Fundação de Amparo a Pesquisa do Estado de Minas Gerais / The present thesis demonstrates, for the first time, the possibility of using of the internal standard method in FIA or BIA systems with amperometric detection. The method allows to obtain accurate results in the presence of fluctuations in the system parameters (injected volume, dispensing rate, ionic strength and accidental insertion of air bubbles) and in the presence of contamination and/or gradual passivation of the working electrode surface. Methods for determination of diclofenac (DCF) using FIA with amperometric detection and simultaneous determination of DCF and codeine (CO) by BIA with amperometric detection have also been developed. The internal standard method was implemented in the flow injection analysis with multiple pulse amperometric detection (FIA-MPA) system by applying a sequence of potential pulses of the working electrode as a function of time. The analyte (+0.80 V/300ms) and internal standard (-0.05 V/400ms) were detected selectively, one at each potential pulse. Significant improvements were obtained in the accuracy of the system when the results before and after normalization (using the internal standard method), were compared. The following errors were calculated, respectively, before and after normalization of the results using the internal standard method. Change in flow rate from 3.0 to 1.0 ml min-1 (-57% e < 1%); volume injection from 300 to 100 &#956;L (-63% and 3%); ionic strength from 0.60 to 0.77 mol L-1 (-44 and 0,2%) and before and after the introduction of air bubbles into the system (-9,6% and 1,7%). A significant improvement was also obtained in the correlation coefficient of the calibration curve in the presence of problems of passivation or contamination of the working electrode. The correlation coefficient was calculated, respectively, as 0,975 and 0,998, before and after the normalization of the results by the IS method. The internal standard method was also applied to correct errors derived from the injection procedure (dispensing rate and injected volume) in batch injection analysis systems (BIA) with amperometric detection when disposable syringes were used. The results obtained before and after normalization of the results by the IS method were the following, respectively: repeatability test (RSD = 6.4 and 1.8%; n = 8), correlation coefficients (R=0.954 and 0.997) and errors obtained in analysis of synthetic samples (E = 9.4 ± 1.5 and 3.5 ± 0.7%; n = 8). The determination of DCF by FIA-MPA was implemented by applying two potential pulses to the BDD electrode using H2SO4 0.1 mol L-1 as supporting electrolyte: (i) + 1.2 V / 50 ms for oxidation and quantification of DCF, and (ii) 0.0 V / 50 ms for constant electrochemical cleaning of the electrode surface. The system showed good stability (RSD = 1.0%, n = 10) and high analytical frequency (135 injections h-1). The method showed a linear response between 5 and 50 &#956;mol L-1 and the detection and quantification limits were as 0.14 and 0.46 mmol L-1, respectively. The proposed method was applied to determine of DCF in pharmaceutical formulations and the obtained results were similar to those obtained by HPLC with a confidence level of 95%. The simultaneous determination of DCF and CO was performed using the following potential pulses: (i) +1.1 V / 50 ms: oxidation and quantification of DCF; (ii) +1.4 V / 50 ms: simultaneous oxidation of DCF and CO, (iii) 0.0 V / 200 ms: for constant electrochemical cleaning of the BDD electrode. The oxidation current from CO was obtained by the difference between currents detected at 1.4 and 1.1 V by using a correction factor. The system showed good stability (RSD = 0.9% and 1.1 for DCF and CO respectively, n = 10) and high analytical frequency (~ 300 injections h-1). The method showed a linear response between 10 and 50 &#956;mol L-1 for DCF and 7.1 and 35.7 &#956;mol L-1 for CO. The LQ and LD were calculated, respectively, at 1.1 and 3.7 &#956;mol L-1, for DCF and 1.0 and 3.3 mol L-1 for CO. The proposed method was applied for simultaneous determination of DCF and CO in pharmaceutical formulations and the obtained results were similar to those obtained by HPLC with a confidence level of 95%. Keywords: Multiple pulse amperometry, FIA, BIA, internal standard, simultaneous analysis, boron doped diamond (BDD), codeine, diclofenac. / Esta tese apresenta, pela primeira vez, a possibilidade de usar o método do padrão interno em sistemas FIA ou BIA com