No description available.

<p>  </p> <p>Infections of dermal wounds is a growing burden for the healthcare industry, with a 2017 market exceeding $17.5 USD. As the number of patients with severe infections continues to increase year after year, there is an alarming downward trend in efficacy for traditional antibiotic treatments. In large part, this is due to the increasing development of antibiotic resistance within common bacteria strains. As microbes evolve to protect themselves from previously effective drugs, there is a growing need for new antimicrobial therapies. While alternatives exist in the market, they are largely impaired by non-selective toxicity which can cause further damage to the cells in the wound bed, as is the case with silver and other strong antiseptics, or the need for high energy, specialized equipment, as with cold atmospheric surface treatments. Gaseous ozone is a promising alternative therapy for treating these wound infections. Because ozone is a strong natural oxidant, it exhibits significant antimicrobial properties, and has also been shown to help stimulate natural wound healing in many cases. Herein is presented the design of a portable system for the topical delivery of gaseous ozone as an antimicrobial treatment for infected dermal wounds. This includes the design and characterization of the portable system and a custom ozone application dressing, the characterization of the safety and efficacy of the system using <em>in vitro</em> and <em>in vivo</em> models, and a disposable system for wound infection monitoring. The system utilizes a portable corona discharge generator to produce gaseous ozone from the ambient environment. The ozone gas is delivered through a dressing engineered to have a hydrophobic interface at the wound bed and disperse the ozone gas across the patch surface for more uniform application. The antimicrobial strength and biocompatibility of the system was optimized at varying ozone output levels. Additionally, an adjunct therapy of topical antibiotics was shown to significantly increase the strength of the treatment without leading to greater cytotoxicity. This synergistic effect between ozone and antibiotics was shown to circumvent natural bacterial resistances to antibiotics, which will have a major impact on the wound care industry. This adjunct treatment was then validated on a porcine animal model for safety and pilot results for efficacy testing. Finally, the pH sensor which can be incorporated with use of the ozone therapy enables objective monitoring of wound condition and is able to signal when appropriate infection therapy should begin. As it stands, this portable ozone wound treatment system shows great promise as an alternative therapy to improve the quality of live for millions of patients.</p>

Submitted by PPGE PPGE (pgedu@ufba.br) on 2015-11-26T16:26:42Z No. of bitstreams: 1 Leonardo Rangel_Versão final.pdf: 2923200 bytes, checksum: e15291f0a586b29fab1ed109127f4d51 (MD5) / Approved for entry into archive by Maria Auxiliadora da Silva Lopes (silopes@ufba.br) on 2015-12-18T14:28:16Z (GMT) No. of bitstreams: 1 Leonardo Rangel_Versão final.pdf: 2923200 bytes, checksum: e15291f0a586b29fab1ed109127f4d51 (MD5) / Made available in DSpace on 2015-12-18T14:28:16Z (GMT). No. of bitstreams: 1 Leonardo Rangel_Versão final.pdf: 2923200 bytes, checksum: e15291f0a586b29fab1ed109127f4d51 (MD5) / A presente dissertação é resultado de reflexões no campo da Educação, desenvolvido no âmbito do Programa de Pós-Graduação em Educação da Universidade Federal da Bahia. O estudo tem a tradição filosófica com base na sua configuração metodológica e emerge em meio aos objetos que vêm construindo o campo específico dos processos formativos. Neste caso, procuramos compreender de que modo a subjetividade vem sendo problematizada e quais suas implicações na vida/formação dos seres humanos. Tal investigação surgiu a partir das inquietações e provocações tencionadas pela problemática que parece a ssumir nos dias de hoje, quase a força de um princípio categórico, e que pode ser traduzida nas expressões: “A vida como obra”, “A vida como projeto”, ou ainda, na expressão mais utilizada no campo da educação: “Ser ou tornar-se autor de sua própria vida”; afirmação que ressalta o caráter de agência dos processos subjetivos e assinala a tendência contemporânea do fenômeno de ampliação dos espaços outrora tidos como da ordem da “interioridade”, “intimidade” ou do “privado”. Pois, a modernidade que se iniciou com o processo da I Revolução Industrial, modificou-se drasticamente com as posteriores revoluções empreendidas no modo de produção capitalista. Em