No description available.

As células estromais mesenquimais (CMMs) se tornaram de grande interesse para a terapia celular devido ao seu potencial de se diferenciar e reconstituir tecidos especializados. Mais recentemente, este interesse tem aumentado significativamente devido à descoberta de que as CMMs são capazes de secretar uma infinidade de mediadores para estimular a regeneração in situ de tecidos lesados. Dessa forma, CMMs podem ser consideradas tanto como um produto terapêutico em si, quanto uma biofábrica de diversas proteínas relevantes do ponto de vista terapêutico. Para atender a estas crescentes demandas, ambas as aplicações requerem o desenvolvimento de processos de expansão celular com alto rendimento, sob condições de cultivo definidas, reprodutíveis, escalonáveis, permitindo a obtenção de produtos com adequada identidade, potência, pureza, segurança e viáveis economicamente. Frente ao exposto, este trabalho teve como objetivos principais o estabelecimento de um processo de expansão de CMMs baseado em biorreatores e a caracterização do secretoma destas células visando aplicações terapêuticas. Para isto, a expansão de CMMs do cordão umbilical (MCUs) foi realizada em frascos multicamadas (MC) e nos biorreatores de leito fixo (LF), tanque agitado com microcarregadores (TA) e fibrasocas (FO). Os resultados mostraram que a taxa de proliferação específica das células foi maior (< tempo de duplicação) no biorreator de FO (36,8 ± 1,7 horas), bem como o fator de expansão (9,8 ± 1,0) e a eficiência na recuperação celular (100%). Um nível similar de produção celular foi observado para o TA, MC e LF com elevado fator de expansão celular (8,8 ± 0,39, 8,7 ± 0,90, 6,9 ± 1,3, respectivamente). No entanto, em termos de eficiência na recuperação celular (%), LF apresentou a menor taxa de recuperação dentre todos os sistemas (18% (± 0,77)), acompanhado pelo TA (61% (± 15,7)). As células mantiveram suas características imunofenotípicas e o potencial de diferenciação em adipócitos, osteócitos e condrócitos em todos os sistemas de cultivo avaliados. Foi também realizada a análise de custos (COG) e avaliação da viabilidade econômica para produção de CMMs visando tratamento da doença do enxerto contra o hospedeiro (DECH) em escala comercial, utilizando os sistemas de cultivo avaliados experimentalmente sob diferentes estratégias de reembolso. Apesar dos resultados experimentais satisfatórios para o biorreator FO, o COG revelou que este sistema tem o maior custo devido aos elevados custos dos consumíveis requeridos e do custo do equipamento. O frasco MC foi considerado como a tecnologia mais rentável e robusta no cenário avaliado e o biorreator TA obteve a segunda posição. O biorreator TA foi escolhido como o mais adequado analisando de maneira conjunta os dados experimentais obtidos, a análise dos custos dos diferentes sistemas de cultivo e a escalonabilidade de cada sistema. Assim, esse biorreator foi eficientemente utilizado para o cultivo de MCUs em condições isentas de SFB e xenoantígenos, sendo possível a produção de uma grande quantidade de células, representando um passo importante no desenvolvimento de um bioprocesso em conformidade com as normas das agências regulatórias. Por fim, com a análise do secretoma das CMMs por espectrometria de massas foi possível a identificação de uma gama enorme de proteínas interessantes (aprox. 