The production of reactive oxygen species by mitochondria is implicated in the mitochondrial dysfunction associated with a range of diseases and ageing. In contrast, reactive oxygen species produced by mitochondria are involved in redox signalling pathways necessary for modulating a number of cell processes. Mitochondrially targeted antioxidants comprised of an antioxidant moiety linked to a lipophilic triphenylphosphonium cation have recently been used to decrease reactive oxygen species-mediated oxidative damage to mitochondria and to investigate the role of mitochondrial reactive oxygen species in redox signalling. These lipophilic cations are selectively accumulated by mitochondria within cells due to the mitochondrial membrane potential. This thesis presents the synthesis and characterization of mitochondrially targeted antioxidant superoxide dismutase and thiol peroxidase mimetics. A mitochondrially targeted derivative of the Mn(II) macrocycle SOD mimetic M40403 (MitoSOD) was synthesized by Mn(II) template synthesis of a chiral tetraamine component and a triphenylphosphonium derivative of 2,6-pyridinedialdehyde. Racemic tetraamine was synthesized by mono-protection of racemic diamine followed by reductive amination of glyoxal and deprotection of di-protected tetraamine but overall this was found to be less efficient than a reported method based on trityl protection. The synthesis of the triphenylphosphonium derivative of 2,6-pyridinedialdehyde involved substitution of protected 4-bromo-2,6-pyridinedialdehyde by the thiolate of 3-mercaptoproanol followed by simultaneous deprotection and alkyl bromide formation, and triphenylphosphine substitution of the thioalkyl bromide substituent. MitoSOD was found to be more lipophilic than M40403 and was kinetically stable to dissociation to Mn(II) and macrocyclic ligand at physiological pH. Pulse radiolysis kinetic studies indicated both MitoSOD and M40403 catalyse the dismutation of superoxide. Fast conductivity and spectrophotometric measurements indicated the mechanism of catalysis involved reaction of the Mn(II) centre with superoxide to give a Mn(III)-peroxide intermediate which reacted with further superoxide to give the parent Mn(II) macrocycle. MitoSOD was significantly accumulated by mitochondria and this was dependent to some extent on the mitochondrial membrane potential. In addition, MitoSOD appeared to react with a product of mitochondrial succinate respiration. A mitochondrially targeted derivative of the organoselenium thiol peroxidase mimetic ebselen (Mitoebselen) was synthesized by O-alkylation of a phenolic ebselen derivative with a triphenylphosphonium derivative of an alkyl iodide. Reaction of excess triphenylphosphine with an ebselen derivative containing an alkyl iodide substituent resulted in substitution of iodide and, unexpectedly, reduction of the isoselenazole moiety to the diselenide redox form. Mitoebselen and its diselenide were both readily reduced to a selenol by an excess of the physiological thiol glutathione. Reaction of the selenol with excess peroxide generated the diselenide, possibly via reaction of unreacted selenol with Mitoebselen formed from a selenenic acid intermediate or with selenenic acid directly. Mitoebselen and its diselenide were both oxidized by excess peroxide to a selenoxide but these reactions were much slower than those between selenol and peroxides, and those between Mitoebselen or its diselenide with glutathione. Together these studies suggested cyclic pathways other than a selenolisoselenazole-selenol cycle could be involved in Mitoebselen or ebselen-catalysed thiol peroxidation.

No description available.

Thesis (M. Phil.)--University of Hong Kong, 2007. / Title proper from title frame. Also available in printed format.

Thesis (M.S. in zoology)--Washington State University, May 2010. / Title from PDF title page (viewed on June 22, 2010). "School of Biological Sciences." Includes bibliographical references (p. 35-39).

Thesis (M. Sc.)--University of Canterbury, 2006. / Typescript (photocopy). Includes bibliographical references (leaves 92-101). Also available via the World Wide Web.

Proefschrift Universiteit Maastricht. / Met bibliogr., lit. opg. - Met een samenvatting in het Nederlands.

