Made available in DSpace on 2015-01-26T13:21:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-08Bitstream added on 2015-01-26T13:30:38Z : No. of bitstreams: 1 000797817.pdf: 449554 bytes, checksum: 7adc756b2ac04e5d1cbc6be34fa7a04e (MD5) / Introdução: A síndrome HELLP é uma complicação severa da pré-eclâmpsia, caracterizada por hemólise, elevação das enzimas hepáticas e trombocitopenia. Apesar de haver padronização dos valores laboratoriais que definem a síndrome HELLP, ainda existe dificuldade para a caracterização da hemólise. OBJETIVO: Avaliar o valor de haptoglobina que determina a hemólise nas pacientes com síndrome HELLP. MÉTODOS: estudo transversal e prospectivo de gestantes e puérperas com de pré-eclâmpsia. Exame laboratorial avaliado: dosagem sérica de haptoglobina. Construção da curva ROC para determinar o valor de corte da haptoglobina para diagnóstico de hemólise na síndrome HELLP. RESULTADOS: O valor da haptoglobina para diagnóstico de hemólise em pacientes com síndrome HELLP foi de 0,26g/L. DISCUSSÃO: A melhor correlação observada foi a haptoglobina com a DHL, indicando que este é o melhor marcador de hemólise intravascular para o diagnóstico da síndrome HELLP. CONCLUSÃO: A dosagem sérica da haptoglobina nos casos de pré-eclâmpsia deve fazer parte dos exames de rotina para diagnóstico de hemólise intravascular da síndrome HELLP / Context: HELLP syndrome is a severe complication of pre-eclampsia, caracterized by hemolysis, elevated liver enzymes and low platelet count. Although there are standardized laboratory values that define the HELLP syndrome, the difficulty still exists for the characterization of hemolysis. PURPOSE: to evaluate the haptoglobin value to diagnose the hemolysis in HELLP syndrome. METHODS: transversal and prospective study of pregnant and postdelivery women with pre-eclampsia. Laboratory tests evaluated: serum haptoglobin. ROC curve to determine the cutoff value of haptoglobin in the diagnosis of hemolysis in HELLP syndrome. RESULTS: The haptoglobin value for hemolysis diagnosis in HELLP syndrome was 0.26 g / L. DISCUSSION: The best correlation was with haptoglobin and DHL, indicating that this is the best marker of intravascular hemolysis for the diagnosis of HELLP syndrome. CONCLUSION: Serum haptoglobin in cases of pre-eclampsia should be part of routine tests for diagnosis of intravascular hemolysis in HELLP syndrome

Orientador: Tomomassa Yano / Dissertação (mestrado) - Universidade Estadual de Campinas,. Instituto de Biologia / Made available in DSpace on 2018-08-14T11:00:37Z (GMT). No. of bitstreams: 1 Thome_JacquelineDarcSilva_M.pdf: 1223668 bytes, checksum: 17699f260f82fb8a09136004fcc1e28a (MD5) Previous issue date: 2006 / Mestrado / Microbiologia / Mestre em Genética e Biologia Molecular

Malaria affects millions of people annually especially in third world countries. Increase in resistance and limited research being conducted adds to the global burden of malaria. Mefloquine, known for unwanted adverse reactions and neurotoxicity, is highly lipophilic and is still used as treatment and prophylaxis. Lipid drug delivery systems are commonly used to increase solubility and efficacy and decrease toxicity. The most generally used lipid drug delivery system is liposomes. The lipid bilayer structure varying in size from 25 nm to 100 μm can entrap both hydrophilic and lipophilic compounds. Similar in structure and size to liposomes, Pheroid™ technology consist of natural fatty acids and is also able to entrap lipophilic and hydrophilic compounds. The aim of this study was to formulate liposomes and Pheroid™ vesicles loaded with mefloquine and evaluate the physiochemical characteristic of the formulations followed by efficacy and toxicity studies. Pheroid™ vesicles and liposomes with and without mefloquine were evaluated in size, morphology, pH and entrapment efficacy during three month accelerated stability testing. Optimization of size determination by flow cytometry lead to accurate determination of size for both Pheroid™ vesicles and liposomes. During the three months stability testing, Pheroid™ vesicles showed a small change in size from 3.07 ± 0.01 μm to approximately 3 μm for all three temperatures. Confocal laser scanning microscopic evaluation of the liposomes showed structures uniform in spherical shape and size. No difference in size or structure between the Pheroid™ vesicles with and without mefloquine were obtained. Significant increase (p=0.027) in size from 6.46 ± 0.01 μm to above 10 μm was observed for liposomes at all the temperatures. Clearly formed lipid bilayer structures were observed on micrographs. With the addition of mefloquine to the liposome