Dissertation, Göttingen. / Lebenslauf. Bibliographical footnotes.

Thesis (Ph. D.)--Columbia University, 1934. / Vita. Bibliography: p. 34-35.

Thesis (Ph. D.)--West Virginia University, 2009. / Title from document title page. Document formatted into pages; contains xx, 242 p. : ill. Includes abstract. Includes bibliographical references (p. 137-145).

Thesis (Ph.D.)--University of Florida, 2005. / Typescript. Title from title page of source document. Document formatted into pages; contains 150 pages. Includes Vita. Includes bibliographical references.

No presente trabalho foram realizados estudos da atividade biológica de complexos imínicos de Cu(II) e Zn(II) derivados da isatina, principalmente frente a células tumorais HeLa. Sabe-se que estes complexos se ligam ao DNA, podendo causar clivagens simples e duplas na sua estrutura, em presença de peróxido de hidrogênio, através de um mecanismo predominantemente oxidativo. Os complexos de Cu(II) geram espécies reativas de oxigênio (EROs) e, além disto, atuam sobre a mitocôndria como agentes desacopladores. Sabe-se ainda que esses compostos interagem com proteínas específicas, como por exemplo a topoisomerase humana IB e as quinases dependentes de ciclinas (CDKs), inibindo significativamente sua atividade, ou a proteassoma, frente à qual apenas uma inibição moderada foi observada. Os ensaios in vitro desenvolvidos nesta tese, permitiram evidenciar a capacidade de os complexos de intercalar no DNA e RNA, além de clivar o primeiro biopolímero. Os resultados obtidos in celullo, evidenciaram a dependência do cobre para a citotoxicidade dos complexos sendo estes compostos capazes de estimular a necrose das células HeLa. Além disso, os resultados demonstraram que estes compostos são capazes de interferir no ciclo celular, interrompendo-o em fase G2/M e de desencadear um desequilíbrio oxidativo. Os principais alvos intracelulares dos complexos, em células HeLa, são a Mitocôndria, pois interferem em seu funcionamento diminuindo o seu potencial de membrana, e o DNA clivando-o. Ademais, pretendeu-se, através de um estudo sistemático por microscopia Raman confocal, identificar danos celulares provocados pelos complexos, especialmente uma grande alteração no núcleo celular de células HeLa após o tratamento com o complexo [Cu(isaepy)]+. Através de ensaios de viabilidade, também foi possível demonstrar que a superexpressão de glicoproteína-P para linhagem MES-AS/Dx5 não resulta na resistência dessas células frente aos compostos testados, e que os complexos não interferem no funcionamento desta glicoproteína. Também foi observado, através dos experimentos com células HaCaT, que o complexo [Zn(isaepy)]2+ é capaz de interferir no ciclo autofágico celular. / In the present work, the synthesis and characterization of oxindolimine complexes of Cu(II) and Zn(II) were carried out. R is known that these complexes bind to DNA and can cause single and double cleavages in their structure, in the presence or absence of hydrogen peroxide, through a predominantly oxidative mechanism. Cu(II) complexes generate reactive oxygen species (ROS) and, in addition, act on mitochondria as decoupling agents. These compounds are also known to interact with specific proteins, such as human topoisomerase IB and cyclin-dependent kinases (CDKs), inhibiting significantly their activity, or the proteasome, against which only moderate inhibition has been observed. The in vitro assays developed in this thesis have demonstrated the ability of these complexes in intercalating at DNA and RNA, in addition to cleaving the first biopolymer. The results in cellulo evidenced the dependence on the copper for the cytotoxicity of the complexes, that can stimulate the necrosis in HeLa cells. In addition, the results demonstrated that these compounds are capable of interfering in the cell cycle, stopping it at G2/M phase and triggering an oxidative imbalance. The main intracellular targets of the complexes, in HeLa cells, are the Mitochondria, where they interfere in its function, diminishing its membrane potential, and the DNA, causing its cleavage. Further, it was intended, through a systematic study by confocal Raman microscopy, to identify cellular damage triggered by the complexes. Particularly, a large change in the cell nucleus of HeLa cells was demonstrated after treatment with the \'[Cu(isaepy)] POT.+\' complex. Through viability assays, it was also demonstrated that overexpression of P-glycoprotein in MES-SA/Dx5 lineage does not result in the resistance of these cells to the tested compounds, and that the complexes do not interfere in the functioning of this glycoprotein. Finally, it was observed, in experiments with HaCaT cells, that \'[Zn(isaepy)] POT.2+\' complex can interfere in the autophagic cell cycle.

