Orientador: Sérgio Santos Mühlen / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-23T00:55:24Z (GMT). No. of bitstreams: 1 Fernandes_LearddeOliveira_M.pdf: 4852939 bytes, checksum: 7a95fc7f08613cf9187cb12b3e4b3f50 (MD5) Previous issue date: 2013 / Resumo: Em experimentos que envolvem estimulação de preparações biológicas e cuja resposta é representada por um ou mais sinais, o controle do processo de estimulação e o monitoramento dos sinais resultantes são essenciais para um melhor entendimento de como o sistema biológico responde à estimulação. Assim, este trabalho foi realizado com o objetivo de desenvolver um instrumento virtual para uso em experimentos com preparações biológicas, capaz de gerar estímulos, adquirir e processar os sinais biológicos decorrentes dos estímulos, a fim de propiciar um único ambiente que atenda ao máximo os requisitos de qualquer protocolo experimental que possa envolver estimulação, aquisição e análise de sinais. Foi então desenvolvida uma plataforma virtual para experimentos em preparações biológicas de modo a automatizar o protocolo experimental de um setup. Todo o desenvolvimento dos sistemas de controle, estimulação, aquisição e processa-mento do sinal do instrumento virtual foi concebido de modo a maximizar a flexibilidade no manuseio e a generalidade nas aplicações. O sistema desenvolvido foi capaz de controlar a geração e a aplicação de estímulos (pulsos de ultras-som de potência) em preparações in vitro de corações de ratos, tornando possível automatizar todo o protocolo experimental de estimulação, adquirir os sinais de temperatura e força de contração da preparação de forma síncrona ao estímulo e pós-processar os sinais adquiridos. O instrumento virtual pode ser aplicado em diferentes setups experimentais de preparações biológicas / Abstract: In experiments involving stimulation of biological preparations and their response expressed by a given acquired signal, control of the stimulation and monitoring of the signal are essential to better understand how the biological system reacts to the stimulation. This study was conducted with the aim of developing a virtual instrument for use in experiments with biological preparations, capable of generating stimuli, acquire and process the biological signals from stimulation, providing a unique environment that meets the maximum requirements from any experimental protocol that might involve stimulation, acquisition and signal analysis. A virtual platform was developed for controlling experiments with biological preparations, making possible to automate the setup. All the development of control, stimulation and acquisition systems, and signal processing of the virtual instrument was designed to maximize the flexibility in the use, and the generality of the applications. The developed system is capable of controlling the generation and application of stimuli (pulses of power ultrasound) in an in vitro preparation of rat hearts, automating the whole experimental protocol of stimulation, acquisition of temperature and contractile force signals synchronized to the stimuli, and post-process the acquired signals. The virtual instrument can be applied in different experimental setups with biological preparations / Mestrado / Engenharia Biomedica / Mestre em Engenharia Elétrica

Thesis (PhD)--Stellenbosch University, 2005. / ENGLISH ABSTRACT: Since their inception, biology and biochemistry have been spectacularly successful in characterising the living cell and its components. As the volume of information about cellular components continues to increase, we need to ask how we should use this information to understand the functioning of the living cell? Computational systems biology uses an integrative approach that combines theoretical exploration, computer modelling and experimental research to answer this question. Central to this approach is the development of computational models, new modelling strategies and computational tools. Against this background, this study aims to: (i) develop a new modelling package: PySCeS, (ii) use PySCeS to study discontinuous behaviour in a metabolic pathway in a way that was very difficult, if not impossible, with existing software, (iii) develop an interactive, web-based repository (JWS