Programa de Pós-Graduação em Ciências Biológicas. Núcleo de Pesquisas em Ciências Biológicas, Pró-Reitoria de Pesquisa e Pós Graduação, Universidade Federal de Ouro Preto. / Submitted by Maurílio Figueiredo (maurilioafigueiredo@yahoo.com.br) on 2015-01-28T20:33:18Z No. of bitstreams: 2 license_rdf: 22190 bytes, checksum: 19e8a2b57ef43c09f4d7071d2153c97d (MD5) TESE_InfluênciaSuplementaçãoCafeina.pdf: 25302484 bytes, checksum: 36134eb93fcec49c878f070d4b0c917a (MD5) / Approved for entry into archive by Gracilene Carvalho (gracilene@sisbin.ufop.br) on 2015-01-30T18:26:56Z (GMT) No. of bitstreams: 2 license_rdf: 22190 bytes, checksum: 19e8a2b57ef43c09f4d7071d2153c97d (MD5) TESE_InfluênciaSuplementaçãoCafeina.pdf: 25302484 bytes, checksum: 36134eb93fcec49c878f070d4b0c917a (MD5) / Made available in DSpace on 2015-01-30T18:26:56Z (GMT). No. of bitstreams: 2 license_rdf: 22190 bytes, checksum: 19e8a2b57ef43c09f4d7071d2153c97d (MD5) TESE_InfluênciaSuplementaçãoCafeina.pdf: 25302484 bytes, checksum: 36134eb93fcec49c878f070d4b0c917a (MD5) Previous issue date: 2012 / Devido à utilização da cafeína como suplemento por atletas; sua relação com o estresse oxidativo gerado pelo exercício físico; e do alto consumo de café no Brasil, de sua importância como fonte de compostos bioativos na dieta, o objetivo deste trabalho foi avaliar se a suplementação de cafeína influencia na performance física de ratas submetidas ao treinamento físico e sua relação com o estresse oxidativo. Para isso, foram realizados 2 experimentos, sendo o primeiro realizado com 40 ratas da linhagem Fischer, divididas em 4 grupos com 10 animais cada. Os grupos foram: C-controle, CC-controle+cafeína, T-treinado, TC-treinado+cafeína. Os animais dos grupos C e T receberam dieta AIN-93M e os animais dos grupos CC e TC, dieta AIN-93M + cafeína (12 mg/Kg/dia). No segundo experimento, foram utilizadas 28 ratas, sendo divididas nos mesmos 4 grupos com 7 animais cada. A dose de cafeína utilizada foi de 15 mg/Kg/dia. Nos 2 experimentos, os animais praticaram natação 5 dias/semana, 30 minutos/dia, durante 7 semanas e com aumento progressivo da carga, de 1 a 5% do peso corporal. Para os animais do 2º experimento, foram realizados dois testes de exaustão, sem carga. O primeiro após 30 dias de dieta suplementada ou não com cafeína e o segundo, no final do experimento. Esses animais foram submetidos ao teste do lactato, onde nadaram com carga de 7,5% do peso corporal. A ingestão alimentar, o ganho de peso foram monitorados semanalmente e após 7 semanas, os animais foram sacrificados, o sangue e os órgãos foram coletados para dosagens bioquímicas e de estresse oxidativo. Os resultados do primeiro experimento mostraram que os efeitos da cafeína foram: aumento da ingestão alimentar; elevação do peso relativo do fígado, do músculo gastrocnêmio, da gordura abdominal e dos níveis de PON (paraoxonase). Os resultados do segundo experimento mostraram que a suplementação da dieta com cafeína não aumentou o tempo de natação e a concentração de lactato sanguíneo não chegou à sua estabilização. Quanto aos seus efeitos, a cafeína foi responsável por elevar o ganho de peso; aumentar a massa muscular; elevar os níveis de TBARS (Substâncias Reativas ao Ácido Tiobarbitúrico) no músculo gastrocnêmio, a catalase no músculo sóleo e a proteína carbonilada no fígado. Quando associada ao grupo controle, a cafeína reduziu a proteína carbonilada no músculo gastrocnêmio e reduziu a atividade da catalase no fígado e rins quando associada ao grupo treinado. Além disso, quando associada ao grupo treinado elevou as concentrações de creatinina e glutationa e a atividade da catalase no coração. A adição de 12 mg de cafeína/kg na dieta de ratas Fischer mostrou um efeito positivo sobre a atividade da PON (paraoxonase) e a dose de 15 mg/kg apresentou uma ação tecido específica para as defesas antioxidantes. __________________________________________________________________________________________ / ABSTRACT: Caffeine has been used as a supplement by athletes and has association with oxidative stress that is generated by exercise, and due the high consumption of coffee in Brazil and its importance as a source of bioactive compounds in the diet, the aim of this study was to evaluate if caffeine supplementation influences on physical performance of rats subjected to physical training and its association with oxidative stress. Two experiments were carried out. The first one used 40 female Fischer rats, divided into 4 groups of 10 animals each. The groups were: C-control, CC-control+caffeine, T-trained, TC-trained+caffeine. The animals in groups C and T were fed with AIN-93M diet and animals of CC and CT groups with AIN-93M+caffeine (12 mg/kg/day) diet. In the second experiment, 28 female Fischer rats were used, divided in the same groups of seven animals each. The caffeine level used in the diet was 15 mg/kg/day. In the two experiments, the animals practiced swimming 5 days/week, 30 min/day during 7 weeks and the load was progressively increasing from 1 to 5% of body weight. In the animals of the second experiment, two exhaustion tests without load were performed. The first test was in 30 days of diet supplemented or not with caffeine, and the second at the end of the experiment. These animals were submitted to the lactate testing, they swam with a load of 7.5% of body weight. Food intake, weight gain was monitored weekly and after seven weeks, the animals were sacrificed, blood and organs were collected for biochemical and oxidative stress assays. The results of the first experiment showed that the effects of caffeine were: increasing food intake, raising the relative weights of liver, gastrocnemius muscle, abdominal fat and levels of PON. The results of the second experiment showed that supplementing the caffeine in the diet did not increase the swimming time and blood lactate concentration did not reach its stabilization. As to its effects, caffeine was responsible for raising the weight gain, increase muscle mass, increase levels of TBARS in the gastrocnemius muscle, catalase in the soleus muscle and protein carbonyl in the liver. When associated to the control group, caffeine reduced the protein carbonyl in gastrocnemius muscle and as well as the activity of catalase in the liver and kidneys when associated with the trained group. Moreover, when associated with the trained group increased creatinine concentrations and glutathione and catalase activity in the heart. The addition of 12 mg caffeine/kg in the diet of female Fischer rats showed a positive effect on the activity of PON and addition of 15 mg/kg showed an response tissue-specific in the antioxidant defenses.