No presente trabalho foram realizados estudos da atividade biológica de complexos imínicos de Cu(II) e Zn(II) derivados da isatina, principalmente frente a células tumorais HeLa. Sabe-se que estes complexos se ligam ao DNA, podendo causar clivagens simples e duplas na sua estrutura, em presença de peróxido de hidrogênio, através de um mecanismo predominantemente oxidativo. Os complexos de Cu(II) geram espécies reativas de oxigênio (EROs) e, além disto, atuam sobre a mitocôndria como agentes desacopladores. Sabe-se ainda que esses compostos interagem com proteínas específicas, como por exemplo a topoisomerase humana IB e as quinases dependentes de ciclinas (CDKs), inibindo significativamente sua atividade, ou a proteassoma, frente à qual apenas uma inibição moderada foi observada. Os ensaios in vitro desenvolvidos nesta tese, permitiram evidenciar a capacidade de os complexos de intercalar no DNA e RNA, além de clivar o primeiro biopolímero. Os resultados obtidos in celullo, evidenciaram a dependência do cobre para a citotoxicidade dos complexos sendo estes compostos capazes de estimular a necrose das células HeLa. Além disso, os resultados demonstraram que estes compostos são capazes de interferir no ciclo celular, interrompendo-o em fase G2/M e de desencadear um desequilíbrio oxidativo. Os principais alvos intracelulares dos complexos, em células HeLa, são a Mitocôndria, pois interferem em seu funcionamento diminuindo o seu potencial de membrana, e o DNA clivando-o. Ademais, pretendeu-se, através de um estudo sistemático por microscopia Raman confocal, identificar danos celulares provocados pelos complexos, especialmente uma grande alteração no núcleo celular de células HeLa após o tratamento com o complexo [Cu(isaepy)]+. Através de ensaios de viabilidade, também foi possível demonstrar que a superexpressão de glicoproteína-P para linhagem MES-AS/Dx5 não resulta na resistência dessas células frente aos compostos testados, e que os complexos não interferem no funcionamento desta glicoproteína. Também foi observado, através dos experimentos com células HaCaT, que o complexo [Zn(isaepy)]2+ é capaz de interferir no ciclo autofágico celular. / In the present work, the synthesis and characterization of oxindolimine complexes of Cu(II) and Zn(II) were carried out. R is known that these complexes bind to DNA and can cause single and double cleavages in their structure, in the presence or absence of hydrogen peroxide, through a predominantly oxidative mechanism. Cu(II) complexes generate reactive oxygen species (ROS) and, in addition, act on mitochondria as decoupling agents. These compounds are also known to interact with specific proteins, such as human topoisomerase IB and cyclin-dependent kinases (CDKs), inhibiting significantly their activity, or the proteasome, against which only moderate inhibition has been observed. The in vitro assays developed in this thesis have demonstrated the ability of these complexes in intercalating at DNA and RNA, in addition to cleaving the first biopolymer. The results in cellulo evidenced the dependence on the copper for the cytotoxicity of the complexes, that can stimulate the necrosis in HeLa cells. In addition, the results demonstrated that these compounds are capable of interfering in the cell cycle, stopping it at G2/M phase and triggering an oxidative imbalance. The main intracellular targets of the complexes, in HeLa cells, are the Mitochondria, where they interfere in its function, diminishing its membrane potential, and the DNA, causing its cleavage. Further, it was intended, through a systematic study by confocal Raman microscopy, to identify cellular damage triggered by the complexes. Particularly, a large change in the cell nucleus of HeLa cells was demonstrated after treatment with the \'[Cu(isaepy)] POT.+\' complex. Through viability assays, it was also demonstrated that overexpression of P-glycoprotein in MES-SA/Dx5 lineage does not result in the resistance of these cells to the tested compounds, and that the complexes do not interfere in the functioning of this glycoprotein. Finally, it was observed, in experiments with HaCaT cells, that \'[Zn(isaepy)] POT.2+\' complex can interfere in the autophagic cell cycle.