detecção amperométrica. O método permite a obtenção de resultados precisos na presença de flutuações em parâmetros dos sistemas (volume injetado, velocidade de injeção, força iônica e inserção acidental de bolhas de ar) e na presença de contaminação e/ou passivação gradual do eletrodo de trabalho. Métodos para determinação de diclofenaco (DCF) usando FIA com detecção amperométrica e simultânea de DCF e codeína (CO) por BIA com detecção amperométrica também foram desenvolvidos. O método do padrão interno foi implementado no sistema de análise por injeção em fluxo com detecção por amperometria de múltiplos pulsos (FIA-MPA) mediante a aplicação de uma sequência de pulsos de potenciais ao eletrodo de trabalho em função do tempo. O analito (+0,80 V/300ms) e o padrão interno (-0,05 V/400ms) foram detectados seletivamente, um em cada pulso de potencial. Melhoras significativas foram obtidas na precisão do método quando os resultados obtidos antes e após a normalização (usando o método do padrão interno) foram comparados. As seguintes variações foram calculadas, respectivamente, antes e após a normalização dos resultados usando o método do padrão interno. Variação na velocidade de vazão de 3,0 para 1,0 mL min-1 (-57% e < 1%); no volume de injeção de 300 para 100 &#956;L (-63% e 3%); força iônica de 0,60 para 0,77 mol L-1 (-44 e 0,2%) e antes e após a inserção de bolhas de ar no sistema (-9,6% e 1,7%). Uma melhora significativa também foi obtida no coeficiente de correlação da curva de calibração na presença de problemas de passivação ou contaminação do eletrodo de trabalho. O coeficiente de correlação foi calculado, respectivamente, em 0,975 e 0,998, antes e após a normalização dos resultados pelo método do PI. O método do padrão interno também foi aplicado para corrigir erros em sistemas de análise por injeção em batelada (BIA) com detecção amperométrica provenientes do procedimento de injeção (velocidade de injeção e volume injetado) quando seringas descartáveis foram utilizadas. Os resultados obtidos antes e após a normalização dos resultados pelo método do PI foram os seguintes, respectivamente: estudo de repetibilidade (DPR=6,4 e 1,8%; n = 8), coeficientes de correlação (R=0,954 e 0,997) e erros obtidos na análise de amostras sintéticas (9,4 ± 1,5% e 3,5 ± 0,7; n = 8). A determinação de DCF por FIA-MPA foi implementada através da aplicação de dois pulsos de potenciais ao eletrodo de BDD em meio de H2SO4 0,1 mol L-1 como eletrólito suporte: (i) +1,2 V / 50 ms para oxidação e quantificação do DCF e, (ii) 0,0 V / 50 ms para constante limpeza eletroquímica do eletrodo de trabalho. O sistema apresentou boa estabilidade (RSD = 1,0%; n=10) e elevada frequência analítica (135 injeções h-1). O método apresentou resposta linear entre 5 e 50 &#956;mol L-1 e os limites de detecção e quantificação foram calculados em 0,14 e 0,46 &#956;mol L-1, respectivamente. O método proposto foi aplicado na determinação de DCF em formulações farmacêuticas e os resultados obtidos foram similares ao obtidos por HPLC a um nível de confiança de 95%. A determinação simultânea de DCF e CO foi realizada usando os seguintes pulsos de potenciais: (i) +1,1 V / 50 ms: oxidação e quantificação de DCF; (ii) +1,4 V / 50 ms: para oxidação simultânea de DCF e CO; (iii) 0,0 V / 200 ms: para constante limpeza eletroquímica do eletrodo de BDD. A corrente de oxidação da CO foi obtida pela subtração entre as correntes detectadas em 1,4 e 1,1 V mediante uso de um fator de correção. O sistema apresentou boa estabilidade (RSD= 1,1 e 0,9% para DCF e CO, respectivamente, n=10) e elevada frequência analítica (~300 injeções h-1). O método apresentou resposta linear entre 10 e 50 &#956;mol L-1 para o DCF e entre 7,1 e 35,7 &#956;mol L-1 para CO. O LD e o LQ foram calculados, respectivamente, em 1,1 e 3,7 &#956;mol L-1, para DCF e 1,0 e 3,3 &#956;mol L-1 para CO. O método proposto foi aplicado na determinação simultânea de DCF e CO em formulações farmacêuticas e os resultados obtidos foram similares ao obtidos por HPLC a um nível de confiança de 95%. / Doutor em Química