grande parte, essas transformações foram impulsionadas pela introdução de novas tecnologias. Deste modo, os processos formativos emergentes apontam para toda uma reconfiguração dos espaços e dos tempos, tanto os exteriores quanto os interiores, e nos levam rumo a novos tipos de embate, rumo a novas formas de se fazer e experimentar a vida, o mundo, o trabalho, a educação, a política etc. Utilizando o cuidado como um dispositivo fundante e um dos modos prioritários de se situar no mundo em nossa atualidade, o presente trabalho tem como escopo uma problematização da relação entre o conhecimento e o modo de subjetivação dominante na cultura ocidental, no intuito de caracterizar suas influências na fundação das perspectivas formativas, com o propósito de levantar alguns impasses e armadilhas legadas pelo projeto iluminista. Ao fim dessa jornada, compreendemos que a referência aos valores constitui hoje, uma das mais importantes fontes de transcendência, e talvez uma das últimas, a nos convidar a viver e reconstruir os sentidos da vida, em suas dimensões individuais e coletivas, em um tipo de formação centrada no cuidado e no princípio de mais vida. / ABSTRACT This dissertation is a result of some thoughts about the education field developed in the Programa de Pós-Graduação em Educação da Universidade Federal da Bahia (Master’s Program of Education at Federal University of Bahia ). The study uses a philosophical tradition as the basis of the theoretical method. It emerges from events which are building the specific research field of formative procedures. At this case, we are looking to understand how subjectivity has been questioned, and what are it life’s implications in the formation of the human being. The investigation has started with questioning issues, about life as a project or a masterpiece. Furthermore, these issues can be known within the Education field by the expressions: “To be or become the author of your own life”. Such an assertion highlights the human agency in the subjective process, and points out the contemporary tendency of the enlargement of the spaces once understood and called “privacy”, “intimacy” and “inner behavior”. Modernity started after the First Industrial Revolution and changed drastically the other capitalist revolution that was about to come. Theses transformations were largely motivated by the introduction and improvement of new technology. Thus, the ruse of formative procedures shows the new set of time and space, both inner and outside processes of development, taking us to new ways of experiencing life, trying different worlds, others concepts of job, education, politics, etc. Using care as a base apparatus, and also as one of the most important ways in which one fits into this world, this research has the purpose to debate the relationship between knowledge and dominant and occidental processes of subjectivity. My goal is to feature the influences in the formative perspectives foundation in order to bring up the traps and impasses of the Enlightment’s legacy. At the end it is possible to realize that the value references compose, nowadays, the most relevant transcendental source, or one of the last invitations to live and rebuild life’s meanings in its collective and individual aspects, focused on care and the principle of more life.

Le méthotrexate (MTX), un agent anti-cancéreux fréquemment utilisé en chimiothérapie, requiert généralement un suivi thérapeutique de la médication (Therapeutic Drug Monitoring, TDM) pour surveiller son niveau sanguin chez le patient afin de maximiser son efficacité tout en limitant ses effets secondaires. Malgré la fenêtre thérapeutique étroite entre l’efficacité et la toxicité, le MTX reste, à ce jour, un des agents anti-cancéreux les plus utilisés au monde. Les techniques analytiques existantes pour le TDM du MTX sont coûteuses, requièrent temps et efforts, sans nécessairement fournir promptement les résultats dans le délai requis. Afin d’accélérer le processus de dosage du MTX en TDM, une stratégie a été proposée basée sur un essai compétitif caractérisé principalement par le couplage plasmonique d’une surface métallique et de nanoparticules d’or. Plus précisément, l’essai quantitatif exploite la réaction de compétition entre le MTX et une nanoparticule d’or fonctionnalisée avec l’acide folique (FA-AuNP) ayant une affinité pour un récepteur moléculaire, la réductase humaine de dihydrofolate (hDHFR), une enzyme associée aux maladies prolifératives. Le MTX libre mixé avec les FA-AuNP, entre en compétition pour les sites de liaison de hDHFR immobilisés sur une surface active en SPR ou libres en solution. Par