1 Abstract Title: Consequences of experimentally induced perinatal cerebral ischemic lesion. Aim: This thesis aims to present the issues and analyze current knowledge in the field of perinatal brain damage, particularly perinatal cerebral ischemic lesions as the most common brain infarction in children. Basic characteristics of the disease, syndromology, pathophysiological mechanisms and risk factors are emphasized. Following the theoretical background the thesis analyzes current trends as well as the limitations of the diagnostics and therapy. The aim is to highlight the need for early diagnosis and emphasize the potential of preventive treatment strategies. The practical part follows the theoretical background and expands the topic on the analysis of motor and behavioral consequences of experimentally induced perinatal ischemic stroke. Method: Cerebral focal ischemia was induced experimentally by photothrombotic method to seven days old rats. Rose Bengal intravenously followed by continual illumination of the senzorimotor cortex using a green laser beam for 10 minutes was induced platelet aggregation and subsequent thrombosis. Eight animals at age of two months were evaluated for 7 days via observational cages PhenoTyper. Results: The theoretical part collected and processed theoretical data on the...

O camundongo mutante denominado bate palmas (BALB/cbapa) originou-se de mutagênese química induzida por N-ethyl-N-nitrosourea (ENU). Suas principais características fenotípicas são alterações posturais com movimentos anormais dos membros posteriores quando suspenso pela cauda, simulando o ato de bater palmas. A partir do padrão de herança a mutação foi identificada como autossômica recessiva. Os resultados do sequenciamento do exoma apontaram como forte candidato o gene lysine (K)-specific methyltransferase 2D (kmt2d, também conhecido como mll2 ou mll4), localizado no cromossomo 15 do camundongo. Essa mutação foi confirmada através de sequenciamento do DNA pelo método de Sanger. A perda da função do gene KMT2D, localizado no cromossomo 12 em humanos, foi descrita como responsável pela síndrome de Kabuki, que é uma anomalia congênita rara, autossômica dominante, também conhecida como síndrome Niikawa-Koruki. O fenótipo clínico da doença é variável, mas algumas características mais comuns são face dismórfica, anormalidades esqueléticas, alterações dermatoglíficas, leve a moderado retardo mental e retardo do crescimento pós-natal. O presente estudo teve como objetivo caracterizar o fenótipo do camundongo mutante bate palmas a partir da análise de parâmetros comportamentais. Para tanto, foram utilizados 30 camundongos mutantes bate palmas- sendo 15 machos e 15 fêmeas- e seus respectivos controles BALB/c, com oito semanas de idade no início dos experimentos. Os testes comportamentais foram realizados em ordem crescente de estresse e ordenados na seguinte sequência 1) observação direta em campo aberto dos parâmetros da atividade geral, sensoriais, psicomotores e os ligados ao sistema nervoso central e autônomo; 2) ansiedade e memória no teste do labirinto em cruz elevada; 3) memória operacional no teste do labirinto em T; 4) coordenação motora no teste da trave elevada e 5) comportamento do tipo depressivo nos testes de suspensão pela cauda e natação forçada. Os dados obtidos no presente estudo demostraram que o mutante bate palmas apresentou aumento na frequência de levantar em ambos os sexos, sugerindo comportamento estereotipado; no entanto, não houve alteração na atividade geral. Na avaliação do sistema sensorial houve redução no reflexo auricular e na resposta de aperto de cauda do mutante bate palmas. Em relação aos parâmetros psicomotores houve diminuição do reflexo de endireitamento e queda do trem posterior, sugerindo deficiência motora, como a hipotonia. Os resultados do teste de labirinto em cruz elevada identificaram menor nível de ansiedade nos mutantes em comparação aos controles. Ainda, as respostas observadas no segundo dia do teste mostraram que não houve perda de memória dos bate palmas. Além disso, o teste do labirinto em T mostrou que não houve alteração na memória espacial dos camundongos mutantes em relação aos BALB/c. Já os resultados dos testes de suspensão pela cauda e de natação forçada foram semelhantes, indicando maior tempo de imobilidade dos mutantes em comparação aos BALB/c. Analisados em conjunto, os resultados desse estudo sugerem que o mutante bate palmas apresentou alterações nos parâmetros sensoriais e psicomotores e possível comportamento estereotipado, relacionadas à mutação do gene kmt2d. / The mutant mouse denominated bate palmas (BALB/cbapa) originated from chemical mutagenesis induced by