Orientador: Solange Maria de Almeida / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-04T06:49:17Z (GMT). No. of bitstreams: 1 Rocha_AnnaSilviaPenteadoSettida_D.pdf: 3382405 bytes, checksum: ac0363e76c4f0a9223b1427cc5dcf489 (MD5) Previous issue date: 2005 / Resumo: Sendo a radiação ionizante causador de efeitos deletérios no processo de reparação tecidual e o selenito de sódio um agente antioxidante, atuando contra os radicais livres no organismo, a realização deste trabalho de pesquisa teve como objetivo avaliar o efeito radioprotetor do selenito de sódio no processo de reparação óssea em tíbias de ratos fêmea (Rattus Norvergicus, Albinus, Wistar). A amostra, constituída por 100 ratos, foi dividida em quatro grupos experimentais: controle, irradiado, selênio e selênio/irradiado. Todos os animais foram submetidos a um ato cirúrgico que teve como finalidade a produção de um defeito ósseo nas tíbias direita e esquerda. Os animais dos grupos selênio e selênio/irradiado receberam dose única, via intraperitoneal, de 1 mg/Kg de peso corpóreo de selenito de sódio, sendo que nos animais do grupo selênio/irradiado, a dose foi injetada 15 horas antes destes serem irradiados. A irradiação para os grupos irradiado e selêniolirradiado foi realizada por um aparelho de Cobalto terapia (C06o) com dose simples de 8 Gy nos membros inferiores, após três dias do procedimento cirúrgico. Transcorridos 7, 14, 21, 28 e 45 dias, o processo de reparação óssea foi avaliado em cortes histológicos corados com Hematoxilina Eosina e Tricrômico de Mallory; pela análise ultra-estrutural por microscopia eletrônica de varredura; e quantitativa mente pela densidade volumétrica. Morfologicamente, observou-se que o grupo selênio/irradiado aos 7 dias apresentava-se mais atrasado que o grupo controle, entretanto aos 28 dias os Efeito do Selenito de Sódio na Reparação Óssea em Tíbias de Ratos Irradiados grupos controle, selênio e selênio/irradiado apresentavam um padrão de reparação óssea semelhante, o que também foi observado pela microscopia eletrônica de varredura, aos 45 dias. Quantitativamente, foi observada diferença estatisticamente significante entre as médias de densidade volumétrica para os grupos selênio e selênio/irradiado aos 7, 14 e 28 dias e entre os grupos controle e selênio aos 14 dias. Assim, concluiu-se que o selenito de sódio, apesar de ter-se mostrado tóxico aos sete dias do processo de reparação tecidual, agiu como um eficaz radioprotetor na reparação óssea de tíbias de ratos / Abstract: Considering that the ionizing radiation may cause deleterious effects on the process of tissue repair and sodium selenite is an antioxidant agent, acting against the free radicals in the organism, this study aimed at evaluating the radioprotective effect of sodium selenite on the process of bone repair in tibiae of Wistar rats. The sample comprised 100 rats and was divided into four experimental groups: control, irradiated, selenium and selenium/irradiated. Ali animals were submitted to a surgical procedure for production of a bane defect in the right and left tibiae. The animals in the selenium and selenium/irradiated groups received a single dose of sodium selenite (1 mg/kg of body weight) via intraperitoneal injection; the animals in the selenium/irradiated group received the injection 15 hours before irradiation. The irradiation for the irradiated and selenium/irradiated groups was applied with a cobalt therapy machine (CO60) with simple dose at 8 Gy on the lower limbs, three days after surgery. After 7, 14, 21, 28 and 45 days, the process of one repair was morphologically evaluated by Hematoxylin Eosin and Mallory Trichrome staining; ultrastructural analysis by scanning electron microscopy; and quantitatively evaluated by the volumetric density. As to morphology, it was observed that the selenium/irradiated group at 7 days presented a Iate repair compared to the control group; however, at 28 days, the control, selenium and selenium/irradiated groups presented a similar pattern of bone repair, which was also revealed by scanning electron microscopy at 45 days. Quantitatively, there was a statistically significant difference between the means of volumetric density for the selenium and seleniumlírradiated groups at 7, 14 and 28 days and between the control and selenium groups at 14 days. Thus, it was concluded that sodium selenite, despite being toxic at the seventh day of tissue repair, was an effective radioprotective agent for bone repair in tibiae of rats / Doutorado / Radiologia Odontologica / Doutor em Radiologia Odontológica