la suite, les FA-AuNP liées au hDHFR fournissent une amplification du signal qui est inversement proportionnelle à la concentration de MTX. La résonance des plasmons de surface (SPR) est généralement utilisée comme une technique spectroscopique pour l’interrogation des interactions biomoléculaires. Les instruments SPR commerciaux sont généralement retrouvés dans les grands laboratoires d’analyse. Ils sont également encombrants, coûteux et manquent de sélectivité dans les analyses en matrice complexe. De plus, ceux-ci n’ont pas encore démontré de l’adaptabilité en milieu clinique. Par ailleurs, les analyses SPR des petites molécules comme les médicaments n’ont pas été explorés de manière intensive dû au défi posé par le manque de la sensibilité de la technique pour cette classe de molécules. Les développements récents en science des matériaux et chimie de surfaces exploitant l’intégration des nanoparticules d’or pour l’amplification de la réponse SPR et la chimie de surface peptidique ont démontré le potentiel de franchir les limites posées par le manque de sensibilité et l’adsorption non-spécifique pour les analyses directes dans les milieux biologiques. Ces nouveaux concepts de la technologie SPR seront incorporés à un système SPR miniaturisé et compact pour exécuter des analyses rapides, fiables et sensibles pour le suivi du niveau du MTX dans le sérum de patients durant les traitements de chimiothérapie. L’objectif de cette thèse est d’explorer différentes stratégies pour améliorer l’analyse des médicaments dans les milieux complexes par les biocapteurs SPR et de mettre en perspective le potentiel des biocapteurs SPR comme un outil utile pour le TDM dans le laboratoire clinique ou au chevet du patient. Pour atteindre ces objectifs, un essai compétitif colorimétrique basé sur la résonance des plasmons de surface localisée (LSPR) pour le MTX fut établi avec des nanoparticules d’or marquées avec du FA. Ensuite, cet essai compétitif colorimétrique en solution fut adapté à une plateforme SPR. Pour les deux essais développés, la sensibilité, sélectivité, limite de détection, l’optimisation de la gamme dynamique et l’analyse du MTX dans les milieux complexes ont été inspectés. De plus, le prototype de la plateforme SPR miniaturisée fut validé par sa performance équivalente aux systèmes SPR existants ainsi que son utilité pour analyser les échantillons cliniques des patients sous chimiothérapie du MTX. Les concentrations de MTX obtenues par le prototype furent comparées avec des techniques standards, soit un essai immunologique basé sur la polarisation en fluorescence (FPIA) et la chromatographie liquide couplée avec de la spectrométrie de masse en tandem (LC-MS/MS) pour valider l’utilité du prototype comme un outil clinique pour les tests rapides de quantification du MTX. En dernier lieu, le déploiement du prototype à un laboratoire de biochimie dans un hôpital démontre l’énorme potentiel des biocapteurs SPR pour utilisation en milieux clinique. / Methotrexate (MTX) cancer therapy requires therapeutic drug monitoring (TDM) for following its levels in a patient during the course of treatment in order to maximize efficacy while minimizing side effects. Despite its narrow therapeutic window, MTX remains until this date, one of the most employed chemotherapy agents. Existing TDM analytical techniques for MTX are costly, time-consuming and labor intensive which are not suitable to promptly generate results within the therapy timeframe. To provide rapid MTX quantification for TDM, a strategy is proposed based on a competitive assay featuring gold nanoparticles and surface plasmonic coupling. More specifically, the inhibition of MTX with its molecular receptor, human dihydrofolate reductase (hDHFR), an enzyme associated with proliferative diseases, is explored. Free MTX mixed with folic acid-functionalized gold nanoparticles (FA-AuNP) are in competition for hDHFR binding sites immobilized on a SPR active surface or free in solution. FA-AuNP binding to hDHFR provides signal amplification which is inversely proportional to the concentration of MTX. Surface plasmon resonance (SPR) is commonly used as a spectroscopic technique for the interrogation of biomolecular interactions. Current commercial SPR instruments are laboratory-based, bulky, expensive, lack sensitivity in complex matrix and have not shown adaptability in clinical settings. In addition, SPR analysis of small molecules such as drugs has not been extensively explored due to lack of sensitivity. The recent advances in materials science and surface chemistry exploiting gold nanoparticle integration for SPR response enhancement