N-ethyl-N-nitrosourea (ENU). Its main phenotypic characteristics are postural alterations with abnormal movements of the hind limbs when suspended by the tail, simulating the act of clap hands. The inheritance pattern of the mutation was identified as autosomal recessive. The potential candidate gene lysine (K) -specific methyltransferase 2D (kmt2d, also known as mll2 or mll4), located on chromosome 15 of the mouse, was identified by the exome sequencing. This mutation was confirmed by DNA sequencing by the Sanger method. The loss of function of KMT2D gene, located on chromosome 12 in humans, has been described as responsible for Kabuki syndrome, which is a rare congenital anomaly, autosomal dominant, also known as Niikawa-Koruki syndrome. The clinical phenotype of the disease is variable, but some common features are dysmorphic face, skeletal abnormalities, dermatoglyphic changes, mild to moderate mental retardation and postnatal growth retardation. The present study aimed to characterize the phenotype of the mutant mouse bate palmas from the analysis of behavioral parameters. Therefore, 30 mutant mice were used - 15 males and 15 females - and their respective BALB/c controls, at eight-wk-old at the beginning of the experiments. Behavioral tests were performed in increasing order of stress and ordered in the following sequence: 1) direct observation in the open field of the parameters linked to general activity, sensorial and psychomotor systems, and those connected to the central and autonomic nervous system; 2) anxiety and memory in the elevated plus maze test; 3) operating memory in the T-maze test; 4) motor coordination in the balance beam test, and 5) depressive-like behavior in the tail suspension and forced swimming tests. The data obtained in the present study demonstrated that the mutant bate palmas presented increase in the rearing frequency in both sexes, suggesting stereotyped behavior; however, there was no change in the general activity. The evaluation of the sensory system demonstrated reduction in the auricular reflex and the tail flick response of the mutant bate palmas. Regarding to the psychomotor parameters, there were observed impairments in the surface-righting reflex and hindquarter fall, suggesting motor deficiency, such as hypotonia. The results of the elevated plus maze test identified a lower level of anxiety in the mutants compared to controls. Still, the responses observed on the second day of the test showed that there was no loss of memory of the bate palmas. In addition, the T-maze test showed that there was no change in the spatial memory of the mutant mice in relation to BALB/c mice. Data of the tail suspension and forced swimming tests were similar, indicating a longer immobilization time of the mutants compared to BALB/c mice. Taken together, the results of this study suggest that the mutant bate palmas showed impairments of the sensorial and psychomotor parameters, and possibly stereotyped behavior related to the mutation of the kmt2d gene.

O camundongo mutante denominado bate palmas (BALB/cbapa) originou-se de mutagênese química induzida por N-ethyl-N-nitrosourea (ENU). Suas principais características fenotípicas são alterações posturais com movimentos anormais dos membros posteriores quando suspenso pela cauda, simulando o ato de bater palmas. A partir do padrão de herança a mutação foi identificada como autossômica recessiva. Os resultados do sequenciamento do exoma apontaram como forte candidato o gene lysine (K)-specific methyltransferase 2D (kmt2d, também conhecido como mll2 ou mll4), localizado no cromossomo 15 do camundongo. Essa mutação foi confirmada através de sequenciamento do DNA pelo método de Sanger. A perda da função do gene KMT2D, localizado no cromossomo 12 em humanos, foi descrita como responsável pela síndrome de Kabuki, que é uma anomalia congênita rara, autossômica dominante, também conhecida como síndrome Niikawa-Koruki. O fenótipo clínico da doença é variável, mas algumas características mais comuns são face dismórfica, anormalidades esqueléticas, alterações dermatoglíficas, leve a moderado retardo mental e retardo do crescimento pós-natal. O presente estudo teve como objetivo caracterizar o fenótipo do camundongo mutante bate palmas a partir da análise de parâmetros comportamentais. Para tanto, foram utilizados 30 camundongos mutantes bate palmas- sendo 15 machos e 15 fêmeas- e seus respectivos controles BALB/c, com oito semanas de idade no início dos experimentos. Os testes comportamentais foram realizados em ordem crescente de estresse e ordenados na seguinte sequência 1) observação direta em campo aberto dos parâmetros da atividade geral, sensoriais, psicomotores e os ligados