京都大学 / 新制・課程博士 / 博士(医学) / 甲第25199号 / 医博第5085号 / 新制||医||1072(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 川口 義弥, 教授 藤田 恭之, 教授 妹尾 浩 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

<p>Diss. (sammanfattning) Umeå : Umeå universitet, 1983, härtill 5 uppsatser</p> / digitalisering@umu

Selenium (Se) has received a lot of attention for its antioxidant and immune modulating properties. Yet, comparably few studies have focused on the horse. Therefore the objectives of this research were to evaluate the influences of Se status on immune function and antioxidant defense in horses. Twenty eight horses were allocated to one of 4 dietary Se treatments: low (LS), adequate (AS), high organic (SP) and high inorganic (SS). First, horses assigned to LS, SP and SS were depleted of Se and received a low Se diet (0.07 ppm Se) for 35 wk, while AS received an adequate Se diet (0.14 ppm Se). During week 28 to 35 immune function was evaluated using a vaccine challenge with keyhole limpet hemocyanin (KLH) and equine influenza as antigens. Then, a 29 wk repletion phase followed. The LS and AS received the same diets described above while SP received an organic Se supplemented diet (0.3 ppm; Sel-Plex, Alltech, Nicholasville, KY) and SS an inorganic Se supplemented diet (0.3 ppm; sodium selenite). Immune function was assessed using a vaccine challenge with ovalbumin (OVA) and equine influenza as antigens during week 22 to 29. Samples collected throughout the depletion and repletion phases were used to assess change in Se status, antioxidant status and oxidative stress. Finally, a mild exercise test served to assess exercise induced oxidative stress. The experimental model responded as hypothesized, evaluated by blood Se and glutathione peroxidase (GPx) activity. Upon vaccination with KLH, antibody response was faster in AS than LS. Antigen specific mRNA expression of T-bet was also higher for AS than LS. Following OVA vaccination humoral and cell-mediated vaccination responses were similar across treatments. However, non-specific stimulation of peripheral blood mononuclear cells indicated suppressed mRNA expression of selected cytokines for LS compared to AS, SP and SS. Antioxidant capacity and oxidative stress were unaffected by change in Se status. A difference in GPx response post exercise was also noted between SP and SS. Low Se status impaired some measures of immune function. Supplementation at 0.3 ppm may benefit horses as indicated by higher GPx activity in idle and exercised horses.

This thesis is concerned with the synthesis and evaluation of various tellurium-based chain-breaking antioxidants. The purpose is to find novel regenerable compounds with improved radical-trapping capacity. In the first part of this work, we explore the possibilities to incorporate tellurium into tocopherols and aromatic amines. Overall, tocopherols carrying alkyltelluro groups are better radical-trapping agents than the corresponding sulfur- and selenium analogues. Among them, 7-octyltelluro δ-tocopherol showed a ca. 17-fold higher reactivity than recorded for α-tocopherol and much better regenerability. Even longer inhibition times were recorded for the corresponding bis(tocopheryl) tellurides. In the aromatic amine series, diphenyl amines carrying alkyltelluro groups were shown to function as efficient radical-quenchers capable of inhibiting peroxidation for 460 min in the presence of N-acetylcysteine. Thiol-consumption experiments suggested that the long inhibition times are due to efficient quenching of in-situ formed alkoxyl radicals in a solvent cage. In the second part of the thesis, we study how the antioxidant properties are affected by variations in the electron density at tellurium and the number of alkyltelluro substituents in the molecule. Evaluation of a series of aryltelluro phenols carrying electron donating and electron withdrawing groups in the para-position of the aryl moiety suggested that a high electron density at the heteroatom prolonged the inhibition time. Among alkyltelluro phenols, alkyltelluro resorcinols and bis(alkyltelluro) phenols, phenols carrying alkyltelluro groups in both ortho positions were superior when it comes to radical-trapping activity and regenerability.