and peptide surface chemistry have shown potential in overcoming the poor sensitivity and surface-fouling limitations for crude biofluids analysis. These novel concepts of SPR technology are incorporated with a miniaturized fully integrated SPR prototype to conduct fast, reliable and sensitive analysis to monitor MTX levels of a patient undergoing chemotherapy. The objective of the thesis is to explore different strategies in improving drug analysis in a complex matrix using SPR biosensors and to put in perspective of the potential of SPR biosensors as a useful TDM tool in clinical laboratories or at a point-of-care situation. To achieve these objectives, a colorimetric solution-based MTX competitive assay is first established with FA-AuNP. Then, the solution-based MTX competitive assay is translated onto a SPR platform. For both developed assays, sensitivity, selectivity, detection limit, dynamic range optimization as well as analysis of methotrexate in complex matrix are inspected. Furthermore, the SPR prototype is validated by its equivalent performance to existing SPR systems and by its utility in executing MTX analysis in actual serum samples from patients undergoing chemotherapy. The concentrations of MTX obtained by SPR biosensing are compared to standard techniques: fluorescence polarization immunoassay (FPIA) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in order to confirm the feasibility of SPR biosensors as a useful clinical tool for performing rapid MTX concentration evaluation. Finally, the successful deployment of the prototype to a hospital laboratory demonstrates enormous prospective of SPR biosensors in clinical use.

Le méthotrexate (MTX), un agent anti-cancéreux fréquemment utilisé en chimiothérapie, requiert généralement un suivi thérapeutique de la médication (Therapeutic Drug Monitoring, TDM) pour surveiller son niveau sanguin chez le patient afin de maximiser son efficacité tout en limitant ses effets secondaires. Malgré la fenêtre thérapeutique étroite entre l’efficacité et la toxicité, le MTX reste, à ce jour, un des agents anti-cancéreux les plus utilisés au monde. Les techniques analytiques existantes pour le TDM du MTX sont coûteuses, requièrent temps et efforts, sans nécessairement fournir promptement les résultats dans le délai requis. Afin d’accélérer le processus de dosage du MTX en TDM, une stratégie a été proposée basée sur un essai compétitif caractérisé principalement par le couplage plasmonique d’une surface métallique et de nanoparticules d’or. Plus précisément, l’essai quantitatif exploite la réaction de compétition entre le MTX et une nanoparticule d’or fonctionnalisée avec l’acide folique (FA-AuNP) ayant une affinité pour un récepteur moléculaire, la réductase humaine de dihydrofolate (hDHFR), une enzyme associée aux maladies prolifératives. Le MTX libre mixé avec les FA-AuNP, entre en compétition pour les sites de liaison de hDHFR immobilisés sur une surface active en SPR ou libres en solution. Par la suite, les FA-AuNP liées au hDHFR fournissent une amplification du signal qui est inversement proportionnelle à la concentration de MTX. La résonance des plasmons de surface (SPR) est généralement utilisée comme une technique spectroscopique pour l’interrogation des interactions biomoléculaires. Les instruments SPR commerciaux sont généralement retrouvés dans les grands laboratoires d’analyse. Ils sont également encombrants, coûteux et manquent de sélectivité dans les analyses en matrice complexe. De plus, ceux-ci n’ont pas encore démontré de l’adaptabilité en milieu clinique. Par ailleurs, les analyses SPR des petites molécules comme les médicaments n’ont pas été explorés de manière intensive dû au défi posé par le manque de la sensibilité de la technique pour cette classe de molécules. Les développements récents en science des matériaux et chimie de surfaces exploitant l’intégration des nanoparticules d’or pour l’amplification de la réponse SPR et la chimie de surface peptidique ont démontré le potentiel de franchir les limites posées par le manque de sensibilité et l’adsorption non-spécifique pour les analyses directes dans les milieux biologiques. Ces nouveaux concepts de la technologie SPR seront incorporés à un système SPR miniaturisé et compact pour exécuter des analyses rapides, fiables et sensibles pour le suivi du niveau du MTX dans le sérum de patients durant les traitements de chimiothérapie. L’objectif de cette thèse est d’explorer différentes stratégies pour améliorer l’analyse des médicaments dans les milieux complexes par les biocapteurs SPR et de mettre en perspective le potentiel des biocapteurs SPR comme un outil