2400) envolvidas em importantes processos biológicos. O futuro monitoramento dessas proteínas em biorreatores poderá representar um método inovador e original de produção de produtos livres de células para uso na terapia celular. / Mesenchymal stem/stromal cells (MSC) have become of great interest for cell therapy because of its potential to differentiate and reconstitute specialized tissues. More recently, such interest has significantly increased due to the discovery that MSC are capable of secreting a plethora of mediators to stimulate the in situ regeneration of injured tissues. Thus, MSC can be considered as a therapeutic product itself and as a biofactory of various relevant therapeutic proteins. To meet these increasing demands, both applications require the development of high-yield, reproducible, scalable and cost-effective bioprocesses under defined culture conditions, obtaining products with proper identity, purity and safety. Based on these, the main goal of this work was the establishment of a MSC expansion process based on bioreactors and secretome characterization of these cells targeting therapeutic applications. The MSC expansion was performed using multi-layered flasks (ML) and fixed bed (PB), stirred tank (STR) and hollow fiber (HF) bioreactors. The results showed that the proliferation rate of the cells was higher (< doubling time) in the HF bioreactor (36.8 ± 1.7 hours), as well as the expansion fold-increase (9.8±1.0) and harvesting efficiency (100%). A similar level of cell production was observed for STR, ML and PB with high fold-increase (8.8±0.39, 8.7±0.90, 6.9±1.3, respectively). However, in terms of harvesting efficiency (%), PB bioreactor presented the lowest retrieval rate across all the technologies (18% (±0.77)), followed by STR (61% (±15.7)). The cells retained their functional properties after culture in all the culture systems evaluated. This study was then extended through the use of a bioprocess economics tool for the evaluation of the economic feasibility of producing MSC-based treatment for acute graft vs. host disease (aGvHD) at commercial scale, using the culture systems experimentally evaluated under different reimbursement strategies. Despite the advantageous experimental results of HF bioreactors, the COG analysis has revealed that this is the least cost effective cell culture system to be used, due to its high consumable and equipment costs. ML flasks ranked first as the most cost effective and robust technology in this scenario and microcarrier-based technologies (STR) ranked in second position. The STR bioreactor was chosen as the most suitable for MSC expansion analyzing the experimental data, COG analysis and scalability of each culture system. Thus, STR bioreactor was efficiently tested for MSC expansion under serum and xeno-free conditions and it was possible to produce a large amount of cells. The development of a scalable microcarrier-based stirred culture system using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Finally, with the MSC secretome analysis by mass spectrometry it was possible to identify a wide range of interesting proteins (approx. 2400) involved in important biological processes. The future monitoring of these proteins in bioreactors may represent a novel and unique method of producing cell-free products for use in cellular therapy.