formulation, a decrease in the amount of bilayer structures and an increase in oil droplets were found. Entrapment efficacy was determined by firstly separating the entrapped drug from the unentrapped drug utilizing a Sephadex®G50 mini column. This was followed by spectrophotometric evaluation by UV-spectrophotometry at 283 nm. Initial entrapment efficacy of both Pheroid™ vesicles and liposomes was above 60%. An increase in entrapment efficacy was observed for Pheroid™ vesicles. The addition of mefloquine to already formulated Pheroid™ vesicles illustrated entrapment efficacy of 60.14 ± 5.59% after 14 days. Formulations loaded with mefloquine resulted in lower pH values as well as a decrease in pH over time. Optimization of efficacy studies utilizing propidium iodide was necessary due to the similarity in size and shape of the drug delivery systems to erythrocytes. A gating strategy was successfully implemented for the determination of the percentage parasitemia. Efficacy testing of mefloquine loaded in Pheroid™ vesicles and liposomes showed a 186% and 207% decrease in parasitemia levels compared to the control of mefloquine. Toxicity studies conducted include haemolysis and ROS (reactive oxygen species) analysis on erythrocytes as well as cell viability on mouse neuroblastoma cells. Pheroid™ vesicles with and without mefloquine resulted in a dose dependent increase in ROS and haemolysis over time. A dose dependent increase in ROS and haemolysis in both liposome formulations were observed, but to a lesser extent. Mefloquine proved to be neurotoxic with similar results obtained when mefloquine was entrapped in liposomes. Pheroid™ vesicles seem to have neuroprotective properties resulting in higher cell viability. Mefloquine could be entrapped successfully in Pheroid™ vesicles and less in liposomes. Pheroid™ vesicles was more stable over a three months accelerated stability testing with more favourable characteristics. The increase in ROS levels of Pheroid™ vesicles could be responsible for the higher efficacy and haemolytic activity. DL-α-Tocopherol in Pheroid™ vesicles possibly acted as a pro-oxidant due to the presence of iron in the erythrocytes. DL-α-Tocopherol showed possible antioxidant properties in the neurotoxicity evaluation resulting in higher cell viability. Even though liposomes illustrated higher efficacy and little haemolysis and ROS production, no difference in neurotoxicity was observed together with unfavourable properties during stability testing makes this drug delivery system less favourable in comparison to Pheroid™ vesicles. Mefloquine was successfully incorporated into Pheroid™ vesicles resulted in high efficacy and showed possible neuroprotection and therefore makes it an ideal system for treatment of malaria. / Thesis (Ph.D. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2011

Malaria affects millions of people annually especially in third world countries. Increase in resistance and limited research being conducted adds to the global burden of malaria. Mefloquine, known for unwanted adverse reactions and neurotoxicity, is highly lipophilic and is still used as treatment and prophylaxis. Lipid drug delivery systems are commonly used to increase solubility and efficacy and decrease toxicity. The most generally used lipid drug delivery system is liposomes. The lipid bilayer structure varying in size from 25 nm to 100 μm can entrap both hydrophilic and lipophilic compounds. Similar in structure and size to liposomes, Pheroid™ technology consist of natural fatty acids and is also able to entrap lipophilic and hydrophilic compounds. The aim of this study was to formulate liposomes and Pheroid™ vesicles loaded with mefloquine and evaluate the physiochemical characteristic of the formulations followed by efficacy and toxicity studies. Pheroid™ vesicles and liposomes with and without mefloquine were evaluated in size, morphology, pH and entrapment efficacy during three month accelerated stability testing. Optimization of size determination by flow cytometry lead to accurate determination of size for both Pheroid™ vesicles and liposomes. During the three months stability testing, Pheroid™ vesicles showed a small change in size from 3.07 ± 0.01 μm to approximately 3 μm for all three temperatures. Confocal laser scanning microscopic evaluation of the liposomes showed structures uniform in spherical shape and size. No difference in size or structure between the Pheroid™ vesicles with and without mefloquine were obtained. Significant increase (p=0.027) in size from 6.46 ± 0.01 μm to above 10 μm was observed for liposomes at all the temperatures. Clearly formed lipid bilayer structures were