O chumbo (Pb) é um metal altamente tóxico no qual os sinais de intoxicação variam bastante ao considerar as diferenças interindividuais. Um dos principais mecanismos de toxicidade do Pb ocorre pela inibição da enzima ácido delta aminolevulínico desidratase (ALAD) no sistema hematopoiético. O Pb também desempenha um importante papel no desbalanço do estado redox, pois sabe-se que ele tem o potencial de aumentar a concentração de espécies reativas de oxigênio (EROS) e inibir enzimas antioxidantes, como por exemplo a glutationa peroxidase (GPx). No entanto, poucos estudos avaliaram estes parâmetros em população ocupacionalmente exposta brasileira. Assim, o presente estudo, objetiva estudar a correlação entre as concentrações de Pb no sangue (Pb-S) de trabalhadores de fábricas de bateria e as atividades das enzimas ALAD e GPx associados com os polimorfismos genéticos da ALAD e GPx. Para tal, foram utilizadas 278 amostras de sangue de trabalhadores expostos ao Pb. As determinações de Pb foram realizadas por espectrometria de massas com plasma acoplado indutivamente (ELAN DRCII Perkin- Elmer). As genotipagens dos polimorfismos genéticos da ALAD e da GPx foram realizadas pela Reação em Cadeia da Polimerase (PCR) em tempo real e as atividades das enzimas ALAD e GPx foram determinadas no sangue por espectrofotometria de UV/VIS. A média da concentração de Pb-S foi de 22,8 ± 14,7 ?g/dL. Foram observadas correlações negativas entre Pb-S e atividade da ALAD (rs -0,24 p<0,01) e Pb-S e atividade de GPx (rs -0,27 p<0,05). Também foi verificada correlação negativa entre a porcentagem de inibição da ALAD e a atividade de GPx (rs -0,21 p<0,01). Em relação aos polimorfismos genéticos, não observamos associação entre os genótipos do ALAD (? -0,19 P>0,05) e GPx (genótipo CT: ? -1,37 P>0,05; genótipo TT: ? -8,37 P>0,05) e a concentração de Pb-S. Não foram observadas associações entre o polimorfismo rs1800668 e a atividade de GPx (genótipo CT: ? -0,016 P>0,05; genótipo TT: ? -0,004 P>0,05). No entanto, foi constatada uma associação entre o genótipo ALAD 1-1 do polimorfismo rs1800435 e a atividade da enzima ALAD (? 3,5 P<0,05). Neste sentido, os resultados deste estudo mostram que o genótipo ALAD 1-1 do polimorfismo rs1800435 do gene ALAD está associado a uma maior atividade da enzima ALAD nos indivíduos expostos ao Pb. Além disso, o polimorfismo rs1800668, localizado no gene que codifica a enzima GPx, não modula a atividade desta enzima nos indivíduos expostos ao metal. Ambos os polimorfismos dos genes ALAD e GPx parecem não influenciar nas concentrações de Pb-S na população estudada. / Lead (Pb) is a highly toxic metal which signs of intoxication vary greatly when considering the interindividual differences. One of the main mechanism of toxicity of Pb is due to inhibition of the enzyme acid delta aminolevulinic dehydratase (ALAD) in the hematopoietic system. Pb also plays an important role in the redox state of unbalance, since it is known that it has the potential of increasing the concentrations of reactive oxygen species (ROS) and inhibit antioxidant enzymes such as glutathione peroxidase (GPx). However, few studies have evaluated these parameters in Brazilian population occupationally exposed. Thus, the present study aims to study the correlation between the concentrations of Pb in the blood (B-Pb) of workers of battery factories and the activities of ALAD and GPx enzymes associated with genetic polymorphisms in ALAD and GPx. Then, blood samples of 278 workers exposed to Pb were collected. Pb was measured by Inductively Coupled Plasma-Mass Spectrometry (Perkin-Elmer ELAN DRCII). The genotyping of genetic