Este trabalho propoe a definicao de uma arquitetura para incorporar jogadores virtuais em uma classe de Sistemas de Apoio a Decisao em Grupos (SADs-G), baseados na utilizacao conjunta de Simulacao Baseada em Multiagentes (MABS) e Jogos de Papeis (RPGs). Estas duas tecnicas, MABS e RPGs, vem sendo utilizadas de forma integrada ha alguns anos devido a capacidade de discussao e aprendizado dos RPG e da capacidade dinamica de MABS. Os jogadores virtuais definidos devem ter a capacidade de tomada de decisao e de comunicacao entre si e com os jogadores reais durante as negociacoes. Os principais aspectos discutidos neste trabalho sao: i) se os jogadores virtuais definidos possuem comportamentos nao-triviais perante aos jogadores reais; ii) se continua ocorrendo o processo de negociacao entre os jogadores, sejam eles reais ou virtuais; iii) se a realizacao de um jogo de forma eletronica (via computador) dificulta a interacao entre os jogadores. De forma a discutir estes aspectos, duas instancias de SADs-G foram implementadas no contexto da Gestao de Recursos Naturais, pois neste dominio o processo de negociacao e extremamente importante e complexo. / This thesis proposes the definition of an architecture to insert virtual players in a particular subclass of Group Decision Support Systems (GDSS), that uses Multi-Agent-Based Simulation (MABS) and Role-Playing Games (RPGs) techniques in an integrated way. These techniques can bring interesting results, since it is possible to join the dynamic capacity of MABS with the discussion and learning capacity of RPGs. The defined virtual players must make decisions and communicate with each other and with the real players during the negotiation process. In this work, the main aspects discussed are the following: i) can virtual players have no-trivial behavior in the face of real players? ii) is the negotiation process between all players (virtual or real) still happening when virtual players are inserted? iii) do electronic games make more difficult the interaction between players? In order to discuss these aspects, two instances of GDSSs were developed in the natural resources management domain. This domain was chosen because its negotiation process is both very important and complex.

The objective of this research is to develop and apply a HILIC UHPLC stationary phase that allows for separation of intact glycoproteins. In Chapter 1 I give an overview of the problems of current glycosylation profiling with regards to biotherapeutics, and my strategy to separate the intact glycoprotein with HILIC. Chapter 2 describes the methods used to produce the nonporous packing material and stationary phase. In Chapter 3 I describe previous work in developing a HILIC polyacrylamide stationary phase, and further improvements I have made. Chapter 4 describes development of an assay in collaboration with Genentech of therapeutic mAb glycosylation. In Chapter 5, I show HILIC-MS of digested ribonuclease B as a beginning step to analyze glycosylated biomarkers.

Este trabalho propoe a definicao de uma arquitetura para incorporar jogadores virtuais em uma classe de Sistemas de Apoio a Decisao em Grupos (SADs-G), baseados na utilizacao conjunta de Simulacao Baseada em Multiagentes (MABS) e Jogos de Papeis (RPGs). Estas duas tecnicas, MABS e RPGs, vem sendo utilizadas de forma integrada ha alguns anos devido a capacidade de discussao e aprendizado dos RPG e da capacidade dinamica de MABS. Os jogadores virtuais definidos devem ter a capacidade de tomada de decisao e de comunicacao entre si e com os jogadores reais durante as negociacoes. Os principais aspectos discutidos neste trabalho sao: i) se os jogadores virtuais definidos possuem comportamentos nao-triviais perante aos jogadores reais; ii) se continua ocorrendo o processo de negociacao entre os jogadores, sejam eles reais ou virtuais; iii) se a realizacao de um jogo de forma eletronica (via computador) dificulta a interacao entre os jogadores. De forma a discutir estes aspectos, duas instancias de SADs-G foram implementadas no contexto da Gestao de Recursos Naturais, pois neste dominio o processo de negociacao e extremamente importante e complexo. / This thesis proposes the definition of an architecture to insert virtual players in a particular subclass of Group Decision Support Systems (GDSS), that uses Multi-Agent-Based Simulation (MABS) and Role-Playing Games (RPGs) techniques in an integrated way. These techniques can bring interesting results, since it is possible to join the dynamic capacity of MABS with the discussion and learning capacity of RPGs. The defined virtual players must make decisions and communicate with each other and with the real players during the negotiation process. In this work, the main aspects discussed are the following: i) can virtual players have no-trivial behavior in the face of real players? ii) is the negotiation process between all players (virtual or real) still happening when virtual players are inserted? iii) do electronic games make more difficult the interaction between players? In order to discuss these aspects, two instances of GDSSs were developed in the natural resources management domain. This domain was chosen because its negotiation process is both very important and complex.