Online) of cellular system models. Three principles that, in our opinion, should form the basis of any new modelling software were laid down: accessibility (there should be as few barriers as possible to PySCeS use and distribution), flexibility (pySCeS should be extendable by the user, not only the developers) and usability (PySCeS should provide the tools we needed for our research). After evaluating various alternatives we decided to base PySCeS on the freely available programming language, Python, which, in combination with the large collection of science and engineering algorithms in the SciPy libraries, would give us a powerful modern, interactive development environment. / AFRIKAANSE OPSOMMING: Sedert hul totstandkoming was biologie en, meer spesifiek, biochemie uiters suksesvol in die karakterisering van die lewende sel se komponente. Steeds groei die hoeveelheid informasie oor die molekulêre bestanddele van die sel daagliks; ons moet onself dus afvra hoe ons hierdie informasie kan integreer tot 'n verstaanbare beskrywing van die lewende sel se werking. Om dié vraag te beantwoord gebruik rekenaarmatige sisteembiologie 'n geïntegreerde benadering wat teorie, rekenaarmatige modellering en eksperimenteeIe navorsing kombineer. Sentraal tot die benadering is die ontwikkeling van nuwe modelle, strategieë vir modellering, en sagteware. Teen hierdie agtergrond is die hoofdoelstelling van hierdie projek: (i) die ontwikkeling van 'n nuwe modelleringspakket, PySCeS (ii) die benutting van PySCeS om diskontinue gedrag in n metaboliese sisteem te bestudeer (iets wat met die huidiglik beskikbare sagteware redelik moeilik is), (en iii) die ontwikkeling vann interaktiewe, internet-gebaseerde databasis van sellulêre sisteem modelle, JWS Online. Ons is van mening dat nuwe sagteware op drie belangrike beginsels gebaseer behoort te wees: toeganklikheid (die sagteware moet maklik bekombaar en bruikbaar wees), buigsaamheid (die gebruiker moet self PySCeS kan verander en ontwikkel) en bruikbaarheid (al die funksionalitiet wat ons vir ons navorsing nodig moet in PySCeS ingebou wees). Ons het verskeie opsies oorweeg en besluit om die vrylik verkrygbare programmeringstaal, Python, in samehang die groot kolleksie wetenskaplike algoritmes, SciPy, te gebruik. Hierdie kombinasie verskaf n kragtige, interaktiewe ontwikkelings- en gebruikersomgewing. PySCeS is ontwikkel om onder beide die Windows en Linux bedryfstelsels te werk en, meer spesifiek, om gebruik te maak van 'n 'command line interface'. Dit beteken dat PySCeS op enige interaktiewe rekenaar-terminaal Python ondersteun sal werk. Hierdie eienskap maak ook moontlik die gebruik van PySCeS as 'n modelleringskomponent in 'n groter sagteware pakket onder enige bedryfstelsel wat Python ondersteun. PySCeS is op 'n modulere ontwerp gebaseer, wat dit moontlik vir die eindgebruiker maak om die sagteware se bronkode verder te ontwikkel. As 'n toepassing is PySCeS gebruik om die oorsaak van histeretiese gedrag van 'n lineêre, eindproduk-geïnhibeerde metaboliese pad te ondersoek. Ons het hierdie interessante gedrag in 'n vorige studie ontdek, maar kon nie, met die sagteware wat op daardie tydstip tot ons beskikking was, hierdie studie voortsit nie. Met PySCeS se ingeboude vermoë om parameter kontinuering te doen, kon ons die oorsake van hierdie diskontinuë gedrag volledig karakteriseer. Verder het ons 'n nuwe metode ontwikkel om hierdie gedrag te visualiseer as 'n interaksie tussen die volledige sisteem se subkomponente. Tydens PySCeS se ontwikkeling het ons opgemerk dat dit baie moeilik was om metaboliese modelle wat in die literature gepubliseer is te herbou en te bestudeer. Hierdie situasie is grotendeels die gevolg van die feit dat nêrens 'n sentrale databasis vir metaboliese modelle bestaan nie (soos dit wel bestaan vir genomiese data of proteïen strukture). Die JWS Online databasis is spesifiek ontwikkel om hierdie leemte te vul. JWS Online maak dit vir die gebruiker moontlik om, via die internet en sonder die installasie van enige gespesialiseerde modellerings sagteware, gepubliseerde modelle te bestudeer en ook af te laai vir gebruik met ander modelleringspakkette soos bv. PySCeS. JWS Online het alreeds 'n onmisbare hulpbron vir sisteembiologiese navorsing en onderwys geword.