observed on micrographs. With the addition of mefloquine to the liposome formulation, a decrease in the amount of bilayer structures and an increase in oil droplets were found. Entrapment efficacy was determined by firstly separating the entrapped drug from the unentrapped drug utilizing a Sephadex®G50 mini column. This was followed by spectrophotometric evaluation by UV-spectrophotometry at 283 nm. Initial entrapment efficacy of both Pheroid™ vesicles and liposomes was above 60%. An increase in entrapment efficacy was observed for Pheroid™ vesicles. The addition of mefloquine to already formulated Pheroid™ vesicles illustrated entrapment efficacy of 60.14 ± 5.59% after 14 days. Formulations loaded with mefloquine resulted in lower pH values as well as a decrease in pH over time. Optimization of efficacy studies utilizing propidium iodide was necessary due to the similarity in size and shape of the drug delivery systems to erythrocytes. A gating strategy was successfully implemented for the determination of the percentage parasitemia. Efficacy testing of mefloquine loaded in Pheroid™ vesicles and liposomes showed a 186% and 207% decrease in parasitemia levels compared to the control of mefloquine. Toxicity studies conducted include haemolysis and ROS (reactive oxygen species) analysis on erythrocytes as well as cell viability on mouse neuroblastoma cells. Pheroid™ vesicles with and without mefloquine resulted in a dose dependent increase in ROS and haemolysis over time. A dose dependent increase in ROS and haemolysis in both liposome formulations were observed, but to a lesser extent. Mefloquine proved to be neurotoxic with similar results obtained when mefloquine was entrapped in liposomes. Pheroid™ vesicles seem to have neuroprotective properties resulting in higher cell viability. Mefloquine could be entrapped successfully in Pheroid™ vesicles and less in liposomes. Pheroid™ vesicles was more stable over a three months accelerated stability testing with more favourable characteristics. The increase in ROS levels of Pheroid™ vesicles could be responsible for the higher efficacy and haemolytic activity. DL-α-Tocopherol in Pheroid™ vesicles possibly acted as a pro-oxidant due to the presence of iron in the erythrocytes. DL-α-Tocopherol showed possible antioxidant properties in the neurotoxicity evaluation resulting in higher cell viability. Even though liposomes illustrated higher efficacy and little haemolysis and ROS production, no difference in neurotoxicity was observed together with unfavourable properties during stability testing makes this drug delivery system less favourable in comparison to Pheroid™ vesicles. Mefloquine was successfully incorporated into Pheroid™ vesicles resulted in high efficacy and showed possible neuroprotection and therefore makes it an ideal system for treatment of malaria. / Thesis (Ph.D. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2011

Orientadores: Antonio Celso Fonseca de Arruda, Aron Jose Pazin de Andrade / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Mecanica / Made available in DSpace on 2018-08-09T06:51:30Z (GMT). No. of bitstreams: 1 Bock_EduardoGuyPerpetuo_M.pdf: 3873649 bytes, checksum: 91eb2ac5543b164e576ca7a5e61d6a45 (MD5) Previous issue date: 2007 / Resumo: Uma bomba centrífuga implantável foi projetada e construída para assistência ventricular de longa duração no tratamento de pacientes portadores de doenças cardiovasculares. A metodologia utilizada foi a de testes de desempenho ¿in vitro¿ compostos de testes de atrito para seleção de materiais para os mancais de apoio, testes de desempenho hidrodinâmico da bomba e testes com sangue humano para a verificação da hemólise causada no bombeamento. Os primeiros testes foram realizados com uma bomba centrífuga não implantável, normalmente utilizada em cirurgias cardíacas, para a determinação do posicionamento do orifício de entrada da bomba. Foram realizadas simulações numéricas por computador para dimensionamento da bomba. Testes de atrito foram feitos para a seleção de materiais do sistema de mancais de apoio proposto para alcançar a durabilidade necessária. Testes de desempenho hidrodinâmico foram utilizados para determinar o melhor desenho para o rotor da bomba. Seu desempenho foi comparado com uma bomba não implantável e com uma bomba implantável similar americana e apresentou melhores resultados que ambas. Testes com sangue foram feitos para quantificar os danos causados aos componentes do sangue pela bomba. O seu índice normalizado de hemólise foi de 0,0054 mg/100L, considerado excelente por se encontrar próximo do limite mínimo encontrado na literatura: entre 0,004 g/ 100L e 0,02 g/ 100L. Como