A doença do sono é causada pelo parasita Tripanosoma brucei. Considerada uma doença negligenciada, mata milhares de pessoas todos os anos na África subsaariana. O T.brucei não apresenta resposta imune pronunciada, o que dificulta o desenvolvimento de vacinas, e os medicamentos disponíveis são escassos. Os tripanossomatídeos são comprovadamente sensíveis ao stress oxidativo causado pelo radical superóxido. Assim, as enzimas superóxido dismutases (SODs) são a primeira linha de defesa contra esse radical. As SODs são metalo enzimas (EC 1.5.1.1) capazes de catalisar a dismutação do superóxido em oxigênio molecular e peróxido de hidrogênio. São classificadas de acordo com o metal incorporado na estrutura: cobre e zinco (CuZnSOD), ferro ou manganês (Fe/MnSOD) e níquel (NiSODs). Neste trabalho de mestrado, a enzima TbFeSODB2 de T.brucei foi, expressa, purificada, cristalizada e teve sua estrutura resolvida. A estrutura cristalográfica da enzima do parasita foi comparada com a enzima humana análoga contendo manganês (HuMnSOD), onde foram evidenciadas as principais diferenças entre as duas estruturas que podem ser exploradas para o desenho do novos inibidores seletivos. Foi realizada uma análise de acoplamento estatístico, onde com base em um alinhamento múltiplo dessas enzimas determinou-se resíduos que são capazes de interferir na seletividade do metal incorporado e estado oligomérico das SODs. / Sleeping sickness, caused by the parasite Tripanosoma brucei, is considered a neglected disease, killing thousands of people every year in subsaharian Africa. T. brucei does not generate a pronounced immune response, difficulting the development of vaccines. Furthermore, available medicines are scarce. Tripanosomatides are known to be sensitive to oxidative stress caused by the superoxide radical. Therefore, the superoxide dismutase enzymes (SODs) are a primary line of defence for the parasites against this radical. SODs are metalloenzymes (EC 1.5.1.1) capable of catalyzing superoxide dismutation into molecular oxygen and hydrogen peroxide. SODs are classified according to the incorporated metal: copper/zinc (CuZnSOD), iron/manganese (Fe or MnSOD) and nickel (NiSODs). In the work presented here, TbFeSODB2 from T. brucei was expressed, purified, crystallized and its 3D structure solved. The crystal structure of the parasite enzyme was compared to the homologous human enzyme containing manganese (HuMnSOD), revealing evidence for differences between both structures which could be exploited in the design of new selective inhibitors. In addition, a statistical coupling analysis was performed on the entire Fe/MnSOD superfamily, based on a multiple sequence alignment. It was shown that this technique was able to identify novel residue determinants of metal selectivity and oligomeric state.