utile pour le TDM dans le laboratoire clinique ou au chevet du patient. Pour atteindre ces objectifs, un essai compétitif colorimétrique basé sur la résonance des plasmons de surface localisée (LSPR) pour le MTX fut établi avec des nanoparticules d’or marquées avec du FA. Ensuite, cet essai compétitif colorimétrique en solution fut adapté à une plateforme SPR. Pour les deux essais développés, la sensibilité, sélectivité, limite de détection, l’optimisation de la gamme dynamique et l’analyse du MTX dans les milieux complexes ont été inspectés. De plus, le prototype de la plateforme SPR miniaturisée fut validé par sa performance équivalente aux systèmes SPR existants ainsi que son utilité pour analyser les échantillons cliniques des patients sous chimiothérapie du MTX. Les concentrations de MTX obtenues par le prototype furent comparées avec des techniques standards, soit un essai immunologique basé sur la polarisation en fluorescence (FPIA) et la chromatographie liquide couplée avec de la spectrométrie de masse en tandem (LC-MS/MS) pour valider l’utilité du prototype comme un outil clinique pour les tests rapides de quantification du MTX. En dernier lieu, le déploiement du prototype à un laboratoire de biochimie dans un hôpital démontre l’énorme potentiel des biocapteurs SPR pour utilisation en milieux clinique. / Methotrexate (MTX) cancer therapy requires therapeutic drug monitoring (TDM) for following its levels in a patient during the course of treatment in order to maximize efficacy while minimizing side effects. Despite its narrow therapeutic window, MTX remains until this date, one of the most employed chemotherapy agents. Existing TDM analytical techniques for MTX are costly, time-consuming and labor intensive which are not suitable to promptly generate results within the therapy timeframe. To provide rapid MTX quantification for TDM, a strategy is proposed based on a competitive assay featuring gold nanoparticles and surface plasmonic coupling. More specifically, the inhibition of MTX with its molecular receptor, human dihydrofolate reductase (hDHFR), an enzyme associated with proliferative diseases, is explored. Free MTX mixed with folic acid-functionalized gold nanoparticles (FA-AuNP) are in competition for hDHFR binding sites immobilized on a SPR active surface or free in solution. FA-AuNP binding to hDHFR provides signal amplification which is inversely proportional to the concentration of MTX. Surface plasmon resonance (SPR) is commonly used as a spectroscopic technique for the interrogation of biomolecular interactions. Current commercial SPR instruments are laboratory-based, bulky, expensive, lack sensitivity in complex matrix and have not shown adaptability in clinical settings. In addition, SPR analysis of small molecules such as drugs has not been extensively explored due to lack of sensitivity. The recent advances in materials science and surface chemistry exploiting gold nanoparticle integration for SPR response enhancement and peptide surface chemistry have shown potential in overcoming the poor sensitivity and surface-fouling limitations for crude biofluids analysis. These novel concepts of SPR technology are incorporated with a miniaturized fully integrated SPR prototype to conduct fast, reliable and sensitive analysis to monitor MTX levels of a patient undergoing chemotherapy. The objective of the thesis is to explore different strategies in improving drug analysis in a complex matrix using SPR biosensors and to put in perspective of the potential of SPR biosensors as a useful TDM tool in clinical laboratories or at a point-of-care situation. To achieve these objectives, a colorimetric solution-based MTX competitive assay is first established with FA-AuNP. Then, the solution-based MTX competitive assay is translated onto a SPR platform. For both developed assays, sensitivity, selectivity, detection limit, dynamic range optimization as well as analysis of methotrexate in complex matrix are inspected. Furthermore, the SPR prototype is validated by its equivalent performance to existing SPR systems and by its utility in executing MTX analysis in actual serum samples from patients undergoing chemotherapy. The concentrations of MTX obtained by SPR biosensing are compared to standard techniques: fluorescence polarization immunoassay (FPIA) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in order to confirm the feasibility of SPR biosensors as a useful clinical tool for performing rapid MTX concentration evaluation. Finally, the successful deployment of the prototype to a hospital laboratory demonstrates enormous prospective of SPR biosensors in clinical use.