Pseudomonas Aeruginosa est un pathogène opportuniste qui adapte son comportement aux molécules présentes dans son milieu. Il augmente en virulence lorsqu’il détecte des peptides natriurétiques dans son environnement. L’isatine est une molécule qui bloque cet effet. Pour mieux comprendre sur quel récepteur l’isatine agit, la conception d’un biocapteur a été menée. Un biocapteur est un outil alliant un bioélément, réagissant spécifiquement avec une cible biologique, et un support physique permettant la transduction du signal pour le rendre mesurable. Les travaux de thèse suivants décrivent la préparation d’un transducteur polymérique, le copolymère d’oléfines cycliques (COC), par greffage de sels de diazonium en surface. La modification de l’isatine par couplage pallado-catalysé compose la partie principale des travaux de synthèse organique effectués. Les méthodes de couplages peptidiques en surface et techniques d’accroche des isatines modifiées sur les surfaces des transducteurs (or ou COC) sont également décrites dans ce manuscrit. Pour finir, l’évaluation des effets biologiques des isatines modifiées et des biocapteurs conçus a été effectuée sur Pseudomonas Aeruginosa et sur d’autres bactéries. / Pseudomonas Aeruginosa is an opportunistic pathogen which can adapt its behavior to the present molecules in its surrounding. It increases in virulence when it detects natriuretics peptides in its environment. Isatin is a molecule which can block this effect. To determine on which receptor isatin acts on, the conception of a biosensor have been conducted. A biosensor is a tool combining a bioelement, reacting specifically with a biological target, and a physical support allowing the transduction of the signal to make it measurable. The following thesis describes the preparation of a polymeric transducer, the cyclic olefin copolymer (COC), by diazonium salts surface grafting. Isatin modification by palladium-catalysed coupling reaction represents the major part of the organic synthesis carried out. Surface peptide coupling methods and techniques to link the modified isatins to the surfaces of transducers (gold or COC) are also described in this dissertation. To finish, biological effects of the modified isatins and designed biosensors have been evaluated on Pseudomonas Aeruginosa and others bacteria.