polymorphisms of ALAD and GPx was performed by Real-Time Polymerase Chain Reaction (PCR) and activity of ALAD and GPx enzymes in the blood was determined by spectrophotometry UV / VIS. The mean concentration of B-Pb was 22.81 ± 14.73 mg / dL. Negative correlation was found between B-Pb and ALAD activity (rs -0.24 p < 0.01) and B-Pb and GPx activity (rs -0.27 p < 0.05). There was also a negative correlation between the percentage of inhibition of ALAD and GPx activity (rs -0.21 p < 0.01). Considering the genetic polymorphisms, no association between ALAD genotypes (? -0.19 P < 0.05) and GPx (CT genotype: ? -1.37 P > 0.05, TT genotype : ? -8.37 P > 0.05 ) and the concentration of B-Pb. No association between rs1800668 polymorphism and GPx activity (CT genotype: ? -0.016 P > 0.05, TT genotype : ? -0.004 P > 0.05). However an association was found between ALAD 1-1 rs1800435 genotype and ALAD enzyme activity (? 3.5 P < 0.05). Accordingly, the results of this study show that the ALAD 1-1 genotype of rs1800435 polymorphism in ALAD gene is associated with increased activity of ALAD enzyme in individuals exposed to Pb. Furthermore the rs1800668 polymorphism located in a gene encoding the enzyme GPx does not modulate the activity of this enzyme in individuals exposed to the metal. Both polymorphisms of ALAD and GPx genes seem to have no influence in the concentrations of B-Pb in the population studied.

O selênio (Se), mineral traço essencial para o ser humano, exerce sua função no organismo por meio de sua participação nas selenoproteínas, dentre as quais destacam-se as glutationas peroxidase (GPx), importantes no sistema de defesa antioxidante. São conhecidas sete isoformas da GPx, sendo a GPx 1 (citosólica) a mais abundante. Como a atividade dessa enzima encontra-se fortemente correlacionada com a concentração sanguínea de Se, este parâmetro é normalmente utilizado como indicador do balanço metabólico do mineral. A concentração de Se nos alimentos depende do local onde foram cultivados e reflete diretamente o teor do mineral no solo, sendo os alimentos da região Norte e Nordeste mais ricos nesse nutriente quando comparados às demais regiões brasileiras. O consumo inadequado de Se provoca uma diminuição da atividade dessa enzima, o que por sua vez pode afetar a proteção antioxidante. Além disso, estudos vêm mostrando que o polimorfismo Pro198Leu no gene da GPx1 também pode diminuir a atividade da GPx. O presente estudo se propôs a verificar a relação entre esse polimorfismo e o estado nutricional relativo ao selênio de uma população adulta residente no município de Fortaleza/Ceará. A população do estudo foi constituída por 176 indivíduos (79 homens e 97 mulheres), com média de idade de 30,4 ± 8,9 anos. Foram realizadas a avaliação da composição corporal por antropometria e a avaliação do consumo alimentar, por meio de três recordatórios de 24h. Os biomarcadores para avaliação do estado nutricional dos indivíduos quanto ao selênio foram: a concentração de Se plasmático e eritrocitário e a atividade da GPx total no eritrócito. Também foi determinado o polimorfismo Pro198Leu no gene que codifica para a GPx 1 por meio de PCR em Tempo Real. Foram encontradas concentrações médias de 62,6 &#181;g/L para Se plasmático e de 101,5 &#181;g/L para Se eritrocitário, não sendo observada correlação entre os dois. A média de ingestão de selênio foi de 76,88 &#181;g/dia, apresentando correlação positiva com o consumo de energia e proteína. Separando por gênero, encontrou-se que as médias de Se plasmático e de Se ingerido dos homens foram significativamente maiores que as das mulheres (p<0,05). A média da atividade da GPx foi