trabalhos futuros, serão feito testes com animais para que a bomba centrífuga implantável possa ser utilizada em pacientes / Abstract: An implantable centrifugal blood pump was developed for long-term ventricular assistance in cardiac patients. In vitro tests were performed, as wear evaluation, performance tests and hemolysis tests in human blood. Preliminary tests were performed with a non-implantable pump in order to properly locate the inlet port. Numerical computational simulations were performed in order to predict its best geometry. Wear evaluations helped to select the best materials for double pivot bearing system proposed to achieve longer durability. Performance tests pointed the best impeller geometry. The implantable centrifugal blood pump was compared with other two blood pumps. One is a centrifugal blood pump for cardiopulmonary bypass and the other is a similar implantable device. The proposed implantable centrifugal blood pump showed the best performance. But, its results showed a strong descendant curve in high flow. Other prototype was manufactured with a different inlet port angle to overcome this problem. The normalized index of hemolysis (NIH) tests was performed with human blood pumped in an In Vitro closed circuit (mock loop) in normalized conditions, as flow of 5 L/min and total pressure ahead of 100 mm Hg. After six hours, NIH measured 0.0054 mg/100L that can be considered excellent since it is close to the minimum found in literature (between 0.004 g/ 100L e 0.02 g/ 100L). As future work, in vivo test will be performed with the implantable centrifugal blood pump looking forward to implant it in patients / Mestrado / Materiais e Processos de Fabricação / Mestre em Engenharia Mecânica

Orientadores: Reinaldo Wilson Vieira, Jorge Humberto Nicola / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Resumo: A circulação extracorpórea (CEC) utiliza-se de um conjunto de máquinas, aparelhos e circuitos, associados a técnicas que permite o isolamento temporário de coração e pulmões. Estudar o comportamento do sangue dentro destes elementos utilizados pela CEC sem alterar substancialmente o seu curso requer técnicas avançadas. A Velocimetria Doppler Laser (VDL) tem se mostrado ideal para estes procedimentos por ter seus sinais de natureza óptica. Dentre as diversas reações suscitadas ao organismo pelo uso da CEC, que produzem alterações do equilíbrio fisiológico de natureza hemodinâmica, fisica e química, ressaltamos, neste estudo, as alterações fisicas geradas pelo trauma mecânico softido pelas células do sangue, principalmente as hemáceas.Objetivos: 1. Estudar e caracterizar os fluxos gerados por bombas centrífugas e de roletes. 2. Estabelecer condições ideais para estudos de turbulência e estagnação do sangue em componente passivo da circulação extracorpórea. 3. Validar a Velocimetria Doppler Laser como método de estudo de componente utilizado em circulação extracorpórea (reservatório de cardioplegia sangüínea), visando a otimização de projeto de tal componente.Material e Método: Foram realizadas medidas não invasivas de velocidades, utilizando um Velocímetro Doppler Laser, de duas dimensões em um equipamento utilizado na CEC denominado reservatório de cardioplegia sangüínea, que nada mais é que um trocador de calor casco e tubo. Utilizamos como fluido a mistura de 36% de solução de glicerina em água, por ser um fluido com características fisicas (viscosidade e densidade) equivalentes ao do sangue. A visualização foi obtida por meio de suspensão de partículas de poliamida de diâmetro de 5 _m no líquido movimentado por uma bomba centrífuga. Resultados e Conclusões : O caráter oscilante do fluxo de uma bomba de roletes é bastante significativo, enquanto que o fluxo produzido por uma bomba centrífuga é bastante constante. Isto foi possível ser verificado por medidas com o velocímetro Doppler laser. Tal resultado permite para experimentos relacionados à turbulência e estagnação em um componente de circulação extracorpórea, a eleição da bomba centrífuga como ideal. A técnica, não invasiva, de velocimetria Doppler laser, devidamente adaptada aos componentes de circulação extracorpórea, associada ainda a defmições e convenções para o fluxo de um líquido hidrodinamicamente semelhante ao sangue, se mostra ideal para estudo de turbulência e estagnação em componentes passivos da circulação extracorpórea. Observamos que não há grandes variações do perfil do escoamento dentro do reservatório de cardioplegia em função dos fluxos utilizados. Por outro lado observamos que as velocidades dentro do reservatório aumentam com o aumento do fluxo, guardando um padrão de semelhança entre as situações