Neste trabalho avaliamos a formação de oxigênio singlete in vitro em fagócitos, (células mononucleares e neutrófilos) isolados de sangue periférico humano, e eosinófilos, de lavado bronco alveolar de camundongos balb/c, ativados por estímulo partículado: zimosan opsonizado contendo o 9,10difenilantraceno (DPA) adsorvido como sonda captadora de 1O2. Por este método, a formação do 1O2 pode ser verificada pela formação do 9,10-difenilantraceno endoperóxido (DPAO2), que é detectado por HPLC. Observamos, que os fagócitos formam 1O2 e que esta formação parece ocorrer de forma diferenciada para os dois tipos celulares (neutrófilos e células mononucleares). Visando ampliar os estudos anteriores sobre o papel da melatonina (MLT) no processo inflamatório, foi testado seu efeito em fagócitos e a relação na produção de 1O2 destas células. Observamos que MLT inibe a formação de 1O2 totalmente no caso de neutrófilos e parcialmente no caso de células mononucleares e eosinófilos. Paralelamente, foi desenvolvida a síntese de um novo captador químico de 1O2, o éster 9,10-antracenil-3-bispropionato de etila (ABPE), cuja finalidade principal é o acúmulo no interior da célula, depois de sofrer hidrólise enzimática. Esta sonda, terá facil acesso ao interior das células em sua forma ester. Este novo captador de 1O2 foi testado em células mononucleares e neutrófilos estimulados de formas diferentes: via receptor independente e dependente. Os resultados demonstraram produção equivalente de 1O2 nestes fagócitos. / In this study, we evaluated the singlet oxygen (1O2) formation in vitro from phagocytes (neutrophils and mononuclear cells) isolated from human blood cells and eosinophils isolated from bronchoalveolar lavage fluid of mice balb/c activated, by opsonized zymosan. To determine whether singlet oxygen is produced by phagocytes, zymosan particles were coated with a specific chemical trap for 1O2, 9,10-diphenylanthracene (DPA). The production of 1O2 was followed using HPLC, to measure its product, 9,10-diphenylanthracene endoperoxide (DPAO2). We also noticed that the 1O2 production occurs at different levels of for two cell types, neutrophils and mononuclear cells. In order to broaden previous studies on the role of melatonin (MLT) in inflammatory processes, its effect was tested in phagocytes was tested in relation to 1O2 formation by these cells. We observed that MLT inhibits the 1O2 formation totallymt neutrophils and partiallym mononuclear cells and eosinophils. At the some time, it was also developed the synthesis of a new probe for 1O2, the 9,10-anthracene-bis-3-ethyl-propionate (ABEP), with the purpose to accumulate inside the cells, after its enzymatic hydrolysis. This probe presents easy acess to the inferior of the cells in its ester form. This new probe for trapping 1O2 was tested in mononuclear cells and neutrophils stimulated in two ways: via independent and dependent receptor. The results showed equivalent production of 1O2 for both cell types.