O veneno do T.nattereri induz uma resposta de memória com a diferenciação de células B B220neg, indicativo de células produtoras de anticorpos de vida longa (ASC). Para avaliar o efeito do veneno na diferenciação de ASCs, camundongos BALB/c foram imunizados e sacrificados nos dias 21, 28, 48, 74 e 120 para avaliação de anticorpos plasmáticos e células B no peritônio, baço e medula óssea. O veneno induziu intensa esplenomegalia, formação de centros germinativos e persistentes níveis de anticorpos específicos IgG1, IgG2a e IgE anafilática. Células B1a e ASC apareceram rapidamente e a população de ASC CD138pos foi dividida em três subtipos (B220highCD43high, B220lowCD43low, e B220negCD43high) que persistiram em diferentes níveis em todos compartimentos. Finalmente, por métodos de neutralização nós sugerimos um papel importante da IL-5 e IL-17 A no desenvolvimento de ASC B220neg e na população B1a e mais ainda, a produção de TNF-a, IL-1b, IL-6, KC bem como a retenção do veneno nas células dendríticas foliculares parece promover os mecanismos para manutenção das ASCs. / T. nattereri fish venom induces a memory immune response with the differentiation of B cells B220neg, an indicative of long-lived antibody-secreting cells - ASC. To assess the effect of the venom on differentiation of ASCs, BALB/c mice were immunized with venom and sacrificed at days 21, 28, 48, 74 and 120 to evaluate plasmatic antibodies and B cell subtypes in peritoneum, spleen and bone marrow. The venom promoted splenomegaly, germinal centers formation and persistent levels of specific antibodies IgG1, IgG2a and anaphylactic IgE. B1a cells and ASC emerged rapidly and CD138pos ASCs can be divided into three subsets (B220high CD43high, B220low CD43low, and B220neg CD43high) that persist at different levels in all compartments. Finally, by neutralization methods we suggested an important role for IL-5 and IL-17A on development of B220neg ASCs and B1a population and moreover the production of TNF-a, IL-1b, IL-6, KC as well as the venom retained in follicular dendritic cells seem to provide mechanisms to explain the maintenance of ASCs.

Die folgende Arbeit befasst sich mit der Alltagstauglichkeit moderner Massendatenverarbeitung, insbesondere mit dem Problem der Assoziationsregelanalyse. Vorhandene Datenmengen wachsen stark an, aber deren Auswertung ist für ungeübte Anwender schwierig. Daher verzichten Unternehmen auf Informationen, welche prinzipiell vorhanden sind. Assoziationsregeln zeigen in diesen Daten Abhängigkeiten zwischen den Elementen eines Datenbestandes, beispielsweise zwischen verkauften Produkten. Diese Regeln können mit Interessantheitsmaßen versehen werden, welche dem Anwender das Erkennen wichtiger Zusammenhänge ermöglichen. Es werden Ansätze gezeigt, dem Nutzer die Auswertung der Daten zu erleichtern. Das betrifft sowohl die robuste Arbeitsweise der Verfahren als auch die einfache Auswertung der Regeln. Die vorgestellten Algorithmen passen sich dabei an die zu verarbeitenden Daten an, was sie von anderen Verfahren unterscheidet. Assoziationsregelsuchen benötigen die Extraktion häufiger Kombinationen (EHK). Hierfür werden Möglichkeiten gezeigt, Lösungsansätze auf die Eigenschaften moderne System anzupassen. Als Ansatz werden Verfahren zur Berechnung der häufigsten $N$ Kombinationen erläutert, welche anders als bekannte Ansätze leicht konfigurierbar sind. Moderne Systeme rechnen zudem oft verteilt. Diese Rechnerverbünde können große Datenmengen parallel verarbeiten, benötigen jedoch die Vereinigung lokaler Ergebnisse. Für verteilte Top-N-EHK auf realistischen Partitionierungen werden hierfür Ansätze mit verschiedenen Eigenschaften präsentiert. Aus den häufigen Kombinationen werden Assoziationsregeln gebildet, deren Aufbereitung ebenfalls einfach durchführbar sein soll. In der Literatur wurden viele Maße vorgestellt. Je nach den Anforderungen entsprechen sie je einer subjektiven Bewertung, allerdings nicht zwingend der des Anwenders. Hierfür wird untersucht, wie mehrere Interessantheitsmaßen zu einem globalen Maß vereinigt werden können. Dies findet Regeln, welche mehrfach wichtig erschienen. Der Nutzer kann mit den Vorschlägen sein Suchziel eingrenzen. Ein zweiter Ansatz gruppiert Regeln. Dies erfolgt über die Häufigkeiten der Regelelemente, welche die Grundlage von Interessantheitsmaßen bilden. Die Regeln einer solchen Gruppe sind daher bezüglich vieler Interessantheitsmaßen ähnlich und können gemeinsam ausgewertet werden. Dies reduziert den manuellen Aufwand des Nutzers. Diese Arbeit zeigt Möglichkeiten, Assoziationsregelsuchen auf einen breiten Benutzerkreis zu erweitern und neue Anwender zu erreichen. Die Assoziationsregelsuche wird dabei derart vereinfacht, dass sie statt als Spezialanwendung als leicht nutzbares Werkzeug zur Datenanalyse verwendet werden kann. / The importance of data mining is widely acknowledged today. Mining for association rules and frequent patterns