equivalentes para os diferentes fluxos, pois há uma pequena impedância no caminho do fluxo, o que nos dá uma idéia da localização de zonas de grande turbulência. Há também confluência de fluxo, representando uma zona de estagnação do fluido, e as demais zonas podem ser definidas como normais ou de baixa turbulência, o que valida a Velocimetria Doppler a Laser como método de estudo para otimização dos componentes descartáveis utilizados em circulação extracorpórea / Made available in DSpace on 2018-08-03T15:30:37Z (GMT). No. of bitstreams: 1 Antunes_Nilson_M.pdf: 10886553 bytes, checksum: 307f8538bb6690d26bbc4fc2e77b7339 (MD5) Previous issue date: 2002 / Abstract: Introduction: Cardiopulmonary bypass ( CBP ) associates techniques that allow a temporary cardiopulmonary isolation through utilization of a set of machines, devices and circuits. Sophisticated techniques must be applied in order to study blood behavior inside cardiopulmonary bypass system without interference. The Laser Doppler Velocimetry (LDV) uses light as source for velocity measurements and therefore, appears to be the ideal method for tOOt kind of study. Among several disturbs induced by CBP that can lead to physiologic, hemodynamic, chemical and physical imbalance, physical alterations created by mechanical trauma over blood cells have received special attention on this study. Objectives: 1. To study and describe the flow generated by centrifugal and roller pumps. 2. To establish ideal conditions for the observation of blood turbulence and stagnation inside passive components of CBP. 3. To validate Laser Doppler Velocimetry as an adequate method of study for CBP components. Material and Method: A Laser Doppler Velocimeter was utilized to measure velocity in two dimensions inside blood cardioplegia reservoir, a shell and tube heat exchanger component of CBP. A blender of 36% glycerin water solution was used because of its similarity with blood. Observation was obtained through dislocation by a centrifugal pump of a 5 µm polyamide partic1es fluid suspension. Results aod Cooclusioos: Flow created by the roller pump has significant oscilattory characteristics. Flow produced by centrifugal pump is linear. The technique of Laser Doppler Velocimetry might permit comprehension of fluid drainage behavior under conditions developed on this study. These results indicate centrifugal pump as the ideal one Non-invasive Laser Doppler Velocimetry, well adapted to CPB components and associated to measure flow of hydrostatically blood-like fluid is ideal for study of fluid turbulence and stagnation inside passive CPB components. No great variations were observed on drainage profile inside cardioplegia reservoir related to type of flow employed. On the other hand, velocity inside the reservoir grew higher as flow increased, maintaining a similar pattern between equivalent situations with different flows, because there is mild impedance on the flow pathway, that leads to identification of severe turbulence zones. There have been observed, in a commercial heat exchanger, zones of flow confluence, representing flow stagnation area, and the remainder were defined as normal and low turbulence zones, which therefore attests Laser Doppler Velocimetry as a method of study for improvement of non reusable components of CBP / Mestrado / Cirurgia / Mestre em Cirurgia

Malaria is currently a huge treat worldwide, as far as infections are concerned, and is responsible for thousands of deaths per annum. The dilemma associated with the development of anti–malarial drug resistance over the past few decades should be addressed as a matter of urgency. Novel drug delivery systems should be developed in order to employ new and existing anti–malarial drugs in the treatment and management of malaria. The aim of these delivery systems should include an improvement in the efficacy, specificity, acceptability and therapeutic index of anti–malarial drugs. Previous studies have suggested that liposomes have the ability to encapsulate, protect and to promote the sustained release of anti–malarial drugs. Two liposome formulations, namely liposomes and chloroquine entrapped in liposomes, were formulated during this thesis and evaluated by conducting a stability study and an in vitro study with the main focus on cell viability. The stability study consisted of a series of stability tests regarding the stability of nine liposome and nine chloroquine entrapped in liposome formulations over a period of twelve weeks. The in vitro study included three assays such as a reactive oxygen species assay, a lipid peroxidation assay and a hemolysis assay. The aims of these studies included the manufacturing