Les neurosciences computationnelles consistent en l’étude du système nerveux par la modélisation et la simulation. Plus le modèle sera proche de la réalité et plus les ressources calculatoires exigées seront importantes. La question de la complexité et de la précision est un problème bien connu dans la simulation. Les travaux de recherche menés dans le cadre de cette thèse visent à améliorer la simulation de modèles mathématiques représentant le comportement physique et chimique de récepteurs synaptiques. Les modèles sont décrits par des équations différentielles ordinaires (EDO), et leur résolution passe par des méthodes numériques. Dans le but d’optimiser la simulation, j’ai implémenté différentes méthodes de résolution numérique des EDO. Afin de faciliter la sélection du meilleur algorithme de résolution numérique, une méthode nécessitant un minimum d’information a été proposée. Cette méthode permet de choisir l’algorithme qui optimise la simulation. La méthode a permis de démontrer que la dynamique d’un modèle de récepteur synaptique influence plus les performances des algorithmes de résolution que la structure cinétique du modèle lui-même. De plus, afin de caractériser des comportements pathogènes, une phase d’optimisation est réalisée. Cependant, certaines valeurs de paramètres rendent le modèle instable. Une étude de stabilité a permis de déterminer la stabilité du modèle pour des paramètres fournis par la littérature, mais également de remonter à des contraintes de stabilité sur les paramètres. Le respect de ces contraintes permet de garantir la stabilité des modèles étudiés, et donc de garantir le succès de la procédure permettant de rendre un modèle pathogène. / Computational Neuroscience consists in studying the nervous system through modeling and simulation. It is to characterize the laws of biology by using mathematical models integrating all known experimental data. From a practical point of view, the more realistic the model, the largest the required computational resources. The issue of complexity and accuracy is a well known problem in the modeling and identification of models. The research conducted in this thesis aims at improving the simulation of mathematical models representing the physical and chemical behavior of synaptic receptors. Models of synaptic receptors are described by ordinary differential equations (ODE), and are resolved with numerical procedures. In order to optimize the performance of the simulations, I have implemented various ODE numerical resolution methods. To facilitate the selection of the best solver, a method, requiring a minimum amount of information, has been proposed. This method allows choosing the best solver in order to optimize the simulation. The method demonstrates that the dynamic of a model has greater influence on the solver performances than the kinetic scheme of the model. In addition, to characterize pathogenic behavior, a parameter optimization is performed. However, some parameter values lead to unstable models. A stability study allowed for determining the stability of the models with parameters provided by the literature, but also to trace the stability constraints depending to these parameters. Compliance with these constraints ensures the stability of the models studied during the optimization phase, and therefore the success of the procedure to study pathogen models.

This dissertation addresses the development of integrative modeling strategies capable of combining deterministic and stochastic, discrete and continuous, as well as multi-scale features. The first set of studies combines the purely deterministic modeling methodology of Biochemical Systems Theory (BST) with a hybrid approach, using Functional Petri Nets, which permits the account of discrete features or events, stochasticity, and different types of delays. The efficiency and significance of this combination is demonstrated with several examples, including generic biochemical networks with feedback controls, gene regulatory modules, and dopamine based neuronal signal transduction. A study expanding the use of stochasticity toward systems with small numbers of molecules proposes a rather general strategy for converting a deterministic process model into a corresponding stochastic model. The strategy characterizes the mathematical connection between a stochastic framework and the deterministic analog. The deterministic framework is assumed to be a generalized mass action system and the stochastic analogue is in the format of the chemical master equation. The analysis identifies situations where internal noise affecting the system needs to be taken into account for a valid conversion from a deterministic to a stochastic model. The conversion procedure is illustrated with several representative examples, including elemental reactions, Michaelis-Menten enzyme kinetics, a genetic regulatory motif, and stochastic focusing. The last study establishes two novel, particle-based methods to simulate biochemical diffusion-reaction systems within crowded environments. These simulation methods effectively simulate and quantify crowding effects, including reduced reaction volumes, reduced diffusion rates, and reduced accessibility between potentially reacting particles. The proposed methods account for fractal-like kinetics, where the reaction rate depends on the local concentrations of the molecules undergoing the reaction. Rooted in an agent based modeling framework, this aspect of the methods offers the capacity to address sophisticated intracellular spatial effects, such as macromolecular crowding, active transport along cytoskeleton structures, and reactions on heterogeneous surfaces, as well as in porous media. Taken together, the work in this dissertation successfully developed theories and simulation methods which extend the deterministic, continuous framework of Biochemical Systems Theory to allow the account of delays, stochasticity, discrete features or events, and spatial effects for the modeling of biological systems, which are hybrid and multiscale by nature.

