surroundings and their attitudes affect most of it so it is main sample of the research. Theoretical part is divided into few chapters. In the initial chapter there are defined basic concepts important for good orientation in the topic - stereotypes, prejudices and attitudes. Its theories, functions and changes are further described more in detail. Next chapters describe epilepsy from medical and social aspects such as Etiology of epilepsy, epileptic seizures classification, its diagnostics and treatment are mentioned there. First aid during epileptic seizures and role of social workers are not missing. Further diploma thesis covers problematics such as social stigmatization and myths connected with epilepsy. From wider perspective impact of epilepsy on social and personal life is revealed to readers. Research part of the diploma thesis was created by quantitative method using questionnaire survey. The aim of...

The steroid substances with effect on nervous tissue are plenteously studied in last years. Their biggest benefits are especially anticonvulsant and anxiolytic effects. This thesis inspects the influence of newly discovered neruactive steroid pregnanolone pyroglutamate synthesized in The Institute of Organic Chemistry and Biochemistry of the Czech Academy of Science (IOCB CAS), which offer potential opportunity of a new therapy of epilepsy from actual pharmacokinetic results. Our goal was to detect, if this substance has any negative effects on locomotor skills of young laboratory rats. In theoretical part of the thesis the development of laboratory rat is briefly descibed, especially its motor skills and sensory functions in comparison with human. The process of the experiments and choice of especially motor skill tests are described in Metogology part. The results of experiment and its comparation with the effect of other neuromediators are introduced and discussed in the practical part of this thesis. There were no serious effects of pregnanolonepyroglutamate applied repeatedly in perinatal period on motor performance. Acute administration of the highest dose of PPG to 12-day-old rat pups resulted in prolongation of time in surface righting and negative geotxis tests. This data are positive for...

Title: Assessment of the attachment style as a part of the differential diagnosis of psychogenic non-epileptic seizures (PNES) Author: Bc. et Bc. Jana Heidingerová Thesis supervisor: doc. PhDr. Lenka Krámská, Ph.D. Pages: 101 Number of sources: 93 The thesis deals with the topic of psychogenic non-epileptic seizures (PNES) and their differential diagnosis in the context of attachment and its measuring. Based on theoretical knowledge, the relation between attachment disorders and the diagnosis of PNES was assumed. This connection was examined by comparing the results of three groups of respondents with different diagnoses (PNES, PNES and epilepsy, epilepsy) and a control group. The theoretical part deals with the problematics of attachment, PNES and epilepsy. The empirical part examines differences between groups of respondents, correlations of the variables with socio-demographic factors and correlations between results of each used method. Three questionnaires, PBI, ECR and ECR-RS were used for the research. The data were collected in the Epilepsy Centre at the Na Homolce Hospital. The research sample consisted of a total of 84 respondents. The results were statistically significant for numerous hypotheses. Research suggest that there are correlations of some subscales with socio-demographic...

Ce travail de thèse explore la mémoire de stimuli émotionnels au cours de quatre études en adoptant une approche pluridisciplinaire chez l’enfant sain et consécutivement à une chirurgie de l’épilepsie du lobe temporal (ELT). Nos objectifs étaient (1) de comparer l’influence des émotions sur la mémoire verbale et non verbale au cours du développement sain et en cas d’ELT, (2) de décrire les bases cérébrales des processus de la mémoire de visages exprimant la peur au cours du développement au moyen de l’IRMf, (3) de s’intéresser à l'impact de l’ELT droit sur ce réseau (4) d’illustrer l’impact d’une chirurgie de l’ELT droite sur la mémoire émotionnelle et les caractéristiques cognitivo-émotionnelles en phase pré- puis post-opératoire. Nos résultats montrent que La reconnaissance mnésique de stimuli émotionnels est perturbée chez les jeunes patients présentant un dysfonctionnement du LTM, sauf pour les visages exprimant la peur. Chez les sujets sains, l’activation de l’amygdale basolatérale qui se met en place au moment de l’adolescence, serait notamment la signature cérébrale du phénomène de modulation émotionnelle des souvenirs associée à la recollection, et la maturation fonctionnelle des structures de la mémoire au sein du lobe temporal médian (LTM) suivrait un gradient caudo-rostral. Des capacités de réorganisation controlatérale sont néanmoins observées chez les patients avec ELT droite, au niveau de l’amygdale et des structures mnésiques du LTM, avec une sur-compensation au niveau du cortex parahippocampique. Ces adaptations permettraient de soutenir la mémoire de visage exprimant la peur sur la base d’un sentiment de familiarité, notamment après le contrôle des crises. Ce travail de thèse a permis de mettre en évidence des résultats novateurs à propos de l’implication du LTM du développement du lien entre mémoire et émotion. / In this multidisciplinary work, four studies were conducted to examine the memory for emotional stimuli in healthy children and post-surgery for temporal lobe epilepsy (TLE). The aims were (1) to compare emotional influences on memory for faces and words in healthy and TLE children, (2) to explore age-related neural networks of fear faces memory with fMRI, (3) to elicit the effect of childhood right-TLE on these developing networks and (4) to illustrate the impact of right-TLE surgery on emotional memory and cognitive-emotional features in a pre- vs. post-surgery case study. Our results show that patients suffering from a MTL dysfunction are impaired in emotional memory, except for fear faces. In heathly participants, emotional modulation of recollected memories is associated with an activation of basolateral amygdala in adolescents and that functional maturation through the mesial temporal lobe (MTL) is characterized by a caudo-rostral gradient. In right-TLE patients, controlateral recovery abilities are nonetheless observed, in amygdala and memory structures in MTL, with an over-activation in parahippocampal cortex. This reorganization would allow sustaining memory for fear faces supported with familiarity process. This thesis highlights new results about MTL involvement in memory-emotions interactions during development.

In its general section, the thesis 'Rehabilitation of Pediatric Patients after Epilepsy Surgery with Postoperative Motor Deficits' discusses epilepsy, treatment of intractable epilepsy and epilepsy surgery. Furthermore, a chapter on possible complications of epilepsy surgery in relation to the locomotor apparatus is included in the general section. The special section deals with rehabilitation in neurology, selected physiotherapy concepts and the relationship between epilepsy and movement activity. The aim of this thesis is to assess the effectiveness of the chosen rehabilitation procedure in five pediatric patients with post-operative motor deficiency. The Gross Motor Function Measure - 88, the Barthel Index and the Box and Blocks Test were used for objective motor assessment.