ao sistema nervoso central e autônomo; 2) ansiedade e memória no teste do labirinto em cruz elevada; 3) memória operacional no teste do labirinto em T; 4) coordenação motora no teste da trave elevada e 5) comportamento do tipo depressivo nos testes de suspensão pela cauda e natação forçada. Os dados obtidos no presente estudo demostraram que o mutante bate palmas apresentou aumento na frequência de levantar em ambos os sexos, sugerindo comportamento estereotipado; no entanto, não houve alteração na atividade geral. Na avaliação do sistema sensorial houve redução no reflexo auricular e na resposta de aperto de cauda do mutante bate palmas. Em relação aos parâmetros psicomotores houve diminuição do reflexo de endireitamento e queda do trem posterior, sugerindo deficiência motora, como a hipotonia. Os resultados do teste de labirinto em cruz elevada identificaram menor nível de ansiedade nos mutantes em comparação aos controles. Ainda, as respostas observadas no segundo dia do teste mostraram que não houve perda de memória dos bate palmas. Além disso, o teste do labirinto em T mostrou que não houve alteração na memória espacial dos camundongos mutantes em relação aos BALB/c. Já os resultados dos testes de suspensão pela cauda e de natação forçada foram semelhantes, indicando maior tempo de imobilidade dos mutantes em comparação aos BALB/c. Analisados em conjunto, os resultados desse estudo sugerem que o mutante bate palmas apresentou alterações nos parâmetros sensoriais e psicomotores e possível comportamento estereotipado, relacionadas à mutação do gene kmt2d. / The mutant mouse denominated bate palmas (BALB/cbapa) originated from chemical mutagenesis induced by N-ethyl-N-nitrosourea (ENU). Its main phenotypic characteristics are postural alterations with abnormal movements of the hind limbs when suspended by the tail, simulating the act of clap hands. The inheritance pattern of the mutation was identified as autosomal recessive. The potential candidate gene lysine (K) -specific methyltransferase 2D (kmt2d, also known as mll2 or mll4), located on chromosome 15 of the mouse, was identified by the exome sequencing. This mutation was confirmed by DNA sequencing by the Sanger method. The loss of function of KMT2D gene, located on chromosome 12 in humans, has been described as responsible for Kabuki syndrome, which is a rare congenital anomaly, autosomal dominant, also known as Niikawa-Koruki syndrome. The clinical phenotype of the disease is variable, but some common features are dysmorphic face, skeletal abnormalities, dermatoglyphic changes, mild to moderate mental retardation and postnatal growth retardation. The present study aimed to characterize the phenotype of the mutant mouse bate palmas from the analysis of behavioral parameters. Therefore, 30 mutant mice were used - 15 males and 15 females - and their respective BALB/c controls, at eight-wk-old at the beginning of the experiments. Behavioral tests were performed in increasing order of stress and ordered in the following sequence: 1) direct observation in the open field of the parameters linked to general activity, sensorial and psychomotor systems, and those connected to the central and autonomic nervous system; 2) anxiety and memory in the elevated plus maze test; 3) operating memory in the T-maze test; 4) motor coordination in the balance beam test, and 5) depressive-like behavior in the tail suspension and forced swimming tests. The data obtained in the present study demonstrated that the mutant bate palmas presented increase in the rearing frequency in both sexes, suggesting stereotyped behavior; however, there was no change in the general activity. The evaluation of the sensory system demonstrated reduction in the auricular reflex and the tail flick response of the mutant bate palmas. Regarding to the psychomotor parameters, there were observed impairments in the surface-righting reflex and hindquarter fall, suggesting motor deficiency, such as hypotonia. The results of the elevated plus maze test identified a lower level of anxiety in the mutants compared to controls. Still, the responses observed on the second day of the test showed that there was no loss of memory of the bate palmas. In addition, the T-maze test showed that there was no change in the spatial memory of the mutant mice in relation to BALB/c mice. Data of the tail suspension and forced swimming tests were similar, indicating a longer immobilization time of the mutants compared to BALB/c mice. Taken together, the results of this study suggest that the mutant bate palmas showed impairments of the sensorial and psychomotor parameters, and possibly stereotyped behavior related to the mutation of the kmt2d gene.