In this thesis, I have studied CMOS dendrites, implemented them on a reconfigurable analog platform and modeled them using MATLAB Simulink. The dendrite model was further used to build a computational model. I implemented a Hidden Markov Model (HMM) classifier to build a simple YES/NO wordspotter. I also discussed the inter-relation between neural systems, CMOS transistors and HMM networks. The physical principles behind the operation of silicon devices and biological structures are similar. Hence silicon devices can be used to emulate biological structures like dendrites. Dendrites are a branched, conductive medium which connect a neurons synapses to its soma. Dendrites were previously believed to be like wires in neural networks. However, recent research suggests that they have computational power. We can emulate dendrites using transistors in the Field Programmable Analog Array (FPAA). Our lab has built the Reconfigurable Analog Signal Processor (RASP) family of FPAAs which was used for the experiments. I analytically compared the mathematical model of dendrites to our model in silicon. The mathematical model based on the device physics of the silicon devices was then used to simulate dendrites in Simulink. An automated tool, sim2spice was then used to convert the Simulink model into a SPICE netlist, such that it can be implemented on a FPAA. This is an easier tool to use for DSP and Neuromorphic engineers who's primary areas of expertise isn't circuit design.

Thesis (Ph. D.)--State University of New York at Binghamton, Thomas J. Watson School of Engineering and Applied Science, Department of Bioengineering, Biomedical Engineering, 2009. / Includes bibliographical references.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / As células dos seres vivos são constantemente ameaçadas por agentes químicos ou físicos que possam causar danos ao DNA. Um dos agentes deste estudo foi o cloreto estanoso (SnCl2), utilizado na medicina nuclear como redutor de um isótopo radioativo do tecnécio, o 99mTc. O SnCl2 é um agente cujo mecanismo de produção de lesões em estruturas celulares envolve a geração de espécies reativas de oxigênio (ERO), tais como o H2O2 e o radical OH. Essas ERO podem causar uma série de doenças, o envelhecimento e até mesmo a morte celular, por apoptose ou necrose. Sendo assim, torna-se importante a pesquisa sobre os efeitos biológicos, tanto desse sal, quanto das outras substâncias que compõem os radiofármacos utilizados em medicina nuclear. Desta forma, nosso objetivo geral foi avaliar a toxicidade do cloreto estanoso, associado, ou não, ao kit 99mTc-MDP, bem como dos demais componentes do kit, em diferentes sistemas biológicos. Eletroforese em gel alcalino de agarose em células de E. coli AB 1157 para a avaliação da genotoxicidade; Ensaio do Cometa em células de sangue total de ratos Wistar para estudar a genotoxicidade; Ensaio do Micronúcleo em células da medula óssea de ratos Wistar para verificar o potencial aneugênico e clastogênico; Ensaio Cometa em células de sangue total e em células mononucleares de sangue periférico humano para estudar a genotoxicidade; Ensaio de Viabilidade com Azul Trypan e citometria de fluxo para analisar a citotoxicidade em PBMC; Ensaio do Micronúcleo em linfócitos humanos para verificar o potencial aneugênico e clastogênico. Em cepas de E. coli AB1157, o SnCl2 e o MDP induziram quebras no DNA genômico, quando isolados; porém quando usados de forma associada, ocorreu uma atenuação do número de quebras. Em ratos Wistar, o 99mTc-MDP não foi genotóxico e também não induziu clastogênese ou aneugênese. Em Sangue total, in vitro, o SnCl2 apresentou efeito dose-resposta. Em PBMC, in vitro, o 99mTc-MDP causou redução da viabilidade celular e apresentou genotoxicidade, porém não induziu clastogênese e nem aneugênese. / Alive cells are constantly threated by chemical and physical agents that can generate DNA damage. Here, the studied agent was stannous chloride (SnCl2), a 99mTc reducing agent employed in nuclear medicine. This salt can produce lesions through generation of reactive oxygen species (ROS) as H2O2 and OH. These ROS can be the origin of several diseases and cell death by apoptosis or necrosis. In this way it is important the research about the biological effects of this salt and the other substances composing the radiopharmaceuticals used in nuclear medicine.The aim of this work was to evaluate, in different biological systems, the stannous chloride toxic potentiality, associated or not to 99mTc-MDP radiopharmaceutical as well as the other kit components. Alkaline electrophoresis agarosis gel of E. coli AB 1157 to evaluate the genotoxicity in prokariotic cells; Comet assay in Wistar rats total blood to evaluate the genotoxicity in eukaryotic cells; Micronucleous assay in Wistar rats bone marrow cells to verify the aneugenic and clastogenic effects; Comet assay in human in peripherical total blood and mononuclear cells to evaluate the genotoxicity; Trypan blue viability assay and flow cytometry to evaluate citotoxicity in PBMC; Micronucleous assay in human lymphocyte cells to verify the aneugenic and clastogenic effects; SnCl2 and MDP when isolated induced breaks in E. coli genomic DNA. But, when used in an associated way it was observed an atenuation of the breaks number. 