Biological activity of IL-2 and IL-7 in vivo is significantly increased when complexed with some of the respective anti-cytokine mAb. Different immune cell subsets can be preferentially stimulated depending on the anti-IL-2 mAb used to complex IL-2. IL-2/anti-IL-2 mAb S4B6 immunocomplexes (IL-2/S4B6) induce preferential expansion of CD122high cells whereas IL-2/anti-IL-2 mAb JES6-1 immunocomplexes (IL-2/JES6-1) highly selectively stimulate CD25high cells in mice. Similarly, IL-7/anti-IL-7 mAb M25 immunocomplexes (IL-7/M25) possess higher stimulatory activity for both naïve and memory CD8+ T cells in vivo in comparison to free IL-7. CTLA-4 and PD-1 molecules are inhibitory receptors which negatively regulate proliferation, survival and effector functions of T cells. Blocking antibodies against these molecules represent promising immunotherapeutic tool for treatment of malignant diseases. We examined possible synergism of IL-2/S4B6 and αCTLA-4 plus αPD-1 mAbs in tumor-bearing mice. We found that the expansion of recently activated CD8+ T cells driven by IL-2/S4B6 was further augmented by αCTLA-4 plus αPD-1 mAbs. However, these two immunotherapeutic approaches did not show synergistic antitumor activity in any mouse tumor model tested. Next, we showed that IL-7/M25 possessed higher biological activity...

Antalet monoklonala antikroppar som används i läkemedel ökar kraftigt. Dessa läkemedel är dyra och risken för förfalskning är stor. Behovet att utveckla en metod för snabb och precis identifiering av monoklonala antikroppar är därför brådskande. För identifiering utfördes analyser med Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-ToF-MS), Capillary Gel Electrophoresis (CGE) and Capillary Zone Electrophoresis (CZE) på nio monoklonala antikroppar. Fokuset var att undersöka huruvida signifikanta fysiokemiska egenskaper och unika aminosyrasekvenser var närvarande och kunde urskiljas. Olika analyser med MALDI-ToF-MS användes till att både separera de monoklonala antikropparna baserat på dess fysiokemiska egenskaper, och annotera aminosyrasekvenser innehållande nyckelfragment. Med metoderna baserade på kapillärelektrofores uppnåddes också separation. CZE föredras framför CGE då mängden data som erhålls från CZE är större och provberedningen är enklare. Sammanfattningsvis utformades ett protokoll för identifieringsprocessen, vilket inleds med MALDI-ToF-MS-analyser av monoklonala antikroppar på reducerad form mot kända referenser. Därefter är en hypotes formulerad utifrån vilka antikroppar som ser mest lika ut. Slutligen analyseras dessa med CZE för fastställning av den monoklonala antikroppens identitet. / The number of monoclonal antibodies used in pharmaceuticals is increasing sharply. These medicines are expensive, and the risk of counterfeiting is high. The need to develop a method for rapid and precise identification of monoclonal antibodies is therefore urgent. For identification, analyses were performed with Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-ToF-MS), Capillary Gel Electrophoresis (CGE) and Capillary Zone Electrophoresis (CZE) on nine monoclonal antibodies. The focus was to investigate whether significant physiochemical features and unique amino acid sequences were present and could be distinguished. Various analyses with MALDI-ToF-MS were used to both separate the monoclonal antibodies based on their physicochemical properties and annotate amino acid sequences containing key fragments. With the methods based on capillary electrophoresis, separation was also achieved. CZE is preferred over CGE as the amount of data obtained from CZE is greater and sample preparation is simpler. In summary, an identification process protocol was designed and is initiated with MALDI-ToF-MS analyses of reduced-form monoclonal antibodies against known references. A hypothesis is then formulated based on which antibodies look the most similar. Finally, these are analysed by CZE to determine the identity of the monoclonal antibody.