As células estromais mesenquimais (CMMs) se tornaram de grande interesse para a terapia celular devido ao seu potencial de se diferenciar e reconstituir tecidos especializados. Mais recentemente, este interesse tem aumentado significativamente devido à descoberta de que as CMMs são capazes de secretar uma infinidade de mediadores para estimular a regeneração in situ de tecidos lesados. Dessa forma, CMMs podem ser consideradas tanto como um produto terapêutico em si, quanto uma biofábrica de diversas proteínas relevantes do ponto de vista terapêutico. Para atender a estas crescentes demandas, ambas as aplicações requerem o desenvolvimento de processos de expansão celular com alto rendimento, sob condições de cultivo definidas, reprodutíveis, escalonáveis, permitindo a obtenção de produtos com adequada identidade, potência, pureza, segurança e viáveis economicamente. Frente ao exposto, este trabalho teve como objetivos principais o estabelecimento de um processo de expansão de CMMs baseado em biorreatores e a caracterização do secretoma destas células visando aplicações terapêuticas. Para isto, a expansão de CMMs do cordão umbilical (MCUs) foi realizada em frascos multicamadas (MC) e nos biorreatores de leito fixo (LF), tanque agitado com microcarregadores (TA) e fibrasocas (FO). Os resultados mostraram que a taxa de proliferação específica das células foi maior (< tempo de duplicação) no biorreator de FO (36,8 ± 1,7 horas), bem como o fator de expansão (9,8 ± 1,0) e a eficiência na recuperação celular (100%). Um nível similar de produção celular foi observado para o TA, MC e LF com elevado fator de expansão celular (8,8 ± 0,39, 8,7 ± 0,90, 6,9 ± 1,3, respectivamente). No entanto, em termos de eficiência na recuperação celular (%), LF apresentou a menor taxa de recuperação dentre todos os sistemas (18% (± 0,77)), acompanhado pelo TA (61% (± 15,7)). As células mantiveram suas características imunofenotípicas e o potencial de diferenciação em adipócitos, osteócitos e condrócitos em todos os sistemas de cultivo avaliados. Foi também realizada a análise de custos (COG) e avaliação da viabilidade econômica para produção de CMMs visando tratamento da doença do enxerto contra o hospedeiro (DECH) em escala comercial, utilizando os sistemas de cultivo avaliados experimentalmente sob diferentes estratégias de reembolso. Apesar dos resultados experimentais satisfatórios para o biorreator FO, o COG revelou que este sistema tem o maior custo devido aos elevados custos dos consumíveis requeridos e do custo do equipamento. O frasco MC foi considerado como a tecnologia mais rentável e robusta no cenário avaliado e o biorreator TA obteve a segunda posição. O biorreator TA foi escolhido como o mais adequado analisando de maneira conjunta os dados experimentais obtidos, a análise dos custos dos diferentes sistemas de cultivo e a escalonabilidade de cada sistema. Assim, esse biorreator foi eficientemente utilizado para o cultivo de MCUs em condições isentas de SFB e xenoantígenos, sendo possível a produção de uma grande quantidade de células, representando um passo importante no desenvolvimento de um bioprocesso em conformidade com as normas das agências regulatórias. Por fim, com a análise do secretoma das CMMs por espectrometria de massas foi possível a identificação de uma gama enorme de proteínas interessantes (aprox. 2400) envolvidas em importantes processos biológicos. O futuro monitoramento dessas proteínas em biorreatores poderá representar um método inovador e original de produção de produtos livres de células para uso na terapia celular. / Mesenchymal stem/stromal cells (MSC) have become of great interest for cell therapy because of its potential to differentiate and reconstitute specialized tissues. More recently, such interest has significantly increased due to the discovery that MSC are capable of secreting a plethora of mediators to stimulate the in situ regeneration of injured tissues. Thus, MSC can be considered as a therapeutic product itself and as a biofactory of various relevant therapeutic proteins. To meet these increasing demands, both applications require the development of high-yield, reproducible, scalable and cost-effective bioprocesses under defined culture conditions, obtaining products with proper identity, purity and safety. Based on these, the main goal of this work was the establishment of a MSC expansion process based on bioreactors and secretome characterization of these cells targeting therapeutic applications. The MSC expansion was performed using multi-layered flasks (ML) and fixed bed (PB), stirred tank (STR) and hollow fiber (HF) bioreactors. The results showed that the proliferation rate of the cells was higher (< doubling time) in the HF bioreactor (36.8 ± 1.7 hours), as well as the expansion fold-increase (9.8±1.0) and harvesting efficiency (100%). A similar level of cell production was observed for STR, ML and PB with high fold-increase (8.8±0.39, 8.7±0.90, 6.9±1.3, respectively). However, in terms of harvesting efficiency (%), PB bioreactor presented the lowest retrieval rate across all the technologies (18% (±0.77)), followed by STR (61% (±15.7)). The cells retained their functional properties after culture in all the culture systems evaluated. This study was then extended through the use of a bioprocess economics tool for the evaluation of the economic feasibility of producing MSC-based treatment for acute graft vs. host disease (aGvHD) at commercial scale, using the culture systems experimentally evaluated under different reimbursement strategies. Despite the advantageous experimental results of HF bioreactors, the COG analysis has revealed that this is the least cost effective cell culture system to be used, due to its high consumable and equipment costs. ML flasks ranked first as the most cost effective and robust technology in this scenario and microcarrier-based technologies (STR) ranked in second position. The STR bioreactor was chosen as the most suitable for MSC expansion analyzing the experimental data, COG analysis and scalability of each culture system. Thus, STR bioreactor was efficiently tested for MSC expansion under serum and xeno-free conditions and it was possible to produce a large amount of cells. The development of a scalable microcarrier-based stirred culture system using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Finally, with the MSC secretome analysis by mass spectrometry it was possible to identify a wide range of interesting proteins (approx. 2400) involved in important biological processes. The future monitoring of these proteins in bioreactors may represent a novel and unique method of producing cell-free products for use in cellular therapy.

