Bibliography VÍTEK, Jiří. The effect of physiotherapy on respiratory muscle strength and functional ability in patients with amyotrophic lateral sclerosis. Prague: Charles University, 2nd Faculty of Medicine, Department of Rehabilitation and Sports Medicine, 2017. 132 p. Supervisor prim. MUDr. Ondřej Horáček, Ph.D. Abstract Objectives To determine the effects of intensive physiotherapeutic care in patients with Amyotrophic lateral sclerosis (ALS), with the focus on objective measurement of respiratory functions and functional status. Methods Patients with ALS (n = 21) were non-randomly assigned into two groups. First group (group R) received a 3-week in-patient rehabilitation programme in the Rehabilitation Hospital Beroun. Second group, control group (group K), received a home exercise programme. Patients from both groups underwent spirometry examination and evaluation of functional skills via the Amyotrophic lateral sclerosis functional rating scale revised questionnaire (ALSFRS-R) three weeks apart. Among the spirometry parameters, forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1) were selected for further evaluation. These parameters are significantly associated with respiratory muscle strength and survival in patients with ALS. Results There were no significant differences in...

Mesenchymal stem cells (MSC) possess a multilineage differentiation potential and have the ability to regulate reactivity of the immune system. They are usually isolated and expanded from the bone marrow, adipose tissue or umbilical cord. MSC represent promising cell population for the treatment of some severe diseases, such as amyotrofic lateral sclerosis (ALS), due to the combination of regenerative and immunomodulatory properties. The aim of this study is to compare MSC from ALS patients and healthy donors in their phenotype, proliferative activity and mainly their immunomodulatory properties. The assessment of impact of the disease on the properties of MSC is important for their autologous use in clinical trials. In this study we used MSC isolated from bone marrow of 14 ALS patients and 15 patients undergoing mostly orthopedic surgery as control group. We also used MSC stimulated for 24 hours by poinflammatory cytokines. Cells were compared in terms of immunophenotype, differentiation in adipocytes and osteoblasts, metabolic activity, expression of selected genes for immunomodulatory molecules and for inhibition of lymphocyte proliferation. Further experiments were focused on evaluation of immunomodulatory properties of MSC. The effect of MSC on peripheral blood mononuclear cells stimulated...

Recent breakthroughs in the RNA quantification of single cells are rapidly transforming the view on biology and medicine. Flexibility and sensitivity of reverse transcription quantitative PCR (RT-qPCR) make it an ideal method for quantification of single-cell material, but its limits had not been yet fully explored. In this thesis, various factors influencing RT-qPCR performance in single-cell application have been assessed, including conditions of sample collection and processing, importance of quality control, performance of reverse transcription, preamplification and role of qPCR assays. We showed that prolonged time for single cell collection as well as repeated freeze-thaw cycles had negligible effect on RT-qPCR data quality. Direct lysis routinely applied for RNA extraction from single cells may be scaled up to 256 cells. The comprehensive comparison of 11 reverse transcriptases in low RNA input conditions identified 2 best-performing enzymes. Decrease in preamplification volume as well as poor primer design resulted in the loss of sensitivity. Finally, the established workflow has been applied to profile gene expression of astrocytes in mouse model of amyotrophic lateral sclerosis (ALS) identifying important components of ALS-induced changes to astrocyte transcriptome. Altogether, the thesis...

Amyotrophic lateral sclerosis (ALS) is a progressive neurological disorder of the central nervous system characterized by loss of motor neurons and voluntary muscle degeneration. Astrocytes play a major role in regulation of the disease onset and progression due to their intimate association with neurons. Regulation of ionic homeostasis is one of their key functions and its failure has been linked to several neurological diseases. The aim of this thesis was to explore differences in membrane properties of astrocytes in ALS. To fulfill this aim, a double transgenic mouse strain with ALS-like phenotype and a specific expression of enhanced green fluorescent protein in astrocytes was generated. To phenotype this strain, two sensorimotor tests, wire grid hang test and rotarod test, were conducted. Immunohistochemistry was used to characterize the strain on a cellular level and to explore changes of specific ion channels. Functional properties of astrocytes were explored using the patch clamp technique. The double transgenic strain has the characteristic ALS-like phenotype and is comparable to the original strain with differences in symptom onset and progression between models and sexes. On the cellular level, there are characteristic ALS features, specifically loss of motor neurons and astrogliosis....

The diploma thesis is focused on the issue of acquired dysarthric and swallowing disorders in amyotrophic lateral sclerosis and on multidisciplinary care provided to patients with this disease. Theoretical part of the thesis is divided into three chapters. The introductory chapter presents a summary of current knowledge about amyotrophic lateral sclerosis (ALS). The following two chapters are dedicated to dysarthria and dysphagia and their specifics in ALS. The practical part of the diploma thesis is represented by the fourth chapter which incorporates a research survey focused on multidisciplinary care provided to patients with ALS. The primary aim of the research is to evaluate the multidisciplinary care provided to two selected patients with ALS with a focus on speech and swallowing disorders. The secondary objectives of the research relate to the evaluation of the extent of acquired dysarthric and swallowing disorders of these patients, providing a comprehensive overview of the course and content of the provided care and gathering information to create information brochure for patients with ALS and caregivers. The research approach to achieve the determined objectives of the research survey is creation of case studies using qualitative methods of data collection, which is a structured interview...

Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...