Traumatic brain injury (TBI) is a leading cause of disability in the United States (Coronado et al., 2011). There is a recognized need for better motor assessments to help mitigate these disabilities. Advances in markerless motion capture and in magnetic resonance imaging (MRI) provide an opportunity to improve clinical assessments, and link them to damage measured in MRI scans. The primary aims of this research were to 1) develop a quantitative motor assessment (QMA), and seed a normative database to enable comparison of impaired behavior to unimpaired, 2) test the sensitivity of the QMA, and 3) link QMA results to underlying TBI damage.The QMA developed in Aim 1 consisted of five tests: finger oscillation, tremor, visually guided movement, reaction time, and balance. We administered the QMA and traditional analgous tests to 132 healthy 18-50 year olds. We later added a coginitive motor integration (CMI) test and a stength-dexterity pinch test, then administered them to 31 (16 male, mean age = 24.7) healthy individuals. We seeded a normative database for the QMA and CMI measures. (A normative database for the pinch test already exists.) Correlations between the QMA and traditional tests were weak but the QMA results followed expected trends.In the second aim, 31 (16 male, mean age = 24.7 years) individuals with TBI completed all of the motor tests, and age- and gender-matched controls completed the CMI and pinch tests. We tested the sensivity of the QMA, the CMI and pinch tests, and traditional tests by their ability to correctly identify TBI subjects based solely on test results. The QMA was more sensitive than the other test groups. In Aim 3, we performed a stepwise regression to evaluate the relationship between motor deficits and brain injury, using motor test results and MRI images from the TBI and control groups. We found significant relationships between deficits in precision and increases in superior lateral ventricular volumes, deficits in pointing tasks and decreases in fractional anisotropy (FA) in the corticospinal tract, deficits in rhythmicity during finger oscillation and decreased FA in the thalamocortical tract. There were also relationships between each of the motor deficit measures and the FA values in the corpus callosum. This was the first step in showing that a quantitative motor assessment using markerless motion capture is feasible. The QMA is sensitive and can be linked to underlying brain damage. Though the QMA is not yet ready for clinical use, this research provides insights that will help address gaps in TBI rehabilitation.

La restauration fonctionnelle du membre supérieur chez l'hémiplégique séquellaire reste toujours l'objectif primordial recherché durant le processus de réhabilitation. De nos jours, les techniques émergentes de rééducation montrent de plus en plus d'efficacité et de réalisme, si elles sont accompagnées par les techniques conventionnelles et classiques. De plus, quelques idées anciennes ont évoluées vers l'utilisation du renforcement musculaire du membre atteint afin d'améliorer sa fonction motrice. En effet, le renforcement musculaire isocinétique constitue une technique potentiellement intéressante de rééducation pour des hémiplégiques. Dans le cadre de notre travail doctoral, nous avons utilisé le concept isocinétique pour réhabiliter les déficits moteurs et diagnostiquer les déficits de coordination du membre supérieur hémiplégique. En premier lieu, nous avons essayé de montrer le rôle de l'isocinétisme pour optimiser la prise en charge du membre supérieur chez l'hémiplégique. Nous avons d'abord présenté une revue de littérature sur le renforcement musculaire isocinétique après hémiplégie suite à une lésion cérébrale. Ensuite, nous avons évalué la faisabilité d'un programme basé sur ce type de renforcement, pour le coude et le poignet chez des hémiparétiques. Ainsi, l'utilité et l'efficacité de ce type de réentraînement isocinétique ont pu être mises en évidence. En second lieu, nous avons utilisé l'isocinétisme pour comprendre l'implication du rôle des afférences dans le déficit de coordination bimanuelle après accident vasculaire cérébral causant une hémiplégie. Dans l'ensemble, ces différents résultats autorisent à recommander l'outil isocinétique pour quantifier les déficits moteur et de coordination du membre supérieur chez l'hémiplégique, en vue d'une meilleure élaboration et mise en place de protocoles de rééducation isocinétique. / Functional restoration of the upper limb in sequelae hemiplegic remains the primary objective during the rehabilitation process. Nowadays, emerging rehabilitation techniques show more efficiency and realism, if they are accompanied by conventional and standard techniques. In addition, some old ideas have evolved to use