A doença do sono é causada pelo parasita Tripanosoma brucei. Considerada uma doença negligenciada, mata milhares de pessoas todos os anos na África subsaariana. O T.brucei não apresenta resposta imune pronunciada, o que dificulta o desenvolvimento de vacinas, e os medicamentos disponíveis são escassos. Os tripanossomatídeos são comprovadamente sensíveis ao stress oxidativo causado pelo radical superóxido. Assim, as enzimas superóxido dismutases (SODs) são a primeira linha de defesa contra esse radical. As SODs são metalo enzimas (EC 1.5.1.1) capazes de catalisar a dismutação do superóxido em oxigênio molecular e peróxido de hidrogênio. São classificadas de acordo com o metal incorporado na estrutura: cobre e zinco (CuZnSOD), ferro ou manganês (Fe/MnSOD) e níquel (NiSODs). Neste trabalho de mestrado, a enzima TbFeSODB2 de T.brucei foi, expressa, purificada, cristalizada e teve sua estrutura resolvida. A estrutura cristalográfica da enzima do parasita foi comparada com a enzima humana análoga contendo manganês (HuMnSOD), onde foram evidenciadas as principais diferenças entre as duas estruturas que podem ser exploradas para o desenho do novos inibidores seletivos. Foi realizada uma análise de acoplamento estatístico, onde com base em um alinhamento múltiplo dessas enzimas determinou-se resíduos que são capazes de interferir na seletividade do metal incorporado e estado oligomérico das SODs. / Sleeping sickness, caused by the parasite Tripanosoma brucei, is considered a neglected disease, killing thousands of people every year in subsaharian Africa. T. brucei does not generate a pronounced immune response, difficulting the development of vaccines. Furthermore, available medicines are scarce. Tripanosomatides are known to be sensitive to oxidative stress caused by the superoxide radical. Therefore, the superoxide dismutase enzymes (SODs) are a primary line of defence for the parasites against this radical. SODs are metalloenzymes (EC 1.5.1.1) capable of catalyzing superoxide dismutation into molecular oxygen and hydrogen peroxide. SODs are classified according to the incorporated metal: copper/zinc (CuZnSOD), iron/manganese (Fe or MnSOD) and nickel (NiSODs). In the work presented here, TbFeSODB2 from T. brucei was expressed, purified, crystallized and its 3D structure solved. The crystal structure of the parasite enzyme was compared to the homologous human enzyme containing manganese (HuMnSOD), revealing evidence for differences between both structures which could be exploited in the design of new selective inhibitors. In addition, a statistical coupling analysis was performed on the entire Fe/MnSOD superfamily, based on a multiple sequence alignment. It was shown that this technique was able to identify novel residue determinants of metal selectivity and oligomeric state.

For trace elements, such as copper (Cu) and zinc (Zn), the bioavailability of these elements, Cu and Zn, in biosolids is important because both are essential elements and both are potential contaminants when biosolids are land applied. A greenhouse study was conducted in factorial experiment based on a completely randomized design (CRD) with four replications on a soil treated with four rates of Cu (0, 50, 100, and 150 mg/kg) and four rates of Zn (0, 150, 300, and 450 mg/kg) on celery plants to investigate the distribution and mobility of these elements as well as growth and antioxidant changes of celery. The results of antioxidant changes were inconclusive due to irregular changes with Zn and Cu applications. However, generally the results show that Cu did not affect superoxide dismutase (SOD) or peroxidase (POD) activities in most of the treatments. On the other hand, Zn stimulated SOD and POD activities in most of the treatments. The photosynthesis rate decreased with the applications of Cu and Zn at the rates above 100 and 300 mg/kg and increased in low Cu concentration (50 mg/kg) compared to S (soil without biosolid).