No description available.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-10-05T11:53:08Z No. of bitstreams: 1 2009 - Leticia Silotti Zampirolli.pdf: 25893124 bytes, checksum: bd44115917dd331e511dc65cd22738a1 (MD5) / Made available in DSpace on 2017-10-05T11:53:08Z (GMT). No. of bitstreams: 1 2009 - Leticia Silotti Zampirolli.pdf: 25893124 bytes, checksum: bd44115917dd331e511dc65cd22738a1 (MD5) Previous issue date: 2009-05-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil. / A series of new dialkylphosphorylhydrazones (phosphorohydrazidic acid, N?-[1,2-dihydro-2- oxo-(R1)-3H-indole-3-iliden]- dialkyl esters was synthesized and characterized by IR, 1H, 13C and 31P NMR and mass spectroscopy. These dialkylphosphorylhydrazones were synthesized in three steps. The first step involved the synthesis of different dialkylphosphites which are obtained by the reaction of PCl3 with three mols of the corresponding alcohols. The second step consisted of the reaction between the dialkylphosphites and hydrazine in a two phase system, leading to the formation of the dialkylphosphorylhydrazines. Finally, the last step was the condensation of these dialkylphosphorylhydrazines with different N-substituted isatins. The analysis of the 1H, 13C and 31P NMR spectra showed the existence of the two possible diastereoisomers E and Z for compounds 1, 2, 6, 10 and 12, while for the remaining compounds only the Z isomer is present. Ten of these compounds were preliminarily tested for their inhibition potential against two protozoa (Trypanosoma cruzi and Leishmania amazonensis). All compounds tested showed cell proliferation inhibition of 98% at 50 ?M for Leishmania amazonensis, whereas for T. cruzi, inhibition of epimastigote cell proliferation was found to be higher than 75% for all compounds tested except 6, which showed a 59% inhibition. These ten compounds were also evaluated against Plasmodium falciparum, affording inhibitions higher than 90% for a 1mM concentration. These compounds were also investigated for their fungicidal activity against phytopatogenic Rhizoctonia solani and Fusarium oxysporum. Compounds 9 and 11 showed a miscelial growth inhibition of 58% for Rhizoctonia solani while compound 12 afforde a 72% inhibition. Compounds 1, 2, 11 and 12 gave Fusarium oxysporum inhibition higher than 52%. Finally, the compounds synthesized were also evaluated for their inhibitory potential against lettuce seed germination and it was observed that the same compounds which showed fungicidal activity were not able to inhibit seed germination. / Uma s?rie de 16 dialquilfosforilidrazonas (?cido fosforoidraz?dico, N? -[1,2-diidro-2-oxo-(R1)- 3H- indol-3-ilideno] -, ?ster de dialquila), sendo todas in?ditas, foram sintetizadas e caracterizadas pelas t?cnicas de espectrometria de IV, RMN de 1H, RMN de 13C, RMN de 31P e massas. As novas dialquilfosforilidrazonas foram sintetizadas em tr?s etapas de rea??o. A primeira etapa consistiu na s?ntese de diferentes fosfitos de dialquila que foram obtidos atrav?s da rea??o do tricloreto de f?sforo (PCl3) com tr?s mols do ?lcool correspondente. Na segunda etapa, a rea??o dos fosfitos de dialquila com a hidrazina, em um sistema bif?sico, levou ? forma??o das dialquilfosforilidrazinas. A ?ltima etapa foi a condensa??o destas dialquilfosforilidrazinas com diferentes isatinas substitu?das. A an?lise dos espectros de RMN de 1H, RMN de 13C, RMN de 31P das dialquilfosforilidrazonas mostraram a coexist?ncia dos dois poss?veis diastereois?meros E e Z, para os compostos 1, 2, 6, 10 e 12, enquanto que para os compostos restantes observou-se apenas o diastereois?mero Z. Dos compostos sintetizados, dez foram avaliados preliminarmente quanto ao potencial inibit?rio de prolifera??o de dois protozo?rios (Trypanosoma cruzi e Leishmania amazonensis). Para Leishmania amazonensis todos os compostos testados apresentaram inibi??o da prolifera??o celular de 98 % a 50 ?M. Enquanto que para T.cruzi verificou-se inibi??o da prolifera??o celular de epimastigotas superior a 75% para todos compostos testados, a exce??o do composto (6) cuja inibi??o foi de 59 %. Esses dez compostos tamb?m foram avaliados frente ao protozo?rio Plasmodium falciparum apresentando inibi??o superior a 90 % para todos os compostos testados, a uma concentra??o de 1mM. Essas dialquilfosforilidrazonas tamb?m tiveram a a??o fungicida avaliada frente aos fungos fitopatog?nicos (Rhizoctonia solani e Fusarium oxysporum). Em Rhizoctonia solani os compostos (9) e (11) apresentaram inibi??o do crescimento miscelial de 58 %, j? o composto (12) apresentou inibi??o de 72%. Para o Fusarium oxysporum destacaram-se os compostos (1, 2, 11 e 12) com inibi??o superior a 52 %. Esses compostos tamb?m foram avaliados quanto ao potencial inibit?rio de germina??o em sementes de alface e verificouse que os mesmos compostos que apresentaram efeitos fungist?ticos, n?o inibiram a germina??o de sementes de alface.

As the principal pro-inflammatory prostanoid, prostaglandin E2 (PGE2) serves as mediator of pain and fever in inflammatory reactions. The biosynthesis of PGE2 starts from arachidonic acid (AA). Cyclooxygenase (COX)-1 and/or COX-2 converts AA to prostaglandin H2 (PGH2), and PGE2 synthases transform PGH2 to PGE2. Current mainstream approach for treating inflammation-related symptoms remains the application of traditional non-steroidal anti-inflammatory drugs (tNSAIDs) and selective COX-2 inhibitors (coxibs). As both categories shut down the biosynthesis of all downstream prostanoids, their application renders several deleterious effects including gastrointestinalulceration and cardiovascular risk. Microsomal prostaglandin E2 synthase-1 (mPGES-1) inhibitors, specifically blocking the production of inflammation-related PGE2, are expected to reduce the adverse effects while retain the anti-inflammation activity. Although several compounds have been reported, only a few have entered clinical trials and none was on the market. Particularly, most of the reported human mPGES-1 inhibitors were not active for wild-type mouse/rat mPGES-1 enzymes, which prevents using the well-established mouse/rat models of inflammation in preclinical studies. Therefore, we expect our designed inhibitors to also be potent against mouse mPGES-1 and thus is suitable for preclinical testing in wild-type mice.