of liposomes, the incorporation of chloroquine into liposomes, the determination of the stability of the formulations as well as the evaluation of the possible in vitro toxicity of liposomes. Results obtained from these studies revealed that liposomes remained more stable over the stability study period in comparison to chloroquine entrapped in liposomes. The entrapment of chloroquine within liposomes was possible, although the initial entrapment efficiency (%) of 14.55 % was much too low. The production of reactive oxygen species occurred to a small extent in the red blood cells and the infected red blood cells. Equal amounts of reactive oxygen species (%) was observed within both the red blood cells and the infected red blood cells with a maximum value of 23.27 % in the presence of the chloroquine entrapped in liposomes at varying concentrations. Red blood cells experienced the highest degree of lipid peroxidation (%) in the presence of chloroquine, at varying concentrations, entrapped in liposomes. The maximum amount of lipid peroxidation (%) was 79.61 %. No significant degree of hemolysis (%) was observed in the red blood cells neither in the presence of the liposomes nor in the presence of the chloroquine entrapped in liposomes at varying concentrations. It can be concluded that liposomes are a more stable formulation and have less toxic effects on red blood cells and infected red blood cells in comparison to the chloroquine entrapped in liposome formulations. Future studies should investigate the possibility of a more stable and less toxic chloroquine entrapped in liposome formulation. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2011.

Malaria is currently a huge treat worldwide, as far as infections are concerned, and is responsible for thousands of deaths per annum. The dilemma associated with the development of anti–malarial drug resistance over the past few decades should be addressed as a matter of urgency. Novel drug delivery systems should be developed in order to employ new and existing anti–malarial drugs in the treatment and management of malaria. The aim of these delivery systems should include an improvement in the efficacy, specificity, acceptability and therapeutic index of anti–malarial drugs. Previous studies have suggested that liposomes have the ability to encapsulate, protect and to promote the sustained release of anti–malarial drugs. Two liposome formulations, namely liposomes and chloroquine entrapped in liposomes, were formulated during this thesis and evaluated by conducting a stability study and an in vitro study with the main focus on cell viability. The stability study consisted of a series of stability tests regarding the stability of nine liposome and nine chloroquine entrapped in liposome formulations over a period of twelve weeks. The in vitro study included three assays such as a reactive oxygen species assay, a lipid peroxidation assay and a hemolysis assay. The aims of these studies included the manufacturing of liposomes, the incorporation of chloroquine into liposomes, the determination of the stability of the formulations as well as the evaluation of the possible in vitro toxicity of liposomes. Results obtained from these studies revealed that liposomes remained more stable over the stability study period in comparison to chloroquine entrapped in liposomes. The entrapment of chloroquine within liposomes was possible, although the initial entrapment efficiency (%) of 14.55 % was much too low. The production of reactive oxygen species occurred to a small extent in the red blood cells and the infected red blood cells. Equal amounts of reactive oxygen species (%) was observed within both the red blood cells and the infected red blood cells with a maximum value of 23.27 % in the presence of the chloroquine entrapped in liposomes at varying concentrations. Red blood cells experienced the highest degree of lipid peroxidation (%) in the presence of chloroquine, at varying concentrations, entrapped in liposomes. The maximum amount of lipid peroxidation (%) was 79.61 %. No significant degree of hemolysis (%) was observed in the red blood cells neither in the presence of the liposomes nor in the presence of the chloroquine entrapped in liposomes at varying concentrations. It can be concluded that liposomes are a more stable formulation and have less toxic effects on red blood cells and infected red blood cells in comparison to the chloroquine entrapped in liposome formulations. Future studies should investigate the possibility of a more stable and less toxic chloroquine entrapped in liposome formulation. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2011.