According to NATO Review, in 2013 it was estimated that over 97% of the world's telecommunications were transmitted via the internet and that this was a growing trend. This essentiality for communication means that information systems have become a natural target and point of attack in military actions and planning. This thesis aims to test the possibility to apply the theory of The enemy as a system and the concentric five-ring modeldeveloped by air force theorist Jon A. Warden III, to the cyber arena. To achieve this aim, the thesis presents a qualitative text analysis of seven different sources describing NotPetya. The targets and effects of the cyber attack are evaluated based upon the concentric five-ring model and the concept of parallel attack. The results of the analysis shows that Warden's theory is highly applicable to the case of NotPetya. This in turn could point to the theory’s applicability to the cyber domain and to strategic operations in cyberspace. However no major conclusions of generalizability can be drawn.

Crocodiles are ectothermic, intermittent breathing reptiles with low metabolic rates. Additionally, they are diving animals that can stay submerged for long periods, which poses special demands on their cardiovascular system. The crocodilian heart is four-chambered with completely separated ventricles, making it unique among other reptiles. It has special anatomical structures that give it the capability to shunt blood away from the lungs, which results in mixing of deoxygenated and oxygenated blood. The purpose of this study was to morphologically describe the heart and review its functions and the significance of its circulatory patterns. Dissection of two crocodilian hearts was performed for morphological characterization of the heart, including the special features that contribute to the shunting: the left aorta, the foramen of Panizza and the cog-teeth-like valves. Obstruction of the pulmonary outflow tract by the cog-teeth-like valves decreases pulmonary blood flow and generates an increased right ventricular pressure, diverting venous blood into the ‘extra’ left aorta and the systemic circulation. Thus, during shunting conditions, venous blood from the left aorta and arterial blood from the right aorta is mixed. The functional significance of this shunting pattern has been extensively discussed and several hypotheses have been proposed. A recent study showed that the absence of the right-to-left shunt does not affect the diving physiology of crocodiles negatively, but it did result in cardiac hypertrophy, indicating that the shunt might have a significance for the circulation and physiology of extant crocodiles.

No description available.

Les maladies neurodégénératives, telles que la maladie d'Alzheimer (AD) et Charcot Marie Tooth (CMT), sont des maladies complexes. Leurs mécanismes pathologiques ne sont toujours pas bien compris et les progrès dans la recherche et le développement de nouvelles thérapies potentielles modifiant la maladie sont lents. Les données catégorielles, comme les échelles de notation et les données sur les études d'association génomique (GWAS), sont largement utilisées dans les maladies neurodégénératives dans le diagnostic, la prédiction et le suivi de la progression. Il est important de comprendre et d'interpréter ces données correctement si nous voulons améliorer la recherche sur les maladies neurodégénératives. Le but de cette thèse est d'utiliser la théorie psychométrique moderne: théorie de la réponse d’item pour analyser ces données catégoriques afin de mieux comprendre les maladies neurodégénératives et de faciliter la recherche de médicaments correspondante. Tout d'abord, nous avons appliqué l'analyse de Rasch afin d'évaluer la validité du score de neuropathie Charcot-Marie-Tooth (CMTNS), un critère important d'évaluation principal pour les essais cliniques de la maladie de CMT. Nous avons ensuite adapté le modèle Rasch à l'analyse des associations génétiques pour identifier les gènes associés à la maladie d'Alzheimer. Cette méthode résume les génotypes catégoriques de plusieurs marqueurs génétiques tels que les polymorphisme nucléotidique (SNPs) en un seul score génétique. Enfin, nous avons calculé l'information mutuelle basée sur la théorie de réponse d’item pour sélectionner les items sensibles dans ADAS-cog, une mesure de fonctionnement cognitif la plus utilisées dans les études de la maladie d'Alzheimer, afin de mieux évaluer le progrès de la maladie. / Neurodegenerative diseases, such as Alzheimer’s disease (AD) and Charcot Marie Tooth (CMT), are complex diseases. Their pathological mechanisms are still not well understood, and the progress in the research and development of new potential disease-modifying therapies is slow. Categorical data like rating scales and Genome-Wide Association Studies (GWAS) data are widely utilized in the neurodegenerative diseases in the diagnosis, prediction and progression monitor. It is important to understand and interpret these data correctly if we want to improve the disease research. The purpose of this thesis is to use the modern psychometric Item Response Theory to analyze these categorical data for better understanding the neurodegenerative diseases and facilitating the corresponding drug research. First, we applied the Rasch analysis in order to assess the validity of the Charcot-Marie-Tooth Neuropathy Score (CMTNS), a main endpoint for the CMT disease clinical trials. We then adapted the Rasch model to the analysis of genetic associations and used to identify genes associated with Alzheimer’s disease by summarizing the categorical genotypes of several genetic markers such as Single Nucleotide Polymorphisms (SNPs) into one genetic score. Finally, to select sensitive items in the most used psychometrical tests for Alzheimer’s disease, we calculated the mutual information based on the item response model to evaluate the sensitivity of each item on the ADAS-cog scale.