de 38,6 U/g Hb. Para o polimorfismo Pro198Leu no gene que codifica para a GPx 1, foram encontrados 96 (54,5%) indivíduos com genótipo selvagem Pro/Pro, 67 (38,1%) indivíduos Pro/Leu e 13 (7,4%) indivíduos Leu/Leu. A frequência encontrada estava em equilíbrio de Hardy-Weinberg. Não foram encontradas diferenças estatísticas significativas nas concentrações de selênio entre os grupos de acordo com o genótipo, no entanto, para a atividade da GPx, houve diferença entre o grupo Pro/Pro e Pro/Leu (p<0,05). No grupo Pro/Pro, foi encontrada correlação positiva entre a concentração de selênio eritrocitário e a atividade eritrocitária total da GPx (p<0,05). Desta forma, podemos concluir que o estado nutricional da população estudada estava adequado com relação ao selênio e que o polimorfismo Pro198Leu apresentou influência sobre a atividade da GPx, que apresentou-se reduzida nos indivíduos que apresentaram alelo variante em seu genótipo. / Selenium (Se) is an essential trace mineral for humans which exerts its function in the body in selenoproteins, among which we highlight the glutathione peroxidase (GPx), important in the antioxidant defense system. There are seven known GPx isoforms and the GPx 1 (cytosolic) is the most abundant. As the activity of this enzyme is strongly correlated with selenium blood concentration this parameter is usually used as an indicator of the mineral metabolic balance. Selenium concentration in food depends on the location where they were grown and directly reflects the soil mineral content. Food in the North and Northeast are richer in selenium when compared to other Brazilian regions. Inadequate consumption of Se causes a decrease in GPx activity, what affects the antioxidant protection. Furthermore, studies have shown that the Pro198Leu polymorphism at GPx1 gene of can also decrease the activity of GPx. Our study aimed to investigate the relationship between this polymorphism and nutritional selenium status in an adult population living in Fortaleza/Ceará/Brazil. The study population consisted of 176 individuals (79 men and 97 women) with a mean age of 30.4 ± 8.9 years. The body composition was assessed by anthropometry and the food intake assessment was done using three 24-hour records. The selenium Se concentration was measured in plasma and erythrocyte and GPx activity was measured in erythrocytes. The Pro198Leu polymorphism at GPx 1 gene was determined by Real Time PCR. We found concentrations of 62.6 &#181;g/L for Se plasma and 101.5 &#181;g/L for erythrocyte. There was no correlation observed between these two markers. The average selenium intake was 76.88 &#181;g/day and it showed positively related to the energy and protein consumption. Separating by gender, men selenium plasma concentration and selenium consumption were significantly higher than for women (p < 0.05). The GPx activity was 38.6 U/g Hb. The Pro198Leu polymorphism at GPx 1 gene frequency were 96 (54.5%) subjects with Pro/Pro, 67 (38.1%) subjects Pro/Leu and 13 (7.4%) individuals Leu/Leu. The rate found was in Hardy -Weinberg equilibrium. There were no statistically significant differences in selenium concentrations between groups according to the genotype. However, for GPx activity we found difference between the group Pro/Pro and Pro/Leu (p< 0.05). We also found positive correlation between Se erythrocyte concentration and GPx activity in Pro/Pro group (p<0.05). Thus, we conclude that the selenium nutritional status of the population studied was adequate and that Pro198Leu polymorphism showed influence on the activity of GPx, which was lower in individuals with the variant allele in their genotype.