Orientador: Eugenio Spano Rosa / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Mecanica / Made available in DSpace on 2018-07-22T03:47:07Z (GMT). No. of bitstreams: 1 Barbosa_MarcosPinotti_D.pdf: 44752191 bytes, checksum: 4001ae207eef045165cfeee88edbd4e8 (MD5) Previous issue date: 1996 / Resumo: A falta de informação básica sobre os mecanismos responsáveis pela hemólise mecânica impede, na maioria dos casos, o uso adequado da bomba centrífuga. Visando a preencher esta lacuna, este trabalho foi conduzido de modo a se estudarem as estruturas básicas do escoamento no interior de uma bomba centrífuga e relacioná-las a um potencial de hemólise. Foram empregadas duas abordagens distintas: simulação numérica de um canal isolado dabomba, com o objetivo de compreender a interação entre as forças inerciais, viscosas e de pressão em diversas situações operacionais, e medição experimental do campo de velocidade nos canais internos da bomba, com a finalidade de se obter uma visão global do escoamento e da interação entre os diferentes canais. O desafio do método numérico foi resolver as equações do movimento escritas em coordenadas ortogonais generalizadas a partir da formulação de fluxo de tensão. O programa computacional foi implementado em FORTRAN, empregando um algoritmo baseado no método dos volumes finitos. As medidas de velocidade foram realizadas por um sistema de anemômetro laser Doppler de última geração, capaz de medir duas componentes simultâneas de velocidade. Desenvolveu-se um protocolo experimental que superou os principais obstáculos associados ao acesso óptico às regiões de interesse no interior da bomba e à similaridade entre as condições de testes e as fisiológicas. A contribuição deste trabalho foi o desenvolvimento de uma metodologia de análise que relaciona as estruturas básicas do escoamento ao potencial de danos às células vermelhas do sangue / Abstract: The lack of basic information about the performance of mechanical hemolysis severely affects centrifugal blood pump handling. The present work faced this problem by studying the flow structures and the way hemolytic potential is influenced by them. Two distinct approaches were employed: numerical simulation of a single channel of the pump, in order to understand the interaction of inertial, viscous and pressure forces in different operational situations, and non-invasive measurements of the velocity field to provide a general picture of the flow field as well as the interaction between the flow generated by each channeI. Solving the Momentum equations, that were written in generalized orthogonal coordinates, using the stress-flux formulation, was the challenge of the numerical method. The computational program was devised by employing the finite volume method and it was implemented in FORTRAN language. The velocity measurements were performed by a laser Doppler anemometer system capable of acquiring and processing two simultaneous orthogonal velocity components. Difficulties arising ftom the optical access to the regions of interest inside the pump and from the similitude between testing and phisiological conditions were overcome by the experimental protocol specially designed for non-intrusive velocity measurements. The main contribution of the present study concems the development of a specific methodology which provides the link between the flow structures and the potential damage to the red blood cells / Doutorado / Termica e Fluidos / Doutor em Engenharia Mecânica