is a central activity in data mining. Three main strategies are available for such mining: APRIORI , FP-tree-based approaches like FP-GROWTH, and algorithms based on vertical data structures and depth-first mining strategies like ECLAT and CHARM. Unfortunately, most of these algorithms are only moderately suitable for many “real-world” scenarios because their usability and the special characteristics of the data are two aspects of practical association rule mining that require further work. All mining strategies for frequent patterns use a parameter called minimum support to define a minimum occurrence frequency for searched patterns. This parameter cuts down the number of patterns searched to improve the relevance of the results. In complex business scenarios, it can be difficult and expensive to define a suitable value for the minimum support because it depends strongly on the particular datasets. Users are often unable to set this parameter for unknown datasets, and unsuitable minimum-support values can extract millions of frequent patterns and generate enormous runtimes. For this reason, it is not feasible to permit ad-hoc data mining by unskilled users. Such users do not have the knowledge and time to define suitable parameters by trial-and-error procedures. Discussions with users of SAP software have revealed great interest in the results of association-rule mining techniques, but most of these users are unable or unwilling to set very technical parameters. Given such user constraints, several studies have addressed the problem of replacing the minimum-support parameter with more intuitive top-n strategies. We have developed an adaptive mining algorithm to give untrained SAP users a tool to analyze their data easily without the need for elaborate data preparation and parameter determination. Previously implemented approaches of distributed frequent-pattern mining were expensive and time-consuming tasks for specialists. In contrast, we propose a method to accelerate and simplify the mining process by using top-n strategies and relaxing some requirements on the results, such as completeness. Unlike such data approximation techniques as sampling, our algorithm always returns exact frequency counts. The only drawback is that the result set may fail to include some of the patterns up to a specific frequency threshold. Another aspect of real-world datasets is the fact that they are often partitioned for shared-nothing architectures, following business-specific parameters like location, fiscal year, or branch office. Users may also want to conduct mining operations spanning data from different partners, even if the local data from the respective partners cannot be integrated at a single location for data security reasons or due to their large volume. Almost every data mining solution is constrained by the need to hide complexity. As far as possible, the solution should offer a simple user interface that hides technical aspects like data distribution and data preparation. Given that BW Accelerator users have such simplicity and distribution requirements, we have developed an adaptive mining algorithm to give unskilled users a tool to analyze their data easily, without the need for complex data preparation or consolidation. For example, Business Intelligence scenarios often partition large data volumes by fiscal year to enable efficient optimizations for the data used in actual workloads. For most mining queries, more than one data partition is of interest, and therefore, distribution handling that leaves the data unaffected is necessary. The algorithms presented in this paper have been developed to work with data stored in SAP BW. A salient feature of SAP BW Accelerator is that it is implemented as a distributed landscape that sits on top of a large number of shared-nothing blade servers. Its main task is to execute OLAP queries that require fast aggregation of many millions of rows of data. Therefore, the distribution of data over the dedicated storage is optimized for such workloads. Data mining scenarios use the same data from storage, but reporting takes precedence over data mining, and hence, the data cannot be redistributed without massive costs. Distribution by special data semantics or user-defined selections can produce many partitions and very different partition sizes. The handling of such real-world distributions for frequent-pattern mining is an important task, but it conflicts with the requirement of balanced partition.