99mTc-MDP was not genotoxic, clastogenic nor aneugenic in Wistar rats In ex vivo human total blood, SnCl2 presented a dose-response effect In ex vivo PBMC; 99mTc-MDP induced a cellular viability reduction and presented genotoxic but not clastogenic or aneugenic effects.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / As células dos seres vivos são constantemente ameaçadas por agentes químicos ou físicos que possam causar danos ao DNA. Um dos agentes deste estudo foi o cloreto estanoso (SnCl2), utilizado na medicina nuclear como redutor de um isótopo radioativo do tecnécio, o 99mTc. O SnCl2 é um agente cujo mecanismo de produção de lesões em estruturas celulares envolve a geração de espécies reativas de oxigênio (ERO), tais como o H2O2 e o radical OH. Essas ERO podem causar uma série de doenças, o envelhecimento e até mesmo a morte celular, por apoptose ou necrose. Sendo assim, torna-se importante a pesquisa sobre os efeitos biológicos, tanto desse sal, quanto das outras substâncias que compõem os radiofármacos utilizados em medicina nuclear. Desta forma, nosso objetivo geral foi avaliar a toxicidade do cloreto estanoso, associado, ou não, ao kit 99mTc-MDP, bem como dos demais componentes do kit, em diferentes sistemas biológicos. Eletroforese em gel alcalino de agarose em células de E. coli AB 1157 para a avaliação da genotoxicidade; Ensaio do Cometa em células de sangue total de ratos Wistar para estudar a genotoxicidade; Ensaio do Micronúcleo em células da medula óssea de ratos Wistar para verificar o potencial aneugênico e clastogênico; Ensaio Cometa em células de sangue total e em células mononucleares de sangue periférico humano para estudar a genotoxicidade; Ensaio de Viabilidade com Azul Trypan e citometria de fluxo para analisar a citotoxicidade em PBMC; Ensaio do Micronúcleo em linfócitos humanos para verificar o potencial aneugênico e clastogênico. Em cepas de E. coli AB1157, o SnCl2 e o MDP induziram quebras no DNA genômico, quando isolados; porém quando usados de forma associada, ocorreu uma atenuação do número de quebras. Em ratos Wistar, o 99mTc-MDP não foi genotóxico e também não induziu clastogênese ou aneugênese. Em Sangue total, in vitro, o SnCl2 apresentou efeito dose-resposta. Em PBMC, in vitro, o 99mTc-MDP causou redução da viabilidade celular e apresentou genotoxicidade, porém não induziu clastogênese e nem aneugênese. / Alive cells are constantly threated by chemical and physical agents that can generate DNA damage. Here, the studied agent was stannous chloride (SnCl2), a 99mTc reducing agent employed in nuclear medicine. This salt can produce lesions through generation of reactive oxygen species (ROS) as H2O2 and OH. These ROS can be the origin of several diseases and cell death by apoptosis or necrosis. In this way it is important the research about the biological effects of this salt and the other substances composing the radiopharmaceuticals used in nuclear medicine.The aim of this work was to evaluate, in different biological systems, the stannous chloride toxic potentiality, associated or not to 99mTc-MDP radiopharmaceutical as well as the other kit components. Alkaline electrophoresis agarosis gel of E. coli AB 1157 to evaluate the genotoxicity in prokariotic cells; Comet assay in Wistar rats total blood to evaluate the genotoxicity in eukaryotic cells; Micronucleous assay in Wistar rats bone marrow cells to verify the aneugenic and clastogenic effects; Comet assay in human in peripherical total blood and mononuclear cells to evaluate the genotoxicity; Trypan blue viability assay and flow cytometry to evaluate citotoxicity in PBMC; Micronucleous assay in human lymphocyte cells to verify the aneugenic and clastogenic effects; SnCl2 and MDP when isolated induced breaks in E. coli genomic DNA. But, when used in an associated way it was observed an atenuation of the breaks number. 99mTc-MDP was not genotoxic, clastogenic nor aneugenic in Wistar rats In ex vivo human total blood, SnCl2 presented a dose-response effect In ex vivo PBMC; 99mTc-MDP induced a cellular viability reduction and presented genotoxic but not clastogenic or aneugenic effects.