Předložená disertační práce se zabývá vývojem algoritmů pro detekci stresu z řečového signálu. Inovativnost této práce se vyznačuje dvěma typy analýzy řečového signálu, a to za použití samohláskových polygonů a analýzy hlasivkových pulsů. Obě tyto základní analýzy mohou sloužit k detekci stresu v řečovém signálu, což bylo dokázáno sérií provedených experimentů. Nejlepších výsledků bylo dosaženo pomocí tzv. Closing-To-Opening phase ratio příznaku v Top-To-Bottom kritériu v kombinaci s vhodným klasifikátorem. Detekce stresu založená na této analýze může být definována jako jazykově i fonémově nezávislá, což bylo rovněž dokázáno získanými výsledky, které dosahují v některých případech až 95% úspěšnosti. Všechny experimenty byly provedeny na vytvořené české databázi obsahující reálný stres, a některé experimenty byly také provedeny pro anglickou stresovou databázi SUSAS.

Two different sets of problems are addressed in this thesis. The first one is on partially observed semi-Markov Games (POSMG) and the second one is on semi-Markov modulated financial market model. In this thesis we study a partially observable semi-Markov game in the infinite time horizon. The study of a partially observable game (POG) involves three major steps: (i) construct an equivalent completely observable game (COG), (ii) establish the equivalence between POG and COG by showing that if COG admits an equilibrium, POG does so, (iii) study the equilibrium of COG and find the corresponding equilibrium of original partially observable problem. In case of infinite time horizon game problem there are two different payoff criteria. These are discounted payoff criterion and average payoff criterion. At first a partially observable semi-Markov decision process on general state space with discounted cost criterion is studied. An optimal policy is shown to exist by considering a Shapley’s equation for the corresponding completely observable model. Next the discounted payoff problem is studied for two-person zero-sum case. A saddle point equilibrium is shown to exist for this case. Then the variable sum game is investigated. For this case the Nash equilibrium strategy is obtained in Markov class under suitable assumption. Next the POSMG problem on countable state space is addressed for average payoff criterion. It is well known that under this criterion the game problem do not have a solution in general. To ensure a solution one needs some kind of ergodicity of the transition kernel. We find an appropriate ergodicity of partially observed model which in turn induces a geometric ergodicity to the equivalent model. Using this we establish a solution of the corresponding average payoff optimality equation (APOE). Thus the value and a saddle point equilibrium is obtained for the original partially observable model. A value iteration scheme is also developed to find out the average value of the game. Next we study the financial market model whose key parameters are modulated by semi-Markov processes. Two different problems are addressed under this market assumption. In the first one we show that this market is incomplete. In such an incomplete market we find the locally risk minimizing prices of exotic options in the Follmer Schweizer framework. In this model the stock prices are no more Markov. Generally stock price process is modeled as Markov process because otherwise one may not get a pde representation of price of a contingent claim. To overcome this difficulty we find an appropriate Markov process which includes the stock price as a component and then find its infinitesimal generator. Using Feynman-Kac formula we obtain a system of non-local partial differential equations satisfied by the option price functions in the mildsense. .Next this system is shown to have a classical solution for given initial or boundary conditions. Then this solution is used to have a F¨ollmer Schweizer decomposition of option price. Thus we obtain the locally risk minimizing prices of different options. Furthermore we obtain an integral equation satisfied by the unique solution of this system. This enable us to compute the price of a contingent claim and find the risk minimizing hedging strategy numerically. Further we develop an efficient and stable numerical method to compute the prices. Beside this work on derivative pricing, the portfolio optimization problem in semi-Markov modulated market is also studied in the thesis. We find the optimal portfolio selections by optimizing expected utility of terminal wealth. We also obtain the optimal portfolio selections under risk sensitive criterion for both finite and infinite time horizon.

Title: Evaluation of postural stability of athletes - a literature review Objectives: Thesis will address retrieval processing rating Dynamic Postural Stability of athletes . Compares analysis or results of research or investigations of different laborers working with this theme. It also mentions tests and machines used in clinical practice. It also investigates in what sports was postural stability tested and how or with what results. The most advanced, currently the most recognized dynamic posturography with impartial value is EquiTest device from NeuroCom. Therefore, the practical part focuses primarily on the results of examination of this device and make statistics about that. Methods: The diploma thesis has descriptive and analytical character. It is elaborate as a literary review . Results: The study includes 142 studies, of which only 11 concern the assessment of postural stability of athletes. Sports disciplines include tennis, football, baseball, tai-chi, taekwondo, gymnastics, basketball, softball. Entire EquiTest testing battery (which contains 7 test in total - Sensory Organization Test, Motor Coordination and Control Test, Adaptation Test, Unilateral Stance Test, Limits Of Stability Test, Rhytmics Weight Shift, Weight Bearing Squat) is almost never used in assessments although it's a...