the affected limb muscle strengthening to improve motor function. Indeed, the isokinetic muscle strengthening is a potentially interesting technique for rehabilitation of hemiplegic patients. As part of our doctoral work, we used the isokinetic concept to rehabilitate motor deficits and diagnose coordination deficits for hemiplegic upper limb. First, we tried to show the role of isokinetic to optimize the management of upper limb in hemiplegic patients. We firstly presented a review of literature on isokinetic muscle strengthening after hemiplegia following a brain injury. Then, we evaluated the feasibility of a training program based on this type of reinforcement for the elbow and wrist in the hemiparetic patients. Thus, the usefulness and effectiveness of this type of isokinetic exercise training have been highlighted. Second, we used the isokinetic to understand the contribution of afference-based processes to the impairment of bimanual coordination after stroke causing hemiplegia. On the whole, these results allow recommending the isokinetic tool to quantify motor and coordination deficits of upper limb in hemiplegic patients, for better development and implementation of isokinetic rehabilitation protocols.

The aim of the study was to evaluate the effect of specific rehabilitation concept on the motor deficit in patients after stroke. This concept is used at the Department of inpatient rehabilitation FN Motol, the main component is exercise on the principles of dynamic neuromuscular stabilization (DNS) and is based on the principles of developmental kinesiology. The study included 12 patients after stroke. Probands participated in the 3 week therapy, exercise was carried out twice a day. Significant (p <0.05) improvement in motor function of the arm, hand, leg and foot was demonstrated by using Student`s paired t-test. There was a statistically significant improvement in postural control and reducing shoulder pain (all assessed using a specific scale: Chedoke McMaster Stroke Assessment). Changes in the degree of spasticity were evaluated by Modified Ashworth scale. A statistically significant improvement in the degree of spasticity was observed in these muscle groups: the elbow flexors , adductors of lower limb, knee extensors and hamstrings. It has been shown that the concept of dynamic neuromuscular stabilization can be advantageously used in selected patients after stroke. It is appropriate to supplement the DNS therapy with training of ADL.

Les manifestations neuro-ophtalmologiques, observées dans la sclérose en plaques sont parfaitement déterminées à l’heure actuelle. Cependant, l’aspect chronique des troubles visuels résultants n’est pas toujours précisément évalué, or de telles lacunes sont un frein à leur prise en charge. Dans une première partie, les caractéristiques des troubles visuels chroniques ont été mesurées. Il s’agissait de quantifier le pourcentage de plaintes visuelles chroniques chez des patients atteints de sclérose en plaques puis de mesurer le degré d’intensité des troubles visuels chroniques, de déterminer leurs origines physiopathologiques et de rendre compte de leur retentissement sur la qualité de vie des patients se plaignant de troubles chroniques. Les voies visuelles afférentes étaient altérées dans 68% des cas. Des troubles oculomoteurs étaient fréquemment observés (89%) dont le nystagmus pendulaire (28%), source de gêne visuelle. Dans une seconde partie, nous nous sommes donc intéressés au nystagmus pendulaire et à sa conséquence fonctionnelle, l’oscillopsie, afin de proposer une prise en charge spécifique. Néanmoins, les mécanismes de ce nystagmus ne sont pas encore bien définis. Il était donc important de développer une hypothèse explicative à partir de l’observation de deux cas particuliers de nystagmus monoculaire et de démontrer que le nystagmus pendulaire de la sclérose en plaques est à distinguer du nystagmus pendulaire du tremblement oculopalatin, souvent confondus. Enfin, nous proposons une méthode évaluant la détection du mouvement (par stimuli de contraste asservis au regard) ainsi qu’un protocole de stimulation optocinétique tentant de réduire ce symptôme / Neuro-ophthalmic manifestations observed in multiple sclerosis are well-known. However, the chronic feature of visual dysfunctions is not always precisely determinated. These imprecision impede the development of specific therapeutic approach. In a first part, the chronic characteristics of visual dysfunctions were assessed. The percentage of chronic visual complaints in multiple sclerosis patients was quantified and then the intensity of chronic visual deficits was measured, their pathophysiologic origins determined and finally their impact on quality of life was taken into account. Visual pathways were