Neste trabalho avaliamos a formação de oxigênio singlete in vitro em fagócitos, (células mononucleares e neutrófilos) isolados de sangue periférico humano, e eosinófilos, de lavado bronco alveolar de camundongos balb/c, ativados por estímulo partículado: zimosan opsonizado contendo o 9,10difenilantraceno (DPA) adsorvido como sonda captadora de 1O2. Por este método, a formação do 1O2 pode ser verificada pela formação do 9,10-difenilantraceno endoperóxido (DPAO2), que é detectado por HPLC. Observamos, que os fagócitos formam 1O2 e que esta formação parece ocorrer de forma diferenciada para os dois tipos celulares (neutrófilos e células mononucleares). Visando ampliar os estudos anteriores sobre o papel da melatonina (MLT) no processo inflamatório, foi testado seu efeito em fagócitos e a relação na produção de 1O2 destas células. Observamos que MLT inibe a formação de 1O2 totalmente no caso de neutrófilos e parcialmente no caso de células mononucleares e eosinófilos. Paralelamente, foi desenvolvida a síntese de um novo captador químico de 1O2, o éster 9,10-antracenil-3-bispropionato de etila (ABPE), cuja finalidade principal é o acúmulo no interior da célula, depois de sofrer hidrólise enzimática. Esta sonda, terá facil acesso ao interior das células em sua forma ester. Este novo captador de 1O2 foi testado em células mononucleares e neutrófilos estimulados de formas diferentes: via receptor independente e dependente. Os resultados demonstraram produção equivalente de 1O2 nestes fagócitos. / In this study, we evaluated the singlet oxygen (1O2) formation in vitro from phagocytes (neutrophils and mononuclear cells) isolated from human blood cells and eosinophils isolated from bronchoalveolar lavage fluid of mice balb/c activated, by opsonized zymosan. To determine whether singlet oxygen is produced by phagocytes, zymosan particles were coated with a specific chemical trap for 1O2, 9,10-diphenylanthracene (DPA). The production of 1O2 was followed using HPLC, to measure its product, 9,10-diphenylanthracene endoperoxide (DPAO2). We also noticed that the 1O2 production occurs at different levels of for two cell types, neutrophils and mononuclear cells. In order to broaden previous studies on the role of melatonin (MLT) in inflammatory processes, its effect was tested in phagocytes was tested in relation to 1O2 formation by these cells. We observed that MLT inhibits the 1O2 formation totallymt neutrophils and partiallym mononuclear cells and eosinophils. At the some time, it was also developed the synthesis of a new probe for 1O2, the 9,10-anthracene-bis-3-ethyl-propionate (ABEP), with the purpose to accumulate inside the cells, after its enzymatic hydrolysis. This probe presents easy acess to the inferior of the cells in its ester form. This new probe for trapping 1O2 was tested in mononuclear cells and neutrophils stimulated in two ways: via independent and dependent receptor. The results showed equivalent production of 1O2 for both cell types.