Platonic Craft and Medical Ethics examines the Platonic theory of craft and shows its application to different ethical problems in medicine, both ancient and modern. I begin by elucidating the Platonic use of the term “craft” or “technē”, using especially the paradigmatic craft of medicine, and explicate a number of important principles inherent in his use of the term. I then show how Plato’s framework of crafts can be applied to two ancient debates. First, I show how Plato’s understanding of crafts is used in discussing the definition of medicine, and how he deals with the issue of “bivalence”, that medicine seems to be capable of generating disease as well as curing it. I follow this discussion into Aristotle, who, though he has a different interpretation of bivalence, has a solution in many ways similar to Plato’s. Second, I discuss the relevance of knowledge to persuasion and freedom. Rhetors like Gorgias challenge the traditional connections of persuasion to freedom and force to slavery by characterizing persuasion as a type of force. Plato addresses this be dividing persuasion between sorcerous and didactic persuasion, and sets knowledge as the new criterion for freedom. Finally, I discuss three modern issues in medical ethics using a Platonic understanding of crafts: paternalism, conclusions in meta-analyses and therapeutic misconceptions in research ethics. In discussing paternalism, I argue that tools with multiple excellences, like the body, should not be evaluated independently of the uses to which the patient intends to put them. In discussing meta-analyses, I show how the division of crafts into goal-oriented and causal parts in the Phaedrus exposes the confusion inherent in saying that practical conclusions can follow directly from statistical results. Finally, I argue that authors like Franklin G. Miller and Howard Brody fail to recognize the hierarchical relationship between medical research and medicine when they argue that medical research ethics should be autonomous from medical ethics per se.

Platonic Craft and Medical Ethics examines the Platonic theory of craft and shows its application to different ethical problems in medicine, both ancient and modern. I begin by elucidating the Platonic use of the term “craft” or “technē”, using especially the paradigmatic craft of medicine, and explicate a number of important principles inherent in his use of the term. I then show how Plato’s framework of crafts can be applied to two ancient debates. First, I show how Plato’s understanding of crafts is used in discussing the definition of medicine, and how he deals with the issue of “bivalence”, that medicine seems to be capable of generating disease as well as curing it. I follow this discussion into Aristotle, who, though he has a different interpretation of bivalence, has a solution in many ways similar to Plato’s. Second, I discuss the relevance of knowledge to persuasion and freedom. Rhetors like Gorgias challenge the traditional connections of persuasion to freedom and force to slavery by characterizing persuasion as a type of force. Plato addresses this be dividing persuasion between sorcerous and didactic persuasion, and sets knowledge as the new criterion for freedom. Finally, I discuss three modern issues in medical ethics using a Platonic understanding of crafts: paternalism, conclusions in meta-analyses and therapeutic misconceptions in research ethics. In discussing paternalism, I argue that tools with multiple excellences, like the body, should not be evaluated independently of the uses to which the patient intends to put them. In discussing meta-analyses, I show how the division of crafts into goal-oriented and causal parts in the Phaedrus exposes the confusion inherent in saying that practical conclusions can follow directly from statistical results. Finally, I argue that authors like Franklin G. Miller and Howard Brody fail to recognize the hierarchical relationship between medical research and medicine when they argue that medical research ethics should be autonomous from medical ethics per se.