impaired in 68% of patients. Ocular motor disorders were frequently observed (89%) including pendular nystagmus (28%), accounted for visual discomfort. In a second part, we focused on pendular nystagmus and its functional consequence, oscillopsia, to propose a specific treatment. First of all, the mechanisms of this nystagmus are not yet well defined. Therefore, we developed some hypothesis from the observation of two patients with monocular nystagmus and demonstrated in a second part that the pendular nystagmus in multiple sclerosis is distinct from the pendular nystagmus of oculopalatal tremor. Finally, we proposed a method evaluating oscillopsia (motion detection by contrast stimuli moving synchronically with gaze) that was tested before and after an optokinetic stimulation protocol aimed to reduce this symptom

Title: Functional consequences of perinatal hypoxia-ischemia in rat Objectives: The aim of this diploma thesis is to design a set of behavioral tests which provide an effective assessment of motor and cognitive-behavioural deficits in adults rats after experimental hypoxic-ischemic insult during the perinatal period (P7). Supposed benefit is to establish a model of motor and cognitive-behavioural abilities of individuals after this procedure. Methods: The present thesis has a theoretical-empirical character. The practical part describes how the experiment was performed. 32 long Evans Rats were randomly devided into two groups: experimental group (HIE) and control group (Ctrl). The method to produce hypoxic-ischemic brain damage in the 7 day-old rats consisted of right common carotid ligation followed by systemic hypoxia by the inhalation of 8% oxygen and 92% nitrogen. The adult animals (55-75 days old) were tested by the following list of behavioral tests: Bar holding test, Rotarod test, Ladder rung walking test, Reaching test, Open field test and Morris water maze test. Sigma Plot and Microsoft Excel 2010 were the programs used for statistical analysis. Results: Results of Open field test, Ladder rung walking test and Morris water maze test confirmed that hypoxic-ischemic insult affects the...

Bibliographic identification Name and surname of the author: Mgr. Jakub Kokštejn Name of the dissertation: Physical activity of children with motor difficulties Workplace: Department of Sports Games Supervisor of the work: Doc. PaedDr. Rudolf Psotta, Ph.D. Year of presentation: 2011 Abstract Objective: Current knowledge of developmental disorders in motor skills of school children suggest potential negative impacts on the development of the personality of the child in the mental, social psychological and behavioural areas and also in the area of competence at school. Very little is known about how an insufficient level of motor skills of a child can limit his/her physical activity. This work was performed to discover whether developmentally determined motor deficit in children of older school age is a risk factor for lower physical activity (PA) and whether the attention of the child has a mediation function in the relationship between the level of motor skills and his/her physical activity. Methods: The research was based on analytical-descriptive examination of the motor skills of children, their physical activity in weekly regimes and the attention level, with subsequent comparison of children with a motor deficit (MD) and children without MD, through correlation analysis and analysis of the mediative...

Bibliographic identification Name and surname of the author: Mgr. Jakub Kokštejn Name of the dissertation: Physical activity of children with motor difficulties Workplace: Department of Sports Games Supervisor of the work: Doc. PaedDr. Rudolf Psotta, Ph.D. Year of presentation: 2011 Abstract Objective: Current knowledge of developmental disorders in motor skills of school children suggest potential negative impacts on the development of the personality of the child in the mental, social psychological and behavioural areas and also in the area of competence at school. Very little is known about how an insufficient level of motor skills of a child can limit his/her physical activity. This work was performed to discover whether developmentally determined motor deficit in children of older school age is a risk factor for lower physical activity (PA) and whether the attention of the child has a mediation function in the relationship between the level of motor skills and his/her physical activity. Methods: The research was based on analytical-descriptive examination of the motor skills of children, their physical activity in weekly regimes and the attention level, with subsequent comparison of children with a motor deficit (MD) and children without MD, through correlation analysis and analysis of the mediative...