A enzima glutationa peroxidase (GPx-1) possui um importante papel na detoxificação de espécies reativas de oxigênio. Neste estudo foi verificada a influência do polimorfismo Pro198Leu da GPx-1 nos níveis de malondialdeído (MDA) e proteínas carboniladas de 84 voluntários expostos ambientalmente ao chumbo (28 homens e 56mulheres; idade: 18 a 60 anos). Os genótipos para o polimorfismo Pro198Leu da GPx-1 foram determinados por PCR seguido por digestão com enzima de restrição. Chumbo no plasma (Pb_P) e chumbo no sangue total (Pb_S) foram determinados por espectrometria de massas com plasma indutivamente acoplado e espectrometria de absorção atômica com forno de grafite, respectivamente. As freqüências alélicas para os alelos T (que codifica o aminoácido prolina) e C (que codifica o aminoácido leucina) da GPx-1 foram 0,73 e 0,27, respectivamente. Os voluntários foram separados em dois grupos, de acordo com a presença ou ausência do alelo que codifica leucina. Não foi encontrada diferença significativa para as concentrações de Pb_P e Pb_S, bem como para os marcadores de estresse oxidativo entre os grupos. Deste modo, os resultados obtidos sugerem que o polimorfismo Pro198Leu da GPx-1 não torna indivíduos ambientalmente expostos ao chumbo, mais suscetíveis aos danos do estresse oxidativo induzidos pelo metal. / Glutathione peroxidase (GPx-1) plays an important role in scavenger reactive oxygen species (ROS). This study was carried out to assess the effects of GPx-1 gene polymorphism (Pro198Leu) on malondialdehyde (MDA) and carbonyl protein ratio in 84 subjects environmentally exposed to lead. Genotypes for the GPx-1 Pro198Leu polymorphism were determined by PCR and restriction fragment length digestion. Lead in plasma (Pb_P) and lead in total blood (Pb_B) were determined by inductively coupled plasma mass spectrometry and by graphite furnace atomic absorption spectrometry, respectively. The allele frequencies for T (which express the proline aminoacid) and C (which express the leucine aminoacid) were 0.73 e 0.27, respectively. The volunteers were divided in two groups, according to the presence or absence of C allele. No significant differences were found in Pb_P and Pb_B, as well as for the other oxidative biomarkers between the groups. Therefore, the obtained results suggest that the GPx-1 Pro198Leu polymorphism do not affect individuals environmentally exposed to lead greater susceptible to oxidative damage induced for this metal.

O selênio (Se) é um mineral essencial para o corpo humano, com ação no sistema imunológico, na glândula tireoide, dentre outras funções. É um nutriente importante na proteção das células contra os efeitos dos radicais livres, já que faz parte da enzima glutationa peroxidase (GPX). O consumo alimentar de Se é dependente da concentração desse elemento nos alimentos que varia em função da concentração do mesmo nos solos. Deste modo, este estudo teve como objetivo geral avaliar o estado nutricional relativo ao selênio de indivíduos adultos da cidade de Manaus-AM, por ser considerada região com solo rico nesse nutriente. Casuística: o estudo foi do tipo transversal baseado em amostra não probabilística de conveniência. A população estudada foi composta por 124 voluntários de ambos os gêneros com idade entre 20 a 59 anos. Critérios de inclusão: não ser atleta de elite, não ser etilista, não apresentar doenças crônicas não transmissíveis e não fazer o uso de suplementos vitamínico-minerais e/ou anti-inflamatórios. Não foram incluídos no estudo os participantes que não entregaram os formulários e questionários solicitados (alimentação e/ou ficha cadastral) e/ou que não compareceram para coleta de sangue. A coleta de sangue foi em jejum de 8 a 12 horas para determinação de selênio no plasma e eritrócitos e da atividade da GPX nos eritrócitos, também foram avaliados o consumo alimentar e as medidas antropométricas. Os resultados antropométricos mostraram que 60,0% dos participantes apresentava excesso de peso, a distribuição dos macronutrientes ficou dentro do recomendado pelas DRIs (Dietary Reference Intakes), a ingestão média de selênio ficou acima do recomendado (72±48,0&#181;g/dia), a concentração média para o selênio plasmático foi de 111,4± 37,0 &#181;g/L, de selênio eritrocitário 211± 62,4 &#181;g/L e a atividade da GPX foi de 73± 21,9 U/g Hb, não sendo observada correlação entre as concentrações obtidas para plasma e eritrócito, porém com correlação positiva para Se eritrocitário e atividade da GPX (r= 0,393). Pode-se, portanto concluir do presente estudo que a boa parte dos indivíduos avaliados, habitantes da cidade de Manaus, foi considerada deficiente, apesar de consumirem quantidade desse mineral consideradas adequadas porém, nenhum apresentando parâmetros bioquímicos de deficiência, a maioria dentro da normalidade, entretanto, com um percentual de indivíduos apresentando risco de toxicidade. / Selenium (Se) is an essential mineral for the human body, with function on the immune system, thyroid glands and other ones. It is an important nutrient in