Orientador: Eneida de Paula / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-07T20:44:09Z (GMT). No. of bitstreams: 1 Prete_PauloSergioCastilho_D.pdf: 3376950 bytes, checksum: fd06e65f64531668e35cf4a9cb817913 (MD5) Previous issue date: 2006 / Resumo: Surfatantes ou detergentes são compostos anfifílicos que, na presença de água, têm a característica de formar agregados micelares. Surfatantes induzem a desestruturação de outros agregados como bicamadas sendo, por isso, usados para ruptura celular ou solubilização de lipídios e proteínas de membrana. A capacidade lítica dos surfatantes resulta de sua estrutura química, que determina o modo de interação dos mesmos com as membranas. Em concentrações mais altas (acima da concentração micelar crítica), os surfatantes desestabilizam as bicamadas lipídicas, levando à formação de micelas-mistas. Ensaios hemolíticos são bons modelos para estudo do efeito lítico de surfatantes em biomembranas. Aplicando em eritrócitos humanos o tratamento quantitativo proposto por Lichtenberg (1985) para estudo da solubilização de bicamadas lipídicas mensurou-se, neste trabalho, as concentrações para início (Csat) e 100% de hemólise (Csol), induzidas por 25 surfatantes clássicos, pertencentes a cinco diferentes famílias. A variação dos valores de Csat, Csol determinada com diferentes hematócritos permitiu o cálculo da constante de ligação surfatante/membrana e da razão surfatante/lipídio de membrana (Re) para início e 100% de hemólise. O parâmetro Re foi usado para classificar os detergentes como fortes, médios ou fracos agentes solubilizantes, com boa correlação com dados da literatura o que nos permitiu propor seu uso para descrever o efeito lítico de surfatantes, como uma alternativa simples e aplicável as membranas biológicas. As transições durante o processo hemolítico foram acompanhadas pela técnica de Ressonância Paramagnética Eletrônica, com uso marcador de spin 5 doxil-estearato (incorporado a 1 mol% nas membranas de eritrócito) e lise induzida pelo surfatante não iônico Triton X100. Concomitante ao aparecimento de hemoglobina e fosfato livres no sobrenadante - indicadores da ruptura da membrana, medidas do parâmetro de ordem daquele marcador de spin permitiram estudar as transições que acontecem durante (membrana:membrana mista) e após (membrana mista:micela mista) a hemólise / Abstract: Surfactants or detergents are amphiphilic compounds that form micellar aggregates in the presence of excess water. Surfactants are able to induce disruption of lamellar aggregates, justifying their use for cell lysis or in the extraction of membrane constituents such as lipids and proteins. The lytic capacity of a given surfactant is determined by its chemical structure, that rules its interaction with the membranes. At high concentration (above the critic micelle concentration), surfactants destabilize lipid bilayer leading to mixed micelle formation. Hemolytic assays are a good model to study the lytic effect of surfactants on biomembranes. In this study we have applied to human erythrocytes the quantitative treatment proposed by Lichtenberg (1985) to describe the solubilization of model lipid membranes. The concentration for onset (Csat) and complete (Csol) hemolysis induced by 25 classic surfactants from five different families were measured. Changes in Csat and Csol values at different hematocrits allowed the determination of the surfactant/membrane lipid molar ratio (Re) for beginning and 100% lysis. The Re arameter was used to classify the surfactants as strong, medium or weak membrane solubilizers. The classification was in good correlation with data in the literature, allowing us to recommend the use of Re parameter to describe the lyric effect of surfactants on biomembranes. The transitions in the hemolytic process were accompanied by Electron Paramagnetic Resonance, using the 5-doxyl-stearate spin-probe (1 mol%, incorporated in the erythrocyte membrane) and the non-ionic surfactant Triton X100. Simultaneously to the appearance of hemoglobin and phosphate released in the supernatant, measurements of the order parameter of the spin probe were used to characterize the transitions that take place during (membrane :mixed membrane) and after (mixed: membrane: mixed micelle) hemolysis / Doutorado / Bioquimica / Doutor em Biologia Funcional e Molecular