protecting cells against the effects of free radicals because it is part of glutathione peroxidase enzyme (GPX). Selenium intake depends on the mineral concentration in aliments which varies based on the mineral content in soils. Thus, this study had as main objective to evaluate the nutritional status related to selenium on adults in Manaus-AM because it is considered as a region of high selenium soil content region. Casuistic: This study was transversal type based in a non-probabilistic convenience sample. The population consisted of 124 volunteers (both genders) aged from 20 to 59 years. Inclusion criteria: To not be an elite athlete or alcoholic, do not have chronic non-communicable diseases and do not use vitamin-mineral supplements and/or anti-inflammatory pills. People who did not submit the requested forms and questionnaires (registration form and/or alimentation) and/or did not attend for blood collection were not included. Blood collection was fasting from 8 to 12 hours for determining selenium in plasma and erythrocytes and the activity of GPX in erythrocytes. Food intake and anthropometric measures were also evaluated. Anthropometric results showed that 60,0% were overweight, macronutrient distribution was within the recommended by DRIs (Dietary Reference Intakes). Average selenium intake was above recommended (72±48,0 &#181;g/day), mean plasmatic selenium concentration was 111,4 ± 37,0 &#181;g/L, for erythrocytes selenium was 211 ± 62,4 &#181;g/L and activity of GPX average was 73 ± 21,9 U/g Hb. No correlation between concentration of Se in plasma and erythrocyte, however, there was positive correlation between GPX activity and Se concentration in erythrocytes (r=0,393). Finally, we conclude that most of the people that were evaluated, citizens of Manaus city, were considered as deficient, although they consumed adequate quantities of this mineral. However, none of them showed biochemical deficiency parameters, most within normality, but with a percentage of them showing toxicity risk.

A scientific and cultural revolution occurred with the sequencing of the human genome. The information provided by this accomplishment has provided tools for researchers to test new ideas in silico and on the bench. In redox biology many of the genes, transcripts, proteins, and redox active species have been well characterized. However, the vast majority have not been quantitated in an absolute manner. This is a necessary step to provide the tools for mathematical modeling and systems biology approaches for predicting changes in the cellular redox environment and the biochemical and biological consequences. Here we demonstrate techniques for the absolute quantitation of human catalase, glutathione peroxidase, peroxiredoxin, thioredoxin, and superoxide dismutase within cells. These techniques can be parsed into two groups: detection of activity and detection of total amount of species. Methods for the absolute quantitation of active catalase, peroxiredoxins, and superoxide dismutase have been developed by utilizing specific characteristics of each enzyme. Catalase generates oxygen in the presence of hydrogen peroxide that can easily be detected with a Clark electrode (oxygen monitor); the data are fit to a single-exponential to determine the observed pseudo-first-order rate constant. From this the effective number of fully active catalase enzymes in the sample can be determined. Peroxiredoxin in the disulfide state can be reduced by thioredoxin; thioredoxin from E. coli loses fluorescence upon oxidation. The loss of fluorescence over time is mathematically fit to a single-exponential to determine the observed pseudo first-order rate constant from which the number of active enzymes can be determined. Using an inhibition assay to detect superoxide dismutase activity along with the rate constants at which superoxide reacts with the dismutase and the competing superoxide-reacting-indicator-molecule, the concentration of active superoxide dismutase can be determined. To detect the total amount of protein of an enzyme in a biological sample, an immunoassay was first implemented. This method utilized Bio-Plex® beads from Bio-Rad; however, it was problematic because the antibodies applied did not perform satisfactorily not allowing sufficient signal-to-noise to be deployed. Quantitative mass spectrometry was then implemented to detect total catalase, glutathione peroxidase 1, peroxiredoxin 2, and thioredoxin 1 in human red blood cells. With the absolute concentration of these enzymes and proteins along with data for oxygen consumption rates and peroxisomal hydrogen peroxide concentration for several cell lines, we hypothesize that a reasonable model of hydrogen peroxide and superoxide flux can be constructed. Quantitative data such as these provide the foundation for the new redox biology of the 21st century. Presented here is a roadmap for the obligatory first steps to dissect quantitatively the cellular and tissue metabolic pathways and redox networks that are the basis of all of biology.